Dissertations / Theses on the topic 'Patched Gene'

BCC is the commonest cancer in European-derived populations and Australia has the highest recorded incidence in the world, creating enormous individual and societal cost in management of this disease. The incidence of this cancer has been increasing internationally, with evidence of a 1 to 2% rise in incidence in Australia per year over the last two decades. The main four epidemiological risk factors for the development of BCC are ultraviolet radiation (UVR) exposure, increasing age, male sex, and inability to tan. The pattern and timing of UVR exposure is important to BCC risk, with childhood and intermittent UVR exposure both associated with an increased risk. The complex of inherited characteristics making up an individual’s ‘sun sensitivity’ is also important in determining BCC risk. Very little is known about population genetic susceptibility to BCC outside of the rare genodermatosis Gorlin syndrome. Mutations in the tumour suppressor gene patched (PTCH) are responsible for this BCC predisposition syndrome and the molecular pathway and target genes of this highly conserved pathway are well described. Derangments in this pathway occur in sporadic BCC development, and the PTCH gene is an obvious candidate to contribute to non-syndromic susceptibility to BCC. The melanocortin 1 receptor (MC1R) locus is known to be involved in pigmentary traits and the cutaneous response to UVR, and variants have been associated with skin cancer risk. Many other genes have been considered with respect to population BCC risk and include p53, HPV, GSTs, and HLAs. There is preliminary evidence for specific familial aggregation of BCC, but very little known about the causes. 56 individuals who developed BCC under the age of 40 in the year 2000 were recruited from the Skin and Cancer Foundation of Australia’s database. This represents the youngest 7 – 8% of Australians with BCC from a database that captures approximately 10% of Sydney’s BCCs. 212 of their first degree relatives were also recruited, including 89 parents and 123 siblings of these 56 probands. All subjects were interviewed with respect to their cancer history and all reports of cancer verified with histopathological reports where possible. The oldest unaffected sibling for each proband (where available) was designated as an intra-family control. All cases and control siblings filled out a questionnaire regarding their pigmentary and sun sensitivity factors and underwent a skin examination by a trained examiner. Peripheral blood was collected from these cases and controls for genotyping of PTCH. All the exons of PTCH for which mutations have been documented in Gorlin patients were amplified using PCR. PCR products were screened for mutations using dHPLC, and all detectable variants sequenced. Prevalence of BCC and SCC for the Australian population was estimated from incidence data using a novel statistical approach. Familial aggregation of BCC, SCC and MM occurred within the 56 families studied here. The majority of families with aggregation of skin cancer had a combination of SCC and BCC, however nearly one fifth of families in this study had aggregation of BCC to the exclusion of SCC or MM, suggesting that BCCspecific risk factors are also likely to be at work. Skin cancer risks for first-degree relatives of people with early onset BCC were calculated: sisters and mothers of people with early-onset BCC had a 2-fold increased risk of BCC; brothers had a 5-fold increased risk of BCC; and sisters and fathers of people with early-onset BCC had over four times the prevalence of SCC than that expected. For melanoma, the increased risk was significant for male relatives only, with a 10-fold increased risk for brothers of people with early-onset BCC and 3-fold for fathers. On skin examination of cases and controls, several phenotypic factors were significantly associated with BCC risk. These included increasing risk of BCC with having fair, easyburning skin (ie decreasing skin phototype), and with having signs of cumulative sun damage to the skin in the form of actinic keratoses. Signs reflecting the combination of pigmentary characteristics and sun exposure - in the form of arm freckling and solar lentigines - also gave subjects a significantly increased risk BCC. Constitutive red-green reflectance of the skin was associated with decreased risk of BCC, as measured by spectrophotometery. Other non-significant trends were seen that may become significant in larger studies including associations of BCC with propensity to burn, moderate tanning ability and an inability to tan. No convincing trend for risk of BCC was seen with the pigmentary variables of hair or eye colour, and a non-significant reduced risk of BCC was associated with increasing numbers of seborrhoeic keratoses. Twenty PTCH exons (exons 2, 3, 5 to 18, and 20 to 23) were screened, accounting for 97% of the coding regions with published mutations in PTCH. Nine of these 20 exons were found to harbour single nucleotide polymorphisms (SNPs), seen on dHPLC as variant melting curves and confirmed on direct sequencing. SNPs frequencies were not significantly different to published population frequencies, or to Australian general population frequencies where SNP database population data was unavailable. Assuming a Poisson distribution, and having observed no mutations in a sample of 56, we can be 97.5% confident that if there are any PTCH mutations contributing to early-onset BCC in the Australian population, then their prevalence is less than 5.1%. Overall, this study provides evidence that familial aggregation of BCC is occurring, that first-degree relatives are at increased risk of all three types of skin cancer, and that a combination of environmental and genetic risk factors are likely to be responsible. The PTCH gene is excluded as a major cause of this increased susceptibility to BCC in particular and skin cancer in general. The weaknesses of the study design are explored, the possible clinical relevance of the data is examined, and future directions for research into the genetics of basal cell carcinoma are discussed.

