Homepage bandeau_genéral

  Director : Fellous Marc (mfellous@pasteur.fr)



A SRY gene partner was identified as TGIF in vivo and in vitro in human or mouse. Yq deletions were characterised in Human infertility. A human Y haplotype 26 was found to be associated with low sperm count leading to male infertility. (Kenneth MCELREAVEY)

Based on whole genome scanning we identified several regions of the human genome associated with autism: 6q23, 6q16-21 and Xpter.( Thomas BOURGERON)

We analysed the genetic of premature ovarian failure in man isolated and syndromic (BPES). We described mutation in FOXL2 gene. FOXL2 gene is also involved in Palled mutation in goat.( Marc FELLOUS)




Sex determination and differentiation: Sex determination is the translation of the chromosomal sex (XX or XY) into the development of the gender-appropriate internal and external reproductive structures. The initial events of sex determination are, therefore, genetically determined. Errors in the process are fairly common and can range in severity from complete sex reversal to minor genital abnormalities. Sex determination, as an early embryological event, is a dynamic morphological process that can help us address basic questions of gene expression, cell-fate determination, and hormone signalling. The mechanisms involved in normal sex differentiation are not yet well defined and, while many genes are known to contribute to the process, the nature and timing of their interactions remain unclear.

Germ cell development and maintenance: Approximately two percent of human males are infertile due to severe defects in sperm production. In 50-60 % of individuals the etiology is unknown. A number of studies have suggested that sperm counts in certain European countries may be declining, the reasons for this decline are unclear but are associated with an increase in other urogenital anomalies such as hypospadias and cryptorchidism. In Paris sperm counts have been declining by two percent per year since the mid 1970's. The genetics of male infertility is complex. Infertile couples have fewer sibs compared with fertile control groups and male infertility shows a distinct pattern of familial aggregation, suggesting that non-Mendelian multifactorial inheritance could play a significant role. Approximately 15% of men with unexplained azoospermia or severe oligozoospermia have microdeletions of the Y chromosome that remove genes that are involved in male germ cell development and maintenance. Despite the recent interest in Y chromosome microdeletions and infertility, there remains a large group of men with unexplained infertility. Premature ovarian failure (POF) has an incidence of approximately 1 in 100. Although immunological disorders are frequently associated with POF, the etiology is unknown in approximately 60 % of cases. Remarkably this relatively common phenotype has received very little genetic investigation. Recent studies suggest that between 30-50% of all POF cases may be familial suggesting an important genetic contribution to the phenotype.

(i) Identify factors involved in human sex determination and differentiation

Active recruitment of familial cases of pathologies associated with human sex determination (XY gonadal dysgenesis, XX ovarian dysgenesis and XX males) is underway. Already this project (collaboration with Dr H. Ostrer NYU) has resulted in the recruitment of >90% of all published familial cases (http://www.med.nyu.edu/genetics/sexdet.html). Additional efforts to recruit families, particularly in the Indian subcontinent are underway (in collaboration with several paediatric centres, both nationally and internationally). Linkage analysis has been performed in some cases and novel loci implicated in human sex determination have been identified.

-Identify novel sex determining genes by the detailed analysis of chromosomal breakpoints associated with sex reversal (eg 9p)

-Identify novel sexes determining/differentiation genes by sequence homology and database mining.

-Identify protein partners of SRY by Far-Western blotting techniques. Several candidates have been identified and are under characterisation (In collaboration with N. Hanley Southhampton)

-Comparative Genome Hybridisation has identified a number of novel loci associated with anomalies of gonadal development. These are currently being characterised (in collaboration with Dr H. Ostrer, NYU and Kurt Hirschhorn, Mount Sinai School of Medicine).

(ii) Identify factors necessary for human germ cell development and maintenance

A large scale effort has been made to identify familial cases of male and female infertility. Currently this involves INSERM U491 (Marseilles, France), and other major European centres of reproductive medicine, which have agreed to participate in this study (Denmark, U.K., Italy, Poland and Germany). Efforts are underway in India to recruit additional families (collaboration with major fertility clinics in Hyderabad, Bombay, Calcutta, Delhi, Ahemabad and Indore).

(iii) Identify genetic risk factors associated with infertility and gonadal cancers in defined populations/ethnic groups.

