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The Journal of Neuroscience, September 15, 1998, 18(18):7127-7137

Involvement of Stretch-Activated Cl Channels in Ramification of Murine Microglia

Claudia Eder, Rolf Klee, and Uwe Heinemann

Department of Neurophysiology, Institute of Physiology, Humboldt University, D 10117 Berlin, Germany

A stretch-activated Cl current (I_Cl) was investigated in cultured murine microglia using the whole-cell configuration of the patch-clamp technique. After application of membrane stretch, a Cl current appeared within seconds, and its amplitude increased further within 3-8 min. I_Cl underwent rundown, which was prevented by addition of 4 mM ATP to the intracellular perfusing solution. The stretch-activated Cl current exhibited outward rectification and did not show any voltage-dependent gating. Lowering the concentration of extracellular Cl from 142 to 12 mM by equimolar substitution of Cl with gluconate shifted the reversal potential of I_Cl by 41.6 ± 1.8 mV in the depolarizing direction. 4,4'-Diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) and 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid (SITS) blocked I_Cl in a voltage- and time-dependent manner. At a test potential of +40 mV, a half-maximal blockade at 16.1 µM DIDS and at 71.0 µM SITS was determined for I_Cl. At a concentration of 200 µM, 5-nitro-2-(3-phenylpropylamino)benzoic acid or flufenamic acid blocked I_Cl by 88% and 75%, respectively. Each of these four Cl channel blockers reversibly inhibited the ramification process of microglia, whereas blockers of voltage-gated Na⁺ and K⁺ channels did not affect the transformation of microglia from their ameboid into the ramified phenotype. It is suggested that in microglia functional stretch-activated Cl channels are required for the induction of ramification but not for maintaining the ramified shape.

Key words: brain macrophages; ramification; stretch-activated Cl current; SITS; DIDS; NPPB; flufenamic acid

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Copyright © 1998 Society for Neuroscience  0270-6474/98/18187127-11$05.00/0

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