Doctor of Philosophy(PhD)
BCC is the commonest cancer in European-derived populations and Australia has the highest recorded incidence in the world, creating enormous individual and societal cost in management of this disease. The incidence of this cancer has been increasing internationally, with evidence of a 1 to 2% rise in incidence in Australia per year over the last two decades. The main four epidemiological risk factors for the development of BCC are ultraviolet radiation (UVR) exposure, increasing age, male sex, and inability to tan. The pattern and timing of UVR exposure is important to BCC risk, with childhood and intermittent UVR exposure both associated with an increased risk. The complex of inherited characteristics making up an individual’s ‘sun sensitivity’ is also important in determining BCC risk. Very little is known about population genetic susceptibility to BCC outside of the rare genodermatosis Gorlin syndrome. Mutations in the tumour suppressor gene patched (PTCH) are responsible for this BCC predisposition syndrome and the molecular pathway and target genes of this highly conserved pathway are well described. Derangments in this pathway occur in sporadic BCC development, and the PTCH gene is an obvious candidate to contribute to non-syndromic susceptibility to BCC. The melanocortin 1 receptor (MC1R) locus is known to be involved in pigmentary traits and the cutaneous response to UVR, and variants have been associated with skin cancer risk. Many other genes have been considered with respect to population BCC risk and include p53, HPV, GSTs, and HLAs. There is preliminary evidence for specific familial aggregation of BCC, but very little known about the causes. 56 individuals who developed BCC under the age of 40 in the year 2000 were recruited from the Skin and Cancer Foundation of Australia’s database. This represents the youngest 7 – 8% of Australians with BCC from a database that captures approximately 10% of Sydney’s BCCs. 212 of their first degree relatives were also recruited, including 89 parents and 123 siblings of these 56 probands. All subjects were interviewed with respect to their cancer history and all reports of cancer verified with histopathological reports where possible. The oldest unaffected sibling for each proband (where available) was designated as an intra-family control. All cases and control siblings filled out a questionnaire regarding their pigmentary and sun sensitivity factors and underwent a skin examination by a trained examiner. Peripheral blood was collected from these cases and controls for genotyping of PTCH. All the exons of PTCH for which mutations have been documented in Gorlin patients were amplified using PCR. PCR products were screened for mutations using dHPLC, and all detectable variants sequenced. Prevalence of BCC and SCC for the Australian population was estimated from incidence data using a novel statistical approach. Familial aggregation of BCC, SCC and MM occurred within the 56 families studied here. The majority of families with aggregation of skin cancer had a combination of SCC and BCC, however nearly one fifth of families in this study had aggregation of BCC to the exclusion of SCC or MM, suggesting that BCCspecific risk factors are also likely to be at work. Skin cancer risks for first-degree relatives of people with early onset BCC were calculated: sisters and mothers of people with early-onset BCC had a 2-fold increased risk of BCC; brothers had a 5-fold increased risk of BCC; and sisters and fathers of people with early-onset BCC had over four times the prevalence of SCC than that expected. For melanoma, the increased risk was significant for male relatives only, with a 10-fold increased risk for brothers of people with early-onset BCC and 3-fold for fathers. On skin examination of cases and controls, several phenotypic factors were significantly associated with BCC risk. These included increasing risk of BCC with having fair, easyburning skin (ie decreasing skin phototype), and with having signs of cumulative sun damage to the skin in the form of actinic keratoses. Signs reflecting the combination of pigmentary characteristics and sun exposure - in the form of arm freckling and solar lentigines - also gave subjects a significantly increased risk BCC. Constitutive red-green reflectance of the skin was associated with decreased risk of BCC, as measured by spectrophotometery. Other non-significant trends were seen that may become significant in larger studies including associations of BCC with propensity to burn, moderate tanning ability and an inability to tan. No convincing trend for risk of BCC was seen with the pigmentary variables of hair or eye colour, and a non-significant reduced risk of BCC was associated with increasing numbers of seborrhoeic keratoses. Twenty PTCH exons (exons 2, 3, 5 to 18, and 20 to 23) were screened, accounting for 97% of the coding regions with published mutations in PTCH. Nine of these 20 exons were found to harbour single nucleotide polymorphisms (SNPs), seen on dHPLC as variant melting curves and confirmed on direct sequencing. SNPs frequencies were not significantly different to published population frequencies, or to Australian general population frequencies where SNP database population data was unavailable. Assuming a Poisson distribution, and having observed no mutations in a sample of 56, we can be 97.5% confident that if there are any PTCH mutations contributing to early-onset BCC in the Australian population, then their prevalence is less than 5.1%. Overall, this study provides evidence that familial aggregation of BCC is occurring, that first-degree relatives are at increased risk of all three types of skin cancer, and that a combination of environmental and genetic risk factors are likely to be responsible. The PTCH gene is excluded as a major cause of this increased susceptibility to BCC in particular and skin cancer in general. The weaknesses of the study design are explored, the possible clinical relevance of the data is examined, and future directions for research into the genetics of basal cell carcinoma are discussed.

