Abstract
Mobile Ca2+ buffers in hair cells have been postulated to play a dual role. On one hand, they carry incoming Ca2+ away from synaptic areas, allowing synapses to be rapidly reset. On the other hand, they limit the spread of free Ca2+ into the cell, preventing cross-talk between different pathways that employ Ca2+ as a second messenger. We have obtained evidence for such mobile Ca2+ buffers in hair cells by comparing the patterns of Ca2+-induced fluo-3 fluorescence under whole-cell and perforated-patch recording conditions. Fluorescent signals under perforated-patch conditions are relatively weak and are limited to the immediate vicinity of the membrane. These observations can be explained by a diffusion-reaction scheme that, in addition to Ca2+ and fluo-3, incorporates endogenous fixed and mobile Ca2+ buffers. Our experiments also suggest that the mobility of the endogenous buffer might be higher than previously thought. A high buffer mobility is expected to enhance the cell's ability to rapidly modulate transmitter release.
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Selected References
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- Allbritton N. L., Meyer T., Stryer L. Range of messenger action of calcium ion and inositol 1,4,5-trisphosphate. Science. 1992 Dec 11;258(5089):1812–1815. doi: 10.1126/science.1465619. [DOI] [PubMed] [Google Scholar]
- Assad J. A., Hacohen N., Corey D. P. Voltage dependence of adaptation and active bundle movement in bullfrog saccular hair cells. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2918–2922. doi: 10.1073/pnas.86.8.2918. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baylor S. M., Hollingworth S. Fura-2 calcium transients in frog skeletal muscle fibres. J Physiol. 1988 Sep;403:151–192. doi: 10.1113/jphysiol.1988.sp017244. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eberhard M., Erne P. Kinetics of calcium binding to fluo-3 determined by stopped-flow fluorescence. Biochem Biophys Res Commun. 1989 Aug 30;163(1):309–314. doi: 10.1016/0006-291x(89)92136-0. [DOI] [PubMed] [Google Scholar]
- Gabso M., Neher E., Spira M. E. Low mobility of the Ca2+ buffers in axons of cultured Aplysia neurons. Neuron. 1997 Mar;18(3):473–481. doi: 10.1016/s0896-6273(00)81247-7. [DOI] [PubMed] [Google Scholar]
- Gross M. D., Gosnell M., Tsarbopoulos A., Hunziker W. A functional and degenerate pair of EF hands contains the very high affinity calcium-binding site of calbindin-D28K. J Biol Chem. 1993 Oct 5;268(28):20917–20922. [PubMed] [Google Scholar]
- Horn R., Marty A. Muscarinic activation of ionic currents measured by a new whole-cell recording method. J Gen Physiol. 1988 Aug;92(2):145–159. doi: 10.1085/jgp.92.2.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hudspeth A. J., Lewis R. S. Kinetic analysis of voltage- and ion-dependent conductances in saccular hair cells of the bull-frog, Rana catesbeiana. J Physiol. 1988 Jun;400:237–274. doi: 10.1113/jphysiol.1988.sp017119. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Issa N. P., Hudspeth A. J. Characterization of fluo-3 labelling of dense bodies at the hair cell's presynaptic active zone. J Neurocytol. 1996 Apr;25(4):257–266. doi: 10.1007/BF02284801. [DOI] [PubMed] [Google Scholar]
- Issa N. P., Hudspeth A. J. Clustering of Ca2+ channels and Ca(2+)-activated K+ channels at fluorescently labeled presynaptic active zones of hair cells. Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7578–7582. doi: 10.1073/pnas.91.16.7578. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Issa N. P., Hudspeth A. J. The entry and clearance of Ca2+ at individual presynaptic active zones of hair cells from the bullfrog's sacculus. Proc Natl Acad Sci U S A. 1996 Sep 3;93(18):9527–9532. doi: 10.1073/pnas.93.18.9527. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jaramillo F. Signal transduction in hair cells and its regulation by calcium. Neuron. 1995 Dec;15(6):1227–1230. doi: 10.1016/0896-6273(95)90003-9. [DOI] [PubMed] [Google Scholar]
- Popov S., Poo M. M. Diffusional transport of macromolecules in developing nerve processes. J Neurosci. 1992 Jan;12(1):77–85. doi: 10.1523/JNEUROSCI.12-01-00077.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pusch M., Neher E. Rates of diffusional exchange between small cells and a measuring patch pipette. Pflugers Arch. 1988 Feb;411(2):204–211. doi: 10.1007/BF00582316. [DOI] [PubMed] [Google Scholar]
- Roberts W. M., Jacobs R. A., Hudspeth A. J. Colocalization of ion channels involved in frequency selectivity and synaptic transmission at presynaptic active zones of hair cells. J Neurosci. 1990 Nov;10(11):3664–3684. doi: 10.1523/JNEUROSCI.10-11-03664.1990. [DOI] [PMC free article] [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]
- Roberts W. M. Spatial calcium buffering in saccular hair cells. Nature. 1993 May 6;363(6424):74–76. doi: 10.1038/363074a0. [DOI] [PubMed] [Google Scholar]
- Rose J. E., Brugge J. F., Anderson D. J., Hind J. E. Phase-locked response to low-frequency tones in single auditory nerve fibers of the squirrel monkey. J Neurophysiol. 1967 Jul;30(4):769–793. doi: 10.1152/jn.1967.30.4.769. [DOI] [PubMed] [Google Scholar]
- Shepherd G. M., Barres B. A., Corey D. P. "Bundle blot" purification and initial protein characterization of hair cell stereocilia. Proc Natl Acad Sci U S A. 1989 Jul;86(13):4973–4977. doi: 10.1073/pnas.86.13.4973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Strautman A. F., Cork R. J., Robinson K. R. The distribution of free calcium in transected spinal axons and its modulation by applied electrical fields. J Neurosci. 1990 Nov;10(11):3564–3575. doi: 10.1523/JNEUROSCI.10-11-03564.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tucker T., Fettiplace R. Confocal imaging of calcium microdomains and calcium extrusion in turtle hair cells. Neuron. 1995 Dec;15(6):1323–1335. doi: 10.1016/0896-6273(95)90011-x. [DOI] [PubMed] [Google Scholar]
- Wu Y. C., Tucker T., Fettiplace R. A theoretical study of calcium microdomains in turtle hair cells. Biophys J. 1996 Nov;71(5):2256–2275. doi: 10.1016/S0006-3495(96)79429-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
- al-Baldawi N. F., Abercrombie R. F. Calcium diffusion coefficient in Myxicola axoplasm. Cell Calcium. 1995 Jun;17(6):422–430. doi: 10.1016/0143-4160(95)90088-8. [DOI] [PubMed] [Google Scholar]