Distribution and activation of intracellular Ca2+ stores in cultured olfactory bulb neurons.
Distribution and activation of intracellular Ca2+ stores in cultured olfactory bulb neurons.
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培养的嗅球神经元细胞内 Ca2 储存的分布和激活。
DOI：
10.1152/jn.1997.78.4.2176
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Keller,A
中科院分区：
文献类型：
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作者：
Carlson,GC;Slawecki,ML;Lancaster,E;Keller,A
卡尔森、格雷格·C、梅丽莎·l·斯劳韦斯基、埃里克·兰开斯特和阿萨夫·凯勒。嗅球神经元细胞内Ca2+储存的分布和激活。中国生物医学工程学报，32(2):387 - 398。在分离细胞培养中研究了嗅球神经元细胞内Ca2+释放通路的存在和分布。使用组织化学技术和Ca2+通量成像来鉴定两种主要的细胞内Ca2+释放机制：肌醇1,4,5-三磷酸受体（IP3R）介导的释放和ryanodine受体介导的释放。用免疫细胞化学方法鉴定培养的神经元特异性标志物β-微管蛋白III。谷氨酸脱羧酶的形态计量学分析和免疫细胞化学表明，培养的神经元群体具有异质表型，其表型与体内嗅球的投射（二尖瓣/簇状）和内在（肾小球周围/颗粒）神经元相对应。IP3R的免疫细胞化学和荧光标记的ryanodine显示，无论细胞类型如何，几乎所有培养的神经元在体细胞和树突中都表达IP3R和ryanodine结合位点。功能成像显示，细胞内Ca2+通量可以在没有外部Ca2+的情况下产生，使用特定于每个细胞内释放途径的激动剂。谷氨酸或半qualate的局部压力应用在名义上Ca2+游离的细胞外溶液中诱发了体细胞和树突中的Ca2+通量，表明存在ip3依赖性Ca2+释放。在谷氨酸受体拮抗剂6-氰基-7-硝基喹啉-2,3-二酮的存在下，这些通量被thapsigargin预孵育阻断，并持续存在。局部应用咖啡因，一种ryanodine受体激动剂，也可以在缺乏细胞外Ca2+的情况下引起细胞内Ca2+通量。这些Ca2+通量被ryanodine预孵育抑制。在所有神经元中，IP3-和ryanodine依赖的释放途径共存，表明它们相互作用以调节细胞内Ca2+浓度。
Carlson, Greg C., Melissa L. Slawecki, Eric Lancaster, and Asaf Keller.Distribution and activation of intracellular Ca2+stores in cultured olfactory bulb neurons.J. Neurophysiol.78: 2176–2185, 1997. The presence and distribution of intracellular Ca2+release pathways in olfactory bulb neurons were studied in dissociated cell cultures. Histochemical techniques and imaging of Ca2+fluxes were used to identify two major intracellular Ca2+release mechanisms: inositol 1,4,5-triphosphate receptor (IP3R)-mediated release, and ryanodine receptor-mediated release. Cultured neurons were identified by immunocytochemistry for the neuron-specificmarker β-tubulin III. Morphometric analyses and immunocytochemistry for glutamic acid-decarboxylase revealed a heterogeneous population of cultured neurons with phenotypes corresponding to both projection (mitral/tufted) and intrinsic (periglomerular/granule) neurons of the in vivo olfactory bulb. Immunocytochemistry for the IP3R, and labeling with fluorescent-tagged ryanodine, revealed that, irrespective of cell type, almost all cultured neurons express IP3R and ryanodine binding sites in both somata and dendrites. Functional imaging revealed that intracellular Ca2+fluxes can be generated in the absence of external Ca2+, using agonists specific to each of the intracellular release pathways. Local pressure application of glutamate or quisqualate evoked Ca2+fluxes in both somata and dendrites in nominally Ca2+free extracellular solutions, suggesting the presence of IP3-dependent Ca2+release. These fluxes were blocked by preincubation with thapsigargin and persisted in the presence of the glutamate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione. Local application of caffeine, a ryanodine receptor agonist, also evoked intracellular Ca2+fluxes in the absence of extracellular Ca2+. These Ca2+fluxes were suppressed by preincubation with ryanodine. In all neurons, both IP3- and ryanodine-dependent release pathways coexisted, suggesting that they interact to modulate intracellular Ca2+concentrations.