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Title: Functionality of NGF-protected PC12 cells following exposure to 6-hydroxydopamine

Abstract

6-Hydroxydopamine (6-OHDA) is often used in models of Parkinson's disease since it can selectively target and kill dopaminergic cells of the substantia nigra. In this study, pre-treatment of PC12 cells with nerve growth factor (NGF) inhibited apoptosis and necrosis by 6-OHDA, including caspase activity and lactate dehydrogenase release. Notably, cells exposed to 6-OHDA in the presence of NGF were subsequently capable of proliferation (when replated without NGF), or neurite outgrowth (with continued presence of NGF). Following 7 days growth in the presence of NGF, expression of {beta}III tubulin and tyrosine hydroxylase and increased intracellular catecholamines was detectable in PC12 cells, features characteristic of functional dopaminergic neurons. NGF-pre-treated PC12 cells retained expression of {beta}III-tubulin and tyrosine hydroxylase, but not catecholamine content following 6-OHDA exposure. These data indicate that NGF-protected cells maintained some aspects of functionality and were subsequently capable of proliferation or differentiation.

Authors:
 [1];  [2];  [1];  [2];  [1];  [3];  [2];  [1];  [2];  [3];  [2];  [4]
  1. Department of Biochemistry, National University of Ireland, Galway (Ireland)
  2. (Ireland)
  3. National Centre for Biomedical Engineering and Science, National University of Ireland, Galway (Ireland)
  4. Department of Biochemistry, National University of Ireland, Galway (Ireland) and National Centre for Biomedical Engineering and Science, National University of Ireland, Galway (Ireland). E-mail: adrienne.gorman@nuigalway.ie
Publication Date:
OSTI Identifier:
20857938
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 351; Journal Issue: 4; Other Information: DOI: 10.1016/j.bbrc.2006.10.104; PII: S0006-291X(06)02371-0; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; APOPTOSIS; CATECHOLAMINES; DOPAMINE; GROWTH FACTORS; HYDROXYLASES; LACTATE DEHYDROGENASE; NECROSIS; NERVE CELLS; NERVOUS SYSTEM DISEASES; TYROSINE

