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1
Wang, Qiufan Claire, and 王秋帆. "Mechanisms of junctional restructuring at the sertoli-sertoli and sertoli-germ cell interfaces during spermatogenesis." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2008. http://hub.hku.hk/bib/B40887686.
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Wang, Qiufan Claire. "Mechanisms of junctional restructuring at the sertoli-sertoli and sertoli-germ cell interfaces during spermatogenesis." Click to view the E-thesis via HKUTO, 2008. http://sunzi.lib.hku.hk/hkuto/record/B40887686.
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Maguire, Sharon Marie. "Germ cell modulation of Sertoli cell function." Thesis, University of Edinburgh, 1994. http://hdl.handle.net/1842/20662.
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The aim of this work was to assess the influence of germ cells on the expression of selected Sertoli cell mRNAs. To this end, adult rats were treated with 650mg/kg methoxyacetic acid (MAA) to induce the specific depletion of >80% of pachytene and later spermatocytes from most tubules, and expression of selected Sertoli cell mRNAs was then assessed at various times after treatment when particular germ cell types were depleted selectively (see Bartlett et al. 1988; Allenby et al., 1991). Studies on the expression of cyclic protein 2(CP-2) mRNA supported the hypothesis that germ cells can influence the cyclic function of Sertoli cells. Expression of CP-2 mRNA was shown by Northern blot analysis to decrease significantly 21 days after MAA treatment. In situ hybridisation showed that CP-2 mRNA expression was decreased or absent from tubules at stages at which CP-2 mRNA is normally expressed (stages IV-VII) when elongate spermatids were depleted selectively from these tubules. This decrease was reflected in loss of CP-2 protein production. These observations lead us to hypothesise that elongate spermatids positively modulate CP-2 expression in the Sertoli cell, with this modulation occurring at the level of transcription. In conclusion, this study demonstrated that germ cells do appear to influence Sertoli cell gene expression. This can occur at the level of transcription as was demonstrated by the effect of elongate spermatids on CP-2 mRNA expression, or may occur post-transcriptionally, as would appear to be the case with ABP.
4
Petersen, Cecilia. "Paracrine regulation of Sertoli cell proliferation /." Stockholm, 2003. http://diss.kib.ki.se/2003/91-7349-443-7/.
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Samy, Eileen Teresa. "Regulation of testin and prostaglandin D2 synthetase expression in sertoli cells: a molecular and cell biologystudy and its implication in sertoli-germ cell interactions." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 1999. http://hub.hku.hk/bib/B3122331X.
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Pearce, Kristen (Kristen Joanne) 1974. "Regulation of adhesion between round spermatids and Sertoli cells in the testis." Monash University, Dept. of Obstetrics and Gynaecology, 2003. http://arrow.monash.edu.au/hdl/1959.1/6606.
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Wong, Ching-hang. "Cell-cell interactions and cell junction dynamics in the mammalian testis." Click to view the E-thesis via HKUTO, 2005. http://sunzi.lib.hku.hk/hkuto/record/B31993084.
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McCabe, Mark James, and markmccabe02@hotmail com. "Hormonal regulation of the testicular Sertoli cell tight junction." RMIT University. Applied Sciences, 2008. http://adt.lib.rmit.edu.au/adt/public/adt-VIT20081212.100348.
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The Sertoli cell tight junction (TJ) of the seminiferous epithelium is important for the developmental process of spermatogenesis as it separates germ cells in the seminiferous tubules from the general circulation in the testicular interstitium. Absence of the TJ leads to spermatogenic arrest and infertility. TJs form at puberty as circulating gonadotrophins luteinising hormone/testosterone and follicle stimulating hormone increase. Several studies have demonstrated hormonal regulation of the two major TJ proteins, claudin-11 and occludin, and also of TJ function in vitro and in vivo. Men with low levels of circulating gonadotrophins exhibit an immature and dysfunctional TJ phenotype, which is reversed upon the exogenous application of gonadotrophins. This thesis hypothesises that claudin-11 and occludin are the major contributors to TJ function, and that gonadotrophins regulate TJ function and structure via these two proteins in several species including humans. This PhD was divided into four separate studies to address these hypotheses. The first study selectively silenced the genetic expression of claudin-11 and occludin with small interfering RNA (siRNA) in cultured immature rat Sertoli cells to determine their contribution to Sertoli cell TJ function in vitro. siRNA treatment against either protein significantly (p less than 0.01) reduced TJ function by ~50% as assessed by transepithelial electrical resistance. Immunocytochemistry displayed marked reductions in the localisation of these proteins to the TJ after siRNA treatment. It was concluded that both proteins significantly contributed to TJ function in vitro. The second and third studies then aimed to study hormonal regulation of the TJ in vivo. Weekly injections of the gonadotrophin releasing hormone antagonist acyline were used to suppress circulating gonadotrophins and spermatogenesis in adult rats. Acyline treatment disrupted i) the localisation of occludin to the TJ and ii) TJ function as shown by permeability to a biotin tracer, which was impermeable to TJs in controls. Short-term hormone replacement partially restored the effects of gonadotrophin suppression. It was concluded that gonadotrophins regulate the maintenance of the TJ in rats in vivo. The third study used the hypogonadal (hpg) mouse, which is a naturally occurring model of gonadotrophin deficiency with inactive spermatogenesis. Claudin-11 in hpg mice was not localised at the TJs, and these were dysfunctional as shown by permeability to biotin. Following hormone treatment, TJs were structurally and functionally competent, demonstrating that gonadotrophins also regulate the formation of TJs in vivo. The fourth study subsequently analysed TJs in gonadotrophin suppressed men, and it was found that claudin-11 staining was reduced from continuous bands in control men, to punctate staining in gonadotrophin-suppressed men, demonstrating that gonadotrophins also regulate the localisation of claudin-11 to the TJ in men in vivo. In summary, it is concluded that the Sertoli cell TJ is hormonally regulated, and that the major contributors to TJ function in vivo and in vitro are claudin-11 and occludin. It is hypothesised that the reduction of claudin-11 localisation to the TJ in men may also result in a loss of human Sertoli cell TJ function, suggesting that the TJ may be a potential target of hormonal contraception in men.
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Gregory, Christopher Wayne. "Biochemical characterization of two hormonally-regulated Sertoli cell proteins /." The Ohio State University, 1993. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487847761308818.
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Wong, Ching-hang, and 黃政珩. "Cell-cell interactions and cell junction dynamics in the mammalian testis." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2005. http://hub.hku.hk/bib/B31993084.
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Wolski, Katja Margrit. "The Sertoli Cell-Spermatid Junctional Complex : a potential avenue for Male contraception." [Tampa, Fla] : University of South Florida, 2006. http://purl.fcla.edu/usf/dc/et/SFE0001747.
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李志恆 and Chi-hang Jonathan Li. "Characterization of a sertoli cell product, rat myotubularin: its involvement in cell-cell interactionsin the testis." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2000. http://hub.hku.hk/bib/B31240550.
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Li, Chi-hang Jonathan. "Characterization of a sertoli cell product, rat myotubularin : its involvement in cell-cell interactions in the testis /." Hong Kong : University of Hong Kong, 2000. http://sunzi.lib.hku.hk/hkuto/record.jsp?B2198170X.
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Opuwari, Chinyerum. "Effect of basella alba and hibiscus macranthus on tm4 sertoli cell functions." Thesis, University of the Western Cape, 2009. http://etd.uwc.ac.za/index.php?module=etd&action=viewtitle&id=gen8Srv25Nme4_3021_1380808051.
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Basella alba (BA) and Hibiscus macranthus (HM) are used by traditional healers in Cameroon to treat male sexual fertility problems. Previous studies showed that in vivo administration of the leaf extracts of both plants caused a significant increase in rat seminal vesicle weight and spermatozoa numbers was accompanied by a significant increase in serum testosterone. The aim of this study was to establish the effects of BA and HM extracts on Sertoli cell functions. TM4 cell line was used in this study as it exhibited properties similar to the Sertoli cells (Mather, 1982). Sertoli cell play a key role in spermatogenesis by regulating and supporting germ cell development. Therefore, any alterations in Sertoli cell physiology or structure may lead to impaired spermatogenesis, germ cell loss and male infertility. Developing germ cells in the seminiferous tubule require a constant supply of lactate and pyruvate (Jutte et al, 1981
1982) and toxicant induced alterations in these nutrients have been shown to induce germ cell necrosis (Monsees et al., 2000). TM4 Sertoli cells were cultured in DMEM/Ham F-12 (M) for one day and exposed to
0.01, 0.1, 1, 10, 100 &mu
g/ml of BA and HM extracts, respectively, for four further days. The extracts were dissolved in 0.5 % DMSO in M, while 0.5 % and 2% DMSO in M were used as negative or positive controls, respectively, and 100mM ethanol as positive control where indicated. Results obtained from the Sertoli cells exposed to BA extracts, showed that the plant extract had no significant effect on the cell viability but induced a significant concentration-dependent increase in lactate (19-67%) and pyruvate levels (39-102%) and a concentration-dependent decrease in the protein content (9-42%). The H&
E histological study confirmed that the BA extract had no cytotoxic effect, as there were no changes in the morphology of the cell. Likewise, apoptotic study using DAPI showed no alteration in the nucleus when compared to the negative control. The HM plant extract significantly enhanced mitochondrial dehydrogenase activity (7fold) in the Sertoli cells but caused only slight alterations in the lactate and pyruvate levels. There was no effect seen in the protein content of the Sertoli cells. H&
E and DAPI staining revealed that there were neither changes in the morphology of the cells nor any alteration regarding the mitotic and apoptotic indices. Thus, the HM extract did not have a cytotoxic effect on the cells. This study demonstrated that the Basella alba methanol extract may enhance spermatogenesis as it stimulated the source of energy required for the development of germ cells without exerting a cytotoxic effect. The Hibiscus macranthus extract stimulated mitochondrial dehydrogenase activities and may thus trigger changes in Sertoli cell physiology. In summary, both plant extracts enhanced certain Sertoli cell
functions and thus might explain the positive in vivo effects of the combined plant extracts on rat spermatogenesis observed by Moundipa et al. (1999).
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Williams, J. "Effects of testicular toxicants on Sertoli cell function in vitro." Thesis, Brunel University, 1987. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.377947.
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Hazra, Rasmani. "Sertoli cell steroid nuclear receptors in testis development and function." Thesis, The University of Sydney, 2014. http://hdl.handle.net/2123/10504.