The Drosophila segment polarity gene, patched (ptc), encodes a transmembrane protein that is the receptor for the hedgehog intercellular signalling molecule. Mutations in the human ptc gene, have been shown to be responsible for Gorlin's syndrome and a predisposition to nevoid basal cell carcinomas. In this study, partial clones of patched homologues were isolated from the long germ band developing house fly, Musca domestica (Diptera), and the intermediate germ band developing house cricket, Acheta domesticus (Orthoptera). Sequence data obtained from the Musca domestica clones showed a high degree of similarity with the Drosophila ptc gene sequence, indicating that the Musca domestica clones contain real ptc homologues. The Acheta ptc fragment was cloned using degenerate PCR, and sequence data has shown it has a high degree of similarity to the comparable regions of other ptc homologues. Two phagemid clones were also isolated from an Acheta cDNA library using a strategy designed to isolate Acheta ptc. One of these clones, PD, was used to create RNA in situ hybridization probes, and its expression was examined during Acheta embryogenesis, although it was later shown that PD was not an Acheta ptc homologue. Expression of the Musca ptc homologue was examined during early development using RNA in situ hybridisation, and immunohistochemistry. These studies have shown that the expression of ptc during Musca domestica development is very similar to ptc expression patterns seen during the early development of Drosophila, suggesting that ptc may be fulfilling a similar role in both species.

O ceratocisto odontogênico (CO) é uma neoplasia benigna que possui um comportamento biológico agressivo. O CO está associado a alterações no gene homologo 1 ao gene patched de Drosophila (PTCH1). Este gene possui diversas isoformas geradas pelo uso alternativo do primeiro exon (1b, 1d e 1e). As isoformas podem possuir diferenças na sua expressão, na capacidade de agir como receptor da proteína sinalizadora Hedgehog (Hh) e na inibição da própria via. Levando em consideração que a via Hh pode estar envolvida na patogênese do CO, o objetivo deste estudo foi avaliar a presença dos diferentes RNAm do gene PTCH1 nesta lesão. Foram incluídos no estudo 40 CO, sendo 12 esporádicos e 28 associados à Síndrome do Carcinoma Basocelular nevóide (SCBN). Destas lesões, 17 eram primárias, 17 eram marsupializadas e 6 eram recorrentes. O estudo da expressão dos exons 1 alternativos do gene PTCH1 foi realizado através da técnica da Transcriptase Reversa PCR (RT PCR). Os resultados demonstram que 90% dos CO, esporádicos e associados à SCBN, expressam o transcrito 1b enquanto nenhum folículo dentário (controle) apresenta a expressão desta isoforma. O RNAm do 1d é expresso na maioria dos CO e em todos os controles. Foi detectada a expressão do transcrito 1b em todas as lesões primárias, porém isso não ocorreu em 4 (24%) lesões marsupializadas. O perfil de expressão dos transcritos na mucosa adjacente à lesão é semelhante ao do CO. Concluímos que a via Hh está superativada no CO e na mucosa adjacente e que a marsupialização pode alterar a expressão das isoformas.

