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A H Sinclair

Publications and source records attributed to A H Sinclair.

At least 19 recordsLinked to original sources
PubMed1999

Temperature-dependent sex determination: upregulation of SOX9 expression after commitment to male development.

In mammals, birds and reptiles the morphological development of the gonads appear to be conserved. This conservation is evident despite the different sex determining switches employed by these vertebrate groups. Mammals exhibit chromosomal sex determination (CSD) where the key sex determining switch is the Y-linked gene, SRY. Although SRY is the trigger for testis determination in mammals, it is not conserved in other vertebrate groups. However, a gene closely related to SRY, the highly conserved transcription factor, SOX9, plays an important role in the testis pathway of mammals and birds. In contrast to the CSD mechanism evident in mammals and birds, many reptiles exhibit temperature dependent sex determination (TSD) where the egg incubation temperature triggers sex determination. Here we examine the expression of SOX9 during gonadogenesis in the American alligator, (Alligator mississippiensis), a reptile that exhibits TSD. Alligator SOX9 is expressed in the embryonic testis but not in the ovary. However, the timing of SOX9 upregulation in the developing testis is not consistent with a role for this gene in the early stages of alligator sex determination. Since SOX9 upregulation in male embryos coincides with the structural organisation of the testis, SOX9 may operate farther downstream in the vertebrate sex differentiation pathway than previously postulated.

Alligators and Crocodiles↗
PubMed1999

Expression of chicken steroidogenic factor-1 during gonadal sex differentiation.

The orphan nuclear receptor, steroidogenic factor-1 (SF-1), regulates steroidogenic enzyme expression and is essential for gonadal and adrenal development in mammals. We have examined expression of the chicken homologue, cSF-1, during gonadal sex differentiation using whole mount in situ hybridisation and RNase protection assays (RPA). In the youngest embryos examined (day 3.5; stages 21-22), cSF-1 transcripts were already detectable by in situ hybridisation in the undifferentiated genital ridge of both sexes. Expression continued in the gonads of both sexes at the time of sexual differentiation (days 5.5-6.5; stages 28-30). Expression then became higher in developing ovaries compared to testes at days 6.5-8. 5 (stages 30-35). At day 13.5 (stage 40), when the gonads are well differentiated, both ovaries and testes showed cSF-1 expression, with higher levels of expression in the left ovary compared to the right (regressing) gonad in females and compared to testes. RPA analysis of isolated gonads confirmed higher expression of SF-1 in differentiating ovaries relative to testes. Expression of cSF-1 in the developing adrenal gland was similar for both sexes at all stages examined. In tissue sections of day 8.5 whole mount gonads, cSF-1 expression was localised in the medulla of the ovary and was weakly detectable in the testis. These observations indicate that SF-1 has a conserved role in early gonadal and adrenal development in vertebrates. However, upregulation of cSF-1 expression during ovarian differentiation is opposite to the pattern seen in mammals, in which SF-1 is downregulated in females. This difference between the birds and mammals may reflect differences in steroidogenic activity of the embryonic ovary versus the testis in the two groups.

Animals↗
PubMed1998

Human sex determination.

Human sex determination is a fascinating topic, particularly at the level of molecular genetics, as it represents an excellent paradigm for mammalian organ development. Recent progress has seen the addition of several new pieces to this developmental jigsaw puzzle. In mammals, the Y chromosome is male determining, and encodes a gene referred to as TDF (testis-determining factor), which induces the indifferent embryonic gonad to develop as a testis. Subsequent male sexual differentiation is largely a consequence of hormonal secretion from the testis. In the absence of the Y chromosome, the testis-determining pathway fails to be initiated, and the embryonic gonad develops as an ovary, resulting in female development. (Ford et al. [1959] Lancet i:711; Jacobs and Strong [1959] Nature, 183:302-303; Jost et al. [1973] Rec. Prog. Horm. Res., 29:1-41).

Animals↗
PubMed1998

Familial Kallmann syndrome: a novel splice acceptor mutation in the KAL gene.

