Developmental changes of cerebral cortical [3H]clonidine binding in rats: influences of guanine nucleotide and...

Cerebral cortical [3H]clonidine binding and influences of GTP and cations were investigated in developing rats. The results from Scatchard plots were compatible with the presence of two populations of binding sites [high-affinity binding (KD = 0.59 nM) and low-affinity binding (KD = 7.12 nM)] in 70-day-old rats but only high-affinity binding (KD = 0.27 nM) on day 1. Low-affinity binding was detectable on day 7. KD values in high- and low-affinity binding were not significantly changed during development after 7 days. Bmax of high-affinity binding reached a peak on day 15, and the value of low-affinity binding gradually increased with age. The addition of 10 microM GTP caused a significant reduction in Bmax of high-affinity binding after day 7. Neither KD nor Bmax of low-affinity binding was affected by 10 microM GTP during development. NaCl (10 and 100 mM) diminished the binding on days 7 and 70. MnCl2 (0.1 and 1.0 mM) markedly increased the binding on days 15 and 70 but not on day 7. It is suggested that: (1) single binding sites of alpha 2-adrenoceptors with higher affinity seem to be present on day 1; (2) low-affinity binding appears on day 7; (3) the number of high-affinity binding sites reaches a peak on day 15, followed by changes in populations of high-affinity as well as low-affinity sites without changing affinity; (4) the regulatory mechanism in alpha 2-receptors by guanine nucleotide reaches functional maturity between days 1 and 7; and (5) the involvement of Na+ and Mn2+ in alpha 2-receptor binding becomes functional by days 7 and 15, respectively.