BENZODIAZEPINE RECEPTOR-SITES IN THE CHICK OPTIC LOBE - DEVELOPMENT AND PHARMACOLOGICAL CHARACTERIZATION

To investigate the interaction between gamma-aminobutyric acid (GABA) and benzodiazepine (BZD) receptor sites during development, the time-course of appearance of flunitrazepam (FNZ) binding sites and their pharmacological characterization were studied in developing chick optic lobe. At the earliest stage examined, embryonic day (Ed) 12, the receptor density was 30.9 % (0.05 +/- 0.01 pmol/mg protein) of that found in the chick optic lobes of adult chicks. The adult value was achieved on Ed 16 (0.16 +/- 0.01 pmol/mg protein). After this stage there was a sharp and transient increase in specific [H-3]FNZ binding of about two-fold reaching a maximal value between hatching and the postnatal day (pnd) 2 (0.33 +/- 0.01 pmol/mg protein). Scatchard analysis at different stages of development revealed the presence of a single population of specific FNZ binding sites. The increase in [H-3]FNZ binding during development was due to a large number of binding sites while their affinity remained unchanged. Competition experiments in the chick optic lobe revealed that the order of potency for displacement of specific [H-3]FNZ binding paralleled the pharmacological potency of the BZDs tested. The IC50s for clonazepam, flunitrazepam, Ro 15-1788 and chlordiazepoxide were 3.02, 4.30, 0.32, and 4778.64 nM respectively. Ro 5-4864, a potent inhibitor of BZD binding to peripheral tissues, had no effect on specific [H-3]FNZ binding indicating that only central BZD binding sites are present in the chick optic lobe. The peak of maximal expression of BZD receptor sites precedes in 5-6 days the peak of GABA receptor sites indicating a precocious development of BZD receptor sites. The different appearance of both peaks may represent important events during development probably related to synaptogenesis.