Mutations in MAP3K1 tilt the balance from SOX9/FGF9 to WNT/β-catenin signaling

In-frame missense and splicing mutations (resulting in a 2 amino acid insertion or a 34 amino acid deletion) dispersed through the MAP3K1 gene tilt the balance from the male to female sex-determining pathway, resulting in 46,XY disorder of sex development. These MAP3K1 mutations mediate this balance by enhancing WNT/β-catenin/FOXL2 expression and β-catenin activity and by reducing SOX9/FGF9/FGFR2/SRY expression. These effects are mediated at multiple levels involving MAP3K1 interaction with protein co-fact

R-spondin1 and FOXL2 act into two distinct cellular types during goat ovarian differentiation

<p>Abstract</p> <p>Background</p> <p>Up to now, two loci have been involved in XX sex-reversal in mammals following loss-of-function mutations, PIS (Polled Intersex Syndrome) in goats and <it>R-spondin1 </it>(<it>RSPO1</it>) in humans. Here, we analyze the possible interaction between these two factors during goat gonad development. Furthermore, since functional redundancy between different <it>R-spondins </it>may influence gonad development, we also studied the expression patterns of <it>RSPO2, 3 </it>and <it>4</it>.</p> <p>Results</p> <p>Similarly to the mouse, <it>RSPO1 </it>shows a sex-dimorphic expression pattern during goat gonad development with higher levels in the ovaries. Interestingly, the PIS mutation does not seem to influence its level of expression. Moreover, using an RSPO1 specific antibody, the RSPO1 protein was localized in the cortical area of early differentiating ovaries (36 and 40 d<it>pc</it>). This cortical area contains the majority of germ cell that are surrounded by FOXL2 negative somatic cells. At latter stages (50 and 60 d<it>pc</it>) RSPO1 protein remains specifically localized on the germ cell membranes. Interestingly, a time-specific relocation of RSPO1 on the germ cell membrane was noticed, moving from a uniform distribution at 40 d<it>pc </it>to a punctuated staining before and during meiosis (50 and 60 d<it>pc </it>respectively). Interestingly, also <it>RSPO2 </it>and <it>RSPO4 </it>show a sex-dimorphic expression pattern with higher levels in the ovaries. Although <it>RSPO4 </it>was found to be faintly and belatedly expressed, the expression of <it>RSPO2 </it>increases at the crucial 36 d<it>pc </it>stage, as does that of <it>FOXL2</it>. Importantly, <it>RSPO2 </it>expression appears dramatically decreased in XX PIS^-/-gonads at all three tested stages (36, 40 and 50 d<it>pc</it>).</p> <p>Conclusion</p> <p>During goat ovarian development, the pattern of expression of <it>RSPO1 </it>is in agreement with its possible anti-testis function but is not influenced by the PIS mutation. Moreover, our data suggest that RSPO1 may be associated with germ cell development and meiosis. Interestingly, another RSPO gene, RSPO2 shows a sex-dimorphic pattern of expression that is dramatically influenced by the PIS mutation.</p

And act into two distinct cellular types during goat ovarian differentiation-2

Stages (36, 40 and 50 d) in XX sex-reversed gonads (XX Males) in comparison with normal males and females ones. GAPDH was used as control. The name of the amplified gene and the number of PCR cycles done is given on the left margin. Samples are from individual fetuses (identified by their number) or from a pool of 3 gonads (N = 3). L: 100 bp DNA ladder molecular weight marker (Bioline).<p><b>Copyright information:</b></p><p>Taken from "and act into two distinct cellular types during goat ovarian differentiation"</p><p>http://www.biomedcentral.com/1471-213X/8/36</p><p>BMC Developmental Biology 2008;8():36-36.</p><p>Published online 2 Apr 2008</p><p>PMCID:PMC2329615.</p><p></p

And act into two distinct cellular types during goat ovarian differentiation-0

EST), two human transcripts and the human gene. Genbank accession numbers of the different sequences are given on the left. A bovine-specific 5' non-coding exon 1 is depicted in red. The goat-specific part of exon 1 is depicted in pink. In goat, the second ATG (ATG) is not conserved in human (ACG). Sp = Signal peptide; Fu = Furin domain; Tsp = Thrombospondin domain; Nls = Nuclear localization signal; AG = conserved acceptor spicing site. b) A Neighbor-Joining tree was constructed with 28 DNA sequences from the four genes belonging to seven mammalian species, plus the putative goat ORF (the corresponding goat sequence is depicted in bold). Confidence values (higher than 50%) after bootstrap test are shown at each node. Genbank accession numbers are given in the methods section.<p><b>Copyright information:</b></p><p>Taken from "and act into two distinct cellular types during goat ovarian differentiation"</p><p>http://www.biomedcentral.com/1471-213X/8/36</p><p>BMC Developmental Biology 2008;8():36-36.</p><p>Published online 2 Apr 2008</p><p>PMCID:PMC2329615.</p><p></p

And act into two distinct cellular types during goat ovarian differentiation-4

is presented alone (left column) or with a DAPI blue nuclear-specific counterstaining (medium and right columns). The right column corresponds to a 5.0 enlargement of the red rectangle depicted on the medium column. At 40 d, RSPO1 is detected in the cortical area (co) of the ovaries where most of c-Kit positive germ cells lies. At this stage both somatic and germ cells are stained (arrows show 2 germinal cells). By contrast, FOXL2 positive cells are in the sub-cortical area (sc) of these early developing ovaries. At 50 d, RSPO1 is detected mainly around the c-Kit positive germ cells easily recognizable by their large and round nuclei (arrows). At this stage, FOXL2 positive somatic cells are located in the two ovarian compartments, cortex and medulla. m = mesonephros; the dotted line delimits both areas (co and sc).<p><b>Copyright information:</b></p><p>Taken from "and act into two distinct cellular types during goat ovarian differentiation"</p><p>http://www.biomedcentral.com/1471-213X/8/36</p><p>BMC Developmental Biology 2008;8():36-36.</p><p>Published online 2 Apr 2008</p><p>PMCID:PMC2329615.</p><p></p

List of CNVs identified with array-CGH in the seven cases with the indication of their code, type, location and size (CanFam2 assembly).

<p>CNVs were checked for occurrence in the Database of Genomic Copy Number Variants in the dog genome (<a href="http://dogs.genouest.org/LUPA.dir/CNV.html" target="_blank">http://dogs.genouest.org/LUPA.dir/CNV.html</a>) and in several papers <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0101244#pone.0101244-Chen1" target="_blank">[29]</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0101244#pone.0101244-Berglund1" target="_blank">[33]</a>.</p

Graphical representation of the SOX9 locus duplications discovered.

<p>The figure shows a 1,6(canFam2 assembly) and magnified views of the two SOX9 duplications detected, by array-CGH, in cases C10 (left) and C44 (right), respectively. The shaded areas indicate a gain in DNA copy number (duplication, average log2 ratios: +0, 5) detected by red dots. Asterisks indicate the 168 bp repeats.</p