Effects of carbachol on rat Sertoli cell proliferation and muscarinic acetylcholine receptors regulation: an in vitro study

• Published in: Life sciences (1973) Vol. 75; no. 14; pp. 1761 - 1773
• Main Authors: Lucas, Thaı́s F.G, Avellar, Maria Christina W, Porto, Catarina S
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Inc 20.08.2004
• Subjects: Analysis of Variance; Animals; Carbachol; Carbachol - pharmacology; Cell Division - drug effects; Cell proliferation (Sertoli cells); Cells, Cultured; Internalization and recycling of receptors; Male; Muscarinic receptors; Rats; Rats, Wistar; Receptors, Muscarinic - drug effects; Scopolamine Hydrobromide - metabolism; Scopolamine Hydrobromide - pharmacology; Sertoli Cells - cytology; Sertoli Cells - drug effects; Temperature; Thymidine - metabolism; Time Factors; Tritium; Internalization and recycling of receptors; Cell proliferation (Sertoli cells); Carbachol; Muscarinic receptors
• ISSN: 0024-3205; 1879-0631
• DOI: 10.1016/j.lfs.2004.05.006

The aim of the present work was to study the effect of muscarinic agonist on cell proliferation and muscarinic acetylcholine receptors (mAChRs) regulation in rat Sertoli cells. Primary cultures of Sertoli cells were obtained from 8-day and 15-day old male Wistar rats. In proliferation assays, [methyl- 3H]thymidine incorporation in Sertoli cells from 8-day and 15-day old rats reached a plateau after 60 min of carbachol incubation and decreased after 120 min of agonist incubation. Binding studies with [N-Methyl- 3H]scopolamine ([ 3H]NMS) indicated a rapid loss of cell surface mAChRs when Sertoli cells from 15-day old rats were incubated with carbachol at 35 °C for 2 min. This effect was temperature-dependent. When the incubation of the cells was prolonged at 35 °C or at 4 °C, after the agonist had been washed away, 94% of mAChRs were present in the cell surface after 120 min incubation at 35 °C. At 4 °C, however, a low percentage of mAChRs was detected in the cell surface. In the presence of cycloheximide, the recycling of mAChRs to the cell surface was not changed, suggesting that the appearance of mAChRs on cell surface was not dependent on de novo receptor synthesis. In conclusion, our studies indicate that the activation of mAChRs may play a role in rat Sertoli cell proliferation. These receptors may be under regulation (internalization and recycling) when cells are exposed to muscarinic cholinergic agonist.