Kallmann syndrome is an inherited disease which is characterised by anosmia (inability to smell) and hypogonadotropic hypogonadism both of which are thought to occur as a result of a failure of correct neuronal migration. To date the only genetic lesions identified are mutations in the X-linked gene, KAL. We conducted a mutation screen of the KAL gene in a family with Kallmann syndrome. This identified a new mutation in the KAL gene which removed an acceptor site at the junction of exon 6/intron 5. Exon 6 of the KAL gene encodes the C-terminal portion of a fibronectin type III domain may be involved in axonal pathfinding. We presume that the described mutation would result in the removal of exon 6 resulting in a frame shift which terminates the protein prematurely. It has been proposed that both mental illness and vesico-ureteric reflux are associated with mutations in the KAL gene. However, results from the family presented here do not show an association between either trait and the KAL gene mutation.

Child↗
PubMed1997

Isolation of rare transcripts by representational difference analysis.

Representational difference analysis (RDA) is a powerful technique for cloning the differences between genomes, and has recently been adapted for cloning differentially expressed genes. RDA, like other PCR-based differential screening methods, is prone to the production of false positives. We have identified a major source of false positives in RDA of cDNA and have introduced improvements which minimise their production. These modifications also significantly increase sensitivity, allowing for the isolation of rare differential transcripts from nanogram amounts of mRNA.

Animals↗
PubMed1997

Sites of estrogen receptor and aromatase expression in the chicken embryo.

Estrogens have been implicated in sexual differentiation of both the gonads and the genitalia of birds. In chicken embryos, the gonads are steroidogenically active from an early age, and the aromatase gene, (cAROM), necessary for estrogen synthesis, is expressed only in females at the time of gonadal sex differentiation. However, no studies have directly demonstrated the distribution of estrogen receptor (cER) transcripts or proteins in the embryonic avian reproductive system. Whole-mount in situ hybridization and immunohistochemistry were used here to identify sites of estrogen receptor expression in the embryonic chicken urogenital system. Estrogen receptor mRNA was observed in both male and female gonads prior to morphological differentiation, at Stage 26 (4.5 days of incubation), and continued until after sexual dimorphism at Stage 32 (7.5 days). Transcripts of cER were also detected in the Müllerian ducts and developing external genitalia of both sexes. Estrogen receptor protein was analysed in the embryonic gonads by immunohistochemistry and found to be most abundant in the cortex of the left ovary, although it was also present in the medulla of both female gonads. No significant cER protein expression was detected in the male gonad by immunohistochemistry. In contrast, the aromatase gene was expressed in the gonads of female embryos from the onset of sexual dimorphism but was not detectable in male gonads at any stage examined. These findings suggest that estrogen involvement in both gonadogenesis and genital development in chickens is mediated by the estrogen receptor.

Animals↗
PubMed1997

Gonadal sex differentiation in chicken embryos: expression of estrogen receptor and aromatase genes.

Estrogen is implicated in sexual differentiation of the avian gonad. Expression of the estrogen receptor and aromatase genes was therefore examined at the time of gonadal sex differentiation in chicken embryos, using reverse transcription and the polymerase chain reaction (RT-PCR). Estrogen receptor (cER) transcripts were detected in the gonads of both presumptive sexes at embryonic days 4.5, 5.5 and 6.5, and in female but not male urogenital tissues at day 3.5. Aromatase (cAROM) transcripts were detected in female but not male gonads from day 6.5 of embryogenesis, and in adult gonads of both sexes. Both female and male embryos thus express cER mRNA before morphological differentiation of the gonads, which begins on day 5, whereas cAROM expression begins at or shortly after the onset of differentiation and is female-specific. Examination of other tissues showed that, in 5.5-day-old embryos, cER expression was limited to the gonads; no transcripts were detected in the mesonephric kidney, liver, brain, hindlimb or heart of either sex. In 9.5-day-old female embryos, cER and cAROM transcripts were present in both the left (ovarian) and the right (regressing) gonads. Altogether, these observations imply that the gonads of both sexes develop the capacity to respond to estrogens early in embryogenesis, before morphological differentiation, whereas the capacity to synthesize estrogens is female-specific and occurs later, at the time of differentiation. These observations are consistent with estrogens having a key role in ovarian development.

Animals↗
PubMed1997

Gonadal dysgenesis: associations between clinical features and sex of rearing.