Doctor of Philosophy
Department of Entomology
James F. Campbell
Thomas W. Phillips
Movement is a fundamental feature of animals that impacts processes across multiple scales in space and time. Due to the heterogeneous and fragmented nature of habitats that make up landscapes, movement is not expected to be random in all instances, and an increase in fitness is an expected consequence for those that can optimize movement to find valuable and scarce recourses. I studied the movement of Rhyzopertha dominica (Coleoptera: Bostrichidae), one of the most important pests of stored grain worldwide, within and between resource patches. At a fine spatial scale, I identified factors that contribute to overall and upward movement in the grain mass. Three-week-old insects tented to stay closer to the surface than one or two-week-old insects. Females tended to be more active and to explore more than males. I also found that males tended to stay closer to the surface than females and that might be related to the ability to attract females from outside the patch since there was no significant difference regarding female’s attraction within the grain patch. Interaction with feeding sites or other individuals of the same sex creates positive feedback and a more clumped spatial pattern of feeding and foraging behavior. On the other hand, interaction with individuals of different sex creates negative feedback and a more random or overdispersed pattern. At a broad spatial scale, I studied the long-term consequence of R. dominica movement on the development of population structure within the U.S. To evaluate population structure, I used reduced representation of the genome followed by direct sequencing of beetles collected from different locations across the U.S where wheat or rice is produced and stored. Ecoregions were more important in explaining structure of R. dominica populations than crop type. I also found significant isolation by distance; however, model selection primarily elected grain production and movement variables to explain population differentiation and diversity. Understanding animal movement is essential to establishing relationships between distribution and surrounding landscape, and this knowledge can improve conservation and management strategies.

