title: Survival role of locally produced acetylcholine in the bovine corpus luteum.
abstract: The present study was conducted to explore the source of acetylcholine (ACH) in the corpus luteum (CL) and to test our hypothesis of an antiapoptotic role of ACH in the bovine CL and, further, to investigate whether nerve growth factor (NGF), insulin-like growth factor 1 (IGF1), and transforming growth factor beta1 (TGFB1) influence the expression of choline acetyltransferase (CHAT), the biosynthetic enzyme of ACH, in cultured bovine luteal cells. Protein expression and immunolocalization of CHAT were carried out at different stages throughout the luteal phase and in cultured luteal and endothelial cells. ACH was measured in luteal tissue at the different luteal stages and in luteal cells cultured for 8 and 24 h. Cell viability and TUNEL assays were performed on cultured midluteal cells treated with or without tumor necrosis factor alpha (TNF)/interferon gamma (IFNG) in the presence of ACH and its muscarinic (atropine) and nicotinic (mecamylamine) receptor antagonists. The CL was devoid of cholinergic nerve fibers. CHAT immunostaining was evident in luteal, endothelial, and stromal cells in luteal tissue sections and in cultured luteal and endothelial cells. CHAT protein was expressed throughout the cycle without any significant changes. ACH concentration in luteal tissue was not changed during the luteal stages but increased over time and with increased cell numbers in luteal cell cultures. ACH increased cell viability and prevented cell death induced by TNF/IFNG. Atropine significantly attenuated ACH action, whereas mecamylamine had no effect. TNF/IFNG treatment downregulated CHAT expression, whereas NGF, IGF1, and TGFB1 upregulated CHAT expression, in cultured luteal cells. The overall findings strongly suggest a nonneural source and antiapoptotic role of ACH in the bovine CL. Locally produced ACH appears to be regulated by NGF, IGF1, and TGFB1.
authors.affiliation: Laboratory of Reproductive Endocrinology, Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan.
authors.forename: M Omar
authors.initials: MO
authors.lastname: Al-Zi'abi
authors.forename: Anom
authors.initials: A
authors.lastname: Bowolaksono
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Okuda
mesh_terms: D000109:Acetylcholine / Q000378:metabolism* / Q000502:physiology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002417:Cattle; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D018680:Cholinergic Antagonists / Q000494:pharmacology; D003338:Corpus Luteum / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D003339:Corpus Luteum Hormones / Q000502:physiology; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D005260:Female; D007371:Interferon-gamma / Q000494:pharmacology; D008184:Luteal Cells / Q000187:drug effects / Q000378:metabolism; D011374:Progesterone / Q000378:metabolism; D011950:Receptors, Cholinergic / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D018680:Cholinergic Antagonists; D003339:Corpus Luteum Hormones; D011950:Receptors, Cholinergic; D014409:Tumor Necrosis Factor-alpha; D011374:Progesterone; D007371:Interferon-gamma; D002795:Choline O-Acetyltransferase; D000109:Acetylcholine