Control Of Sex Determination Research Articles

Molecular analysis and developmental expression of the Sex-lethal gene of Sciaraocellaris (Diptera order, Nematocera suborder)

This paper reports the cloning and characterization in Sciara ocellaris of the gene homologous to Sex-lethal ( Sxl) of Drosophila melanogaster. This gene plays the key role controlling sex determination and dosage compensation in the latter species. The Sciara Sxl gene produces a single transcript encoding a single protein in both males and females. This protein, found inside the nucleus, is expressed in all tissues. Both Sciara and Drosophila Sxl proteins are highly conserved at their two RNA-binding domains. In both Sciara sexes, the Sxl protein co-localizes with transcription-active regions dependent on RNA polymerase II but not on RNA polymerase I. It would appear that in Sciara, Sxl does not appear to play the key discriminative role in controlling sex determination and dosage compensation that it plays in Drosophila.

The involvement of ovarian tumour in the intracellular localization of Sex-lethal protein.

The Drosophila ovarian tumour gene is required at multiple times in the germline for oogenesis. A second gene, Sex-lethal, controls sex determination in the soma and also has a separate germline function affecting similar oogenic stages as ovarian tumour. We demonstrate that ovarian tumour is not required for early Sex-lethal gene expression in the female germline, as had been previously reported. Instead, we provide evidence that ovarian tumour has a specific role in the developmentally regulated accumulation of SEX-LETHAL protein within the cytoplasm and nucleus. Furthermore, the examination of nurse cell polytene chromosomes produced by certain ovarian tumour mutations showed that SEX-LETHAL protein can associate with discrete chromosomal sites in the germline and that this pattern appears to change as the egg chamber matures. This is the first indication that SEX-LETHAL is capable of direct physical interactions with chromosomes (albeit in a mutant background) and is consistent with the developmentally regulated nuclear localization of SEX-LETHAL being important for oogenesis.

Sex determination in flies, fruitflies and butterflies.

Sex determination mechanisms, differing in their modality, are widely represented in all the various animal taxa, even at the intraspecific level. Within the highly diversified Class Insecta, Drosophila has been used to unravel the mechanistic molecular and genetic interactions that are involved in sex determination. Indeed, the molecularly characterized genes of the Drosophila sex determination hierarchy X:A > Sxl > tra > dsx have been fruitful starting points in the cloning of homologous genes from other insect species. This cascade seems to control sex determination in all Drosophila species. However, no sex-specific regulatory Sxl homologues have been isolated from the Mediterranean fruitfly (medfly), Ceratitis capitata, the housefly, Musca domestica, Chrysomya rufifacies nor from the distantly related phorid fly Megaselia scalaris. Moreover, all these other species use primary signals different from the intricate X:A counting system of Drosophila. However, dsx homologues isolated from these and other dipteran species as well as from the silkmoth, Bombyx mori, share a conserved sex-specific regulation based on alternative splicing. An understanding of the sex determination mechanisms in insects that are of agricultural or public health importance may help in the development of improved methods for their control using the sterile insect technique.

Molecular and evolutionary analysis of a plant Y chromosome

Plants have evolved a great diversity of sex determination systems. Among these, the XY system, also found in mammals, is one of the most exciting since it gives the opportunity to compare the evolution of sex chromosomes in two different kingdoms. Whereas genetic and molecular mechanisms controlling sex determination in drosophila and mammals, have been well studied, very little is known about such processes in plants. White campion ( Silene latifolia) is an example of plant with X and Y chromosomes. What is the origin of the X and Y chromosomes? How did they evolve from a pair of autosomes? In our laboratory, we have isolated the first active genes located on a plant Y chromosome. We are using them as markers to trace the origin and evolution of sex chromosomes in the Silene genus.