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Steroid action is vital for the development and function of male gonads. Androgen and the androgen receptor (AR) are critical for the initiation and maintenance of complete spermatogenesis and male fertility. Despite this fundamental role, the precise AR-regulated pathways controlling spermatogenic development have yet to be resolved. A major focus of this research was to use a unique gain-of-function transgenic (Tg) mouse model to determine the temporal role of Sertoli cell-specific AR expression in testicular development. The Sertoli cell-specific rat Abpa promoter directed human Tg AR [Tg Sertoli cell (SC)-specific AR (TgSCAR)] expression, providing strong premature postnatal AR immunolocalized to Sertoli cell nuclei. This novel transgenic model provides unique opportunity to directly differentiate vital AR-regulated genes and regulatory pathways involved in optimal Sertoli cell maturation, and meiotic-postmeiotic germ cell development. The current research has also been investigated the functional role of the Sertoli cell glucocorticoid receptor (GR) in vivo using a Tg Cre-loxP approach to conditionally disrupt GR expression. Sertoli cell GR knock-out (SCGRKO) was shown by absent Sertoli cell-specific GR immunolocalization. This novel SCGRKO model has revealed that Sertoli cell-mediated GR actions support normal testicular function. Sertoli cell GR is required to maintain normal testicular Sertoli/germ cell numbers and circulating gonadotrophin levels, as well as optimal Leydig cell maturation and steroidogenesis, providing new insight into gluocorticoid-mediated impact on male reproduction.
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Cowan, Gillian. "Fetal germ cell differentiation and the impact of the somatic cells." Thesis, University of Edinburgh, 2009. http://hdl.handle.net/1842/4164.
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Specification of a germ cell lineage and appropriate maturation are essential for the transfer of genetic information from one generation to the next. Germ cells form from pluripotent precursor cells that migrate into the gonadal ridge and undergo commitment to either the female or male lineage. In the fetal ovary, germ cells enter meiotic prophase I, then arrest at the diplotene stage; in the testis germ cells do not begin meiosis until puberty. Abnormal differentiation of germ cells can result in malignant transformation. Somatic cells play a key role in modulating the developmental fate of the germ cells. Research into germ cell development during fetal life has almost exclusively focused on studies in rodents, but we, and others, have reported several fundamental differences in the expression of germ cell specific markers in the human compared with the mouse. The studies described in this thesis have investigated germ cell-specific gene expression and the possible impact of the somatic cells during development. This was achieved by studying human fetal gonads obtained during the 1st and 2nd trimesters of pregnancy and through the use of both wild-type and mutant mouse ES cell lines. Studies on germ cells in the human fetal testis have extended the findings of others, and confirmed that germ cell populations at different stages of maturation co-exist in the human fetal testis, a situation that is in contrast to that in rodents. For example expression of M2A and AP2γ was restricted to the OCT4-positive gonocyte population, while VASA and NANOS1 were localised exclusively to the to the OCT4-negative prespermatogonia. DAZL was expressed in both populations. Analysis also revealed that both the gonocyte and prespermatogonial populations proliferate throughout the 2nd trimester. Recent studies have implicated retinoic acid (RA) in the control of meiotic entry in germ cells of the fetal mouse ovary. In this study we demonstrated for the first time that two genes implicated in the action of RA in mouse gonad, STRA8 and NANOS2, are also expressed in a similar sexspecific- manner in the human fetal gonads, and that the RA receptors are present in both somatic and germ cells suggesting that RA may regulate germ cell function in the human as well as the mouse. However, whilst the mesonephros appears to be the primary site of RA synthesis in the mouse our initial studies indicate that in the human the gonad itself may be a more likely site of RA biosynthesis. In the fetal mouse testis, RA is degraded by the enzyme Cyp26b1 present in the somatic cells and germ cells do not enter meiosis, our novel findings suggest that CYP26B1 is more abundant in the human fetal ovary than the testis, suggesting that meiotic entry may be controlled by an alternative signalling pathway in the human. One of the methods that can aid our understanding of somatic cell gene expression in the gonad is in vitro culture. To date, there have been no published reports of the successful in vitro culture of somatic cells from the human fetal testis. In the current study, populations of human somatic cells were dissociated and maintained in vitro and characterised. Analysis demonstrated that cells expressing mRNAs characteristic of Sertoli cells, Leydig cells and peritubular myoid (PTM) cells were present initially, but long-term culture resulted in downregulation in expression of mRNAs specific for Sertoli cells and Leydig cells, suggesting that these cells either failed to survive or underwent alterations to their phenotype. In contrast PTM/fibroblast cells proliferated in vitro and initially maintained androgen receptor expression. These cultures therefore hold promise for studies into the signalling or cell-cell interactions in testicular somatic cells especially those relevant to the PTM population. Several studies have claimed differentiation of putative germ cells from ES cells. In the current study, analysis of mouse ES cell lines has expanded on results showing that ES cells and early germ cells express a number of genes in common. Kit signalling was shown to be important for ES cell survival as they differentiate although expression of Kit was heterogeneous. We also demonstrated that ES cells that did not express Kit displayed a decreased expression of the early germ cell genes Blimp1, Fragilis and Stella, implicating Kit signalling in the control of germ cell-associated gene expression in ES cells. This may be important to future studies optimising germ cell derivation from ES cells. In conclusion, this study has demonstrated important differences in protein expression patterns in germ cells of the human fetal testis compared to the mouse, and has raised questions about whether the proposed mechanism controlling meiotic entry of germ cells in the mouse can be applied to the human. The establishment of a system for culturing human fetal gonadal somatic cells may lead to further understanding of gene expression and development in the human fetal testis, and data suggest that the Kit/Kitl signalling system may influence germ cell gene expression in mouse ES cells.
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Igdoura, Suleiman Abdalla. "Biogenesis, sorting and trafficking of sulfated glycoprotein-1 (SGP-1) in rat Sertoli cells." Thesis, McGill University, 1994. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=28467.
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Sulfated Glycoprotein-1 (SGP-1) secreted by Sertoli cells is a homologue of human prosaposin known to be processed into four 15 kDa saposins (A,B,C and D). Saposins activate lysosomal enzymes which break down membrane glycolipids. The objectives of the present investigation were to examine the developmental expression of SGP-1 and its targeting to lysosomes of Sertoli cells, to characterize the lysosomal form(s) of SGP-1, to examine SGP-1's delivery to Sertoli cells phagosomes and finally to assess the effects of androgens on SGP-1 expression.SGP-1 exists in two forms, a 65 kDa and a 70 kDa form. Labeling experiments demonstrated that the 65 kDa form of SGP-1 is synthesized first then post-translationally modified to the 70 kDa form. Western blot analysis of purified Sertoli cell lysosomes revealed a 65 kDa protein and 15 kDa proteins. The latter were isolated and shown to be saposins which were possibly derived from the 65 kDa precursor. Immunocytochemical studies using anti SGP-1 antibodies demonstrated that lysosomes deliver SGP-1 or/and saposins to phagosomes containing residual bodies. Thus, besides a secretory route of SGP-1 there appears to be a lysosomal pathway that may play a significant role in the hydrolysis of glycolipids of spermatid membranes present in phagocytosed residual bodies. Permeabilization of purified Sertoli Golgi fractions revealed that the 65 kDa protein is strongly associated with Golgi membrane. This association was resistant to low pH, resistant to excess free mannose 6-phosphate and independent of N-linked glycosylation, but sensitive to EDTA treatment. In contrast, the 70 kDa form of SGP-1 was released from permeabilized Golgi Fractions at pH 7.4 and 5.4 but retained at pH 6.4 indicating that it is a soluble protein with biochemical characteristics of a regulated secretory protein. In vivo treatment with tunicamycin resulted in an increase in the density of lysosomal SGP-1 suggesting that N-linked carbohydrate residues are not essential for the targeting of this protein to the lysosomes of Sertoli cells. In contrast, in the nonciliated cells of the efferent duct, tunicamycin treatment resulted in a significant reduction in the labeling density of lysosomal SGP-1 suggesting a sugar dependent lysosomal transport mechanism.
Thus, this study demonstrates a unique lysosomal form of SGP-1 (65 kDa). The targeting of the 65 kDa form to the lysosomes and eventually to the phagosomes involves an early association with Golgi membranes independent of mannose 6-phosphate receptors. The trafficking of the 70 kDa form of SGP-1 to the tubular lumen may depend on its selective segregation into secretion granules within the Golgi apparatus of Sertoli cells. Finally, the expression of SGP-1 by Sertoli cells is modulated by androgens.
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Mruk, Dolores Dorothy. "A study on the dynamics of sertoli-germ cell interactions : new perspectives on male fertility control /." Thesis, Hong Kong : University of Hong Kong, 1999. http://sunzi.lib.hku.hk/hkuto/record.jsp?B22079002.
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Hooley, Robert P. "Studies on Sertoli cell function using in vivo and in vitro models." Thesis, University of Edinburgh, 2008. http://hdl.handle.net/1842/24705.
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SK11 cells had detectable expression of ERβ mRNA, and activated an ERE-luciferase reporter construct in response to a range of oestrogenic ligands. In contrast endogenous Ar expression was low, but transfection with mouse Ar cDNA resulted in sufficient Ar expression to activate a luciferase reporter under control of the Rhox5 proximal promoter in response to androgen treatment. However transfected SK11 cells could not stimulate endogenous Rhox5 mRNA expression under similar conditions. Infection of SK11 cells with an advanced construct in vitro resulted in efficient transgenic expression in 80-100% of cells, and was associated with excellent cell survival. In vivo infection in mouse testes with an adenoviral vector containing a GFP transgene, performed by injection via the efferent ductules, resulted in SC-only transgene expression. When a dose of 4x108 pfu was introduced the seminiferous tubule’s architecture was severely damaged, invasion by macrophages and neutrophils occurred, plus expression of markers of hypoxia and apoptosis was detected. Infection with lower doses (1x105 – 1x107 pfu) resulted in disruption to the seminiferous epithelium consisting of loss of pachytene germ cells and formation of intra-epithelial vacuoles. Formation of vacuoles may be due to interaction of adenovirus with the coxsackie/adenoviral receptor (CAR) in SC-SC junctions. Androgen-depletion in rats using a single dose of EDS to ablate the Lc population that synthesise testosterone caused reduced expression of Ar, Rhox5, espin and β3-tubulin mRNA and Ar protein 6 days after treatment, but the expression and distribution of the junctional proteins espin, Cx43, zona occludins 1, and N-cadherin were unaffected. We can conclude that SK11 cells lack vital factor/s required for activation of response elements by androgens in their endogenous promoter regions, which raises the possibility that association with other testicular cell types (GC or PTM) may be required for normal SC function to be maintained. Adenoviral vectors appear good for efficient introduction of transgenes into SC in vitro but not for use in vivo. Finally, testosterone depletion using the EDS-rat model revealed reduced mRNA expression of putative androgen responsive genes identified in previous array studies, and provides a valuable model for validation of other putative targets identified in other studies.
