VOLUME REGULATION OF SINGLE ASTROGLIAL CELLS IN PRIMARY CULTURE
VOLUME REGULATION OF SINGLE ASTROGLIAL CELLS IN PRIMARY CULTURE
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DOI：
10.1016/0304-3940(92)90264-8
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发表时间：
1992-08-31
影响因子：
2.5
通讯作者：
HANSSON, E
中科院分区：
文献类型：
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作者：
ERIKSSON, PS;NILSSON, M;HANSSON, E
以Fura-2/AM的荧光探针2，7-双（羧乙基）-5，6-羧基荧光素（BCECF/AM）为荧光探针，用显微荧光法检测星形胶质细胞相对体积的变化。在他们的等色点。450 nm和358 nm，探针是离子不敏感的，发射的荧光信号仅与细胞内染料浓度有关。通过改变激发波长，细胞内pH值或Ca 2+瞬变的变化可以与单个细胞的相对体积变化同时记录。暴露于低渗缓冲液后，1型星形胶质细胞在30 s内肿胀，随后发生调节性体积减少（RVD）。当暴露于高渗缓冲液中时，星形胶质细胞萎缩并表现出调节性体积增加（RVI）。1 mM谷氨酸在60秒内诱导星形胶质细胞体积增加，并诱发胞质Ca 2+瞬变，但不改变细胞内pH值。
Relative volume variations in cultured astrocytes were examined by microspectrofluorimetry after loading the cells with the highly fluorescent intracellular probes 2,7-bis(carboxyethyl)-5,6-carboxyfluorescein (BCECF/AM) of fura-2/AM. At their isosbestic points. 450 nm and 358 nm, respectively, the probes were ion-insensitive and the fluorescent signals emitted related only to the intracellular dye concentration. By varying the excitation wavelengths, changes in intracellular pH or Ca2+ transients could be recorded simultaneously with the relative volume variations of the individual cells. After exposure to a hypotonic buffer, type 1 astrocytes swelled within 30 s and subsequently underwent regulatory volume decrease (RVD). When exposed to a hypertonic buffer, the astrocytes shrunk and exhibited regulatory volume increase (RVI). One mM glutamate induced an increase in astrocyte volume in 60 sec and evoked cytosolic Ca2+ transients but did not change intracellular pH.