Analysis of the sex ratio in Bradysia matogrossensis (Diptera, Sciaridae)

In sciarid flies, the control of sex determination and of the progeny sex ratio is exercised by the parental females, and is based on differential X-chromosome elimination in the initial stages of embryogenesis. In some species, the females produce unisexual progenies (monogenic females) while in others, the progenies consist of males and females (digenic females). The sex ratio of bisexual progenies is variable, and departs considerably from 1:1. Bradysia matogrossensis shows both monogenic and digenic reproduction. In a recently established laboratory strain of this species, 15% of the females were digenic, 10% produced only females, 13% produced only males, and 62% produced progenies with one predominant sex (33% predominantly of female and 29% predominantly male progenies). These progeny sex ratios were maintained in successive generations. Females from female-skewed progenies yielded female- and male-producing daughters in a 1:1 ratio. In contrast, daughters of females from male-skewed progenies produce bisexual or male-skewed progenies. The X-chromosome of B. matogrossensis shows no inversion or other gross aberration. These results suggest that the control of the progeny sex ratio (or differential X-chromosome elimination) involves more than one locus or, at least, more than one pair of alleles. The data also suggest that, in sciarid flies, monogeny and digeny may share a common control mechanism.

SRY protein is expressed in ovotestis and streak gonads from human sex-reversal

In mammals, a master gene located on the Y chromosome, the testis-determining gene SRY, controls sex determination. SRY protein is expressed in the genital ridge before testis determination, and in the testis it is expressed in Sertoli and germ cells. Completely sex-reversed patients are classified as either 46,XX males or 46,XY females. SRY mutations have been described in only 15% of patients with 46,XY complete or partial gonadal dysgenesis. However, although incomplete or partial sex-reversal affects 46,XX true hermaphrodites, 46,XY gonadal dysgenesis, and 46,XX/46,XY mosaicism, only 15% of the 46,XX true hermaphrodites analyzed have the SRY gene. Here, we demonstrate that the SRY protein is expressed in the tubules of streak gonads and rete testis, indicating that the SRY protein is normally expressed early during testis determination. Based on these results, we propose that some factors downstream from SRY may be mutated in these 46,XY sex-reversal patients. We have also analyzed SRY protein expression in the ovotestis from 46,XX true hermaphrodites and 46,XX/46,XY mosaicism, demonstrating SRY protein expression in both testicular and ovarian portions in these patients. This suggests that the SRY protein does not inhibit ovary development. These results confirm that other factors are needed for complete testis development, in particular, those downstream of the SRY protein.

Conservation of a sex-determining gene.

Vertebrates exhibit a surprising array of sex-determining mechanisms, including X- and Y-chromosome heterogametes in male mammals, Z- and W-chromosome hetero-gametes in female birds, and a temperature-dependent mechanism in many reptiles1. The Y-chromosome-linked SRY gene initiates male development in mammals2,3, but other vertebrates lack SRY and the genes controlling sex determination are largely unknown. Here we show that a gene implicated in human testis differentiation, DMRT1, has a gonad-specific and sexually dimorphic expression profile during embryogenesis in mammals, birds and a reptile (Alligator mississippiensis). Given the different sex-determining switches in these three groups, this gene must represent an ancient, conserved component of the ver-tebrate sex-determining pathway.

Expression of Dmrt1 in the Genital Ridge of Mouse and Chicken Embryos Suggests a Role in Vertebrate Sexual Development

