Abstract
Calcium dynamics associated with a single action potential were studied quantitatively in the calyx of Held, a large presynaptic terminal in the rat brainstem. Terminals were loaded with different concentrations of high- or low-affinity Ca2+ indicators via patch pipettes. Spatially averaged Ca2+ signals were measured fluorometrically and analyzed on the basis of a single compartment model. A single action potential led to a total Ca2+ influx of 0.8-1 pC. The accessible volume of the terminal was about 0.4 pl; thus the total calcium concentration increased by 10-13 microM. The Ca(2+)-binding ratio of the endogenous buffer was about 40, as estimated from the competition with Fura-2, indicating that 2.5% of the total calcium remained free. This is consistent with the peak increase in free calcium concentration of about 400 nM, which was measured directly with MagFura-2. The decay of the [Ca2+]i transients was fast, with time constants of 100 ms at 23 degrees C and 45 ms at 35 degrees C, indicating Ca2+ extrusion rates of 400 and 900 s-1, respectively. The combination of the relatively low endogenous Ca(2+)-binding ratio and the high rate of Ca2+ extrusion provides an efficient mechanism for rapidly removing the large Ca2+ load of the terminal evoked by an action potential.
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- Adler E. M., Augustine G. J., Duffy S. N., Charlton M. P. Alien intracellular calcium chelators attenuate neurotransmitter release at the squid giant synapse. J Neurosci. 1991 Jun;11(6):1496–1507. doi: 10.1523/JNEUROSCI.11-06-01496.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Agard D. A., Hiraoka Y., Shaw P., Sedat J. W. Fluorescence microscopy in three dimensions. Methods Cell Biol. 1989;30:353–377. doi: 10.1016/s0091-679x(08)60986-3. [DOI] [PubMed] [Google Scholar]
- Augustine G. J., Charlton M. P. Calcium dependence of presynaptic calcium current and post-synaptic response at the squid giant synapse. J Physiol. 1986 Dec;381:619–640. doi: 10.1113/jphysiol.1986.sp016347. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Augustine G. J., Neher E. Neuronal Ca2+ signalling takes the local route. Curr Opin Neurobiol. 1992 Jun;2(3):302–307. doi: 10.1016/0959-4388(92)90119-6. [DOI] [PubMed] [Google Scholar]
- Barnes-Davies M., Forsythe I. D. Pre- and postsynaptic glutamate receptors at a giant excitatory synapse in rat auditory brainstem slices. J Physiol. 1995 Oct 15;488(Pt 2):387–406. doi: 10.1113/jphysiol.1995.sp020974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Blaustein M. P. Calcium transport and buffering in neurons. Trends Neurosci. 1988 Oct;11(10):438–443. doi: 10.1016/0166-2236(88)90195-6. [DOI] [PubMed] [Google Scholar]
- Borst J. G., Helmchen F., Sakmann B. Pre- and postsynaptic whole-cell recordings in the medial nucleus of the trapezoid body of the rat. J Physiol. 1995 Dec 15;489(Pt 3):825–840. doi: 10.1113/jphysiol.1995.sp021095. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Borst J. G., Sakmann B. Calcium influx and transmitter release in a fast CNS synapse. Nature. 1996 Oct 3;383(6599):431–434. doi: 10.1038/383431a0. [DOI] [PubMed] [Google Scholar]
- Brinley F. J., Jr Calcium buffering in squid axons. Annu Rev Biophys Bioeng. 1978;7:363–392. doi: 10.1146/annurev.bb.07.060178.002051. [DOI] [PubMed] [Google Scholar]
- Escobar A. L., Monck J. R., Fernandez J. M., Vergara J. L. Localization of the site of Ca2+ release at the level of a single sarcomere in skeletal muscle fibres. Nature. 1994 Feb 24;367(6465):739–741. doi: 10.1038/367739a0. [DOI] [PubMed] [Google Scholar]
- Fogelson A. L., Zucker R. S. Presynaptic calcium diffusion from various arrays of single channels. Implications for transmitter release and synaptic facilitation. Biophys J. 1985 Dec;48(6):1003–1017. doi: 10.1016/S0006-3495(85)83863-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Forsythe I. D. Direct patch recording from identified presynaptic terminals mediating glutamatergic EPSCs in the rat CNS, in vitro. J Physiol. 1994 Sep 15;479(Pt 3):381–387. doi: 10.1113/jphysiol.1994.sp020303. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Friauf E. Transient appearance of calbindin-D28k-positive neurons in the superior olivary complex of developing rats. J Comp Neurol. 1993 Aug 1;334(1):59–74. doi: 10.1002/cne.903340105. [DOI] [PubMed] [Google Scholar]
