Detection of Relaxin Release by Porcine Luteal Cells Using a Reverse Hemolytic Plaque Assay: Effect of Prostaglandins E2 and F2α, Human Chorionic Gonadotropin, and Oxytocin1

The release of relaxin from cultured porcine luteal cells derived from pregnant sows was detected by a reverse hemolytic plaque assay. In this assay, luteal cells are cocultured in monolayers with protein-A-coupled ovine erythrocytes. In the presence of porcine relaxin antiserum and complement, a zone of hemolysis--a plaque--develops around relaxin-releasing luteal cells. Treatment with prostaglandin E2 (10(-8) and 10(-6) M) significantly accelerated the rate of plaque formation; in contrast, human chorionic gonadotropin (10-1,000 IU/ml) inhibited the rate of plaque formation. Oxytocin (10(-8) to 10(-4) M) had no detectable effect on relaxin release. However, none of these treatments or long-term preexposure to prostaglandin F2 alpha increased the total proportion of large luteal cells that released relaxin, which remained at about 50%. These results are consistent with the idea that prostaglandins of uterine and/or luteal origin and pituitary luteinizing hormone may contribute, alone or perhaps in combination, to the overall regulation of ovarian relaxin release during pregnancy in the sow. In addition, the results indicate that the effects of prostaglandins are restricted to a subpopulation of large luteal cells that release detectable amounts of relaxin in culture.