title: Very early activation of m-calpain in peripheral nerve during Wallerian degeneration.
abstract: Peripheral nerve injury results in a series of events culminating in degradation of the axonal cytoskeleton (Wallerian degeneration). In the time period between axotomy and cytoskeletal degradation (24-48 h in rodents), there is calcium entry and activation of calpains within the axon. The precise timing of these events during this period is unknown. In the present study, antibodies were generated to three distinct peptide epitopes of m-calpain, and a fusion protein antibody was generated to the intrinsic calpain inhibitor calpastatin. These antibodies were used to measure changes in these proteins in mouse sciatic nerves during Wallerian degeneration. In sciatic nerve homogenates and cultured dorsal root ganglion (DRG) neurites, m-calpain protein was significantly reduced in transected nerves very early after nerve injury, long before axonal degeneration occurred. Levels of m-calpain protein remained low as compared to control nerves for the remainder of the 72-h time course. No changes in calpastatin protein were evident. Systemic treatment of animals with the protease inhibitor leupeptin partially prevented the rapid loss of calpain protein. Removal of calcium in DRG cultures had the same effect. These data indicate that m-calpain protein is lost very early after axonal injury, and likely reflect activation and degradation of this protein long before the cytoskeleton is degraded. Calpain activation may be an early event in a proteolytic cascade that is initiated by axonal injury and culminates with axonal degeneration.
authors.affiliation: Department of Neurology, Emory University School of Medicine, 1639 Pierce Drive, Suite 6000, Atlanta, GA 30322, USA. jglass03@emory.edu
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Glass
authors.forename: Deborah G
authors.initials: DG
authors.lastname: Culver
authors.forename: Allan I
authors.initials: AI
authors.lastname: Levey
authors.forename: Norman R
authors.initials: NR
authors.lastname: Nash
mesh_terms: D000818:Animals; D001369:Axons / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology; D002135:Calcium-Binding Proteins / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D002154:Calpain / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism*; D015536:Down-Regulation / Q000187:drug effects / Q000502:physiology; D007976:Leupeptins / Q000494:pharmacology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010447:Peptide Hydrolases / Q000187:drug effects / Q000378:metabolism; D010525:Peripheral Nerves / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D010523:Peripheral Nervous System Diseases / Q000378:metabolism* / Q000503:physiopathology; D011480:Protease Inhibitors / Q000494:pharmacology; D011817:Rabbits; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013997:Time Factors; D014404:Tubulin / Q000378:metabolism; D014855:Wallerian Degeneration / Q000378:metabolism* / Q000503:physiopathology
chemical_list: D002135:Calcium-Binding Proteins; D007976:Leupeptins; D011480:Protease Inhibitors; D014404:Tubulin; C033668:calpastatin; D010447:Peptide Hydrolases; D002154:Calpain; C449000:m-calpain