Golgi apparatus is involved in intracellular Ca2+ regulation in epithelial LLC-PK1 cells.
Several lines of evidence suggest that the Golgi apparatus is involved in Ca2+ regulation in renal epithelial LLC-PK1 cells. Laser scanning confocal microscopy (LSCM) was employed to establish that a prominent perinuclear region is occupied mainly by the Golgi apparatus in this cell line. LSCM measurements in individual cells with the ionized Ca2+ indicator calcium green revealed that stimulation of LLC-PK1 cells with arginine vasopressin (AVP) resulted in the elevation of ionized Ca2+ levels. However, the vasopressin-induced rise in ionized Ca2+ was attenuated if the Golgi apparatus was disassembled by pretreating the cells with brefeldin A (BFA). Subcellular measurements of total Ca2+ with ion microscopy in cryogenically prepared cells indicated that 1) within 1 min of AVP treatment significant quantities of sequestered Ca2+ were released from the perinuclear Golgi region and 2) the BFA treatment reduced the total Ca2+ stored in the Golgi region. These observations indicate that the Golgi apparatus is sensitive to hormonal stimulation and may play important roles in intracellular Ca2+ regulation in LLC-PK1 cells.
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Department of Chemistry, Baker Laboratory, Cornell University, Ithaca, New York 14853-1301, USA.
Chandra
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Ridsdale
A J
AJ
Morrison
G H
GH
D000818:Animals; D020126:Brefeldin A; D002118:Calcium / Q000378:metabolism*; D003517:Cyclopentanes / Q000494:pharmacology; D006056:Golgi Apparatus / Q000502:physiology*; D007425:Intracellular Membranes / Q000378:metabolism*; D018374:LLC-PK1 Cells / Q000187:drug effects / Q000378:metabolism*; D018613:Microscopy, Confocal; D013347:Subcellular Fractions / Q000378:metabolism; D013552:Swine; D014667:Vasopressins / Q000494:pharmacology
D003517:Cyclopentanes; D014667:Vasopressins; D020126:Brefeldin A; D002118:Calcium