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Nerve growth factor treatment alters Ca2+ pump levels in PC12 cells

Keller, Dan1; Grover, Ashok K.1,2

Developmental Neuroscience

Nerve growth factor (NGF) treatment converts rapidly dividing PC12 cells into a neuronal phenotype. To understand the Ca2+ sequestration mechanisms accompanying this differentiation, we examined the endoplasmic reticulum Ca2+ (SERCA) pump levels using two different assays: ATP-dependent azide insensitive oxalate stimulated 45Ca2+ uptake by PC12 cells permeabilized with saponin, and Western blots using a monoclonal antibody which reacts with all the SERCA isoforms. We also examined the reaction to an antibody against the plasma membrane Ca2+ (PMCA) pump. NGF treatment decreased the SERCA pump expression but it increased the PMCA pump level. These results are consistent with a greater role of PMCA pumps in neuronal cells than in most other cells and with an increased role of SERCA pumps during cell proliferation.

1Department of Medicine, McMaster University, Hamilton, Canada L8N 3Z5

2Corresponding Author: Ashok K. Grover

Acknowledgements: We thank S.E. Samson for culturing the PC12 cells, C.M. Misquitta for helping with the Western blots, Dr M. Coughlin for NGF and Dr L.T. Young for critical review of the manuscript. Part of this work constituted the undergraduate thesis of D.K. This work was supported by MRC. A.K.G. is a Career Investigator of the Heart and Stroke Foundation of Ontario.

Received 28 September 1999 accepted 14 October 1999

© 2000 Lippincott Williams & Wilkins, Inc.