Sex-determining mechanisms are highly variable between phyla. Only one example has been found in which structurally and functionally related genes control sex determination in different phyla: the sexual regulators mab-3 of Caenorhabditis elegans and doublesex of Drosophila both encode proteins containing the DM domain, a novel DNA-binding motif. These two genes control similar aspects of sexual development, and the male isoform of DSX can substitute for MAB-3 in vivo, suggesting that the two proteins are functionally related. DM domain proteins may also play a role in sexual development of vertebrates. A human gene encoding a DM domain protein, DMRT1, is expressed only in the testis in adults and maps to distal 9p24.3, a short interval that is required for testis development. Earlier in development we find that murine Dmrt1 mRNA is expressed exclusively in the genital ridge of early XX and XY embryos. Thus Dmrt1 and Sry are the only regulatory genes known to be expressed exclusively in the mammalian genital ridge prior to sexual differentiation. Expression becomes XY-specific and restricted to the seminiferous tubules of the testis as gonadogenesis proceeds, and both Sertoli cells and germ cells express Dmrt1. Dmrt1 may also play a role in avian sexual development. In birds the heterogametic sex is female (ZW), and the homogametic sex is male (ZZ). Dmrt1 is Z-linked in the chicken. We find that chicken Dmrt1 is expressed in the genital ridge and Wolffian duct prior to sexual differentiation and is expressed at higher levels in ZZ than in ZW embryos. Based on sequence, map position, and expression patterns, we suggest that Dmrt1 is likely to play a role in vertebrate sexual development and therefore that DM domain genes may play a role in sexual development in a wide range of phyla.

SlY1, the first active gene cloned from a plant Y chromosome, encodes a WD-repeat protein.

Unlike the majority of flowering plants, which possess hermaphrodite flowers, white campion (Silene latifolia) is dioecious and has flowers of two different sexes. The sex is determined by the combination of heteromorphic sex chromosomes: XX in females and XY in males. The Y chromosome of S.latifolia was microdissected to generate a Y-specific probe which was used to screen a young male flower cDNA library. We identified five genes which represent the first active genes to be cloned from a plant Y chromosome. Here we report a detailed analysis of one of these genes, SlY1 (S.latifolia Y-gene 1). SlY1 is expressed predominantly in male flowers. A closely related gene, SlX1, is predicted to be located on the X chromosome and is strongly expressed in both male and female flowers. SlY1 and SlX1 encode almost identical proteins containing WD repeats. Immunolocalization experiments showed that these proteins are localized in the nucleus, and that they are most abundant in cells that are actively dividing or beginning to differentiate. Interestingly, they do not accumulate in arrested sexual organs and represent potential targets for sex determination genes. These genes will permit investigation of the origin and evolution of sex chromosomes in plants.

The MurineFem1Gene Family: Homologs of theCaenorhabditis elegansSex-Determination Protein FEM-1

The pathway controlling sex determination in the nematodeCaenorhabditis elegansis a model for the genetic control of cell-fate determination. We report here the cloning and characterization of a new mouse gene family with homology to FEM-1, a signal-transducing regulator in theC. eleganssex-determination pathway. This gene family consists of two known members, designatedFem1aandFem1b.The highest degree of homology between the two mouse proteins and the nematode protein is in a domain that encodes seven sequential ANK repeats. TheFem1agene localizes to chromosome 17 and is highly expressed in adult heart and skeletal muscle. TheFem1bgene localizes to chromosome 9 and is highly expressed in adult testis. Both genes are expressed during embryogenesis. The existence of FEM-1 homologs in the mouse raises the possibility that evolutionary conservation of ancient FEM-1 signaling interactions may play a role in vertebrate cell-fate determination.

The evolutionary dynamics of sex determination.

REVIEW There is substantial cytogenetic data indicating that the process of sex determination can evolve relatively rapidly. However, recent molecular studies on the evolution of the regulatory genes that control sex determination in the insect Drosophila melanogaster, the nematode Caenorhabditis elegans, and mammals suggest that, although certain sex determination regulatory genes have evolved relatively rapidly, other sex determination regulatory genes are quite conserved. Thus, studies of the evolution of sex determination, a process that appears to have elements that undergo substantial evolutionary change and others that may be conserved, could provide substantial insights into the kinds of forces that both drive and constrain the evolution of developmental hierarchies.

