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Conservation of the function of DMRT1 regulatory sequences in mammalian sex differentiation
Author(s) -
Boyer Alexandre,
Dornan Saffron,
Daneau Isabelle,
Lussier Jacques,
Silversides David W.
Publication year - 2002
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/gene.10158
Subject(s) - biology , transgene , green fluorescent protein , cre recombinase , microbiology and biotechnology , embryonic stem cell , gonadal ridge , genetically modified mouse , embryo , transgenesis , gene , embryogenesis , genetics , reproductive technology
Abstract Summary: Among genes involved in sex determination and differentiation, DMRT1 is the only one characterized to date containing a domain (the DM domain) that is conserved between phyla. To study DMRT1 transcriptional regulation within mammalian phyla, we generated transgenic mice that express green fluorescent protein (GFP) or Cre‐recombinase (Cre) under the control of 2.6 kb of pig DMRT1 5′ flanking sequences (pDMRT1p‐GFP and pDMRT1p‐Cre, respectively). Within the pDMRT1p‐GFP positive mice, GFP expression was observed in the XY genital ridge by embryonic day 11.5 (e11.5) and remained detectable during testis embryonic development to birth. GFP expression was restricted within testis cords as soon as cords were detectable. No fluorescence was observed in developing ovaries, although more sensitive RT‐PCR analysis revealed transgene expression in embryonic ovaries from e13.5 to e15.5. RT‐PCR performed on fluorescent activated cell sorter (FACS)‐purified GFP cells from e14.5, e17.5, and e19.5 developing testis showed that GFP expression was restricted to cells expressing the endogenous mouse Dmrt1 . GFP cells also expressed Mis and Oct4 , showing that the transgene is expressed in both Sertoli cell and germ cell compartments. In postnatal testis, transgene expression was detectable by GFP fluorescence from P0 to P21 in mice heterozygous for the transgene and through adulthood in mice homozygous for the transgene. In pDMRT1p‐Cre positive mice, Cre expression was detected within the genital ridges of both XY and XX embryos. We conclude that DMRT1 regulatory mechanisms during sexual differentiation are functionally conserved across mammalian evolution. The transgenic mouse lines described should provide useful marker systems for studies involving Dmrt1 gene expression during sex differentiation. genesis 34:236–243, 2002. © 2002 Wiley‐Liss, Inc.