La symbiose entre les légumineuses et les rhizobia est d'une importance majeure dans les écosystèmes terrestres du fait de sa capacité à fixer l'azote atmosphérique. Le dialogue moléculaire entre les deux partenaires, qui mène finalement au développement de nodosités hébergeant les bactéries fixatrices, peut être initié par la liaison des facteurs Nod (NF) sécrétés par les rhizobia aux récepteurs à NF de la membrane plasmatique (PM) des poils racinaires de la légumineuse. Cela déclenche un influx de Ca2+ dans la cellule, suivi d'une cascade d'événements de signalisation ionique, impliquant des changements dans les flux de H+, K+ et Cl- à travers la MP. Mon objectif a été de caractériser les mécanismes moléculaires qui sous-tendent ces premiers événements de signalisation ionique chez la légumineuse modèle Medicago truncatula. En utilisant la technique du patch-clamp sur des protoplastes obtenus par digestion enzymatique ou ablation laser assistée de la paroi cellulaire de poils en croissance, j'ai contribué à caractériser plusieurs conductances ioniques de la MP de ces cellules. Je me suis particulièrement concentrée sur une conductance cationique activée par l'hyperpolarisation membranaire (HACC), particulièrement perméable à Ca2+. Cette conductance est rapidement activée (en moins d'une minute) par l'addition de NF à une concentration physiologique et son activation est dépendante de la présence de récepteurs NFP ("Nod Factor Perception") fonctionnels. Ces résultats suggèrent que c'est cette conductance qui conduit l'influx précoce de Ca2+ déclenché par la perception des NF. Je me suis aussi intéressée à des systèmes de transport cationiques membranaires exprimés dans les poils absorbants de M. truncatula, appartenant aux familles HKT et GLR (Glutamate receptor-like), en tant qu'acteurs potentiels des premiers événements ioniques de signalisation. L'analyse par génétique inverse du rôle de 3 GLR fortement exprimés suggère que ces gènes ne jouent pas un rôle majeur dans la mise en place de la conductance HACC et ne sont pas indispensables à la nodulation. D'autre part, les transporteurs HKT, qui se sont révélés sélectifs de Na+, sont exprimés dans les nodosités, suggérant un rôle dans la symbiose
Symbiosis between legumes and rhizobia is of major importance in terrestrial ecosystems due to its ability to fix atmospheric nitrogen. The molecular dialogue between the two partners, which ultimately leads to development of nodules hosting the N2 fixing bacteria, can be initiated by the binding of Nod factors (NF) secreted by the rhizobial partner on NF receptors at the legume root hair plasma membrane (PM). This triggers a Ca2+ influx through the PM, followed by a cascade of ionic signaling events, involving changes in H+, K+, and Cl- fluxes at the PM. My objective was to characterize molecular mechanisms underlying these early ionic signaling events in the legume model Medicago truncatula. By using the patch-clamp technique on protoplasts obtained either by cell wall enzymatic digestion or laser-assisted ablation from growing root hairs, I have contributed to characterize several ion conductances from this cell type. I especially focused on a cationic conductance activated by membrane hyperpolarization (HACC), uniquely found to be most permeable to Ca2+. This conductance was quickly activated (within less than 1 minute) following NF addition at physiological concentration. Its activation was dependent on the presence of functional NFP (“Nod Factor Perception”) receptors, which suggested that this conductance mediates the early Ca2+ influx triggered by NF perception. In addition, cationic transport systems expressed in M. truncatula root hairs and belonging to the HKT and Glutamate receptor-like (GLR) families were investigated as potential contributors to the early ionic signaling events. Loss-of-function mutant analysis for 3 highly expressed GLRs suggested that these genes did not play major roles in the expression/activity of the HACC conductance, and were not indispensable for nodulation. On the other hand, the HKT transporters, which were found to be Na+-selective, were expressed in nodules, which suggested a role in symbiosis

Yiu Wai Han.
"June 2003."
Thesis (Ph.D.)--Chinese University of Hong Kong, 2003.
Includes bibliographical references (p. 146-156)
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Mode of access: World Wide Web.
Abstracts in English and Chinese.

Includes bibliographical references (leaves 204-230).
xvi, 230 leaves : ill. ; 30 cm.
Investigates the behaviour and ecology of the two major larval parasitoids of the diamondback moth, Plutella xylostella; Cotesia plutellae and Diadegma semiclausum.
Thesis (Ph.D.)--University of Adelaide, Dept. of Applied and Molecular Ecology, 2001

No
The ability of neurons to modulate synaptic strength underpins synaptic plasticity, learning and memory, and adaptation to sensory experience. Despite the importance of synaptic adaptation in directing, reinforcing, and revising the behavioral response to environmental influences, the cellular and molecular mechanisms underlying synaptic adaptation are far from clear. Brain-derived neurotrophic factor (BDNF) is a prime initiator of structural and functional synaptic adaptation. However, the signaling cascade activated by BDNF to initiate these adaptive changes has not been elucidated. We have previously shown that BDNF activates mitogen- and stress-activated kinase 1 (MSK1), which regulates gene transcription via the phosphorylation of both CREB and histone H3. Using mice with a kinase-dead knock-in mutation of MSK1, we now show that MSK1 is necessary for the upregulation of synaptic strength in response to environmental enrichment in vivo. Furthermore, neurons from MSK1 kinase-dead mice failed to show scaling of synaptic transmission in response to activity deprivation in vitro, a deficit that could be rescued by reintroduction of wild-type MSK1. We also show that MSK1 forms part of a BDNF- and MAPK-dependent signaling cascade required for homeostatic synaptic scaling, which likely resides in the ability of MSK1 to regulate cell surface GluA1 expression via the induction of Arc/Arg3.1. These results demonstrate that MSK1 is an integral part of a signaling pathway that underlies the adaptive response to synaptic and environmental experience. MSK1 may thus act as a key homeostat in the activity- and experience-dependent regulation of synaptic strength.