Ploidy induction and sex control in fish

Ploidy induction and sex control in fish

Novel functions for `nuclear factors' in the cytoplasm: theSex-lethalparadigm

In recent years, novel functions for a number of nuclear factors have been uncovered in the cytoplasm, mainly at the level of translation. These factors behave as multifunctional regulators of gene expression and many play key roles in cell differentiation and development. One of the best characterized examples is that of Sex-lethal (SXL), an RNA-binding protein that is expressed in female Drosophila flies and controls sex determination and dosage compensation. Recent findings indicate that SXL, a paradigmatic regulator of splicing, also controls translation of target mRNAs. This review attempts to summarize this evidence and provide an overview of `nuclear factors' with roles in translation.

Sex organ determination and differentiation in the dioecious plant Melandrium album ( Silene latifolia ): a cytological and histological analysis

Melandrium album (Silene alba) is a dioecious plant with heteromorphic sex chromosomes (XY system). Sexual dimorphism is a result of developmental blocks in male or female reproductive organ formation within young bipotential flower buds. Progress in understanding the genetic and molecular mechanisms controlling sex determination in this species relies on a detailed description of developmental timing in the two sexes, with emphasis on those early stages during which sexual dimorphism is established. We used a combination of histological and scanning electron microscopy analysis to refine the comparative study and description of the staging of male compared to female flower development. We show that (1) female dimorphism results from modifications in flower meristem organisation, namely a sudden arrest of cell divisions in whorl 4 of male flowers at the time when meristem partitioning is achieved between whorls 3 and 4, and (2) male dimorphism is part of the stamen differentiation process corresponding to stamen arrest at the early sporogenous stage in female flowers. Thus, Melandrium is a natural double ”mutant” that is affected in very early and distinct processes of reproductive organ differentiation. Our results are used to discuss the most likely nature of the specific functions controlling sexual dimorphism in Melandrium.

Sex determination and flower development in dioecious plants

Abstract This talk briefly describes the different modes of sexuality in flowering plants (hermaphroditism, monoecy, dioecy), and the control of hermaphroditic flower development by a small set of interacting genes. The problem is stated of the necessity of an additional control in unisexual flower development. Genetic control of sex determination in dioecious species is then considered.

Ceratopteris: A Model System for Studying Sex-Determining Mechanisms in Plants

The homosporous fern Ceratopteris richardii is a model system for studying the genetic and epigenetic factors that are involved in sex-determining processes in plants. The sex of the Ceratopteris gametophyte is determined by antheridiogen, a gibberellin-like pheromone that is secreted by the hermaphroditic gametophyte and promotes male development of sexually immature gametophytes. Abscisic acid blocks the antheridiogen response in Ceratopteris. Several mutations that affect sex expression have been isolated and characterized; these mutations define at least some of the major regulatory genes that are involved in determining the sex of the Ceratopteris gametophyte. By studying the epistatic interactions among these mutants, it is possible to determine how these genes interact with one another to form a regulatory genetic hierarchy that controls sex determination and expression in Ceratopteris. While a genetic approach is useful in identifying genes that are involved in sex determination, other molecular approaches have been successfully employed in isolating sex-specific genes in Ceratopteris as well as genes involved in other aspects of its reproductive development.

The fog-3 gene and regulation of cell fate in the germ line of Caenorhabditis elegans.

In the nematode Caenorhabditis elegans, germ cells normally adopt one of three fates: mitosis, spermatogenesis or oogenesis. We have identified and characterized the gene fog-3, which is required for germ cells to differentiate as sperm rather than as oocytes. Analysis of double mutants suggests that fog-3 is absolutely required for spermatogenesis and acts at the end of the regulatory hierarchy controlling sex determination for the germ line. By contrast, mutations in fog-3 do not alter the sexual identity of other tissues. We also have characterized the null phenotype of fog-1, another gene required for spermatogenesis; we demonstrate that it too controls the sexual identity of germ cells but not of other tissues. Finally, we have studied the interaction of these two fog genes with gld-1, a gene required for germ cells to undergo oogenesis rather than mitosis. On the basis of these results, we propose that germ-cell fate might be controlled by a set of inhibitory interactions among genes that specify one of three fates: mitosis, spermatogenesis or oogenesis. Such a regulatory network would link the adoption of one germ-cell fate to the suppression of the other two.