This has already been performed on an infertile Danish population and is being extended to include a larger sample size and to include testicular cancer (TC) patients. Further major studies of Swedish, Finnish, German and Austrian populations (both infertility and TC). Other studies in Iran and in India are being performed to access the genetic risk in these non-European populations. Epidemiological evidence indicates that the genetics of unexplained infertility and unexplained ambiguous genitalia are non-Mendelian. We have begun to identify some of these factors in one European population. This analysis will be extended. The aim of this project is to recruit familial cases in France of unexplained ambiguous genitalia. Active recruitment will be for three years, following which; linkage or association studies will be performed depending on the population structure.

(iv) The relation between environmental factors and problems of gonad development and function. In collaboration with Dr P Saunders and Pr R Sharpe (Edinburgh) we have been examining the expression of cell cycle proteins and the estrogen in testicular cancer. Preliminary results suggest a leading role for estrogen in the initiation of testicular cancer. Studies are ongoing. This project, involving groups from Denmark, Finland, Germany and Holland, is focused on genetic susceptibility to TC, identification of genes involved in the development and progression of TC (linkage analysis, CGH), analysis of cell cycle proteins and the analysis of genomic instability (both Y chromosome and autosomal loci).



Role of sexual dimorphism on brain development and psychiatric diseases

Identification and characterisation of new candidate genes for schizophrenia and autism

Based on whole genome scan results, several regions of the human genome were studied on chromosome 6, X and Y.

Chromosome 6p23

On chromosome 6p23, five independent studies have localised a locus for predisposition to schizophrenia between markers D6S274 and D6S285. Within this 2 cM genetic interval, we isolated and characterised two novel genes Autosomal Highly Conserved Protein (FAM8A1) and KIF13A that we are currently testing for association in patients with schizophrenia.

Family with sequence similarity 8 (FAM8A1) : At the beginning of this study (1998), FAM8A1 was the first gene identified between D6S274 et D6S285. This gene code for a putative membrane protein, specific to metazoans. We also identified the conserved orthologous genes in C. elegans et D. melanogaster. FAM8A1 does not contain any known motif and its function is still not elucidated. FAM8A1 is ubiquitously expressed in the adult with a testis specific transcript. During mouse development, the expression is restricted to the central nervous system (CNS). There are five unexpressed pseudogenes of FAM8A1 (FAM8A2P-A6P) in the human genome ) each one inserted within a human endogenous retrovirus (HERV). The capture of the FAM8A1 cellular mRNA by ERV occurred during primate evolution and resemble to the transduction of cellular oncogene. After the capture, this retroelement (ERV + FAM8A1) was transposed in several place of the genome on human chromosome 2, 6p21, 6q16, 11 et Y. Before FAM8A1, this transduction mechanism has never been observed in the human. The coding regions of the functional gene were tested by SSCP in 73 schizophrenic patients and no association was observed.

KIF13A : We identified this gene within the same genetic interval than FAM8A1. KIF13A is a new member of the unc-104/KIF1A kinesin family involved in the anterograde transport (from the cell body to the synapses) of cellular organites and synaptic vesicules (cargos). Mutations in the unc-104 from C. elegans results in abnormal behaviour (unc = uncoordinated) and a decrease of synaptic vesicules. During early mouse development, KIF13A expression is restricted to the CNS. Several single nucleotide polymorphisms (SNPs) within KIF13A will be tested in schizophrenic patients.

Chromosome 6q16-21

A genome scan was previously performed and pointed chromosome 6q21 as a candidate region for autism. This region contains the glutamate receptor 6 (GluR6 or GRIK2) gene, a functional candidate for the syndrome. Glutamate is the principal excitatory neurotransmitter in the brain and is directly involved in cognitive functions such as memory and learning. We used two different approaches, the affected sib-pair (ASP) method and the transmission disequilibrium test (TDT), to investigate the linkage and association between GluR6 and autism. The ASP method, conducted on 59 families, showed a significant excess of allele sharing, generating an elevated multipoint maximum LOD score (GENEHUNTER-PLUS NPL= 3.28; P = 0.0005). Using 107 additional families with a single affected child, a significant maternal transmission disequilibrium was observed (TDT linkage P = 0.0004). Furthermore, TDT (with only one affected proband per family) and Haplotype Relative Risk (HRR) analyses showed significant association between GluR6 and autism (TDT association P = 0.008; HRR P = 0.01). In contrast to maternal transmission, paternal transmission of GluR6 alleles was as expected in the absence of linkage, suggesting a maternal effect such as imprinting. Mutation screening was performed in 33 affected individuals, revealing several nucleotide polymorphisms (SNPs), including one amino acid change (M867I) found in 8% of the autistic subjects. This change takes place in a highly conserved domain of the protein and seems to be more maternally transmitted than expected to autistic males (P = 0.007). Presently, we are testing for the imprinting status of GluR6 and in collaboration with C. Mulle at the University of Bordeaux, we are testing the functional consequence of the amino acid change (M867I) on the receptor properties.

