Calcium‐activated proteolysis of neurofilament proteins in goldfish Mauthner axons

Tim D. Raabe, Tri Nguyen, George D. Bittner

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

We have examined the proteolytic breakdown of neurofilament proteins (NFPs) in isolated Mauthner axoplasm (M‐axoplasm). Documentation of proteolytic breakdown of NFPs in M‐axoplasm is important because NFPs are not degraded in distal segments of severed Mauthner axons (M‐axons) maintained in vivo for up to 62 days at 20°C. By incubating M‐axoplasm with 2 mM calcium in vitro, we have demonstrated that M‐axoplasm contains an endogenous calcium‐activated neutral protease that degrades NFPs. This calcium‐activated proteolysis of M‐axoplasm NFPs produced novel bands on silver‐stained gels. These novel bands were presumed to be NFP breakdown products because they reacted with antibodies to the α‐intermediate filament antigen (anti‐IFA) on immunoblots from these gels. Incubations of M‐axoplasm with 2 mM calcium plus exogenous calpain produced novel bands similar to those observed for M‐axoplasm incubated with 2 mM calcium. Incubations of M‐axoplasm with 2m M calcium plus calpain inhibitors did not produce these novel bands. These in vitro data indicate that M‐axoplasm contains calpain that degrades NFPs and produces novel bands similar to those observed from distal segments of severed M‐axons maintained in vivo longer than 62 days postseverance. Factors that affect the activity of calpain or affect the ability of calpain to degrade NFPs could account for the delayed degradation of NFPs in distal segments of severed M‐axons maintained in vivo. © 1995 John Wiley & Sons, Inc.

Original languageEnglish (US)
Pages (from-to)253-261
Number of pages9
JournalJournal of Neurobiology
Volume26
Issue number2
DOIs
StatePublished - Feb 1995
Externally publishedYes

Keywords

  • axonal degeneration
  • calpain
  • neurofilaments
  • proteolysis

ASJC Scopus subject areas

  • Neuroscience(all)
  • Cellular and Molecular Neuroscience

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