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Cunningham, James M. "Maturation of Sertoli cell sectretory function during sexual development of the lamb." Thesis, University of Reading, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.255976.
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Fenelon, Miriam Claire. "Regulation of gene expression in the immature Sertoli cell : a role for oestrogens?" Thesis, University of Edinburgh, 2001. http://hdl.handle.net/1842/23344.
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In the first part of the thesis changes in the temporal and spatial expression of Sertoli cell mRNA and proteins were investigated in the postnatal rat testis. Novel results regarding ERb expression during this time of development were reported. In addition patterns of Musashi-1, GATA-1 and GATA-4 Sertoli cell expression were described including new information demonstrating changes in Musashi-1 and GATA-1 protein localisation in the Sertoli cell during the transition from proliferating cells to mature differentiated Sertoli cells. Artificially elevated levels of oestrogen in the neonatal testis delayed Sertoli cell maturation and subsequently delayed expression of GATA-1 protein demonstrating the restriction of GATA-1 protein expression to non-proliferating Sertoli cells which has not previously been described. A primary rat Sertoli cell culture system was established in which Sertoli cells retained hormonal sensitivity consistent with the expression of androgen receptor and ERb proteins. New preliminary data describing upregulation of ERb protein levels by oestrogen in cultured rat Sertoli cells are reported. Steroid regulation of GATA-4 expression in cultured rat Sertoli cells was not detected in the present study at the level of either mRNA or protein. Problems obtained sufficient numbers of Sertoli cells in culture during this investigation was overcome by employing an immortalised mouse Sertoli cell line from the H-2Kb-tsA58 transgenic mouse and conformation of results obtained using primary cultures of rat Sertoli cells was carried out. In conclusion, the studies outlined in this thesis have described expression patterns of markers of Sertoli cell functional maturation and preliminary data regarding their steroid regulation. Such new data can be employed in future studies to further evaluate direct effects of oestrogen and oestrogen-like chemicals on Sertoli cell behaviour in the postnatal rat testis with a view to elucidating the mechanisms involved in modification of testicular function by oestrogens.
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Pearce, Keenau Mark. "The effect of cissampelos capensis extract on prostate cancer, sertoli and leydig cell function." University of the Western Cape, 2014. http://hdl.handle.net/11394/4307.
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Magister Scientiae (Medical Bioscience) - MSc(MBS)This study investigated the effect of the C. capensis extract (0.001, 0.01, 0.1, 1, 10, 100 1000 μg/ml) on LNCaP prostate cancer, TM3 Leydig and TM4 Sertoli cells for 24 and 96 hours. The following parameters were investigated: morphology, cell viability (XTT), testosterone modulation, DNA fragmentation (TUNEL), lactate dehydrogenase activity (LDH), testosterone production, anti-cancer drug combination. In a separate set of experiments, parameters affecting the initiation, progression and metastasis of cancer were investigated. These included the ability of the aqueous C. capensis rhizome extract to inhibit of reactive oxygen species (ROS) and reactive nitrogen species (RNS) production, and to inhibit collagenase and elastase activity.
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Holdcraft, Robert Wesley. "Regulation of spermatogenesis by androgen receptor : effect of hypomorphic and cell-specific mutations /." Thesis, Connect to this title online; UW restricted, 2003. http://hdl.handle.net/1773/10249.
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Keeping, Lyndon E. "The seasonal and stage-related cycles of lipid droplets in Sertoli cells in the seasonal breeding mink Mustela vison." Thesis, University of Ottawa (Canada), 1991. http://hdl.handle.net/10393/7953.
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The relationship between Sertoli cell lipid inclusions and spermatogenic activity was investigated using light microscopy and morphometric analysis of the lipid droplet content of Sertoli cells and spermatids from the seasonal breeding mink. Tissue was obtained and analyzed each month during a twelve month period to establish the presence of seasonal and stage-related cycles of lipid droplet content in these cells. It is concluded that the amount of lipid inclusions in Sertoli cells varies with the degree of spermatogenic activity being lowest after the release of mature spermatids in the lumen during the active spermatogenic phase and at the time of maximal testicular regression. Furthermore, cholesterol esters are a component of these lipid inclusions and the seasonal changes in lipid inclusions of Sertoli cells are reflected by seasonal changes in the amount of Sertoli cell cholesterol esters. (Abstract shortened by UMI.)
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Alcock, Joëlle. "A role for Id2 in maintenance of sertoli and germ cell function in murine spermatogenesis." Thesis, University of Nottingham, 2005. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.423470.
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MONTEIRO, FILHO Waldo Oliveira. "Intervenção farmacológica sobre o sistema serotoninérgico durante o desenvolvimento testicular pré-natal e neonatal de ratos Wistar utilizando cloridrato de fluoxetina." Universidade Federal Rural de Pernambuco, 2009. http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/5884.
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Currently, selective serotonin reuptake inhibitors (SSRIs) are the most frequently prescribed antidepressants for the treatment of depression and major side effects in adults related to sexual dysfunction. Depressive disorders are common both during the prenatal period and early months after birth and fluoxetine is widely used to treat depression in this period. Until the present moment, no further study on the pharmacological intervention in the serotonin system during the critical period for testicular development in neonatal rats and the effects on the spermatogenic process in sexually mature rats was performed. We carried out 3 experiments using fluoxetine hydrochloride. In the first experiment observed the effect of direct administration of fluoxetine during postnatal testicular development in rats, but with assessment at 150 days of age. In this paper, we observed reduction in testicular weight gross and net volume of tubules and seminiferous epithelium. The tubular diameter was reduced in animals treated with 20 mg/Kg which caused an increase in total length of seminiferous tubules in this group. There were no histological changes in the population of Sertoli cells, daily sperm production and efficiency of the spermatogenic process. The second experiment studied the effects of transplacental transfer of fluoxetine and breast milk on the testicular development of prepubertal rats. In this experiment, were observed changes in the body weight development, in the testicular volumetric parameters and total length of seminiferous tubules, mainly in higher doses of fluoxetine. There was a trend of reduction of Sertoli cells population and delay in the appearance tubular lumen in animals exposed to higher dose. Inthe third experiment was used the same protocol of the second experiment, but the testis were performed in rats sexually mature. According to the results was observed that fetal and neonatal contact with fluoxetine during the critical period of Sertoli cells proliferation influenced the somatic development of animals and induced changes in testicular parameters, such as the weight of the epididymis and seminal gland, volumes of the seminiferous epithelium, the total Leydig cells volume, total length of seminiferous tubules and a trend of reduction in sperm production per testis. Although fluoxetine has interfered with the Leydig cells volumetry, were not observed changes in plasma levels of testosterone and spermatogenesis. Most of the changes were related to the group treated with the highest dosage of the antidepressant, 20 mg/kg, implying that this dosage fluoxetine may actually cause changes in somatic development and reproductive function.
Atualmente, os inibidores da recaptação seletiva da serotonina (ISRS) são os antidepressivos mais freqüentemente prescritos para o tratamento da depressão, sendo o principal efeito colateral em adultos relacionado às disfunções sexuais. Distúrbios depressivos são comuns tanto durante o período pré-natal quanto nos meses iniciais após o nascimento e a fluoxetina é largamente utilizado no tratamento da depressão neste período. Até o presente momento, nenhum estudo mais pormenorizado sobre a intervenção farmacológica do sistema serotoninérgico durante o período crítico do desenvolvimento testicular em ratos neonatos e os seus reflexos no processo espermatogênico em ratos maturos sexualmente foi realizado. Neste trabalho foram realizados 3 experimentos utilizando cloridrato de fluoxetina. No primeiro experimento foi observado o efeito da administração direta da fluoxetina durante o período pós-natal do desenvolvimento testicular de ratos Wistar porém, com avaliação aos 150 dias de idade. Neste trabalho se observou redução o peso testicular bruto e líquido, volume de túbulos e epitélio seminíferos. O diâmetro tubular foi reduzido nos animais tratados com 20mg/Kg o que gerou aumento no comprimento total de túbulos seminíferos neste grupo. Não foram observadas alterações morfométricas na população de células de Sertoli, produção espermática diária e eficiência do processo espermatogênico. No segundo experimento foram estudados os efeitos da transferência de fluoxetina transplacentária e pelo leite materno sobre o desenvolvimento testicular de ratos Wistar pré-buberes. Neste experimento se observou alterações no desenvolvimento ponderal de peso corporal, em parâmetros volumétricos testiculares e comprimento total de túbulos seminíferos em dosagensmais elevadas de fluoxetina. Notou-se tendência de redução da população total de células de Sertoli por testículo e atraso no aparecimento do lume tubular nos animais expostos a maior dose. No terceiro experimento se utilizou o mesmo protocolo do segundo experimento, porém as análises foram realizadas em animais maturos sexualmente. De acordo com os resultados observados o contato fetal e neonatal com fluoxetina, no período crítico de proliferação das células de Sertoli influenciou no desenvolvimento somático dos animais e ocasionou alterações em parâmetros testiculares, tais como o peso de epidídimo e glândula seminal, volumetria do epitélio seminífero, volumetria total das células de Leydig, comprimento total dos túbulos seminíferos e uma tendência de redução na produção espermática por testículo. Ainda que a fluoxetina tenha interferido na volumetria das células de Leydig, não se constatou alteração nos níveis plasmáticos de testosterona e da espermatogênese. A maior parte das alterações está relacionada ao grupo tratado com a maior dosagem do antidepressivo, 20mg/Kg, inferindo que nessa dosagem a fluoxetina pode, de fato, causar alterações no desenvolvimento somático e do aparelho reprodutivo.
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Wong, Wai-pung Elissa. "Regulation of spermatogenesis in the microenvironment of the rat seminiferous epithelium the roles of cell polarity proteins /." Click to view the E-thesis via HKUTO, 2009. http://sunzi.lib.hku.hk/hkuto/record/B4357239X.
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Banco, B. "ESPRESSIONE DI MARKERS IMMUNOISTOCHIMICI IN GONADI NORMALI E PATOLOGICHE DI CANE." Doctoral thesis, Università degli Studi di Milano, 2012. http://hdl.handle.net/2434/168075.
