Neuritogenesis and the Nerve Growth Factor-induced Differentiation of PC-12 Cells Requires Annexin II-mediated Plasmin Generation

• Published in: The Journal of biological chemistry 2001-12, Vol.276 (52), p.49350-49358
• Main Authors: Jacovina, Andrew T., Zhong, Fengming, Khazanova, Elena, Lev, Emil, Deora, Arunkumar B., Hajjar, Katherine A.
• Format: Article
• Language: English
• Subjects: Aminocaproic Acid - pharmacology; Animals; Annexin A2 - genetics; Annexin A2 - metabolism; Antibodies; Cell Differentiation; Cell Membrane - metabolism; Cells, Cultured; Endothelium, Vascular - cytology; Endothelium, Vascular - metabolism; Fibrinolysin - metabolism; Gene Expression Regulation; Humans; Immunohistochemistry; Nerve Growth Factor - pharmacology; Neurites - drug effects; Neurites - physiology; Neurons - cytology; PC12 Cells; Peptides - immunology; Peptides - metabolism; plasminogen; Proto-Oncogene Proteins p21(ras) - genetics; Proto-Oncogene Proteins p21(ras) - metabolism; Rats; Tissue Plasminogen Activator - immunology; Up-Regulation - physiology; Urokinase-Type Plasminogen Activator - immunology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M106289200

One of the key morphological changes associated with the nerve growth factor (NGF)-induced differentiation of rat adrenal pheochromocytoma (PC-12) cells is the growth of axon-like processes called neurites. A growing body of evidence suggests that this process may be dependent upon plasmin, a serine protease generated from plasminogen (Plg) by either urokinase Plg activator (u-PA) or tissue Plg activator (t-PA). Prior work in our laboratory has identified annexin II (Ann-II) as a co-receptor for Plg and t-PA that promotes and localizes plasmin generation near the cell surface. In the present study, we report a 3–9-fold increase in Ann-II protein and message levels in NGF-treated PC-12 cells. Message stability and nuclear run-on assays suggest that this induction occurs at the level of gene transcription. Neurite outgrowth assays on and within a three-dimensional matrix demonstrate the inhibition of NGF-induced PC-12 cell differentiation by polyclonal and monoclonal antibodies directed against Ann-II as well as by the overexpression of antisense Ann-II mRNA. Neuritogenesis is also impaired by α2-plasmin inhibitor, antibodies directed against t-PA and u-PA, and ε-aminocaproic acid, a lysine analog that inhibits Plg activation and the binding of Plg to Ann-II. Plasmin generation assays reveal a 2-fold increase in plasmin production on NGF-treated PC-12 cells, which can be blocked by a polyclonal antibody directed against the tail region of Ann-II. From these data, we conclude that Ann-II is transcriptionally up-regulated by NGF and that Ann-II-mediated plasmin generation may play an important role during neurite development in the differentiating PC-12 cell.