TY - JOUR
T1 - Aminoglycosides increase intracellular calcium levels and ERK activity in proximal tubular OK cells expressing the extracellular calcium-sensing receptor
AU - Ward, DT
AU - McLarnon, SJ
AU - Riccardi, D
PY - 2002/6
Y1 - 2002/6
N2 - Aminoglycoside antibiotics (AGAs) are nephrotoxic, with most of the damage confined to the proximal tubule, but the mechanism for cellular toxicity is not clear. It has been previously shown that the extracellular-calcium sensing receptor (CaR) is expressed in intact rat proximal tubule and can be stimulated by the AGA neomycin. To investigate whether CaR could contribute to AGA-induced nephrotoxicity, the acute responses to various AGAs in the proximal tubule-derived opossum kidney (OK) cell line were examined. The presence in OK cells of CaR-related transcripts and protein was demonstrated by northern analyses, reverse transcriptase–PCR, immunocytochemistry, and immunoblotting. OK cells responded to elevated extracellular calcium (Ca2+o) and neomycin but also to gentamicin and tobramycin with an increase in cytosolic [Ca2+]. Ca2+o, neomycin, and gentamicin also activated the extracellular signal-regulated kinases, ERK1 and ERK2. Neomycin-induced ERK activation was both dose- and time-dependent and was attenuated by inhibitors of phosphatidylinositol 3-kinase, phosphatidylinositol bisphosphate (PIP2)–specific phospholipase C, and MEK1, but not of protein kinase C. Thus, proximal tubular OK cells express a CaR that mediates Ca2+i mobilization and PIP2-PLC-dependent ERK activation in response to AGAs and thus could play a role in AGA-induced nephrotoxicity.
AB - Aminoglycoside antibiotics (AGAs) are nephrotoxic, with most of the damage confined to the proximal tubule, but the mechanism for cellular toxicity is not clear. It has been previously shown that the extracellular-calcium sensing receptor (CaR) is expressed in intact rat proximal tubule and can be stimulated by the AGA neomycin. To investigate whether CaR could contribute to AGA-induced nephrotoxicity, the acute responses to various AGAs in the proximal tubule-derived opossum kidney (OK) cell line were examined. The presence in OK cells of CaR-related transcripts and protein was demonstrated by northern analyses, reverse transcriptase–PCR, immunocytochemistry, and immunoblotting. OK cells responded to elevated extracellular calcium (Ca2+o) and neomycin but also to gentamicin and tobramycin with an increase in cytosolic [Ca2+]. Ca2+o, neomycin, and gentamicin also activated the extracellular signal-regulated kinases, ERK1 and ERK2. Neomycin-induced ERK activation was both dose- and time-dependent and was attenuated by inhibitors of phosphatidylinositol 3-kinase, phosphatidylinositol bisphosphate (PIP2)–specific phospholipase C, and MEK1, but not of protein kinase C. Thus, proximal tubular OK cells express a CaR that mediates Ca2+i mobilization and PIP2-PLC-dependent ERK activation in response to AGAs and thus could play a role in AGA-induced nephrotoxicity.
UR - http://europepmc.org/abstract/med/12039977
U2 - 10.1097/01.ASN.0000015623.73739.B8
DO - 10.1097/01.ASN.0000015623.73739.B8
M3 - Article
C2 - 12039977
SN - 1046-6673
VL - 13
SP - 1481
EP - 1489
JO - Journal of the American Society of Nephrology
JF - Journal of the American Society of Nephrology
IS - 6
ER -