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This study aimed to determine the immunophenotypical characterization of canine Sertoli cells (SCs) and related tumors and canine ovarian neoplasms.
In veterinary literature histological and immunohistochemical aspects of canine Sertoli cell tumor (SCT) are poorly investigated and data on normal testes of mature and immature dogs are absent.
In human medicine, numerous studies have shown that during maturation from fetal to adult testis, SCs undergo a process of differentiation whereby the expression of some markers is maintained, while other markers disappear and others are acquired.
The comparison between the expression of cellular markers in normal and neoplastic testes may make clearer the temporal sequence of markers appearance in the dog, important for understanding the mechanisms underlying their re-expression in disorders of the adult testis. The analogies observed between canine and human species might clarify if the dog could be considered a good animal model for human testicular cancer.
To evaluate the expression of Sertoli cell markers in normal and neoplastic canine testes, formalin fixed and paraffin embedded sections of testes from 2 fetuses, 15 newborns (from 0 to 20 days old), 5 puppies (from 43 days to 6 months old), 14 adult dogs and 21 canine SCTs (20 benign and 1 metastatic malignant SCT) were processed for histology and immunohistochemistry (ABC method) with antibodies against vimentin (1:1000), CKAE1/AE3 (1:3000), desmin (1:300), INH α (1:40) and AMH (1:30000).
Histologically, 13/20 benign SCTs were classified as classical SCTs while 7/20, showing intracytoplasmic vacuoles, were considered “lipid rich”. The malignant SCT, largely necrotic, was composed of large cystic tubules lined by numerous layers of poorly-differentiated neoplastic SCs characterized by marked anisocytosis and anisokaryosis.
The mitotic index of the SCTs ranged from 0 to1.6, with the highest value recorded in the malignant SCT.
Immunohistologically, vimentin was always expressed by normal SCs from puppies and adults and by all SCTs taken in consideration. On the other hand, only 1 foetus and 4 newborns were positive immunolabelled.
INH α was present exclusively in fetal and neonatal testes and in 13/21 SCTs, while desmin and CKAE1/AE3 were never present within normal testes but exclusively expressed by 7/21 and 5/21 SCTs, respectively. Finally, AMH was always expressed by foetal and neonatal testes, by the youngest puppies (43-45 days old) and by all the SCTs taken in consideration.
We speculate that the expression of CKAE1/AE3, INH α, desmin and AMH observed in SCTs and not in mature testes may be a manifestation of de-differentiation. In fact, neoplastic SCs re-express foetal and neonatal makers, normally lost with maturation, demonstrating an immature phenotype. The re-expression of immunophenotypical markers of immaturity has been already described in several testicular pathologic conditions and the identification of the mechanisms via which this “cellular reversal maturation” occurs should be further investigated.
Regarding ovarian canine epithelial tumours, given the variety of histological features that characterize this lesion and that can cause diagnostic difficulties or misinterpretation, the aim of this study was to investigate the reliability of the marker HBME-1 in canine normal ovaries, granulosa cell tumors, and epithelial ovarian neoplasms to determine whether this marker could be included in an immunohistochemical panel for differential diagnoses of canine ovarian tumors.
Samples were obtained from 4 normal ovaries, 10 granulosa cell tumors, and 18 epithelial ovarian tumors. After formalin fixation and paraffin embedding, tissue sections were stained with hematoxylin and eosin and probed immunohistochemically for the HBME-1 marker. Granulosa cells and related tumors were consistently negative for HBME-1. Normal ovarian surface epithelium and 17 out of 18 ovarian epithelial tumors were positive for HBME-1. The results suggested that HBME-1 would be a useful marker for the differential diagnosis of ovarian tumors in the dog.
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Zabida, Omer Saleh. "The effect of methamphetamine on the blood-testis barrier." University of the Western Cape, 2018. http://hdl.handle.net/11394/6775.
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>Magister Scientiae - MScIntroduction The blood-testis barrier (BTB) is formed by tight junctions between adjacent Sertoli cells. The barrier formed by these tight junction helps to create a specialized environment for spermatogenesis and provide an immunological barrier to protect developing germ cells. Methamphetamine (Meth) is known as neurotoxin however, its effects on the male reproductive system, especially on Sertoli cells and, the BTB are not well established. Therefore, this study aimed to determine the effects of Meth on the TM4 mouse testis Sertoli cell line and on the integrity of the BTB permeability. Materials and Methods This study investigated the effect of selected concentrations of Meth (0.1 μM, 1 μM, 10 μM, 20 μM and 100 μM) on TM4 mouse testis Sertoli cell line for 24 until 96 hours, using two treatments: an “acute” study (24 hrs exposure) and a “chronic” study, where treatment occurred on a daily basis over 96 hrs. The following parameters were investigated: viability, cell proliferation, mitochondrial activity, monolayer permeability.
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Plotton, Ingrid. "Implication de facteurs sertoliens dans la physiologie et la physiopathologie de la spermatogenèse : étude des ARNm de la clusterine et du stem cell factor." Lyon 1, 2006. http://www.theses.fr/2006LYO10079.
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La cellule de Sertoli a un rôle fondamental dans le contrôle des différentes étapes de la spermatogenèse. Le stem cell factor et la clusterine sont produits par la cellule de Sertoli. Nous avons mesuré par RT-PCR quantitative leurs ARNm au cours du développement et après avoir induit un trouble de la spermatogenèse chez le rat et dans des biopsies testiculaires de patients présentant une azoospermie soit obstructive (groupe contrôle) soit par trouble de la spermatogenèse. Les résultats suggèrent que la différenciation des cellules de Sertoli est altérée dans les troubles de la spermatogenèse constitutifs ou idiopathiques chez l'homme, contrairement aux troubles de la spermatogenèse acquis chez l'homme ou induit chez le rat adulteThe sertoli cell is essential in the control of the different stages in the spermatogenisis. Stem cell factor and clusterin are product by the Sertoli Cell. We mesured by quantitative RT PCR their mRNA during the developpement and after induction of spermatogenisis failure in the rat and in the testicular biopsies from men with either obstructive azoospermia (control group) or spermatogenic failure. The results suggest that Sertoli cell differentiation is altered in cases of constitutive or idiopathic spermatogenic failure in human, in opposite of acquired spermatogenesic failure in human or induced spermatogenic failure in adult rat
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Taylor, Jacqueline Susan. "The Effect of Pyrethroid Compounds on the Expression of Estrogen Receptors in Mouse Sertoli Cells and Implications for Male Infertility." Thesis, University of Canterbury. Biological Sciences, 2006. http://hdl.handle.net/10092/1482.
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Male fertility is largely controlled by the hypothalamic-pituitary axis, a careful balance between stimulating and suppressing gene expression and the secretion of hormones. The critical factors for male fertility have in the past been thought to be limited to testosterone and the gonadotropins. Estrogen has only recently been demonstrated to be both a crucial requirement for fertility and a cause of infertility. Reports in the early 1990s demonstrated a decrease in mean sperm counts over the last 50 years. A hypothesis for this observation is the increase of xenoestrogens in the environment that are able to mimic and potential disrupt the natural estrogens involvement in fertility. Although the mechanisms of estrogens involvement are not yet defined, the Sertoli cells are a potential sites of action as they possess receptors for the hormone and are able to locally produce it. Sertoli cells both act to protect and provide for the male germ cells and the developing spermatozoa. Pyrethroids are common synthetic insecticides of which some have previously shown estrogenic activity. Therefore this investigation examined the effects of pyrethoids, whose estrogenicity was confirmed via the yeast assay, on the estrogen receptor expression in mouse Sertoli cells as a model for general effects of estrogenic chemicals on male fertility. The results first confirmed the estrogenicity of some pyrethroids and these pyrethroids when exposed to mouse Sertoli cells effected estrogen receptor mRNA expression however in a different way to the natural ligand 17β-estradiol.
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Dias, Tânia Isabel Rodrigues Amaral. "Evaluation of cell death markers and reproductive parameters in models of diabetes mellitus." Master's thesis, Universidade da Beira Interior, 2012. http://hdl.handle.net/10400.6/1121.
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Diabetes mellitus (DM) represents one of the greatest threats to modern global health and its incidence is rapidly rising worldwide. It describes a metabolic disorder characterized by hyperglycaemia resulting from defective insulin secretion, resistance to insulin action, or both. There are two types of DM, type-1 DM (T1DM) and type-2 DM (T2DM), both associated with male infertility. T1DM is associated with insulin deprivation and although the overall in vivo effects in the reproductive function is well known, there is a lack of studies concerning about the insulin control over the physiological functions of cells from the reproductive system. Importantly, the in vivo studies are often focused after the disease is fully establish, but it is known that a prediabetic state, which is characterized by insulin resistance, precedes the development of DM, especially T2DM. With our work, we aimed to further investigate the association between DM and male infertility by analyzing several apoptotic markers and reproductive parameters. To do so, we simulated type 1 DM in cultured rat Sertoli cells and analyzed the mRNA and protein expression levels of several cellular markers involved in the mitochondrial apoptotic pathway. We also developed an animal model of prediabetes to evaluate the effect of this pathological state in the reproductive parameters as well as in the mitochondrial apoptotic pathway.
Our results lead us to suggest that insulin interferes with the interaction between pro and anti-apoptotic proteins. As the interaction of these proteins decide the cell fate and exert a strict control over the apoptotic signaling, insulin has a key role in the maintenance of the spermatogenesis. Our rat model shared many of the clinical and metabolic characteristics of the prediabetic state observed in humans such as glucose resistance and progression from normoglycaemia/normoinsulinemia to moderate hyperglycaemia/hyperinsulinemia due to food intake. This prediabetic state induced important alterations in cauda epididymis spermatozoa morphology showing that these animals may develop subfertility or fertility problems. The apoptotic signalling in cauda epididymis spermatozoa of the high-energy (HED) fed animals presented lower Bax mRNA levels and lower cytC protein levels although the apoptotic endpoint, caspase-3 activity, was not altered. This suggests that the apoptotic process may be controlled by other mechanisms rather than the mitochondrial pro-apoptotic proteins, such as the anti-apoptotic cellular systems.
