Specific calcium antagonist binding sites in brain.

The binding of the calcium antagonist [3H] nitrendipine ([3H] NDP) to brain and heart is described and the brain site is characterized. The binding is saturable, specific and of very high affinity with KD values of 0.16 nM in brain and 0.21 nM in heart. Our kinetic results are similar to those recently reported by two other groups (1, 2), indicating a saturable, high affinity binding site in brain. In brain the binding sites are enriched in crude nuclear and synaptosomal fractions. The highest levels of binding are seen in the hippocampus, caudate and cerebral cortex with much lower levels in the cerebellum and pons. Calcium has a marked stimulatory effect on [3H] NDP binding at 10(-4) M. Addition of 0.5 mM CaCl2 to EDTA treated membranes nearly doubles the number of binding sites. Of the many drugs and neurotransmitters tested only other calcium antagonists, i.e. verapamil, inhibit binding (IC50 = 250 nM). The inhibition of [3H] NDP binding by verapamil is apparently non-competitive and not complete, suggesting that [3H] NDP binds to several sites, only some of which are inhibited by verapamil. The [3H] NDP binding site is probably a protein since it is very sensitive to trypsin, heat and sulfhydryl reagents.