Citation Formats

Kavanagh, Edel T., National Centre for Biomedical Engineering and Science, National University of Ireland, Galway, Loughlin, John P., Department of Physiology, National University of Ireland, Galway, Herbert, Kate Reed, Dockery, Peter, Department of Anatomy, National University of Ireland, Galway, Samali, Afshin, National Centre for Biomedical Engineering and Science, National University of Ireland, Galway, Doyle, Karen M., Department of Physiology, National University of Ireland, Galway, and Gorman, Adrienne M. Functionality of NGF-protected PC12 cells following exposure to 6-hydroxydopamine. United States: N. p., 2006. Web. doi:10.1016/j.bbrc.2006.10.104.
Kavanagh, Edel T., National Centre for Biomedical Engineering and Science, National University of Ireland, Galway, Loughlin, John P., Department of Physiology, National University of Ireland, Galway, Herbert, Kate Reed, Dockery, Peter, Department of Anatomy, National University of Ireland, Galway, Samali, Afshin, National Centre for Biomedical Engineering and Science, National University of Ireland, Galway, Doyle, Karen M., Department of Physiology, National University of Ireland, Galway, & Gorman, Adrienne M. Functionality of NGF-protected PC12 cells following exposure to 6-hydroxydopamine. United States. doi:10.1016/j.bbrc.2006.10.104.
Kavanagh, Edel T., National Centre for Biomedical Engineering and Science, National University of Ireland, Galway, Loughlin, John P., Department of Physiology, National University of Ireland, Galway, Herbert, Kate Reed, Dockery, Peter, Department of Anatomy, National University of Ireland, Galway, Samali, Afshin, National Centre for Biomedical Engineering and Science, National University of Ireland, Galway, Doyle, Karen M., Department of Physiology, National University of Ireland, Galway, and Gorman, Adrienne M. Fri . "Functionality of NGF-protected PC12 cells following exposure to 6-hydroxydopamine". United States. doi:10.1016/j.bbrc.2006.10.104.
@article{osti_20857938,
title = {Functionality of NGF-protected PC12 cells following exposure to 6-hydroxydopamine},
author = {Kavanagh, Edel T. and National Centre for Biomedical Engineering and Science, National University of Ireland, Galway and Loughlin, John P. and Department of Physiology, National University of Ireland, Galway and Herbert, Kate Reed and Dockery, Peter and Department of Anatomy, National University of Ireland, Galway and Samali, Afshin and National Centre for Biomedical Engineering and Science, National University of Ireland, Galway and Doyle, Karen M. and Department of Physiology, National University of Ireland, Galway and Gorman, Adrienne M.},
abstractNote = {6-Hydroxydopamine (6-OHDA) is often used in models of Parkinson's disease since it can selectively target and kill dopaminergic cells of the substantia nigra. In this study, pre-treatment of PC12 cells with nerve growth factor (NGF) inhibited apoptosis and necrosis by 6-OHDA, including caspase activity and lactate dehydrogenase release. Notably, cells exposed to 6-OHDA in the presence of NGF were subsequently capable of proliferation (when replated without NGF), or neurite outgrowth (with continued presence of NGF). Following 7 days growth in the presence of NGF, expression of {beta}III tubulin and tyrosine hydroxylase and increased intracellular catecholamines was detectable in PC12 cells, features characteristic of functional dopaminergic neurons. NGF-pre-treated PC12 cells retained expression of {beta}III-tubulin and tyrosine hydroxylase, but not catecholamine content following 6-OHDA exposure. These data indicate that NGF-protected cells maintained some aspects of functionality and were subsequently capable of proliferation or differentiation.},
doi = {10.1016/j.bbrc.2006.10.104},
journal = {Biochemical and Biophysical Research Communications},
number = 4,
volume = 351,
place = {United States},
year = {Fri Dec 29 00:00:00 EST 2006},
month = {Fri Dec 29 00:00:00 EST 2006}
}
  • Rat PC12 pheochromocytoma and human A875 melanoma cells express nerve growth factor (NGF) receptors on their surfaces. Covalent crosslinking of bound /sup 125/I-NGF to PC12 or A875 intact cells or plasma membrane-enriched fractions resulted in labelling of a peptide doublet at Mr . 110,000 and a single labelled peptide at Mr . 200,000 for each of the cell and membrane preparations. PC12 cells exhibited biphasic binding properties with two apparent binding sites: KD . 5.2 nM sites and KD . 0.3 nM sites. The high-affinity PC12 binding sites were trypsin resistant, and /sup 125/I-NGF dissociated slowly from them. A875 cellsmore » exhibited sites with homogeneous properties (KD . 1.0 nM), all binding sites were trypsin sensitive, and /sup 125/I-NGF dissociated rapidly in the presence of unlabelled NGF. Membrane-enriched fractions from either cell type contained binding sites with a uniform low affinity (KD . 3 nM) that were trypsin sensitive, and /sup 125/I-NGF rapidly dissociated from them. Sixty to 80 percent of binding sites in membranes could be converted to the high-affinity, trypsin-resistant state by addition of wheat germ agglutinin (WGA). The loss of high-affinity, trypsin-resistant sites from PC12 cells during preparation of plasma membrane fractions does not appear to be the result of selective isolation of low-affinity sites or proteolytic degradation since there is a loss of /sup 125/I-NGF binding immediately after cell lysis which is not blocked by protease inhibitors. The differences between receptor properties on PC12 cells and on A875 cells apparently are the result of differences in the respective intracellular environments. Thus, cell components other than the binding unit of the NGF receptor may be responsible for the different properties of receptor.« less
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