Those two experimental models led us to conclude that the subfertility/infertility problems caused by DM may be mediated by insulin, which has an important effect in the regulation of the interaction between pro and anti-apoptotic proteins and special attention must be taken in the prediabetic state where crucial alterations in rat spermatozoa occurred. Due to the rising incidence and associated complications of DM and male infertility it is crucial to further investigate in these two systems to isolate possible mechanisms and evaluate the overall effects as a strategy to develop possible therapeutics.A diabetes mellitus (DM) representa uma das maiores ameaças à saúde global moderna e a sua incidência está a aumentar rapidamente em todo o mundo. Esta doença consiste numa desordem metabólica, caracterizada por hiperglicemia, resultante de uma secreção defeituosa de insulina, resistência à ação da insulina ou ambas. Existem dois tipos de DM, tipo 1 e tipo 2, ambas as condições relacionadas com a infertilidade masculina. A DM tipo 1 está associada com a privação de insulina e embora os efeitos globais in vivo na função reprodutiva sejam bem conhecidos, há uma falta de estudos relativamente ao controlo da insulina sobre as funções fisiológicas das células do sistema reprodutivo. Sobretudo, os estudos in vivo são frequentemente feitos depois de a doença estar completamente estabelecida, mas sabe-se que há um estado pré-diabético, caracterizado por resistência à insulina, que antecede o desenvolvimento da DM, especialmente da DM tipo 2. Com o nosso trabalho, pretendemos investigar mais profundamente a ligação entre a DM e a infertilidade masculina, através da análise de vários marcadores de morte celular e de parâmetros reprodutivos. Para isso, simulámos o estado de DM tipo 1 humano em células de Sertoli de rato e analisámos os níveis de expressão de mRNA e proteína de vários marcadores de morte celular envolvidos na via mitocondrial. Por outro lado, também desenvolvemos um modelo animal de pré-diabetes de modo a reproduzir esta condição patológica e avaliar alterações produzidas nos parâmetros reprodutivos, assim como nos níveis de expressão de mRNA e proteína de marcadores de morte celular envolvidos na via mitocondrial. Os resultados obtidos levam-nos a sugerir que a insulina interfere com a interação entre proteínas pró- e anti-apoptóticas. Uma vez que esta interação pode decidir o destino celular e exercer um controlo rigoroso sobre a sinalização apoptótica, a insulina terá um papel chave na manutenção da espermatogénese. O modelo de rato utilizado compartilhava muitas das características clínicas e metabólicas do estado pré-diabético observado em humanos, tais como resistência à glucose e a progressão de normoglicemia/normoinsulinemia para hiperglicemia/hiperinsulinemia moderada devido à ingestão de alimentos. Este estado prédiabético induziu alterações significativas na morfologia dos espermatozoides da cauda do epidídimo, mostrando que esses animais poderão desenvolver problemas de subfertilidade ou de fertilidade. Na sinalização apoptótica na cauda do epidídimo, os animais sujeitos à dieta de alta energia (HED) apresentaram níveis mais baixos de mRNA Bax e da proteína citocromo C, embora a avaliação quantitativa do endpoint apoptótico, a atividade da caspase-3, não tenha evidenciado quaisquer alterações entre a situação HED e controle. Isto sugere que o processo apoptótico pode ser controlado por outros mecanismos que não somente os proteicos pró-apoptóticos mitocondriais, como por exemplo os sistemas anti-apoptóticos celulares. Os resultados obtidos com estes dois modelos experimentais levam-nos a concluir que os problemas de subfertilidade/infertilidade causados pela DM podem ser mediados pela insulina, que tem um efeito importante na regulação da interação entre proteínas pró e antiapoptóticas e, por esse motivo, deverá ser dedicada uma atenção especial às disfunções ocorridas no estado pré-diabético, onde observámos alterações cruciais na morfologia de espermatozoides epididimais de ratos. Devido à crescente incidência da DM e às complicações associadas ao nível da infertilidade masculina é fundamental aprofundar o conhecimento nestes dois sistemas, de modo a isolar possíveis mecanismos envolvidos e a avaliar os efeitos globais, como uma estratégia para desenvolver possíveis abordagens terapêuticas.
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Gerber, Jonathan [Verfasser]. "Investigation of the Junctional Complex at the Blood-Testis Barrier in SCCx43KO mice and Establishment and Functional Characterization of a Murine Sertoli Cell Line Deficient of Connexin43 / Jonathan Gerber." Hannover : Bibliothek der Tierärztlichen Hochschule Hannover, 2015. http://d-nb.info/1073848302/34.
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Gawriluk, Thomas R. "Targeted Knockout of Beclin-1 Reveals an Essential Function in Ovary and Testis." UKnowledge, 2014. http://uknowledge.uky.edu/biology_etds/19.
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An estimated 12% of couples worldwide are infertile. The contributing factor is approximately equal between men and women with nearly 25% diagnosed as idiopathic. Despite the increasing numbers of couples seeking assistance from infertility clinics, few molecular mechanisms have been identified for treatment. Autophagy is an evolutionarily conserved cellular process for bulk degradation and recycling of cytosolic components through the lysosome to maintain homeostasis. Several studies have observed increased levels of autophagy during ovarian folliculogenesis and gonadal steroidogenesis; however, no genetic studies to determine the significance of autophagy exist.
To investigate the function of autophagy in the ovary and testis, a directed genetic knockout approach was used to independently knockout two key autophagy genes, Becn1 and Atg7. Chapter 2 reports that deficiency of Becn1 results in 56% fewer primordial follicles at postnatal day 1. In addition, Atg7 knockout mice do not have identifiable primordial follicles, suggesting that autophagy is necessary for survival of female germ cells during embryogenesis. Chapter 3 presents that Becn1 is necessary to sustain pregnancy and the deficiency of Becn1 in granulosa cells is a novel genetic model to study preterm labor due to impaired corpora lutea function. The results indicate that Becn1 is necessary for lipid droplet formation and subsequent progesterone production in luteal cells. In contrast, Atg7 is not necessary and deficiency results in overproduction of progesterone throughout pregnancy, suggesting that the defect in Becn1 conditional knockout mice is additional to autophagy. Chapter 4 presents that Sertoli cell expression of Becn1 is required for spermatogenesis after 8 weeks of age. Beyond 9-weeks-old, Becn1 conditional knockout mice are unable to sire a litter due to a failure of spermatogenesis and a Sertoli-cell-only phenotype in a majority of the seminiferous tubules. Atg7 was also identified as a necessary factor for spermatogenesis beyond 26-weeks-old. Together the data presented in Chapter 4 suggests that autophagy is necessary for adult Sertoli cell function. Primarily, this dissertation presents data from the first functional studies on autophagy in the reproductive tract. The results demonstrate an understanding of the functional significance for Becn1 and Atg7 in both the ovary and testis.
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Yance, Viviane dos Reis Vieira. "Contribuição das características clínicas, hormonais e radiológicas para o diagnóstico diferencial dos tumores de ovário produtores de andrógenos e hipertecose do estroma ovariano em mulheres na pós-menopausa." Universidade de São Paulo, 2016. http://www.teses.usp.br/teses/disponiveis/5/5135/tde-24102016-143113/.
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Introdução: Hiperandrogenemia associada a sinais clínicos de virilização na mulher após a menopausa é uma condição rara e pouco estudada. Os tumores ovarianos secretores de andrógenos (TOSA) e a hipertecose do estroma ovariano (HPT) são as etiologias mais frequentes de hiperandrogenismo nesta faixa etária. A diferenciação entre estas duas condições é difícil, pois as manifestações clínicas são semelhantes e caracterizadas por hirsutismo, alopecia androgênica, clitoromegalia, hipertrofia muscular e agravamento da voz. O perfil hormonal das mulheres pós-menopausadas com TOSA e HPT pode não ser um parâmetro ideal para discriminar estas duas condições. Além disso, os estudos de imagem podem não caracterizar com precisão estas lesões ovarianas. Devido às dificuldades, em estabelecer o diagnóstico diferencial entre TOSA e HPT, a ooforectomia bilateral é a terapêutica indicada para as mulheres menopausadas com diagnóstico de hiperandrogenismo de origem ovariana; embora na HPT o tratamento clínico com análogo do hormônio liberador de gonadotrofinas (aGnRH) possa ser uma opção terapêutica eficaz. Objetivos: O nosso objetivo foi avaliar a contribuição das características clínicas, do perfil hormonal e dos exames radiológicos para o diagnóstico diferencial entre TOSA e HPT em mulheres na pós-menopausa. Métodos: Trinta e quatro mulheres pós-menopausadas, na faixa etária de 52 a 80 anos de idade, que foram encaminhadas à Unidade de Endocrinologia do Hospital das Clinicas da Faculdade de Medicina da Universidade de São Paulo, entre 1999 e 2013 por hiperandrogenismo clínico e com diagnóstico histológico de TOSA (13 mulheres) e HPT (21 mulheres) foram avaliadas retrospectivamente. Os diagnósticos histológicos foram revisados e confirmados por um único patologista com experiência em patologia ginecológica. Os dados clínicos de hiperandrogenismo, o perfil hormonal (T, E2, LH, FSH) e as imagens radiológicas pélvicas (Ultrassom transvaginal e Ressonância Magnética, RM) foram obtidos a partir da revisão de prontuários médicos. Resultados: Em relação aos dados da história clínica, não houve diferença significativa entre os dois grupos de pacientes para nenhuma das variáveis clínicas analisadas, exceto para o número de gestações, que foi significantemente maior no grupo com TOSA. Os sinais clínicos de hiperandrogenismo, especialmente agravamento da voz (p < 0,001) e hipertrofia muscular (p = 0,01), foram mais prevalentes no grupo de pacientes com TOSA do que o grupo de HPT. Embora na análise dos parâmetros hormonais, os pacientes do grupo com TOSA tenham apresentado níveis mais elevados de T e E2 e níveis mais baixos de gonadotrofinas (p < 0,01 e p <0,01, respectivamente) do que o grupo de pacientes com HPT, uma grande sobreposição nos níveis hormonais foi observada entre os pacientes dos dois grupos. A RM de pelve apresentou uma boa acurácia para diferenciar os TOSAs da HPT em mulheres pós-menopausadas com hiperandrogenismo. Conclusão: Neste grupo de pacientes, as características que mais contribuíram para o diagnóstico diferencial entre TOSA e HPT foram o agravamento da voz e a hipertrofia muscular, os níveis séricos de testosterona e gonadotrofinas e a presença de nódulo ovariano na RM de pelve. Embora a associação das características clínicas, hormonais e radiológicas contribua para a elaboração de uma hipótese diagnóstica fundamentada, a análise histopatológica continua a ser o padrão ouro para o diagnóstico diferencial de hiperandrogenismo de origem ovariana em mulheres na pós-menopausaIntroduction: The presence of virilizing signs associated to high serum of androgen levels in postmenopausal women is a rare and poorly understood condition. Virilizing ovarian tumors (VOT) and ovarian stromal hyperthecosis (OH) are the most common hyperandrogenism etiologies in the postmenopausal women. The differential diagnosis between the two conditions is often difficult, because they present similar clinical features such as hirsutism, androgenic alopecia, clitoromegaly, muscle hypertrophy and deepening of the voice. The hormonal profile of postmenopausal women with VOT and OH may not be the optimal discriminating factor between these two conditions. Moreover, imaging may not accurately characterize these ovarian lesions. Due to the difficulties in establishing the differential diagnosis between VOT and OH, bilateral oophorectomy is the treatment of choice in postmenopausal women with hyperandrogenism of ovarian origin. However, the treatment with gonadotropin-releasing hormone analogue (GnRHa) might be an effective therapy in women with OH. Objectives: Our aim was to evaluate the contribution of clinical features, hormonal profile and radiological studies in the differential diagnosis between VOT and OH in postmenopausal women. Methods: Thirty-four postmenopausal women ranging from 52 to 80 years of age with clinical hyperandrogenism referred to the Endocrinology Unity of Hospital das Clinicas da Faculdade de Medicina da Universidade de São Paulo, between 1999 and 2013, with diagnosis of VOT (13 women) and OH (21 women) were evaluated retrospectively. Histological diagnoses were reviewed and confirmed by a single pathologist with expertise in gynecologic pathology. Clinical hyperandrogenism data, hormonal status (T, E2, LH, FSH) and the pelvic images (Transvaginal sonography and Magnetic Resonance Image- MRI) findings were obtained from medical records. Results: No clinical data evaluated in the study was significantly different between the two groups of patients. A higher number of pregnancies in the VOT group was observed, which was statistically different from the OH group. The clinical signs of hyperandrogenism, especially deepening of the voice (p < 0.001) and muscle hypertrophy (p = 0.01), were more prevalent in the VOT\'s than OH\'s group. Although, the VOT\'s group showed higher T and E2 levels and lower gonadotropins levels than the OH\'s group (p < 0.01 and p < 0.01, respectively), a great overlap in the hormone levels occur between VOT and OH patients. Pelvic MRI presented a good accuracy to differentiate these two conditions in hyperandrogenic postmenopausal women. Conclusion: In this group of patients, the main features to the differential diagnosis between VOT and OH were deepening of the voice and muscle hypertrophy, serum levels of testosterone and gonadotropins and presence of ovarian nodule in the pelvic MRI. Although the association of clinical, hormonal and radiological features contributes to the differential diagnosis between these two conditions, histopathological analysis remains the gold standard for the differential diagnosis of ovarian hyperandrogenism in post menopausal women
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Erasmus, Nicolete. "Investigations on the in vitro effects of aqueous Eurycoma longifolia Jack extract on male reproductive functions." Thesis, University of Western Cape, 2012. http://etd.uwc.ac.za/index.php?module=etd&action=viewtitle&id=gen8Srv25Nme4_2238_1375971626.
