PMID- 35772748 OWN - NLM STAT- MEDLINE DCOM- 20220704 LR - 20230703 IS - 1525-3163 (Electronic) IS - 0021-8812 (Print) IS - 0021-8812 (Linking) VI - 100 IP - 7 DP - 2022 Jul 1 TI - Stimulatory effects of TGFalpha in granulosa cells of bovine small antral follicles. LID - 10.1093/jas/skac105 [doi] LID - skac105 AB - Intraovarian growth factors play a vital role in influencing the fate of ovarian follicles. They affect proliferation and apoptosis of granulosa cells (GC) and can influence whether small antral follicles continue their growth or undergo atresia. Transforming growth factor-alpha (TGFalpha), an oocyte-derived growth factor, is thought to regulate granulosa cell function; yet its investigation has been largely overshadowed by emerging interest in TGF-beta superfamily members, such as bone morphogenetic proteins (BMP) and anti-Mullerian hormone (AMH). Here, effects of TGFalpha on bovine GC proliferation, intracellular signaling, and cytokine-induced apoptosis were evaluated. Briefly, all small antral follicles (3-5 mm) from slaughterhouse specimens of bovine ovary pairs were aspirated and the cells were plated in T25 flasks containing DMEM/F12 medium, 10% FBS, and antibiotic-antimycotic, and incubated at 37 degrees C in 5% CO2 for 3 to 4 d. Once confluent, the cells were sub-cultured for experiments (in 96-, 12-, or 6-well plates) in serum-free conditions (DMEM/F12 medium with ITS). Exposure of the bGC to TGFalpha (10 or 100 ng/mL) for 24 h stimulated cell proliferation compared to control (P < 0.05; n = 7 ovary pairs). Proliferation was accompanied by a concomitant increase in mitogen-activated protein kinase (MAPK) signaling within 2 h of treatment, as evidenced by phosphorylated ERK1/2 expression (P < 0.05, n = 3 ovary pairs). These effects were entirely negated, however, by the MAPK inhibitor, U0126 (10uM, P < 0.05). Additionally, prior exposure of the bGC to TGFalpha (100 ng/mL) failed to prevent Fas Ligand (100 ng/mL)-induced apoptosis, as measured by caspase 3/7 activity (P < 0.05, n = 7 ovary pairs). Collectively, the results indicate TGFalpha stimulates proliferation of bGC from small antral follicles via a MAPK/ERK-mediated mechanism, but this action alone fails to prevent apoptosis, suggesting that TGFalpha may be incapable of promoting their persistence in follicles during the process of follicular selection/dominance. CI - (c) The Author(s) 2022. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com. FAU - Lundberg, Allie L AU - Lundberg AL AD - Orna Therapeutics, Cambridge, MA 02139, USA. FAU - Jaskiewicz, Nicole M AU - Jaskiewicz NM AD - Department of Animal and Veterinary Sciences, The University of Vermont, Burlington, VT 05405, USA. FAU - Maucieri, Abigail M AU - Maucieri AM AD - Department of Animal and Veterinary Sciences, The University of Vermont, Burlington, VT 05405, USA. FAU - Townson, David H AU - Townson DH AD - Department of Animal and Veterinary Sciences, The University of Vermont, Burlington, VT 05405, USA. LA - eng GR - #2013-67016-21071/U.S. Department of Agriculture/ GR - #DGE 1450271/National Institute of Food and Agriculture/ GR - #DGE 1450271/National Science Foundation/ PT - Journal Article PL - United States TA - J Anim Sci JT - Journal of animal science JID - 8003002 RN - 0 (Intercellular Signaling Peptides and Proteins) RN - 0 (Transforming Growth Factor alpha) RN - 0 (Transforming Growth Factor beta) RN - 80497-65-0 (Anti-Mullerian Hormone) SB - IM MH - Animals MH - Anti-Mullerian Hormone/metabolism MH - Cattle MH - Female MH - Granulosa Cells/metabolism MH - Intercellular Signaling Peptides and Proteins/metabolism MH - Oocytes MH - *Ovarian Follicle/physiology MH - *Transforming Growth Factor alpha/metabolism/pharmacology MH - Transforming Growth Factor beta/metabolism/pharmacology PMC - PMC9246657 OAB - A variety of hormones regulate ovarian function in the cow, thus influencing fertility. One such hormone, transforming growth factor-alpha, TGFalpha, is expressed by the oocyte (egg) of the bovine ovary; yet little other information about the actions of this molecule on ovarian cells is available. In this study, we determined that although TGFalpha directly stimulates growth and proliferation of cells of the bovine ovary, and does so via specific signaling mechanisms, it fails to prevent immune-mediated programmed cell death. The latter observation diminishes the importance of TGFalpha relative to other oocyte-derived hormones in terms of ovarian function and overall animal fertility. OABL- eng OTO - NOTNLM OT - apoptosis OT - bovine OT - follicle OT - granulosa cell OT - mitogen-activated protein kinase (MAPK) signaling OT - transforming growth factor-alpha EDAT- 2022/07/01 06:00 MHDA- 2022/07/06 06:00 PMCR- 2023/07/01 CRDT- 2022/06/30 20:12 PHST- 2022/01/27 00:00 [received] PHST- 2022/03/30 00:00 [accepted] PHST- 2022/06/30 20:12 [entrez] PHST- 2022/07/01 06:00 [pubmed] PHST- 2022/07/06 06:00 [medline] PHST- 2023/07/01 00:00 [pmc-release] AID - 6620783 [pii] AID - skac105 [pii] AID - 10.1093/jas/skac105 [doi] PST - ppublish SO - J Anim Sci. 2022 Jul 1;100(7):skac105. doi: 10.1093/jas/skac105.