Differential expression of GDF-9 and BMP- 15 duringfollicular development in canine ovaries evaluated by flowcytometry
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Growth differentiation factor 9 (GDF-9) and bone morphogenetic protein 15 (BMP-15) playimportant functions in follicular and oocyte development in many species. This study eval-uated the dynamic expression of GDF-9 and BMP-15 in canine follicles cells using flowcytometry analysis. Follicular cells were removed from three sizes of antral follicles (small,medium and large) from ovaries of bitches throughout the estrus cycle. Cells were incu-bated with anti-human GDF-9 polyclonal and anti-mouse BMP-15 monoclonal antibodies.A size and complexity discriminatory gate was used for the cytometryc analysis in theinitial dot plot and, additionally, a CD45 marker for leukocyte and propidium iodide (PI)were used for erythrocyte and debris discrimination. The evidence corroborated the pres-ence of both proteins in canine follicle cells, but these proteins were not expressed equallyduring follicular development. The results analyzed by ANOVA showed that GDF-9 expres-sion decreased (P < 0.05) during follicular growth in anestrus and proestrous/estrous, butincreased in diestrus (P < 0.05). The expression levels of BMP-15 rose (P < 0.05) from small tomedium sizes in anestrous without changing at diestrus. Small antral follicles expressed thehighest values of GDF-9 at anestrus while only BMP-15 showed higher value in small antralfollicles at proestrous-estrus compared to diestrus and anestrus. Both proteins decreasedin proestrous/estrous (P < 0.05) with increasing follicle size, registering the lowest levels inlarge follicles. The flow cytometric assay was able to assess GDF-9 and BMP-15 expressionin canine follicular cells, showing that these proteins were differentially expressed duringfollicular development, possibly related to the special features of canine reproduction.
Artículo de publicación ISI
Artículo de publicación SCOPUS
DOI: DOI: 10.1016/j.anireprosci.2016.02.005
Quote ItemAnimal Reproduction Science 167 (2016) 59–67
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