PMID: 11341965May 9, 2001Paper

Establishment of variant PC12 subclones deficient in stimulation-secretion coupling

Biochimica Et Biophysica Acta
Y Shoji-KasaiM Takahashi

Abstract

Clonal rat pheochromocytoma (PC12) cells have been widely used to study the molecular mechanism of exocytosis. We have isolated variant PC12 subclones with deficiencies in stimulation-secretion coupling, by a single cell recloning, and investigated the defects. PC12-1G2 hardly released dopamine following high-K(+)-induced depolarization, but normal release was evoked by the Ca(2+)-ionophore, ionomycin. Fura-2 fluorometry indicated that a nicardipine-sensitive component of Ca(2+) influx was missing, suggesting that PC12-1G2 has defects in L-type Ca(2+) channel function. PC12-2B3 was not responsive to high-K(+)-induced depolarization and ionomycin, and voltage-dependent Ca(2+) entry was identical to that of the normal clone. Electron microscopy revealed that the number of vesicles adjacent or directly attached to the plasma membrane was decreased in PC12-2B3. The expression of presynaptic proteins was analyzed by immunoblotting using a panel of antibodies. Syntaxin 1, VAMP-2, SNAP-25, Munc18, Rab3C and Sec-6 were decreased compared to the control clone and that of synaptophysin was extremely low. PC12-D60 synthesized and released dopamine normally, but had almost lost its catecholamine-uptake activity. These results show that mul...Continue Reading

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Citations

Mar 21, 2003·Brain Research·Edward D SchillerDouglas P Dohrman
Jan 17, 2002·Neurochemistry International·S BiegerD Maysinger
Dec 17, 2002·The Journal of Physiology·Ian M FearonColin A Nurse
Sep 20, 2005·Journal of Neuroscience Methods·Kiyoshi OhnumaMakoto Asashima
Oct 11, 2016·ACS Chemical Neuroscience·Nihar KinarivalaPaul C Trippier

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