4 research outputs found
Recombinant BMP15 inhibition of progesterone secretion.
<p>Granulosa cells from small antral (1–3 mm in diameter) ovine follicles were cultured for 96 h in serum-free conditions. Cultures were performed in the absence (control) or in the presence of increasing dose (10, 50, 200ng/ml) of wild type recombinant human BMP15<sup>wt</sup> or BMP15<sup>Y235C</sup> mutant. Each treatment was tested in triplicate in each of 5 independent experiments. The results represent the amount of progesterone secreted by 50 000 cells between 48 h and 96 h of culture (mean ±SEM). Dose effect was analyzed by one-way ANOVA by comparing each dose to control (**, p<0.01; ***, p<0.001). Mutation effect was analyzed by Student t-test within each dose ($, p<0.05).</p
BMP15 positive selection pressure among mammals.
<p>Phylogenetic tree of BMP15 from 24 mammals species reconstructed using TreeBeST and rooted by minimizing with the number of duplications and losses. Bootstrap values are given when nodes are strongly supported (>80%). Branches or species that have a significant LRT for positive selection calculation are indicated in bold (threshold of <i>q</i>= 5% of false positives). The scale represents the substitution rate. </p
BMP15 evolution among TGFß/BMP family members.
<p>Phylogenetic tree of thirteen members of the TGFβ/BMP superfamily expressed by the mammalian ovary reconstructed using maximum likelihood. Bootstrap values are given when nodes are strongly supported (>80%). The scale represents the substitution rate.</p
Functional analysis of amino acids under positive selection in human BMP15.
<p>A, Human BMP15 full sequence showing prodomain, cleavage site (underlined) and mature peptide (framed sequence); known human variants for BMP15 found in POI patients (framed grey highligthed); and identified amino acids under positive selection (framed colored highlighted): BMP15<sup>F146</sup>, BMP15<sup>L189</sup>, BMP15<sup>Y235</sup>. B, <i>In </i><i>vitro</i> reporter luciferase assay from COV434 granulosa cells transiently transfected with empty vector +/- 100ng recombinant human BMP15 (mock +/- BMP15) or wild type human BMP15 expressing vector (WT) or the different BMP15 variants vectors obtained by directed-mutagenesis of residues under positive selection. Results are expressed as the mean (±SD) of 4 independent experiments. Differences between means were analyzed by one-way ANOVA by comparing each condition to WT (*, p<0.05; **, p<0;01; ***, p<0.001).</p