Release of chicken luteinising hormone-releasing hormone-I (cLHRH-I) by mediobasal hypothalamus in the cockerel: validation of an incubation system and effect of excitatory amino acids

Abstract

Summary ― An in vitro system for the incubation of mediobasal hypothalami (MBH) of cockerels and a radioimmunoassay for chicken luteinising hormone-releasing hormone-I (cLHRH-1) were developed. The size of the hypothalamic fragment (MBH including the median eminence) and the incubation conditions used (40°C, under constant shaking and gassing) preserved the physiological properties of the tissue. It was possible to maintain the MBH in vitro and to study the LHRH release for several hours. The assay proved sensitive enough (ED BO = 0.794 pmol/tube, ie 4.59 pg/ml) and sufficiently precise (within-assay coefficient of variation = 4.4% and between-assay coefficient of variation = 10.2%) to measure the amounts of peptide released in the incubation medium. The use of this incubation system provided the first evidence of the stimulating effect of the excitatory amino acids glutamate, NMDA and kainate on the secretion of cLHRH-1 in birds. Our results suggest that the effect on the NMDA receptor is predominant.