Sry Transgene Research Articles

Independent contribution of gonads and sex chromosomes to sex differences in bone mass and strength in the four-core genotypes mouse model.

Vertebrate sexual dimorphism is ascribed to the presence of testes or ovaries, and, hence, to the secretion of gonad-specific hormones. However, mounting evidence indicates that sex differences in tissues and organs also stem from the presence of sex chromosomes (XX or XY). To tease out the contribution of gonads from sex chromosomes to the musculoskeletal system, we used the Four-Core Genotypes (FCG) mouse model, in which the Sry gene, which dictates testis formation, was either deleted from the Y chromosome, resulting in XY mice with ovaries (XY-SryO), or overexpressed in XX mice, resulting in XX mice with testes (XXT), together with gonadal males with XY-SryT (Sry deletion and overexpression of the Sry transgene in chromosome 3) and females with XXO. The FCG mice are generated by crossing XXO with XY-SryT mice, all of C57BL/6J background. We now show that the musculoskeletal phenotype of 2- to 4-mo-old FCG mice varies based on both gonads and sex chromosomes, depending on the age and the organ/tissue/cell analyzed. The effect of sex chromosomes on body weight, fat and lean/skeletal muscle mass, and bone mass and structure is minor in 2-/3-mo-old mice, soon after sexual maturation. The contribution of sex chromosomes (XX vs XY-Sry in mice with the same gonads and sex hormones) to several of our measurements becomes apparent in adult 4-mo-old mice. The contribution of 1X and 1Y-Sry vs 2X chromosomes varies among different measurements in gonadal males or females, and mice with XY-Sry chromosomes might have higher or lower values that XX mice. Our study shows XX vs XY-Sry chromosome contribution to the musculoskeletal phenotype, which becomes more evident as the animals reach peak bone mass, suggesting that although gonadal sex has a major role, sex chromosomes are also an unrecognized contributor to musculoskeletal mass and bone strength.

Generation and mutational analysis of a transgenic mouse model of human SRY.

SRY is the Y-chromosomal gene that determines male sex development in humans and most other mammals. After three decades of study, we still lack a detailed understanding of which domains of the SRY protein are required to engage the pathway of gene activity leading to testis development. Some insight has been gained from the study of genetic variations underlying differences/disorders of sex determination (DSD), but the lack of a system of experimentally generating SRY mutations and studying their consequences in vivo has limited progress in the field. To address this issue, we generated a mouse model carrying a human SRY transgene able to drive testis determination in XX mice. Using CRISPR-Cas9 gene editing, we generated novel genetic modifications in each of SRY's three domains (N-terminal, HMG box, and C-terminal) and performed a detailed analysis of their molecular and cellular effects on embryonic testis development. Our results provide new functional insights unique to human SRYand present a versatile and powerful system in which to functionally analyze variations of SRY including known and novel pathogenic variants found in DSD.

Alterations of sex determination pathways in the genital ridges of males with limited Y chromosome genes†.

We previously demonstrated that in the mouse only two Y chromosome genes are required for a male to produce an offspring with the help of assisted reproduction technologies (ART): testis determinant Sry and spermatogonial proliferation factor Eif2s3y. Subsequently, we have shown that the function of these genes can be replaced by transgenic overexpression of their homologs, autosomally encoded Sox9 and X-chromosome encoded Eif2s3x. Males with Y chromosome contribution limited to two (XEif2s3yOSry), one (XEif2s3yOSox9 and XOSry,Eif2s3x), and no genes (XOSox9,Eif2s3x) produced haploid germ cells and sired offspring after ART. However, despite successful assisted reproductive outcome, they had smaller testes and displayed abnormal development of the seminiferous epithelium and testicular interstitium. Here we explored whether these testicular defects originated from altered pro-testis and pro-ovary factor signaling in genital ridges at the time of sex determination. Timed pregnancies were generated to obtain transgenic XEif2s3yOSry, XEif2s3yOSox9, XOSry,Eif2s3x, XOSox9,Eif2s3x, and wild-type XX and XY fetuses at 12.5 days post coitum. Dissected genital ridges were assessed for their morphology and anatomy, and expression of pro-testis and pro-ovary transcripts. All transgenic males displayed incomplete masculinization of gonadal shape, impaired development of testicular cords and gonadal vasculature, and decreased expression of factors promoting male pathway. Fetal gonad masculinization was more effective when sex determination was driven by the Sry transgene, in the presence of Y chromosome genes, and to a lesser extent a double dosage of X genes. The study adds to the understanding of the role of Y chromosome genes and their homologs during sex determination.

