Immunocytochemical visualization of luteinizing hormone-human chorionic gonadotropin (LH-hCG) receptors in Leydig cells in primary culture.

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Abstract
Visualization of human chorionic gonadotropin (hCG) binding sites was obtained by immunocytochemical reaction on Leydig cells cultured in chemically defined medium. After a short in vitro incubation with hCG or luteinizing hormone (LH), the cells were fixed and the bound molecules were revealed using anti-hCG or anti-LH antisera. In both cases the immunocytochemical reaction appeared as granulations at the cell surface. After prolonged (48 hr) culture in the presence of 0.5, 5, or 50 ng/ml of hCG, the hormone receptor complex is still visible. Similarly, following short exposure to hCG (0.5 or 50 ng/ml) and one or two days of culture without hCG, the immunocytochemical reaction is still present. These observations suggest that the half-life of the bound hormone is very long. This in vitro system in which the amount and time of hormone exposure are precisely defined provides arguments in favor of a long-term maintenance of the receptor complexes at the cell surface.