- Groden D. L., Guan Z., Stokes B. T. Determination of Fura-2 dissociation constants following adjustment of the apparent Ca-EGTA association constant for temperature and ionic strength. Cell Calcium. 1991 Apr;12(4):279–287. doi: 10.1016/0143-4160(91)90002-v. [DOI] [PubMed] [Google Scholar]
- Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
- Helmchen F., Imoto K., Sakmann B. Ca2+ buffering and action potential-evoked Ca2+ signaling in dendrites of pyramidal neurons. Biophys J. 1996 Feb;70(2):1069–1081. doi: 10.1016/S0006-3495(96)79653-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jackson M. B., Konnerth A., Augustine G. J. Action potential broadening and frequency-dependent facilitation of calcium signals in pituitary nerve terminals. Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):380–384. doi: 10.1073/pnas.88.2.380. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jaffe D. B., Johnston D., Lasser-Ross N., Lisman J. E., Miyakawa H., Ross W. N. The spread of Na+ spikes determines the pattern of dendritic Ca2+ entry into hippocampal neurons. Nature. 1992 May 21;357(6375):244–246. doi: 10.1038/357244a0. [DOI] [PubMed] [Google Scholar]
- Kao J. P., Tsien R. Y. Ca2+ binding kinetics of fura-2 and azo-1 from temperature-jump relaxation measurements. Biophys J. 1988 Apr;53(4):635–639. doi: 10.1016/S0006-3495(88)83142-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Konishi M., Berlin J. R. Ca transients in cardiac myocytes measured with a low affinity fluorescent indicator, furaptra. Biophys J. 1993 Apr;64(4):1331–1343. doi: 10.1016/S0006-3495(93)81494-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Llinás R., Sugimori M., Silver R. B. Microdomains of high calcium concentration in a presynaptic terminal. Science. 1992 May 1;256(5057):677–679. doi: 10.1126/science.1350109. [DOI] [PubMed] [Google Scholar]
- Llinás R., Sugimori M., Simon S. M. Transmission by presynaptic spike-like depolarization in the squid giant synapse. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2415–2419. doi: 10.1073/pnas.79.7.2415. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lohmann C., Friauf E. Distribution of the calcium-binding proteins parvalbumin and calretinin in the auditory brainstem of adult and developing rats. J Comp Neurol. 1996 Mar 25;367(1):90–109. doi: 10.1002/(SICI)1096-9861(19960325)367:1<90::AID-CNE7>3.0.CO;2-E. [DOI] [PubMed] [Google Scholar]
- Martinez-Serrano A., Blanco P., Satrústegui J. Calcium binding to the cytosol and calcium extrusion mechanisms in intact synaptosomes and their alterations with aging. J Biol Chem. 1992 Mar 5;267(7):4672–4679. [PubMed] [Google Scholar]
- Mintz I. M., Sabatini B. L., Regehr W. G. Calcium control of transmitter release at a cerebellar synapse. Neuron. 1995 Sep;15(3):675–688. doi: 10.1016/0896-6273(95)90155-8. [DOI] [PubMed] [Google Scholar]
- Morest D. K. The growth of synaptic endings in the mammalian brain: a study of the calyces of the trapezoid body. Z Anat Entwicklungsgesch. 1968 Nov 4;127(3):201–220. doi: 10.1007/BF00526129. [DOI] [PubMed] [Google Scholar]
- Neher E., Augustine G. J. Calcium gradients and buffers in bovine chromaffin cells. J Physiol. 1992 May;450:273–301. doi: 10.1113/jphysiol.1992.sp019127. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Neher E. The use of fura-2 for estimating Ca buffers and Ca fluxes. Neuropharmacology. 1995 Nov;34(11):1423–1442. doi: 10.1016/0028-3908(95)00144-u. [DOI] [PubMed] [Google Scholar]
- Neher E., Zucker R. S. Multiple calcium-dependent processes related to secretion in bovine chromaffin cells. Neuron. 1993 Jan;10(1):21–30. doi: 10.1016/0896-6273(93)90238-m. [DOI] [PubMed] [Google Scholar]
- Oliva C., Cohen I. S., Mathias R. T. Calculation of time constants for intracellular diffusion in whole cell patch clamp configuration. Biophys J. 1988 Nov;54(5):791–799. doi: 10.1016/S0006-3495(88)83017-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Regehr W. G., Atluri P. P. Calcium transients in cerebellar granule cell presynaptic terminals. Biophys J. 1995 May;68(5):2156–2170. doi: 10.1016/S0006-3495(95)80398-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Regehr W. G., Delaney K. R., Tank D. W. The role of presynaptic calcium in short-term enhancement at the hippocampal mossy fiber synapse. J Neurosci. 1994 Feb;14(2):523–537. doi: 10.1523/JNEUROSCI.14-02-00523.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Regehr W. G., Tank D. W. The maintenance of LTP at hippocampal mossy fiber synapses is independent of sustained presynaptic calcium. Neuron. 1991 Sep;7(3):451–459. doi: 10.1016/0896-6273(91)90297-d. [DOI] [PubMed] [Google Scholar]