The purpose of this study was to review the phenotypic and endocrine features or a series of patients with ambiguous genitalia or sex-reversal due to gonadal dysgenesis (GD) and to analyse the impact of these on the decision about sex of rearing. This study is a retrospective analysis of 22 patients with GD treated between 1964 and 1994. We assessed external genitalia, internal genitalia, internal genital structures, gonadal morphology (n = 22), basal and human CG (hCG) stimulated serum testosterone levels (n = 11) and serum gonadotropin levels (n = 13) in patients with GD. Basal and hCG stimulated testosterone levels were also measured for 43 control patients. There were no significant associations or correlations between internal or external genital phenotype, endocrine function and gonadal morphology. There was a significant association between sex of rearing and external genitalia (P = 0.03). Patients with gonadal dysgenesis had significantly lower stimulated/basal testosterone levels than the controls (P = 0.0001). Given that the clinical features of various forms of GD overlap considerably, gonadal biopsy should remain the investigation of choice when attempting to define the pathology.

Adolescent↗
PubMed1996

A novel germ line mutation in SOX9 causes familial campomelic dysplasia and sex reversal.

Mutations in the gene SOX9 result in the syndrome of campomelic dysplasia (CD) which includes sex-reversal in 75% of 46,XY affected individuals. These mutations only affect a single allele of SOX9 suggesting a dominant mode of inheritance for this syndrome. Consequently, CD and autosomal sex reversal may result from haploinsufficiency of SOX9. The SOX9 gene maps to the long arm of human chromosome 17 and translocations in this region also result in CD. We report a family in which there were three affected patients, two of whom showed 46,XY sex-reversal. Interestingly, despite all three patients being heterozygous for a familial mutation in SOX9 (Insertion of a cytosine residue at nucleotide position 1096), their gonadal phenotypes varied widely. The proband was found to have 46,XY true hermaphroditism with ambiguous genitalia. The other two sibs were 46,XY and 46,XX, and both had bilateral ovaries with normal female genitalia. The somatic cells in both parents revealed wild-type SOX9 nucleotide sequences. However, mutational analysis of the SOX9 gene in the father's germ cells revealed they were mosaic for mutant and wild-type sequences. This family is particularly informative as it demonstrates that the same SOX9 mutation can produce very different 46,XY gonadal phenotypes. The range of gonadal morphologies observed may be explained by several possible mechanisms such as variable penetrance of the mutation, increased activity of the non-mutant SOX9 allele or stochastic environmental factors. These results also demonstrate that paternal germ cell mosaicism of a mutant SOX9 sequence can result in a CD phenotype amongst his offspring.

Disorders of Sex Development↗
PubMed1996

A male-specific role for SOX9 in vertebrate sex determination.

Mutation analyses of patients with campomelic dysplasia, a bone dysmorphology and XY sex reversal syndrome, indicate that the SRY-related gene SOX9 is involved in both skeletal development and sex determination. To clarify the role SOX9 plays in vertebrate sex determination, we have investigated its expression during gonad development in mouse and chicken embryos. In the mouse, high levels of Sox9 mRNA were found in male (XY) but not female (XX) genital ridges, and were localised to the sex cords of the developing testis. Purified fetal germ cells lacked Sox9 expression, indicating that Sox9 expression is specific to the Sertoli cell lineage. Sex specificity of SOX9 protein expression was confirmed using a polyclonal antiserum. The timing and cell-type specificity of Sox9 expression suggests that Sox9 may be directly regulated by SRY. Male-specific expression of cSOX9 mRNA during the sex determination period was also observed in chicken genital ridges. The conservation of sexually dimorphic expression in two vertebrate classes which have significant differences in their sex determination mechanisms, points to a fundamental role for SOX9 in testis determination in vertebrates. Sox9 expression was maintained in the mouse testis during fetal and adult life, but no expression was seen at any stage by in situ hybridisation in the developing ovary. Male-specific expression was also observed in the cells surrounding the Müllerian ducts and in the epididymis, and expression in both sexes was detected in the developing collecting ducts of the metanephric kidney. These results suggest that SOX9 may have a wider role in the development of the genitourinary system.

Animals↗
PubMed1995

New genes for boys.