Four core genotypes mouse model: localization of the Sry transgene and bioassay for testicular hormone levels.

BackgroundThe “four core genotypes” (FCG) mouse model has emerged as a major model testing if sex differences in phenotypes are caused by sex chromosome complement (XX vs. XY) or gonadal hormones or both. The model involves deletion of the testis-determining gene Sry from the Y chromosome and insertion of an Sry transgene onto an autosome. It produces XX and XY mice with testes, and XX and XY mice with ovaries, so that XX and XY mice with the same type of gonad can be compared to assess phenotypic effects of sex chromosome complement in cells and tissues.FindingsWe used PCR to amplify the Sry transgene and adjacent genomic sequences, to resolve the location of the Sry transgene to chromosome 3 and confirmed this location by fluorescence in situ hybridization (FISH) of the Sry construct to metaphase chromosomes. Using quantitative PCR, we estimate that 12–14 copies of the transgene were inserted. The anogenital distance (AGD) of FCG pups at 27–29 days after birth was not different in XX vs. XY males, or XX vs. XY females, suggesting that differences between XX and XY mice with the same type of gonad are not caused by difference in prenatal androgen levels.ConclusionThe Sry transgene in FCG mice is present in multiple copies at one locus on chromosome 3, which does not interrupt known genes. XX and XY mice with the same type of gonad do not show evidence of different androgen levels prenatally.

Abstract 249: Male Mice with XX Sex Chromosome Complement Exhibit Reduced Angiotensin II-Induced Hypertension and Atherosclerosis

Objective: Hypertension and atherosclerosis exhibit sexual dimorphism, partially ascribed to effects of sex hormones. However, recent studies suggest that sex chromosome complement may also contribute to sexual dimorphism of these diseases. The renin-angiotensin system has been suggested to contribute to sexual dimorphism of hypertension and atherosclerosis. However, the relative contribution of sex hormones, versus chromosomes, in the regulation of angiotensin II (AngII)-mediated responses is unknown. We hypothesized that the presence of an XX chromosome complement in male hyperlipidemic mice would reduce AngII-induced hypertension and atherosclerosis. Methods and Results: Male Ldlr-/- mice with an Sry mutation but with an Sry transgene on autosomes were bred to Ldlr-/- females to generate XY or XX males. Male XY or XX mice were segregated to sham-operated or orchiectomized (ORC) groups and fed a high fat diet (42% kcal from fat). Two weeks later, mice were infused with AngII (1,000 ng/kg/min) for 4 weeks. In the presence of sex hormones (sham groups), systolic blood pressure (SBP) was significantly lower in AngII-infused XX than XY males (XY, 136 ± 4; XX, 119 ± 4 mmHg, p=0.024). In XY males, ORC had no significant effect on SBP compared to sham (ORC XY, 129 ± 6 mmHg; p=0.48). However, following castration of XX males, SBP significantly increased (ORC XX 135 ± 3 mmHg; p =0.002). In XY males, ORC resulted in a significant increase in atherosclerotic lesion surface area in the aortic arch (sham, 19 ± 4; ORC, 32 ± 7%; p=0.04). In contrast, ORC had no significant effect on lesion surface area in XX males (sham, 17 ± 3; ORC, 15 ± 2%; p=0.53). Moreover, in the absence of sex hormones, lesion surface area was markedly increased in XY compared to XX males (XY, 32 ± 7; XX, 15 ± 2%; p=0.002). Conclusions: These results demonstrate that effects of testosterone to regulate AngII-induced hypertension and atherosclerosis are dependent on sex chromosome complement. In XY males, testosterone protects against AngII-induced atherosclerosis. In XX males, testosterone protects against AngII-induced hypertension. Moreover, there is pronounced effect of XY chromosome complement to promote AngII-induced atherosclerosis in males.