- Reuter H., Porzig H. Localization and functional significance of the Na+/Ca2+ exchanger in presynaptic boutons of hippocampal cells in culture. Neuron. 1995 Nov;15(5):1077–1084. doi: 10.1016/0896-6273(95)90096-9. [DOI] [PubMed] [Google Scholar]
- Roberts W. M. Localization of calcium signals by a mobile calcium buffer in frog saccular hair cells. J Neurosci. 1994 May;14(5 Pt 2):3246–3262. doi: 10.1523/JNEUROSCI.14-05-03246.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sala F., Hernández-Cruz A. Calcium diffusion modeling in a spherical neuron. Relevance of buffering properties. Biophys J. 1990 Feb;57(2):313–324. doi: 10.1016/S0006-3495(90)82533-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schneggenburger R., Zhou Z., Konnerth A., Neher E. Fractional contribution of calcium to the cation current through glutamate receptor channels. Neuron. 1993 Jul;11(1):133–143. doi: 10.1016/0896-6273(93)90277-x. [DOI] [PubMed] [Google Scholar]
- Schweizer F. E., Betz H., Augustine G. J. From vesicle docking to endocytosis: intermediate reactions of exocytosis. Neuron. 1995 Apr;14(4):689–696. doi: 10.1016/0896-6273(95)90213-9. [DOI] [PubMed] [Google Scholar]
- Simon S. M., Llinás R. R. Compartmentalization of the submembrane calcium activity during calcium influx and its significance in transmitter release. Biophys J. 1985 Sep;48(3):485–498. doi: 10.1016/S0006-3495(85)83804-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sinha S. R., Wu L. G., Saggau P. Presynaptic calcium dynamics and transmitter release evoked by single action potentials at mammalian central synapses. Biophys J. 1997 Feb;72(2 Pt 1):637–651. doi: 10.1016/s0006-3495(97)78702-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith S. J., Augustine G. J. Calcium ions, active zones and synaptic transmitter release. Trends Neurosci. 1988 Oct;11(10):458–464. doi: 10.1016/0166-2236(88)90199-3. [DOI] [PubMed] [Google Scholar]
- Stuart G. J., Dodt H. U., Sakmann B. Patch-clamp recordings from the soma and dendrites of neurons in brain slices using infrared video microscopy. Pflugers Arch. 1993 Jun;423(5-6):511–518. doi: 10.1007/BF00374949. [DOI] [PubMed] [Google Scholar]
- Stuenkel E. L. Regulation of intracellular calcium and calcium buffering properties of rat isolated neurohypophysial nerve endings. J Physiol. 1994 Dec 1;481(Pt 2):251–271. doi: 10.1113/jphysiol.1994.sp020436. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tank D. W., Regehr W. G., Delaney K. R. A quantitative analysis of presynaptic calcium dynamics that contribute to short-term enhancement. J Neurosci. 1995 Dec;15(12):7940–7952. doi: 10.1523/JNEUROSCI.15-12-07940.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ukhanov K. Y., Flores T. M., Hsiao H. S., Mohapatra P., Pitts C. H., Payne R. Measurement of cytosolic Ca2+ concentration in Limulus ventral photoreceptors using fluorescent dyes. J Gen Physiol. 1995 Jan;105(1):95–116. doi: 10.1085/jgp.105.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wagner J., Keizer J. Effects of rapid buffers on Ca2+ diffusion and Ca2+ oscillations. Biophys J. 1994 Jul;67(1):447–456. doi: 10.1016/S0006-3495(94)80500-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wu L. G., Saggau P. Pharmacological identification of two types of presynaptic voltage-dependent calcium channels at CA3-CA1 synapses of the hippocampus. J Neurosci. 1994 Sep;14(9):5613–5622. doi: 10.1523/JNEUROSCI.14-09-05613.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zhao M., Hollingworth S., Baylor S. M. Properties of tri- and tetracarboxylate Ca2+ indicators in frog skeletal muscle fibers. Biophys J. 1996 Feb;70(2):896–916. doi: 10.1016/S0006-3495(96)79633-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zhou Z., Misler S. Action potential-induced quantal secretion of catecholamines from rat adrenal chromaffin cells. J Biol Chem. 1995 Feb 24;270(8):3498–3505. [PubMed] [Google Scholar]