Explore the source record for details and available documents.

DNA-Binding Proteins↗
PubMed1994

SRY protein enhances transcription of Fos-related antigen 1 promoter constructs.

In mammals, testis determination is under the control of the chromosome Y-linked SRY gene. Sry is expressed in the fetal mouse just before development of the testis and shows germ-cell-dependent expression in the adult mouse. SRY protein contains a high-mobility-group (HMG)-box DNA-binding domain, and potential target sequences have been identified. The fos-related antigen 1 (fra-1) gene is closely related to the protooncogene c-fos and encodes a component of transcription factor AP-1. Fra-1 is expressed during spermatogenesis, and the promoter of the rat fra-1 gene contains several potential binding sites for members of the HMG-box family of DNA-binding proteins. We demonstrate that purified SRY protein binds strongly to one of the putative fra-1 HMG-box response elements and that SRY enhances the transcription of rat fra-1 promoter constructs in cotransfection experiments. These results suggest that the function of HMG-box transcription factors may be mediated, in part, by activation of members of the AP-1 transcription factor family.

Amino Acid Sequence↗
PubMed1994

Phosphoglycerate kinase pseudogenes in the tammar wallaby and other macropodid marsupials.

Phosphoglycerate kinase (EC 2.7.2.3; PGK) exists in two forms in marsupials. PGK1 is an X-linked house-keeping enzyme, and PGK2 is a mainly testis-specific enzyme under autosomal control. We have used PGK1 probes derived from two closely related species of macropodid marsupials (kangaroos and wallabies) to demonstrate the existence of a large family of pseudogenes in the tammar wallaby (Macropus eugenii). Over 30 fragments are detectable after Taq digestion. We estimate that there are 25-30 copies per genome. Most are autosomally inherited and are apparently not closely linked. Only two restriction fragments that appeared to be sex linked could be detected. Varying degrees of hybridization of fragments to the probes suggest different levels of homology, and hence different ages of origin. The existence of two PGK1 homologous restriction fragments from the X and a large number from the autosomes was also demonstrated by somatic cell hybridization for two other macropodid species, the wallaroo (M. robustus) and the red kangaroo (M. rufus). These results are compared with those from human and mouse, and it is suggested that the propensity of PGK1 to form pseudogenes is an ancient (approximately 130 MYR BP) characteristic of mammals. The high level of polymorphism detected in the tammar makes these PGK1 probes potentially useful for measuring genetic variability in this species and other macropodids.

Animals↗
PubMed1993

The human SRY transcript.

SRY encodes the Y-linked testis-determining factor in humans. A predominant 900 bp transcript originates from a single exon and encompasses the putative SRY coding sequence. We show that in human adult testis SRY transcription involves multiple start sites. In addition to a previously defined major initiation site, transcripts originating at least 410 bp upstream of this site were detected. Using a cDNA specific RT-PCR assay, embryonic and adult human tissues were screened for SRY expression. In humans, SRY transcription is not restricted to the presumptive and the mature gonadal tissues in the embryo and the adult respectively but can be detected in a range of other locations. Two human cell lines, NTERA-2 cl.D1 (NT2/D1) and Hep G2, have been identified which express SRY at similar levels to adult testis. The NT2/D1 SRY transcripts appear to have the same structure as those in adult testis. HMBA-induced differentiation of NT2/D1 cells results in a diminution of SRY mRNA, while transcription of SRY in retinoic acid differentiated NT2/D1 is unaffected.

Adult↗
PubMed1993

Fertile females of the mole Talpa occidentalis are phenotypic intersexes with ovotestes.

We investigated the origin of XX sex reversal in the insectivorous mole Talpa occidentalis. Cytogenetic, histological and hormonal studies indicate that all XX individuals analyzed from two different populations are true hermaphrodites, with ovotestes. This suggests that XX sex reversal may be the norm in this species. The intersexes are functional fertile females and the trait is transmitted and maintained in the population. Intersexes lack the Y chromosome gene SRY (sex determining region Y gene), shown to be the testis determining gene. These results suggest that XX intersex moles may have arisen from a mutation of a gene located downstream from SRY/TDY in the testis determining pathway.

Animals↗