Abstract 175: The Role of ER-α, ER-ß, and AR in the FCG Model of Ischemic Brain Injury

Background: Ischemic stroke is a sexually dimorphic disease. Females are protected from ischemic brain injury throughout most of the lifespan, but the contribution of hormones in this “ischemia resistant” phenotype remains unclear. The “four core genotype” (FCG) model is used to differentiate the effect of the chromosomal complement (XX vs. XY) from an animal’s gonadal (estrogen vs. testosterone producing) sex through examination of five genotypes XYMwt, XY-M, XXM, XXF, and XYF. XY-M FCG mice are made by the translocation of Sry from the Y chromosome to an autosome. Stroke sex differences have been largely attributed to the neuroprotective effects of estrogen. However, it remains unclear whether hormonal receptors can be influenced at the chromosomal level, which could further explain sexual dimorphism in stroke phenotypes. We hypothesize that the sex chromosome complement and transgene Sry (the testis determining gene found on the Y chromosome) contribute to the sexually dimorphic stroke phenotype. Methods: FCG animals were gonadectomized at 3 weeks of age. Animals were subjected to stroke at 8-12 weeks of age. Cytosolic samples were evaluated for androgen receptor (AR) and estrogen receptor (ER-α and ER-β) levels in sham and stroke samples. RT-PCR for Sry on the stroke hemisphere (delta-delta-CT) was performed on the FCG mice and wild type males. Results: Western blot analysis showed that XXM and XXF mice had significant stroke induced reduction in AR expression (p<0.01). No significant difference in the expression of ER-α was seen in sham and stroke mice among genotypes. ER-β expression increased significantly in XXM post stroke (p<0.05), but not in the other genotypes. RT-PCR analysis showed approximately 90-fold increase of mRNA expression of Sry in the phenotypes with the exogenous Sry (XYM and XXM) compared to WTM mice with endogenous Sry (p<0.05). Conclusion: XXM and XYM mice show an increase in Sry expression compared to WTM suggesting insertional and positional effects of Sry transgene. AR expression in XXM and XXF mice is influenced at the chromosomal level, suggesting effects of X chromosome dosage on AR expression. Stroke induced ER-β expression in XXM mice may be an effect of the interaction between transgene Sry and other hormonal axes.

Homeoproteins Six1 and Six4 Regulate Male Sex Determination and Mouse Gonadal Development

The Y-linked gene Sry regulates mammalian sex determination in bipotential embryonic gonads. Here, we report that the transcription factors Six1 and Six4 are required for male gonadal differentiation. Loss of Six1 and Six4 together, but neither alone, resulted in a male-to-female sex-reversal phenotype in XY mutant gonads accompanied by a failure in Sry activation. Decreased gonadal precursor cell formation at the onset of Sry expression and a gonadal size reduction in both sexes were also found in mutant embryos. Forced Sry transgene expression in XY mutant gonads rescued testicular development but not the initial disruption to precursor growth. Furthermore, we identified two downstream targets of Six1/Six4 in gonadal development, Fog2 (Zfpm2) and Nr5a1 (Ad4BP/Sf1). These two distinct Six1/Six4-regulated pathways are considered to be crucial for gonadal development. The regulation of Fog2 induces Sry expression in male sex determination, and the regulation of Nr5a1 in gonadal precursor formation determines gonadal size.

Ubiquitous expression of Sry induces embryonic lethality related to suppression of Tie2/Tek expression

Sry (sex-determining region on the Y chromosome) is a mammalian sex-determining gene on the Y chromosome. In mice, the transient expression of Sry in supporting cell precursor cells between 10.5 and 12.5 days post-coitus (dpc) triggers the differentiation of Sertoli cells from granulosa cells. The importance of the strict regulation of Sry expression remains unknown. Thus, we attempted to produce a Sry ubiquitous-expressing transgenic (Tg) mouse in which foreign Sry is driven by the CAG (cytomegalovirus immediate-early enhancer, chicken beta-actin promoter, and the fusion intron of chicken beta-actin and rabbit beta-globin)-Sry gene for ubiquitous expressing Sry. A low rate (2/127) of Tg pups was observed, whereas the rate of early-stage transgenic embryos before birth was 19.2% (5/26). The Sry ubiquitous-expressing embryos showed abnormal development. The results suggest that ubiquitous expression of Sry exerts a negative effect on embryonic development. One of the two adult Tg mice showed low levels of Sry expression. The other Tg mouse showed high Sry transgene expression, but was mosaic for the transgene. Developmental analysis of transgenic F1 embryos produced from the mosaic Tg mouse revealed that ubiquitous expression of Sry had a lethal effect on embryonic development around 12.5 dpc. The histological data indicated that ubiquitous expression of Sry induced abnormal cardiovascular development, resulting in embryonic death. Enhanced expression of Sry suppressed endogenous Tie2/Tek (tyrosine kinase with Ig and EGF homology domains 2/tunica interna endothelial cell kinase) expression in Sry-transfected primary cultured cells from wild type embryonic hearts. The results indicate that the tissue-specific and stage-specific expression of Sry is essential for normal embryogenesis.

