Abstract

Potassium-stimulated 45Ca entry into rat brain synaptosomes was measured at times ranging from 1 to 60 s. The K-rich solutions were used to depolarize the synaptosomes. Backflux of 45Ca from the synaptosomes was negligible during the first 10-20 s of incubation. An initial ("fast") phase of K-stimulated Ca entry, lasting from 1 to 2 s was observed. This phase was inhibited by low concentrations of La (KI approximately equal to 0.3 microM). It was also abolished ("inactivated") by incubating the synaptosomes in depolarizing solutions (containing veratridine, gramicidin, or elevated [K]o) before the addition of 45Ca. An additional long lasting ("slow") phase of K-stimulated Ca entry was also detected. This "slow" Ca entry was much less sensitive to La (KI > 100 microM) and was not affected by depolarizing the synaptosomes before the addition of 45Ca. The rate of influx during the fast phase was about four times the rate of Ca influx during the slow phase. Neither the fast nor slow phase of Ca entry was sensitive to tetrodotoxin (10 microM), a potent blocker of Na channels, but both phases were inhibited by Ni, Mn, Mg, and other agents that block Ca channels. The data are consistent with the presence of two distinct populations of voltage-regulated, divalent cation-selective pathways for Ca entry in presynaptic brain nerve endings.

Highlights

  • Voltage-regulated pathways for divalent cations (Ca channels) control the entry of calcium in m a n y cells

  • Ca entry into nerve endings is of special interest because it triggers transmitter release (Katz and Miledi, 1965 and 1969; Katz, 1969; Llinas et al, 1972; Blaustein, 1975); the small dimensions of mammalian presynaptic terminals preclude the use of impalement or injection methods

  • In some experiments either Tris-maleate or 2,(N-morpholino)ethane sulfonic acid-hydroxyethylpiperazine-N'-2-ethane sulfonic acid (HEPES) buffer was substituted for the TrisHEPES in the various solutions; no differences in the results were observed

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Summary

Introduction

Voltage-regulated pathways for divalent cations (Ca channels) control the entry of calcium in m a n y cells (see reviews by Reuter [1973], Baker and Glitsch [1975], and Hagiwara [1975]). These Ca channels share some characteristics: they are blocked by polyvalent cations (e.g., La, Ni, Co, and Mg) but are not affected by tetrodotoxin (TTX), a selective inhibitor of Na channels (Narahashi, 1974). Ca entry into nerve endings is of special interest because it triggers transmitter release (Katz and Miledi, 1965 and 1969; Katz, 1969; Llinas et al, 1972; Blaustein, 1975); the small dimensions of mammalian presynaptic terminals preclude the use of impalement or injection methods. GEN.PHYSIOL. The RockefellerUniversityPress 90022-1295/80/12/0709/20 $1.00 709 Volume 76 December 1980 709-728

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