Relaxin triggers calcium transients in human granulosa-lutein cells

Although the peptide hormone relaxin is synthesized by the human corpus luteum in vivo, its potential to serve as a local factor in the regulation of luteal function is not clear. Using an enzyme-linked immunosorbent assay for human relaxin, we detected relaxin in the culture medium of human granulosa-lutein cells as early as after 6 days in culture. Moreover, 1 x 10(5) IU/l human chorionic gonadotropin stimulated relaxin release about fourfold during a 48-h incubation on culture days 6-8 (and 7-9), but not earlier (on days 1, 3 and 4). The stimulatory action of human chorionic gonadotropin on progesterone release was not influenced by relaxin, and relaxin alone was without stimulatory effect. However, human recombinant relaxin (between 0.1 and 12.5 micrograms/l) increased intracellular free Ca2+ basal levels to maximal peak levels exceeding 1000 nmol/l in about 64% of all tested cells (N = 168) with no obvious dependency on the culture day. The relaxin-induced Ca2+ signal was not affected by removal of extracellular Ca2+. As depletion of intracellular Ca2+ stores by ionomycin rendered the cells unresponsive to relaxin or diminished their ability to respond, these results point to an intracellular source of the Ca2+ signal. In summary, our data indicate the presence of a functional relaxin receptor on human granulosa-lutein cells, which is linked to Ca2+ release from intracellular stores.