Perifused pituitary cultures: a model for LHRH regulation of LH secretion

In this paper, a dispersed anterior pituitary cell system that is continuously perifused and suitable for the study of luteinizing hormone-releasing hormone (LHRH)-anterior pituitary physiology is described. Adult male Sprague-Dawley rats were decapitated and the anterior pituitaries removed, dispersed using collagenase, mixed with Biogel P2, and packed into 0.9-cm columns using sterile technique. All experiments were conducted using Krebs-Ringer-bicarbonate-glucose buffer, and a stable base line was achieved. A constant dose of LHRH (50 ng) administered several times at 1-h intervals stimulated the same quantal release of LH. LH secretion is linear in response to LHRH over the range of 1-50 nM and to rat hypothalamic extracts over a range of 1/32 to 1/2 hypothalamic equivalents, regardless of whether the extracts are administered in ascending or descending order. These results indicate that perifusion of dispersed pituitary cells provides a useful model for the study of LHRH physiology, combining the advantages of the long-term viability of pituitary cells in culture with the dynamic nature afforded by perifusion.