BMP15 prevents cumulus cell apoptosis through CCL2 and FBN1 in porcine ovaries.
BACKGROUND
Bone morphogenetic protein-15 (BMP15) is a maternal gene necessary for mammalian reproduction. BMP15 expression increased in oocytes accompanied by follicle growth and development. The function and regulation mechanism of BMP15 in porcine cumulus cell apoptosis process is still unclear now.


METHODS
In this study, flow cytometry (FCM) was used to analyze the effects of BMP15 with different concentrations to cumulus cell apoptosis. High-throughput sequencing technology was carried out to screen regulatory genes linked closely with BMP15. In order to confirm the function of (MCP-1)/CCL2 and FBN1 in cumulus cell apoptosis, RNA interference (RNAi) method was used to inhibit the expression of (MCP-1)/CCL2 and FBN1. Apoptosis and proliferation of cumulus cell treated with siRNA transfection technology were measured by FCM, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, quantitative real time-PCR (RT-qPCR) and western blotting.


RESULTS
The results showed that the apoptosis levels of cumulus cell treated by BMP15 decreased significantly in a dose-dependent manner. The expression of related genes protein 1 (MCP-1)/CCL2 and fibrillin1 (FBN1) were both regulated by BMP15. After transfection, the proliferation of porcine cumulus cells increased significantly and apoptosis of cumulus cells was prevented while FBN1 was silenced after BMP15 treatment. The proliferation of cumulus cells decreased significantly and apoptosis rate of cumulus cells increased significantly while CCL2 was silenced.


CONCLUSIONS
The results obtained in this study firstly demonstrated that CCL2 and FBN1 are important regulatory factors of BMP15 in preventing cumulus cell apoptosis in porcine ovaries.
College of Animal Science, Jilin University, Changchun 130062, China.
Bo
B
Zhai
Huiyu
H
Liu
Xu
X
Li
Lisheng
L
Dai
Yan
Y
Gao
Changhong
C
Li
Lianjiang
L
Zhang
Yu
Y
Ding
Xianfeng
X
Yu
Jiabao
J
Zhang
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D055430:Bone Morphogenetic Protein 15 / Q000494:pharmacology*; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000235:genetics / Q000378:metabolism*; D054885:Cumulus Cells / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D005260:Female; D000071838:Fibrillin-1; D000071837:Fibrillins; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000187:drug effects; D020868:Gene Silencing; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D010053:Ovary / Q000187:drug effects*; D013552:Swine; D054392:Adipokines
D055430:Bone Morphogenetic Protein 15; D018932:Chemokine CCL2; D000071838:Fibrillin-1; D000071837:Fibrillins; D008840:Microfilament Proteins; C111835:CCL2 protein, human; C000605722:FBN1 protein, human; D054392:Adipokines