Chromosomes X et Y

Due to the gender difference in autism (male to female sex ratio 4:1), sexual chromosomes are also investigated. Several regions of the X and the Y chromosome were associated with autism on the bases of cytogenetic rearrangements. Chromosome Xp22.3 closed to the pseudo autosomal region (PAR) was found deleted in three girls with autism. This region is also the most significant region identified by the whole genome scan of the PARIS study. Several candidate genes are localised within this interval and are currently tested. In order to determine a Y chromosome effect in the predisposition of the male to the syndrome, we analysed the Y chromosome haplogroup frequencies in 107 autistic subjects from France, Sweden and Norway. No significant difference in Y-haplotype distribution between the affected and control groups was observed. Although this study cannot exclude the presence of a Y susceptibility gene, our results are not suggestive of a Y chromosome effect in autism.



Mutations in FOXL2, a forkhead transcription factor gene, are the major cause of type I and Ii blepharophimosis-ptosis-epicanthus inversus syndrome (BPES), a rare genetic disorder. In BPES type I a complex eyelid malformation is associated with premature ovarian failure (POF), while in BPES type II the eyelid defect occurs as an isolated entity. We have dentified novel mutations in the FOXL2 gene in BPES type I and II families, in sporadic BPES patients, and in BPES families where the type could not be established. Mutations were detected in 67% of the patients studied. In total twenty-one mutations were identified, seventeen of which are novel, and one microdeletion. Thirteen of these FOXL2 mutations are unique. We demonstrate that there is a genotype-phenotype correlation for both types of BPES by the finding that mutations predicted to result in a truncated protein both lacking or containing the forehead domain lead to type I BPES. In contrast, duplications within or downstream of the forehead domain, and a frameshift downstream of them, all predicted to result in an extended protein, cause type Ii BPES. In addition, in thirty unrelated patients with isolated POF no causal mutations were identified in FOXL2. This study provides further evidence that FOXL2 haploinsufficiency may cause BPED type I and II by the effect of a null allele and a hypomorphic allele, respectively. Furthermore, we propose that in a fraction of the BPES patients the genetic defect may not reside within the coding region of the FOXL2 gene and may be caused by a position effect. IN goats, FOXL2 expression was studied and found sexually dimorphic in the gonads at all developmental stages, from the beginning of genital ridges formation (30 days post coïtum) until adulthood? Its expression pattern, studied on RT samples by Real time PCR (TaqMan), revealed a higher level in ovaries than in testis (in a ratio varying from 30 to 1300 fold, depending on the stage). Moreover, we demonstrated that FOXL2 gene expression is directly affected by the polled intersex Syndrome (PIS) mutation, shown to be a 11.7 kb deletion in collaboration with INRA of Jouy en Josas. This deletion drastically decreased FOXL2 expression in the XX sex reversed gonads at all developmental stages. This study carried out on the goat species, indicates that in addition to its role in folliculogenesis, FOXL2 seems to be a critical factor for the first steps of ovarian development. Therefore, this result brings to light an interesting candidate gene for female to male sex reversal in humans.



puce Publications of the unit on Pasteur's references database


  Office staff Researchers Scientific trainees Other personnel

Michèle AOSAKA - aosaka@pasteur.fr

Dauphine DE CROZES - dauphine@pasteur.fr

Marc FELLOUS - mfellous@pasteur.fr

Kenneth McELREAVEY IP - kenmce@pasteur.fr

Giovanna VINCI CR1 INSERM - vinci@pasteur.fr

Thomas BOURGERON M.C. Université P7 - thomasb@pasteur.fr

Lluis QUINTANA-MURCI Chercheur CNRS - quintana@pasteur.fr

Reiner VEITIA-SARMIENTO M.C. Université P7 - rveitia@pasteur.fr

Stéphane JAMAIN Etudiant en thèse (prévue 01/06/2002) sjamain@pasteur.fr

Julie COQUET Etudiante en DEA à partir du 01/11/01

Pascal LEONARDI Etudiant en thèse

Brigitte LEMERCIER Technicienne IP - blemer@pasteur.fr

Hélène QUACH Technicienne - hquach@pasteur.fr

Fatima TALEB Technicienne INSERM en remplacement de Mr CLERGUE (vacations) (départ 15 décembre)


Page Top research Institut Pasteur homepage

If you have problems with this Web page, please write to rescom@pasteur.fr.