Yann Guiguen

Teleost fish are important models for human biology, health, and disease. Because genome duplication in a teleost ancestor (TGD) impacts the evolution of teleost genome structure and gene repertoires, we must discriminate gene functions that are shared and ancestral from those that are lineage-specific in teleosts or tetrapods to accurately apply inferences from teleost disease models to human health. Generalizations must account both for the TGD and for divergent evolution between teleosts and tetrapods after the likely two rounds of genome duplication shared by all vertebrates. Progress in sequencing techniques provides new opportunities to generate genomic and transcriptomic information from a broad range of phylogenetically informative taxa that facilitate detailed understanding of gene family and gene function evolution. We illustrate here the use of new sequence resources from spotted gar (Lepisosteus oculatus), a rayfin fish that diverged from teleosts before the TGD, as well...

Within the framework of a genomics project on livestock species (AGENAE), we initiated a high-throughput DNA sequencing program of Expressed Sequence Tags (ESTs) in rainbow trout, Oncorhynchus mykiss. We constructed three cDNA libraries including one highly complex pooled-tissue library. These libraries were normalized and subtracted to reduce clone redundancy. ESTs sequences were produced, and 96,472 ESTs corresponding to high quality sequence reads were released on the international database, currently representing 42.5% of the overall sequence knowledge in this species. All these EST sequences and other publicly available ESTs in rainbow trout have been included on a publicly available Website (SIGENAE) and have been clustered into a total of 52,930 clusters of putative transcripts groups, including 24,616 singletons. 57.1% of these 52,930 clusters are represented by at least one Agenae EST and 14,343 clusters (27.1%) are only composed by Agenae ESTs. Sequence analysis also revea...

In fish, estrogens are well known for their involvement in ovarian differentiation and have been shown to be very potent feminizing agents when administrated in vivo during early development. However, the mechanism of action of exogenous estrogens is poorly understood. We report here on the feminizing effects of estrogen treatment on the testicular levels of some steroidogenic enzyme messenger RNAs [mRNAs; cholesterol side-chain cleavage (P450scc), 17-hydroxylase/lyase (P450c17), 3beta-hydroxysteroid dehydrogenase (3betaHSD), 11beta-hydroxylase (P45011beta), and aromatase (P450aro)] in the rainbow trout, Oncorhynchus mykiss. Treatment was carried out by dietary administration of 17beta-estradiol (E(2); dosage of 20 mg/kg diet) to a genetically all male population. Steroidogenesis in the differentiating testis was demonstrated to be strongly altered by E(2), as this treatment resulted in considerable decrease in P450c17, 3betaHSD, and P45011beta mRNAs after only 10 days of treatment....

ABSTRACT The genetic and environmental factors underlying sex determination and differentiation in teleost fish are still a matter of active debate. Nevertheless, the current hypothesis is that ovarian aromatase (cyp19a) expression plays a key role in orienting the embryonic gonad towards the female phenotype. On another hand, it is now well known that the brain of teleosts has a very high aromatase activity and that brain aromatase (cyp19b) expression occurs early during development. However, the potential role of aromatase in brain sexual differentiation is totally unknown. In this study, we monitor, for the first time in teleosts, the expression of the main steroidogenic enzymes in genetic all-male and all-female populations. Brains were dissected from male and female larvae around the period of sexual differentiation between 37 and 54 days post-fertilization at 10°C. At each of these time points, samples were pooled (n=30 to 50 depending on the stage) and mRNA were extracted for real-time RT-PCR gene expression analysis of cyp19b (cytochrome P450 aromatase B), cyp11a1 (cholesterol side chain cleavage), hsd3b1 (3β-hydroxysteroiddehydrogenase), cyp17a1 (17-hydroxylase) and sf1 (nr5a1b, steroidogenic factor 1). Very interestingly, a clear sexual dimorphism was observed between males and females which, in general, showed expression levels lower than in males. This was particularly obvious in the case of cyp19b and cyp11a1, but also for cyp17a1, at least at the youngest stages. Of particular interest is the fact that measurements performed on the gonads of the same animals showed opposite results, i.e., higher gene expression of most steroidogenic enzymes in females than males. Strikingly, brain aromatase (cyp19b) expression is much higher in the brain of males than in the brain of females, whereas expression of gonadal aromatase (cyp19a) is much higher in the differentiating female gonad. These results demonstrate that, at the time of gonadal differentiation, the brain is already sexually differentiated with at least some genes encoding steroidogenic enzymes exhibiting a clear sexual dimorphism. This early brain molecular sex differentiation may actually be more pronounced as in addition to these results, we also identified using a global microarray approach some other genes with a clear sexually dimorphic expression.

