PC12 Cell Mutants That Possess Low- but Not High-Affinity Nerve Growth Factor Receptors Neither Respond to nor Internalize Nerve Growth Factor

• Published in: The Journal of cell biology Vol. 102; no. 3; pp. 830 - 843
• Main Authors: Green, Steven H., Rydel, Russell E., Connolly, James L., Greene, Lloyd A.
• Format: Journal Article
• Language: English
• Published: New York, NY Rockefeller University Press 01.03.1986; The Rockefeller University Press
• Subjects: Animals; Biological and medical sciences; Bucladesine - pharmacology; Cell Division; Cell growth; Cell Line; Cell lines; Cell physiology; Fundamental and applied biological sciences. Psychology; Glycoproteins - analysis; Growth factor receptors; Human growth; Internalization; Molecular and cellular biology; Neoplasm Proteins - analysis; nerve growth factor; Nerve Growth Factors - metabolism; Nerve Growth Factors - pharmacology; Nerves; Neurites; Neurons; Ornithine Decarboxylase - analysis; PC12 cells; Pheochromocytoma - genetics; Pheochromocytoma - metabolism; Pheochromocytoma - ultrastructure; pheochromocytoma cells; Phosphoproteins - analysis; Rats; Receptors; Receptors, Cell Surface - genetics; Receptors, Cell Surface - metabolism; Receptors, Nerve Growth Factor; Responses to growth factors, tumor promotors, other factors; Cell culture; Vertebrata; Membrane receptor; Mammalia; Cell line; Rat; Internalization; Rodentia; Nerve growth factor; Mutation
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.102.3.830

Four mutant PC12 pheochromocytoma cell lines that are nerve growth factor (NGF)-nonresponsive (PC12nnr) have been selected from chemically mutagenized cultures by a double selection procedure: failure both to grow neurites in the presence of NGF and to survive in NGF-supplemented serum-free medium. The PC12nnrcells were deficient in all additional NGF responses surveyed: abatement of cell proliferation, changes in glycoprotein composition, induction of ornithine decarboxylase, rapid changes in protein phosphorylation, and cell surface ruffling. However, PC12nnrcells closely resembled non-NGF-treated PC12 cells in most properties tested: cell size and shape; division rate; protein, phosphoprotein, and glycoprotein composition; and cell surface morphology. All four PC12nnrlines differed from PC12 cells in three ways in addition to failure of NGF response: (a) PC12nnrcells failed to internalize bound NGF by the normal, saturable, high-affinity mechanism present in PC12 cells. (b) The PC12nnrcells bound NGF but entirely, or nearly entirely, at low-affinity sites only, whereas PC12 cells possess both high- and low-affinity NGF binding sites. (c) The responses to dibutyryl cyclic AMP that were tested appeared to be enhanced or altered in the PC12nnrcells compared to PC12 cells. Internalization of, and responses to, epidermal growth factor were normal in the PC12nnrcells ruling out a generalized defect in hormonal binding, uptake, or response mechanisms. These findings are consistent with a causal association between the presence of high-affinity NGF receptors and of NGF responsiveness and internalization. A possible relationship is also suggested between regulation of cAMP responses and regulation of NGF responses or NGF receptor affinity.