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Eurycoma longifolia (Tongkat Ali
TA) is a Malaysian shrub used to treat various illnesses including male infertility. Considering that TA is also used to improve male fertility and no report
regarding its safety has been published, this study investigated the effects of a patented, aqueous TA extract on various sperm and testicular functions. Materials and Methods This study
encompasses two parts (part 1: on spermatozoa
part 2: on TM3-Leydig and TM4-Sertoli cells). Part 1: Semen samples of 27 patients and 13 fertile donors were divided into two groups,
washed and swim-up prepared spermatozoa, and incubated with different concentrations of TA (1, 10, 20, 100, 2000 &mu
g/ml) for 1 hour at 37°
C. A sample without addition of TA served as control. After incubation with TA,
the following parameters were evaluated: viability (Eosin-Nigrosin test), total and progressive motility (CASA), acrosome reaction (triple stain technique), sperm production of reactive oxygen
species (ROS
dihydroethidium test
DHE), sperm DNA fragmentation (TUNEL assay) and mitochondrial membrane potential (&Delta
&psi
m) (Depsipher kit). Part 2: TM3-Leydig and TM4-Sertoli cells
incubated with different concentrations of TA (0.4, 0.8, 1.6, 3.125, 6.25, 12.5, 25, 50 &mu
g/ml) and control (without extract) for 48 and 96 hours. After incubation with TA, the following parameters were
evaluated: viability (XTT), cell proliferation (protein assay), testosterone (testosterone ELISA test) and pyruvate (pyruvate assay). Results Part 1: For washed spermatozoa, significant
dose-dependent trends were found
for viability, total motility, acrosome reaction and sperm ROS production. However, these trends were only significant if the highest concentrations were included in the calculation. In the swim-up spermatozoa, ROS production of spermatozoa showed a biphasic relationship with its lowest percentage at 10 &mu
g/ml, yet, no significance could be
observed (P=0.9505). No influence of TA could be observed for sperm DNA fragmentation nor &Delta
&psi
m.
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Vija, Lavinia. "Androgen Signaling in Sertoli Cells." Thesis, Paris 11, 2014. http://www.theses.fr/2014PA11T031/document.
Full textAbstract:
Les cellules de Sertoli (CS) jouent des rôles essentiels pour la régulation de la spermatogenèse, via la signalisation modulée par le récepteur aux androgènes (RA). Les objectifs de ce travail ont été d’identifier les mécanismes moléculaires de la régulation androgénique et des rôles des partenaires moléculaires dans la régulation androgénique des cellules de Sertoli pendant le développement testiculaire, du fœtus à l’âge adulte, pendant différentes stades du développement. Nous avons caractérisé et étudié une nouvelle lignée immortalisée, mature, de CS, ST38c, présentant une expression substantielle de RA endogène, une activation transcriptionnelle du RA induite par les androgènes, ainsi qu’une régulation et une stabilisation de la protéine RA par des mécanismes post traductionnels. Ce modèle a été utilisé afin de tester l’hypothèse que la suppression de l’hormone antimüllérienne (AMH) est modulée directement par les androgènes, via la RA, dans les cellules matures de Sertoli.En parallèle, nous avons testé l’hypothèse que la résistance physiologique aux androgènes du nouveau-né est liée à l’expression différentielle de quelques corégulateurs du RA. Ainsi, nous avons analysé l’expression et la contribution de deux corégulateurs du RA (SRC-2 et HBO1) pendant l’ontogenèse testiculaire humaine et pour des pathologies liées au disfonctionnement de l’action des androgènes ou du RA (syndromes d’insensibilité aux androgènes, hypogonadisme hypogonadotrophique congénital).Nous avons démontré, après des essais de transfection in vitro, que SRC-2 est un coactivateur, alors que HBO1 est in corepresseur du RA dans un modèle de cellules de Sertoli. Nous avons cartographié l’expression testiculaire humaine de SRC-2 et HBO1, pendant différentes stades du développement pré et postnatal et nous avons démontré que SRC-2 présente une expression stable, contrastant avec le profil d’expression différentielle, progressive avec l’âge de RA dans la cellule de Sertoli, suggérant que l’expression du SRC-2 est indépendante de la signalisation androgènique. Par contre, HBO1 et le RA présentent un profil de maturation et d’expression temporelle corrélé, suggérant que l’expression du HBO1 serait liée à une signalisation du RA fonctionnelle dans les cellules de Sertoli. Nous avons aussi démontré que l’expression du HBO1 est induite par les androgènes en présence du RA. Contrairement au SRC-2, HBO1 est non seulement exprimé dans les cellules de Sertoli, mais aussi dans les spermatogonies, pouvant représenter un marqueur potentiellement intéressant des cellules germinales.Enfin, nous avons aussi étudié l’immuno-expression testiculaire de RA et AMH chez des patients post pubères avec un syndrome de déficit en 5α-réductase type 2, et insensibilité minime aux androgènes (MAIS), afin d’étudier la contribution des androgènes (testostérone versus dihydrotéstosterone) pour la spérmatogenèse, la répression de l’AMH ainsi que pour mieux comprendre les perspectives de fertilité chez ces patientsSertoli cells (SC) have essential roles in the androgen regulation of spermatogenesis, via the androgen receptor (AR)-mediated signaling. This work aimed at identifying the molecular mechanisms related to the androgenic regulation of the AR and its molecular partners in Sertoli cells during different testicular developmental stages. We first characterized and studied a novel murine mature immortalized Sertoli cell line, called ST38c, which harbors substantial expression of endogenous AR, conserves its androgen-dependent transcriptional activation and exhibits agonist-dependent transcriptional and posttranslational regulation, as well as posttranslational stability.We used this cellular model in order to test the hypothesis that anti Müllerian Hormone (AMH) suppression in mature Sertoli cells would be directly androgen and AR mediated.Next, we hypothesized that the physiological androgen resistance in the neonate would also be related to the differential expression of several AR coregulators. Therefore, we analysed the differential expression and contribution of two AR co-regulators (SRC-2 and HBO1) during human testicular ontogeny, as well as in pathologies associated with androgen action or AR impairment (such as androgen insensitivity syndromes, congenital hypogonadotropic hypogonadism).Using in vitro transfection assays, we showed that SRC-2 is an AR coactivator while HBO1 is an AR corepressor in Sertoli cell models. We provided the cartography of SRC-2 and HBO1 expression during human testicular postnatal different stages and showed that SRC-2 presented a stable expression contrasting with the progressive evolution profile of the AR signaling, suggesting that Sertoli SRC-2 expression was independent of the androgen signaling. Interestingly, HBO1 and AR presented a temporal and positively correlated maturation profile, suggesting that HBO1 expression would be related to a functional AR signaling in the Sertoli cell. Moreover HBO1 expression is induced by androgens in the presence of the AR. Unlike SRC-2, HBO1 is not only expressed in Sertoli cells, but also in spermatogonia, being an interesting germ cell marker.Finally, we also studied AR and AMH immunoexpression in posptubertal cases of 5-α reductase type 2 deficiency and minimal androgen receptor resistance, in respect with the spermatogenesis status of seminiferous tubules, and androgen induced AMH suppression in order to assess the differential contribution of testosterone versus dihydrotestosterone and gather more information about the fertility perspectives in this particular pathologies
39
Tréfier, Aurelie. "Le traductome induit par le récepteur FSH et l'implication des B-arrestines dans le contrôle de la traduction des ARNm 5' TOP." Thesis, Tours, 2017. http://www.theses.fr/2017TOUR4040.