Identification of a RAPD marker linked to sex determination in Pistacia vera using bulked segregant analysis

The Random Amplified Polymorphic DNA (RAPD) technique was used to amplify DNA segments, with the objective of finding markers linked to sex determination in the dioecious species, Pistacia vera. Progenies from two female parents pollinated by a common male parent were studied. Two bulks of DNA were made in each cross, one from males and one from females, by pooling an equal weight of fresh leaves from each individual contributing to the bulk prior to DNA extraction. DNA was extracted from each bulked sample and from each of the contributing individuals. DNA was also extracted from 14 cultivars of P. vera and from 94 open-pollinated, fewweeks-old P. vera seedlings of unknown sex. Seven hundred different decamer oligonucleotide primers were used to perform DNA amplification, with 1 of these (OPO08) producing a 945 bp amplification band that was present only in the bulked female samples and absent in the bulked male samples of the two crosses. The relationship between band presence and female sex expression was conserved in every individual obtained from the two crosses and in the 14 cultivars unrelated to the crosses. We propose that this band is tightly linked to the gene(s) that control sex determination in pistachio. The OPO08945 RAPD marker could be used in a breeding program to screen the gender of pistachio plants long before they reach reproductive maturity, resulting in considerable savings of time and economic resources. In order to verify that assumption we screened 94 additional seedlings with the OPO08 primer and obtained results consistent with a 1∶1 male:female ratio.

Sex-determining genes in the homosporous fern Ceratopteris

Haploid Ceratopteris gametophytes are either hermaphroditic or male. The determinate of sex type is the pheromone antheridiogen (ACE) which is secreted by the meristic hermaphrodite and promotes ameristic male development of sexually undetermined gametophytes. Several mutations effecting the sex of the haploid gametophyte have been isolated and are described. The hermaphroditic (her) mutants are insensitive to ACE and develop as meristic hermaphrodites. These mutations effect ameristic male development in the presence of ACE but have no effect on hermaphroditic development. While most her mutations also have no effect on diploid sporophyte development, some partially ACE-insensitive her mutations have profound effects on sporophyte development. The transformer (tra) mutation effects both meristem and archegonia formation and causes the gametophyte to be an ameristic male under conditions that promote hermaphroditic development. The feminization (fem) mutation effects antheridia development in both male and hermaphroditic gametophytes and causes the gametophyte to develop as a meristic female in the absence or presence of the pheromone. The her1 tra1 double mutant is male in the absence or presence of ACE, indicating that tra1 is epistatic to her1. The phenotypes of her1, tra1 and fem1 single gene mutant phenotypes and the her1 tra1 double mutant phenotype are used to deduce a model suggesting how the products of these genes might interact in a regulatory pathway to control sex determination.

230 MOLECULAR MARKERS FOR EARLY DETERMINATION OF SEX OF PISTACHIO SEEDLINGS

The Random Amplified Polymorphic DNA (RAPD) technique was used to develop molecular markers linked to sex expression in Pistacia vera, a dioecious species. Progenies from two female parents (`Lassen' and `Kerman') pollinated by a common male parent (`Peters') were studied. Two bulks of DNA were made in each cross, one from males and one from females. DNA was extracted from each bulked sample as well as from each of the contributing individuals and from 14 additional P. vera cultivars. Twelve hundred decamer oligonucleotide primers were used to perform DNA amplification on the bulk DNA. This analysis led to the identification of one primer (OPO08) that produces a 945 bp. amplification band present only in females and absent in males. The relationship between band presence and female sex expression was conserved in every individual obtained from the two crosses and in 14 cultivars unrelated to the crosses. This band, which we propose is tightly linked to the gene(s) controlling sex determination, provides a reliable marker for sex of pistachio seedlings and should be a useful tool in pistachio breeding.