A critical time window ofSryaction in gonadal sex determination in mice

In mammals, the Y-linked sex-determining gene Sry cell-autonomously promotes Sertoli cell differentiation from bipotential supporting cell precursors through SRY-box containing gene 9 (Sox9), leading to testis formation. Without Sry action, the supporting cells differentiate into granulosa cells, resulting in ovarian development. However, how Sry acts spatiotemporally to switch supporting cells from the female to the male pathway is poorly understood. We created a novel transgenic mouse line bearing an inducible Sry transgene under the control of the Hsp70.3 promoter. Analysis of these mice demonstrated that the ability of Sry to induce testis development is limited to approximately 11.0-11.25 dpc, corresponding to a time window of only 6 hours after the normal onset of Sry expression in XY gonads. If Sry was activated after 11.3 dpc, Sox9 activation was not maintained, resulting in ovarian development. This time window is delimited by the ability to engage the high-FGF9/low-WNT4 signaling states required for Sertoli cell establishment and cord organization. Our results indicate the overarching importance of Sry action in the initial 6-hour phase for the female-to-male switching of FGF9/WNT4 signaling patterns.

Effects of sex chromosome aneuploidy on male sexual behavior.

Incidence of sex chromosome aneuploidy in men is as high as 1:500. The predominant conditions are an additional Y chromosome (47,XYY) or an additional X chromosome (47,XXY). Behavioral studies using animal models of these conditions are rare. To assess the role of sex chromosome aneuploidy on sexual behavior, we used mice with a spontaneous mutation on the Y chromosome in which the testis-determining gene Sry is deleted (referred to as Y(-)) and insertion of a Sry transgene on an autosome. Dams were aneuploid (XXY(-)) and the sires had an inserted Sry transgene (XYSry). Litters contained six male genotypes, XY, XYY(-), XXSry, XXY(-)Sry, XYSry and XYY(-)Sry. In order to eliminate possible differences in levels of testosterone, all of the subjects were castrated and received testosterone implants prior to tests for male sex behavior. Mice with an additional copy of the Y(-) chromosome (XYY(-)) had shorter latencies to intromit and achieve ejaculations than XY males. In a comparison of the four genotypes bearing the Sry transgene, males with two copies of the X chromosome (XXSry and XXY(-)Sry) had longer latencies to mount and thrust than males with only one copy of the X chromosome (XYSry and XYY(-)Sry) and decreased frequencies of mounts and intromissions as compared with XYSry males. The results implicate novel roles for sex chromosome genes in sexual behaviors.

Down-Regulation of Endogenous Wt1 Expression by Sry Transgene in the Murine Embryonic Mesonephros-Derived M15 Cell Line

Wt1 is one of numerous candidate genes comprising the hypothetical chain of gene expression essential for male sex differentiation of the bipotential indifferent gonads during embryogenesis. However, the evidence in the literature is ambivalent regarding the position of Wt1 relative to Sry in this scheme; Wt1 might act either upstream or downstream of Sry. In the present study, the effects of Sry expression upon Wt1 were investigated using M15 cells (XX karyotype), which are derived from murine embryonic mesonephros and express endogenous Wt1. In 3 stably-transformed Sry-expressing M15 cell lines, we showed that the expression levels of the mRNAs coding for all 4 isoforms of the WT1 proteins were down-regulated. Similarly, Wnt 4 expression was down-regulated in these cell lines. Silencing of Sry in the transformed cell lines using ribozymes or short hairpin RNAs (shRNAs) resulted in elevated levels of Wt1 and Wnt4 expression. These results strongly indicate that Wt1 might be under the control of Sry during gonadal differentiation in the mouse. In electrophoretic mobility shift assays (EMSA), we demonstrated that the 3.7 kb 5'-upstream DNA stretch of Wt1 containing potential Sry binding sites was capable of forming molecular complexes with nuclear protein(s) from Sry expressing cells but not with those from control non-Sry expressing cells. In summary, our present results support the notion that Wt1 is located downstream of Sry and down-regulated by the sex determining gene. Although the precise biological meaning of the present findings have yet to be clarified, it is possible that Wt1 plays a dual role during gonadal differentiation, i. e., turning on Sry expression on one hand, and being down-regulated by its product, Sry, on the other, possibly forming a type of negative feed-back mechanism. Further work is needed to substantiate this view.

Sex Steroids and Sex Chromosomes at Odds?

Sex Steroids and Sex Chromosomes at Odds?