Full textAbstract:
La FSH est une des hormones clés qui régule la reproduction chez les mammifères. Chez le mâle, elle cible les cellules de Sertoli, qui expriment le RFSH. La cellule de Sertoli a un rôle trophique important pour le bon développement de la spermatogenèse. Dans cette thèse, nous avons établi le premier traductome, c’est-à-dire l’ensemble des ARNm en cours de traduction, dépendant du RFSH. La traduction de certains ARNm significativement modulés par la FSH exercerait un rétrocontrôle sur la signalisation FSH-dépendante. L’analyse du protéome nous a permis de valider ce traductome au niveau systémique. Nous avons également démontré l’implication des β-arrestines dans la traduction d’ARNm dépendante de la FSH. Les β-arrestines forment un assemblage moléculaire avec le module de traduction p70S6K/rpS6. Cet assemblage est impliqué dans la traduction des ARNm 5’TOP, qui encodent la machinerie traductionnelle. C’est l’activation FSHdépendante des protéines G qui promeut l’activation de p70S6K au sein du module β-arrestines/ p70S6K/ rpS6. Ce travail constitue une nouvelle avancée sur les mécanismes grâce auxquels la FSH exerce sa fonction biologique de dans ses cellules-cibles naturelles de la gonade mâleFSH is one of the key hormones that regulate the reproductive function in mammals. In the male, FSH targets Sertoli cells, which express the FSHR. Sertoli cells play an important trophic role in the development of spermatogenesis. Here, we have provided the first FSHR-induced translatome, that encompasses all the mRNA being actively translated. The translation of some mRNAs significantly modulated by FSH may exert a feedback control on FSH-dependent signaling. The analysis of the proteome has validated the FSHR translatome at the systems level. We also demonstrated the involvement of β-arrestins in the FSH-stimulated translation of mRNA. β-arrestins form a molecular assembly with the p70S6K / rpS6 translation module. This molecular assembly is involved in the translation of 5'TOP mRNA, which encode proteins of the translational machinery. FSH-activated G proteins leads to p70S6K activation within the β-arrestins/ p70S6K/ rpS6 module. This work provides new advance on the mechanisms whereby FSH exerts its biological function in its natural target cells of the male gonad
40
Kadkhodamohammadi, Abdolrahim. "Counting Sertoli Cells in Thin Testicular Tissue." Thesis, Uppsala universitet, Institutionen för informationsteknologi, 2012. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-175237.
Full textAbstract:
This master thesis develops a novel system to model the tubular structure in thin sections of testicular tissue and count the Sertoli cells. A three-phase method is proposed to model the tubular structure in microscopic images of the tissue, the model is deployed to detect the cells. In the first phase, the germ-mass, which represents the inside layer of tubules, are detected. All cells are detected by radial symmetry transform and then the graph cut algorithm is used to separate the germ cells. Each region covered by a compact set of germ cells is considered as the germ-mass. In the second phase, all bright areas in the image are detected and used to adjust the germ-mass regions. In the last phase, all edges that are line-like are identified and straight lines are fitted to the edges. The lines are later connected to compensate for the broken parts of the tubules' boundaries. The closest cells to the germ-mass are chosen as the Sertoli cell candidates. The approximate boundary of tubules and the angle between the candidate cells are used to detect the Sertoli cells. Our experimental results show that our system is able to detect the tubule and the Sertoli cells with reasonable accuracy. If the method can not find enough edges to approximate the tubule's boundary, detecting Sertoli cells is complicated; the system can report those situations to the experts. Since we use the symmetry attribute of the cells to detect them, the method is quite robust against noise, artifacts, and non-uniform illumination. The method is able to capture all tubules, even tubules that do not have any bright region in the middle (lumen). To the best of my knowledge, no one has proposed a method to model tubular structure without lumen. The border approximation method can work well even for tubules that are partially in the image. It should be mentioned that the proposed method could be applied to model any tubular structure with one or more cells types.
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Amlani, Shahira R. "Intermediate filaments in Sertoli cells : distribution and possible function." Thesis, University of British Columbia, 1987. http://hdl.handle.net/2429/27790.
Full textAbstract:
The distribution of vimentin intermediate filaments in Sertoli cells during spermatogenesis was observed with immunofluorescence, and was confirmed with electron microscopy.
The distribution of vimentin filaments within Sertoli cells changes with changes in the germ cell population. At stages where elongate spermatids reside in crypts located deep within the seminiferous epithelium, groups of eight to twelve intermediate filaments were consistently found at the convex surface of the spermatid heads. Here the filaments are in close association with ectoplasmic specializations. At later stages of spermatogenesis, intermediate filaments are not found in crypt areas. Because of their association with particular stages of developing germ cells, intermediate filaments in Sertoli cells may be involved in the attachment and positioning of developing germ cells within the seminiferous epithelium.
Intermedate filaments in general are thought to be involved in anchoring the nucleus and cytoplasmic organelles within a cell. In order to test this hypothesis, acrylamide, a specific perturbant of intermediate filaments in vitro, was perfused through rat and ground squirrel testes in order to perturbate the intermediate filament system within Sertoli cells. No effects of acrylamide on intermediate filaments were observed in vivo at either the light microscopic or ultrastructural level. However, toxic effects were observed upon treatment with high concentrations of acrylamide, indicating that Sertoli cells and associated germ cells were indeed exposed to the perturbant.
Based on these studies, one can conclude that: (1) vimentin filaments in Sertoli cells change their distribution during spermatogenesis; (2) vimentin filaments are closely associated with specific stages of developing germ cells, and may be involved in the positioning and attachment of these cells to Sertoli cells within the seminiferous epithelium, and (3) acrylamide has no effect on vimentin filaments in Sertoli cells in vivo.Medicine, Faculty of
Graduate
42
Hayes, Marianne Kay. "Bovine testicular cells in vitro: establishment of primary cultures and investigations of secretory functions : a thesis presented for the degree of Doctor of Philosophy in the University of Adelaide." Title page, contents and summary only, 1986. http://web4.library.adelaide.edu.au/theses/09PH/09phh4178.pdf.
Full textAbstract:
Includes bibliographical references (leaves 98-128). Investigates protein secretion by bovine Sertoli cells in culture. Cultures were obtained from bulls at all stages of post natal development and from sexually mature animals.
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Pfeiffer, David Carl. "Actin-related intercellular adhesion junctions in vertebrate Sertoli cells." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1998. http://www.collectionscanada.ca/obj/s4/f2/dsk3/ftp05/nq27225.pdf.
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Jesus, Tito Miguel Boléo Teles de. "Aquaporins as molecular partners of CFTR in Sertoli Cells." Master's thesis, Universidade da Beira Interior, 2013. http://hdl.handle.net/10400.6/1621.
Full textAbstract:
The establishment of the adequate ionic and water environment within the lumen of
the seminiferous tubules (SFT), with the secretion of its numerous components, is vital for
the normal occurrence of spermatogenesis. Specific transporters for HCO3
-, a mobile
physiological buffer that plays a crucial role in the maintenance of intracellular and
extracellular pH, have been described in the SFTs. Additionally, water transporters known as
Aquaporins have been identified in the SFT, being most probably involved in the secretion of
the luminal fluid. Within the SFT, Sertoli cells (SCs) play a crucial role in the establishment of
the luminal fluid. Thus, it is imperative to unravel HCO3
- and water transport dynamics in SCs
to identify and counteract possible alterations related with reduced male fertility caused by
pathological conditions associated with altered water and HCO3
- transport, such as cystic
fibrosis (CF).
So the first objective of this work was to evaluate mRNA and protein expression of the
cystic fibrosis transmembrane conductance regulator (CFTR) in SCs, by RT-PCR and
immunoblot, respectively, using primary cultures obtained from 20-day old rats. The second
objective was to identify the expression of specific aquaporin (AQP) isoforms (AQP0-AQP9),
using a similar approach. Finally, the third objective was to investigate the possible physical
interaction between CFTR and specific AQP isoforms in SCs, using the co-immunoprecipitation
technique.
We were able to detect in cultured SCs both, the expression of mRNA transcripts and
of proteins of the bicarbonate transporter CFTR and AQP isoforms 4, 8 and 9. The presence of
the proteins AQP6 and AQP7 was also detected although no mRNA transcript correspondent to
these AQPs isoforms was observed. Furthermore, in the present study, we observed a direct
interaction between AQP4 and CFTR, using the co-immunoprecipitation technique.
Thus, in the SFTs, ion and water secretion, driven by specific SCs membrane
transporters, is an important step that helps to control the final osmolarity and fluidity of the
luminal content. Our results allowed us to identify the expression of various AQP isoforms in
rat SCs, particularly AQP4, AQP8 and AQP9, highlighting the importance of water transport in
these cells and suggesting that the different AQPs may serve more than one particular
function. Additionally, our results indicate that AQP4 physically interacts with CFTR,
evidencing a possible role for this HCO3
- transporter in the regulation of AQPs and water
homeodynamics in rat SCs. Defective water transport might be the leading cause of the
obstructive pathologies, followed by atrophy and infertility, which are observed in men with
CF. So it is possible that disruption of a functional complex involving AQP4 and CFTR might
contribute to the pathogenesis of male infertility/subfertility in CF.O estabelecimento da composição adequada no lúmen dos túbulos seminíferos (SFTs), com a secreção dos vários componentes, é vital para a normal ocorrência da espermatogênese. Transportadores específicos de HCO3 - , que atua como um tampão fisiológico celular que desempenha um papel crucial na manutenção do pH intracelular e extracelular, foram descritos nos SFTs. Além disso, a presença de transportadores de água conhecidos como Aquaporinas, tem também sido sugerida nos SFTs, estando estes provavelmente envolvidos na secreção do fluido luminal. Nos SFTs, as células de Sertoli (SCs) têm um papel crucial no estabelecimento do fluido luminal. É por isso imperativo compreender a dinâmica do transporte de água e HCO3 - em SCs, a fim de identificar e neutralizar possíveis alterações nesses sistemas, relacionadas com a redução da fertilidade masculina causadas por condições patológicas associadas com alterações no transporte de água e HCO3, como acontece na fibrose cística (CF). Assim, o primeiro objetivo deste trabalho foi avaliar a expressão de mRNA e proteína do regulador de condutância transmembranar da fibrose cística (CFTR) em SCs, por RT-PCR e imunodetecção (respetivamente), utilizando culturas primárias obtidas a partir de ratos de 20 dias de idade. O segundo objetivo foi identificar a expressão de isoformas específicas de aquaporinas (AQP0-AQP9), utilizando uma abordagem similar. Finalmente, o terceiro objetivo foi investigar a possível interação física entre o CFTR e isoformas de AQP específicas em SCs, usando a técnica de co-imunoprecipitação. Fomos capazes de detetar a expressão de transcritos de mRNA e de proteínas do transportador CFTR e das isoformas AQP4, AQP8 e AQP9 em SCs de rato. A presença das proteínas AQP6 e AQP7 também foi detetada, embora não se tenha observado a presença dos transcritos correspondentes. Além disso, no presente estudo, observou-se uma interação direta entre AQP4 e CFTR, usando a técnica de co-imunoprecipitação. Assim, a secreção de iões e de água nos SFTs, impulsionada por transportadores de membrana específicos presentes nas SCs, é um evento importante no controle da osmolaridade e fluidez do conteúdo luminal. Os nossos resultados permitiram identificar a expressão de várias isoformas de AQP em SCs rato, particularmente AQP4, AQP8 e AQP9, destacando a importância do transporte de água nessas células e sugerindo que as diferentes AQPs podem desempenhar mais de que uma função particular. Além disso, os nossos resultados indicam também que a AQP4 interage fisicamente com o CFTR, sugerindo um possível papel para este transportador na regulação das AQPs e na homeodinâmica do transporte de água em SCs de rato. Disfunções no transporte de água podem ser a causa de patologias obstrutivas, associadas com a atrofia e infertilidade observadas em homens com CF. É por isso possível que a rutura de um complexo funcional envolvendo a AQP4 e o CFTR possa contribuir para a infertilidade/subfertilidade masculina subjacente à patogénese da CF.
