title: Human and pig SRY 5' flanking sequences can direct reporter transgene expression to the genital ridge and to migrating neural crest cells.
abstract: Mechanisms for sex determination vary greatly between animal groups, and include chromosome dosage and haploid-diploid mechanisms as seen in insects, temperature and environmental cues as seen in fish and reptiles, and gene-based mechanisms as seen in birds and mammals. In eutherian mammals, sex determination is genetic, and SRY is the Y chromosome located gene representing the dominant testes determining factor. How SRY took over this function from ancestral mechanisms is not known, nor is it known what those ancestral mechanisms were. What is known is that SRY is haploid and thus poorly protected from mutations, and consequently is poorly conserved between mammalian species. To functionally compare SRY promoter sequences, we have generated transgenic mice with fluorescent reporter genes under the control of various lengths of human and pig SRY 5' flanking sequences. Human SRY 5' flanking sequences (5 Kb) supported reporter transgene expression within the genital ridge of male embryos at the time of sex determination and also supported expression within migrating truncal neural crest cells of both male and female embryos. The 4.6 Kb of pig SRY 5' flanking sequences supported reporter transgene expression within the male genital ridge but not within the neural crest; however, 2.6 Kb and 1.6 Kb of pig SRY 5' flanking sequences retained male genital ridge expression and now supported extensive expression within cells of the neural crest in embryos of both sexes. When 2 Kb of mouse SRY 5' flanking sequences (-3 to -1 Kb) were placed in front of the 1.6 Kb of pig SRY 5' flanking sequences and this transgene was introduced into mice, reporter transgene expression within the male genital ridge was retained but neural crest expression was lost. These observations suggest that SRY 5' flanking sequences from at least two mammalian species contain elements that can support transgene expression within cells of the migrating neural crest and that additional SRY 5' flanking sequences can extinguish this expression.
authors.affiliation: Faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, Quebec, Canada.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Boyer
authors.forename: Nicolas
authors.initials: N
authors.lastname: Pilon
authors.forename: Diana L
authors.initials: DL
authors.lastname: Raiwet
authors.forename: Jacques G
authors.initials: JG
authors.lastname: Lussier
authors.forename: David W
authors.initials: DW
authors.lastname: Silversides
mesh_terms: D024506:5' Flanking Region / Q000235:genetics*; D000818:Animals; D002465:Cell Movement / Q000235:genetics; D005260:Female; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009432:Neural Crest / Q000166:cytology / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D019849:Sex Determination Processes; D051876:Sex-Determining Region Y Protein / Q000235:genetics*; D013552:Swine / Q000235:genetics*
chemical_list: D008164:Luminescent Proteins; C497745:SRY protein, human; D051876:Sex-Determining Region Y Protein