Influence on spatiotemporal patterns of a male-specific Sox9 activation by ectopic Sry expression during early phases of testis differentiation in mice

Testis induction is associated with gonadal Sry and Sox9 expression in mammals. This study investigated whether Sry expression directly induces male-specific Sox9 activation during early phases of testis differentiation. We have established an XX sex-reversal mouse line carrying the Sry transgene driven by a weak basal promoter of the Hsp70.3 gene ( Hsp-Sry), whereby the transgene was activated in the gonads along the entire anteroposterior axis from earlier stages. The effects of misexpression and overexpression of Sry on the spatiotemporal pattern of Sox9 expression were examined using both XX and XY gonads of Hsp-Sry transgenic embryos. It was shown that ectopic expression of Sry transcripts in the entire gonadal area from earlier stages promotes neither any advance in the timing nor any appreciable ectopic activation of endogenous Sox9 expression. Immediately after the onset of Sox9 activation, however, both the level of Sox9 expression and the number of SOX9-positive cells were significantly enhanced in Hsp-Sry/XY gonads, as compared with those in wild-type/XY and Hsp-Sry/XX gonads. These findings suggest that, although Sry is capable of up-regulating Sox9 expression dose-dependently, Sry mRNA expression alone is not likely to provide positional or timing information needed for male-specific Sox9 activation in developing XY gonads.

Analysis of sex differences in EGC imprinting

Sex-specific differences are apparent in the methylation patterns of H19 and Igf2 imprinted genes in embryonic germ cells (EGCs) derived from 11.5 or 12.5 days post coitum (dpc) primordial germ cells (PGCs). Here we studied whether these differences are associated either with the sex chromosome constitution of the EGCs or with the sex of the genital ridge (testis versus ovary) from which the PGCs were isolated. For this purpose we derived pluripotent EGC lines from sex-reversed embryos, either XY embryos deleted for Sry (XY Tdym1 ) or XX embryos carrying an Sry transgene. Southern blotting of the EGC DNA was used to analyze the differentially methylated regions of Igf2 and H19. The analysis revealed that both genes were more methylated in EGCs with an XY sex chromosome constitution than in those with an XX sex chromosome constitution, irrespective of the phenotypic sex of the genital ridge from which the EGCs had been derived. We conclude that the sex-specific methylation is intrinsic and cell-autonomous, and is not due to any influence of the genital ridge somatic cells upon the PGCs.

Neonatal mice possessing an Sry transgene show a masculinized pattern of progesterone receptor expression in the brain independent of sex chromosome status.

To assess the relative roles of sex chromosome genes and gonadal steroid hormones in producing sex differences in progesterone receptor (PR) expression in the forebrain of neonatal mice, we used mice in which the Sry gene had been deleted from the Y-chromosome and inserted as a transgene on an autosome in both XX and XY genotypes. Levels of PR immunoreactivity (PRir) in the anteroventral periventricular nucleus, the medial preoptic nucleus, and the ventromedial nucleus were significantly higher in mice that possessed an Sry transgene compared with mice that lacked an Sry transgene, regardless of their complement of sex chromosomes (XX vs. XY). This result suggests that sexual differentiation of PR expression in these regions is likely controlled mainly by gonadal hormones, not by the genetic sex of the brain cells. No differences in PRir were detected between wild-type XY mice with the Sry gene on the Y-chromosome and XY mice with the Sry transgene, suggesting that testicular hormones produced in these two genotypes have comparable effects on neural tissue.

A Model System for Study of Sex Chromosome Effects on Sexually Dimorphic Neural and Behavioral Traits

We tested the hypothesis that genes encoded on the sex chromosomes play a direct role in sexual differentiation of brain and behavior. We used mice in which the testis-determining gene (Sry) was moved from the Y chromosome to an autosome (by deletion of Sry from the Y and subsequent insertion of an Sry transgene onto an autosome), so that the determination of testis development occurred independently of the complement of X or Y chromosomes. We compared XX and XY mice with ovaries (females) and XX and XY mice with testes (males). These comparisons allowed us to assess the effect of sex chromosome complement (XX vs XY) independent of gonadal status (testes vs ovaries) on sexually dimorphic neural and behavioral phenotypes. The phenotypes included measures of male copulatory behavior, social exploration behavior, and sexually dimorphic neuroanatomical structures in the septum, hypothalamus, and lumbar spinal cord. Most of the sexually dimorphic phenotypes correlated with the presence of ovaries or testes and therefore reflect the hormonal output of the gonads. We found, however, that both male and female mice with XY sex chromosomes were more masculine than XX mice in the density of vasopressin-immunoreactive fibers in the lateral septum. Moreover, two male groups differing only in the form of their Sry gene showed differences in behavior. The results show that sex chromosome genes contribute directly to the development of a sex difference in the brain.