45
Danahey, Daniel Gerard. "A biochemical and immunohistochemical study of rat sertoli cells /." The Ohio State University, 1990. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487682558445099.
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Sneddon, Sharon F. "Oestrogen regulation of gene expression in male germ cells and Sertoli cells." Thesis, University of Edinburgh, 2005. http://hdl.handle.net/1842/29373.
Full textAbstract:
The aims of this study were to investigate the role of steroid hormones, in particular oestrogens, in murine spermatogenesis. A major focus of these investigations was the role played by ERβ in the modulation of germ cell and somatic cell function. Studies were conducted both using a transformed murine Sertoli cell line (SK11), which has maintained a differentiated Sertoli cell phenotype and spermatogonial stem cells, which were successfully isolated and characterised. ERβ mRNA and protein were shown to be expressed in the SK11 cells both in the undifferentiated and differentiated states. Transient transfections using ERE or ARE-luciferase reporter constructs and stimulation with steroid ligands revealed that the cells contained functional steroid hormone receptors. Knockdown of ERβ mRNA and protein was achieved in the cells after targeted deletion using a short hairpin RNAi containing vector; this blunted the ability of the cells to respond to oestrogen. Isolation of spermatogonial stem cells was carried out using immunomagnetic beads. The stem cell population were shown to express Oct-4 and GFRα-1 mRNAs, both of which are stem cell markers, but not c-kit, which is a marker of differentiated germ cells. Taqman Q-RT-PCR demonstrated that the stem cell population expressed ERβ. Oct-4 mRNA expression was shown to be reduced by RNAi; this induced the cells to undergo differentiation in vitro characterised by increased expression of c-kit. In conclusion, the current studies have extended our understanding of the impact of steroid hormones on testicular function and have revealed for the first time that spermatogonial stem cells are ERβ positive. The SK11 cell line has been found to provide a suitable model system for the study of steroid regulation of Sertoli cell function. In addition, the use of RNAi has provided and exciting new avenue by which to manipulate gene expression levels in testicular germ and somatic cells.
47
Martins, Ana Catarina Dias. "Effects of sex steroid hormones on sertoli cells metabolic pathways." Master's thesis, Universidade da Beira Interior, 2012. http://hdl.handle.net/10400.6/1126.
Full textAbstract:
Developing germ cells use lactate, derived from glucose metabolism of Sertoli cells (SCs), as their main energy source. Androgens and estrogens have been implicated in the modulation of testicular cells energy metabolism, particularly in SCs. The goal of the present study was to shed light on the effects of sex steroid hormones on glucose metabolic pathways in rat SCs. The mRNA levels of glucose transporters 1 and 3 (GLUT1 and GLUT3), phosphofructokinase 1 (PFK1) and lactate dehydrogenase chain C (LHD C) were analyzed by RT-PCR, and protein levels of GLUTs, PFK1, LDH and monocarboxylate transporter 4 (MCT4) were analyzed by Western Blot, in enriched primary cultures of immature rat SCs treated with 17β-estradiol (E2) or 5α-dihydrotestosterone (DHT). Our results show that both E2 and DHT downregulated the gene transcript levels of PFK-1, GLUT1 and GLUT3. However, only DHT-treated cells presented a downregulation of LDH C gene transcript levels. Interestingly, the protein levels of these enzymes and transporters remained unaltered except in DHT-treated cells that presented a significant decrease on GLUT1 protein levels evidencing a possible pathway for the regulation of glucose metabolism in SCs by androgens. Taken together, these results demonstrated a relationship between the action of sex steroid hormones and energy metabolism of SCs, providing evidences for the mechanisms by which E2 and DHT exert their function as modulators of rat SCs glucose metabolism.As células germinativas em desenvolvimento utilizam lactato, um produto do metabolismo da glicose das células de Sertoli (SCs), como a principal fonte de energia. O papel dos androgénios e estrogénios na modulação do metabolismo energético das células testiculares tem vindo a ser estudado, particularmente nas SCs. O presente estudo tem como objetivo explorar o efeito de hormonas esteróides sexuais sobre as vias envolvidas no metabolismo da glicose em SCs de ratos. Foram analisados os níveis de mRNA de transportadores de glicose (GLUT1 e GLUT3), fosfofrutoquinase-1 (PFK1) e lactato desidrogenase isoforma C (LHD C) por RT-PCR, e por Western Blot foram analisados os níveis proteicos de GLUTs, PFK-1, LDH e transportador de ácidos monocarboxílicos 4 (MCT4). Foram utilizadas para este estudo culturas primárias de SCs de ratos imaturos tratadas com 17βestradiol (E2) ou 5α -dihidrotestosterona (DHT). Os resultados obtidos demonstram que tanto o E2 como o DHT regulam os níveis de transcrição da PFK1, GLUT1 e GLUT3. No entanto, apenas as células tratadas com DHT apresentam uma diminuição nos níveis de transcrição da LDH C. Curiosamente, os níveis de proteína destas enzimas e transportadores permaneceram inalterados, exceto em células tratadas com DHT que apresentaram uma diminuição significativa nos níveis proteicos de GLUT1, pondo em evidência uma possível via para a regulação do metabolismo da glicose em SCs por androgénios. Em conjunto, estes resultados demonstraram uma relação entre a ação das hormonas esteróides sexuais e metabolismo energético das SCs, facultando novas evidências sobre os mecanismos através dos quais o E2 e a DHT exercem a sua função como moduladores do metabolismo da glicose em SCs de rato.
48
Wang, Xiaoying [Verfasser]. "Reduced immunogenicity of induced pluripotent stem cells derived from Sertoli cells / Xiaoying Wang." Aachen : Hochschulbibliothek der Rheinisch-Westfälischen Technischen Hochschule Aachen, 2014. http://d-nb.info/1065413998/34.
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Zhu, Li. "Identification and characterization of physically interacting partners of retinoic acid receptor alpha in sertoli cells." Pullman, Wash. : Washington State University, 2009. http://www.dissertations.wsu.edu/Dissertations/Spring2009/l_zhu_042009.pdf.
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Ribeiro, Mariana Antunes. "Reconstrução de redes regulatórias gênicas em células de Sertoli humanas expostas ao 2,3,7,8-Tetraclorodibenzo-p-dioxina (TCDD)." Botucatu, 2017. http://hdl.handle.net/11449/151523.
Full textAbstract:
Orientador: Wellerson Rodrigo ScaranoResumo: A fertilidade masculina e a espermatogênese estão diretamente ligadas à capacidade das células de Sertoli em produzir fatores associados ao desenvolvimento das células germinativas. As células de Sertoli expressam receptores para FSH e testosterona e são os principais reguladores da espermatogênese. Aproximadamente 60-70% dos casos de infertilidade masculina são considerados idiopáticos, devido aos mecanismos moleculares envolvidos na espermatogênese ainda serem desconhecidos. Estudos recentes relatam que os microRNAs (miRNAs), são capazes de modular a função testicular durante a espermatogênese e sua expressão alterada pode estar envolvida na infertilidade masculina. miRNAs podem desempenhar papel importante na resposta aos xenobióticos que têm todas as consequências adversas para a saúde. Um grupo importante de compostos orgânicos com potencial tóxico são as dioxinas, como o 2,3,7,8-tetraclorodibenzo-p-dioxina (TCDD). Modelos experimentais de exposição ao TCDD, em camundongos, demonstraram que sua exposição provoca baixa contagem de espermatozóides e atraso na puberdade. Neste estudo, analisamos o efeito do TCDD nas células de Sertoli humanas in vitro após 72h a uma dose de 10nM. Nossos resultados mostraram que as enzimas antioxidantes catalase, superóxido dismutase e glutationa peroxidase diminuíram sua atividade e confirmaram o estresse oxidativo causado pelo TCDD nesse tipo celular. 78 miRNAs apresentaram expressão alterada, com regulação positiva de 73 e regulação negat... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Male fertility and spermatogenesis are directly linked to the ability of Sertoli cells to produce factors associated with the development of germ cells. Sertoli cells express receptors for FSH and testosterone, and are the major regulators of spermatogenesis. Approximately 60-70% of male infertility cases are considered idiopathic, due to the molecular mechanisms involved in spermatogenesis are still unknown. Recent studies report that microRNAs (miRNAs) are capable of modulating spermatogenesis in testicular function and its altered expression may be involved in male infertility. miRNAs may play a role in response to xenobiotics that have all the adverse consequences for health. An important group of organic compounds that are potentially toxic are the dioxins such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Experimental models of exposure to TCDD in mice showed that its exposure causes low sperm count and delayed puberty. In this study, we analyzed the effect of TCDD on human Sertoli cells after a exposure of 72h in vitro at a dose of 10nM. Our results showed that the antioxidant enzymes catalase, superoxide dismutase and glutathione peroxidase decreased their activity and confirmed the oxidative stress caused by TCDD in this cell type. 78 miRNAs showed altered expression with upregulation of 73 miRNAs and downregulation of 5 miRNAs compared to the control group. Regarding the gene expression profile, 51 genes showed deregulated, of which 46 genes with upregulation and d... (Complete abstract click electronic access below)
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