... Address all correspondence and requests for reprints to: Dr. Yann Guiguen, Laboratoire Institut National de la Recherche Agronomique-Station Commune de Recherches en Ichtyophysiologie, Biodiversité et Environement, Campus de Beaulieu, 35042 Rennes Cedex, France. ...

The reproductive function of fish, which is very sensitive to the variations of environmental factors, appears also to be particularly vulnerable to the presence of xenobiotics in the aquatic medium. Many physiological processes can be impaired, from sexual differentiation to female and male gametogenesis, due to disruptions among complex neuro-endrocrine, endocrine or paracrine regulations. This paper describes the main regulation

Sex hormone-binding globulin (SHBG) binds androgens and estrogens in the blood of many vertebrates, including teleost fish. In mammals, SHBG is synthetized in the liver and secreted into the blood. In fish, shbga also exhibits a hepatic expression. In salmonids, in which the gene has been duplicated, the recently discovered shbgb gene exhibits a predominantly ovarian expression. The present work aimed at gaining new insight into shbgb gene structure and expression during gonadal sex differentiation, a steroid-sensitive process, and Shbgb protein structure and binding characteristics; specifically, rainbow trout (Oncorhynchus mykiss) shbgb was analyzed. shbgb structure was analyzed in silico while expression was characterized during gonadal sex differentiation using all-male and all-female populations. We observed that shbgb gene and cognate-protein structures are similar to homologs previously described in zebrafish and mammals. The shbgb gene is predominantly expressed in differentiating female gonads, with increased expression around the end of ovarian differentiation. In the ovary, shbgb mRNA was detected in a subset of somatic cells surrounding the ovarian lamellae. Furthermore, Shbgb binds steroids with a higher selectivity than Shbga, exhibiting a higher affinity for estradiol compared to Shbga. In conclusion, Shbgb binding characteristics are clearly different from those of Shbga. Shbgb is expressed in the differentiating ovary during a period when the synthesis and action of testosterone and estradiol must be tightly regulated. This strongly suggests that Shbgb participates in the regulation of steroid metabolism and/or mediation, that is, needed during early gonadal development in rainbow trout.

In this review, we describe recent results concerning the genetics of sex determination in mammals. Particularly, we developed the study of the FOXL2 gene and its implication in genetic anomalies in goats (PIS mutation) and humans (BPES). We present the expression of FOXL2 in the ovaries of different species.

Sex determination is known to be male heterogametic in the rainbow trout, Oncorhynchus mykiss; however, scattered observations that deviate from this rather strict genetic control have been reported. Here, we provide a detailed morphological and histological characterization of the gonadal differentiation and development (from 43 days postfertilization to 11 months of age) in an all-female (XX) population with a genetically governed masculinization phenotype. In comparison with control males and females, the gonadal differentiation in these animals was characterized by many perturbations, including significantly fewer germ cells. This decrease in germ cells was confirmed by the significantly decreased expression of 2 germ cell maker genes (vasa and sycp3) in the masculinized XX populations as compared with the control females and control males. Although only a proportion of the total adult population was partially or fully masculinized, this early differentiating phenotype affected nearly all the sampled animals. This suggests that the adult masculinization phenotype is the consequence of an early functional imbalance in ovarian differentiation in the entire population. We hypothesize that the lower number of germ cells that we observed in this population could be one cause of their masculinization.