Calcium-activated proteolysis of neurofilament proteins in goldfish Mauthner axons.
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 degrees 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 alpha-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 2mM 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.
Raabe
T D
TD
Department of Zoology, University of Texas at Austin 78712-1064, USA.
Nguyen
T
T
Bittner
G D
GD
D000818:Animals; D001369:Axons / Q000378:metabolism / Q000502:physiology; D002118:Calcium; D002154:Calpain / Q000502:physiology; D004586:Electrophoresis; D006054:Goldfish / Q000502:physiology*; D015151:Immunoblotting; D009419:Nerve Tissue Proteins / Q000502:physiology*; D016900:Neurofilament Proteins / Q000378:metabolism*
D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; D002154:Calpain; D002118:Calcium