RHOG-DOCK1-RAC1 Signaling Axis Is Perturbed in DHEA-Induced Polycystic Ovary in Rat Model.
The function of RHOG, a RAC1 activator, was explored in the ovary during ovarian follicular development and pathological conditions. With the help of immunoblotting and immunolocalization, we determined the expression and localization of RHOG in normal (estrous cycle) and polycystic ovaries using Sprague Dawley (SD) rat model. Employing polymerase chain reaction and flow cytometry, we analyzed the transcript and expression levels of downstream molecules of RHOG, DOCK1, and RAC1 in the polycystic ovarian syndrome (PCOS) ovary along with normal antral follicular theca and granulosa cells after dehydroepiandrosterone (DHEA) supplementation. The effect of RHOG knockdown on DOCK1, VAV, and RAC1 expression was evaluated in the human ovarian cells (SKOV3), theca cells, and granulosa cells from SD rats with the help of flow cytometry. Oocyte at secondary follicles along with stromal cells showed optimal expression of RHOG. Immunoblotting of RHOG revealed its maximum expression at diestrus and proestrus, which was downregulated at estrus stage. Mild immunostaining of RHOG was also present in the theca and granulosa cells of the secondary and antral follicles. Polycystic ovary exhibited weak immunostaining for RHOG and that was corroborated by immunoblotting-based investigations. RHOG effectors DOCK1 and ELMO1 were found reduced in the ovary in PCOS condition/DHEA. RHOG silencing reduced the expression of DOCK1 and RAC1 in the theca and granulosa cells from SD rat antral follicles and that was mirrored in the human ovarian cells. Collectively, RHOG can mediate signaling through downstream effectors DOCK1 and RAC1 during ovarian follicular development (theca and granulosa cells and oocyte), but DHEA downregulated them in the PCOS ovary.
1 Endocrinology Division, CSIR-Central Drug Research Institute, Lucknow, India.
Vaibhave
V
Ubba
1 Endocrinology Division, CSIR-Central Drug Research Institute, Lucknow, India.
Upendra Kumar
UK
Soni
1 Endocrinology Division, CSIR-Central Drug Research Institute, Lucknow, India.
Sangappa
S
Chadchan
1 Endocrinology Division, CSIR-Central Drug Research Institute, Lucknow, India.
Vineet Kumar
VK
Maurya
1 Endocrinology Division, CSIR-Central Drug Research Institute, Lucknow, India.
Vijay
V
Kumar
1 Endocrinology Division, CSIR-Central Drug Research Institute, Lucknow, India.
Ruchika
R
Maurya
1 Endocrinology Division, CSIR-Central Drug Research Institute, Lucknow, India.
Himanshu
H
Chaturvedi
1 Endocrinology Division, CSIR-Central Drug Research Institute, Lucknow, India.
Rajender
R
Singh
1 Endocrinology Division, CSIR-Central Drug Research Institute, Lucknow, India.
Anila
A
Dwivedi
1 Endocrinology Division, CSIR-Central Drug Research Institute, Lucknow, India.
Rajesh Kumar
RK
Jha
D000818:Animals; D002460:Cell Line; D003687:Dehydroepiandrosterone; D004195:Disease Models, Animal; D030762:Estrous Cycle; D005260:Female; D020558:GTP Phosphohydrolases / Q000378:metabolism*; D006801:Humans; D006080:Ovarian Follicle / Q000378:metabolism* / Q000473:pathology; D011085:Polycystic Ovary Syndrome / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D011627:Puberty; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D020744:rac GTP-Binding Proteins / Q000378:metabolism*; D020830:rac1 GTP-Binding Protein / Q000378:metabolism*
C489578:Rhog protein, rat; D003687:Dehydroepiandrosterone; D020558:GTP Phosphohydrolases; C499134:Rac1 protein, rat; D020744:rac GTP-Binding Proteins; D020830:rac1 GTP-Binding Protein
DHEA; DOCK1; ELMO1; PCOS; RAC1; RHOG; SKOV3; ovary; theca and granulosa cells