C57BL/6J-T-associated sex reversal in mice is caused by reduced expression of a Mus domesticus Sry allele.

C57BL/6J-T-associated sex reversal (B6-TAS) in XY mice results in ovarian development and involves (1) hemizygosity for Tas, a gene located in the region of Chromosome 17 deleted in T(hp) and T(Orl), (2) homozygosity for one or more B6-derived autosomal genes, and (3) the presence of the AKR Y chromosome. Here we report results from experiments designed to investigate the Y chromosome component of this sex reversal. Testis development was restored in B6 T(Orl)/+ XY(AKR) mice carrying a Mus musculus Sry transgene. In addition, two functionally different classes of M. domesticus Sry alleles were identified among eight standard and two wild-derived inbred strains. One class, which includes AKR, did not initiate normal testis development in B6 T(Orl)/+ XY mice, whereas the other did. DNA sequence analysis of the Sry ORF and a 5' 800-bp segment divided these inbred strains into the same groups. Finally, we found that Sry is transcribed in B6 T(Orl)/+ XY(AKR) fetal gonads but at a reduced level. These results pinpoint Sry as the Y-linked component of B6-TAS. We hypothesize that the inability of specific M. domesticus Sry alleles to initiate normal testis development in B6 T(Orl)/+ XY(AKR) mice results from a biologically insufficient level of Sry expression, allowing the ovarian development pathway to proceed.

Evidence that the testis determination pathway interacts with a non-dosage compensated, X-linked gene.

In a number of mammals, including mouse and man, it has been shown that at equivalent gestational ages, males are developmentally more advanced than females, even before the gonads form. In mice, although some strains of Y chromosome exert a minor accelerating effect in pre-implantation development, it is a post-implantation effect of the difference in X chromosome constitution that is the major cause of the male/female developmental difference. Thus XX females are retarded in their development by about 1.5 h relative to X(M)O females or XY males; however, they are more advanced than X(P)O females by about 4 h. It has been suggested that this early developmental difference between XX and XY embryos may "weight the dice" in favour of ovarian and testicular development, respectively, although expression of Sry will normally overcome any such bias. Here we test this proposal by comparing the relative frequencies of female, hermaphrodite and male development in X(P)O, XX and X(M)O mice that carry an incompletely penetrant Sry transgene. The results show that testicular tissue develops more frequently in XX,Sry transgenics than in either of the two types of XO transgenics. Thus the incidence of testicular development is affected by X dosage rather than by the developmental hierarchy. This implies there is a non-dosage compensated gene (or genes) on the X chromosome, which interacts with the testis-determining pathway. Since the pseudoautosomal region (PAR) is known to escape X-inactivation, penetrance of the Sry transgene was also assessed in X(M)Y(*X) mice that have two doses of the PAR but have a single dose of all genes proximal to the distal X marker Amel. These mice showed similar levels of testicular development to X(M)O mice with the transgene; thus the non-dosage compensated X gene maps outside the PAR.

Mouse homologues of the human AZF candidate gene RBM are expressed in spermatogonia and spermatids, and map to a Y chromosome deletion interval associated with a high incidence of sperm abnormalities.

An RNA-binding motif (RBM) gene family has been identified on the human Y chromosome that maps to the same deletion interval as the 'azoospermia factor' (AZF). We have identified the homologous gene family (Rbm) on the mouse Y with a view to investigating the proposal that this gene family plays a role in spermatogenesis. At least 25 and probably >50 copies of Rbm are present on the mouse Y chromosome short arm located between Sry and the centromere. As in the human, a role in spermatogenesis is indicated by a germ cell-specific pattern of expression in the testis, but there are distinct differences in the pattern of expression between the two species. Mice carrying the deletion Yd1, that maps to the proximal Y short arm, are female due to a position effect resulting in non-expression of Sry ; sex-reversing such mice with an Sry transgene produces males with a high incidence of abnormal sperm, making this the third deletion interval on the mouse Y that affects some aspect of spermatogenesis. Most of the copies of Rbm map to this deletion interval, and the Yd1males have markedly reduced Rbm expression, suggesting that RBM deficiency may be responsible for, or contribute to, the abnormal sperm development. In man, deletion of the functional copies of RBM is associated with meiotic arrest rather than sperm anomalies; however, the different effects of deletion are consistent with the differences in expression between the two species.