Heat-shock proteins in axoplasm: high constitutive levels and transfer of inducible isoforms from glia.
To characterize heat-shock proteins (HSPs) of the 70-kDa family in the crayfish medial giant axon (MGA), we analyzed axoplasmic proteins separately from proteins of the glial sheath. Several different molecular weight isoforms of constitutive HSP 70s that were detected on immunoblots were approximately 1-3% of the total protein in the axoplasm of MGAs. To investigate inducible HSPs, MGAs were heat shocked in vitro or in vivo, then the axon was bathed in radiolabeled amino acid for 4 hours. After either heat-shock treatment, protein synthesis in the glial sheath was decreased compared with that of control axons, and newly synthesized proteins of 72 kDa, 84 kDa, and 87 kDa appeared in both the axoplasm and the sheath. Because these radiolabeled proteins were present in MGAs only after heat-shock treatments, we interpreted the newly synthesized proteins of 72 kDa, 84 kDa, and 87 kDa to be inducible HSPs. Furthermore, the 72-kDa radiolabeled band in heat-shocked axoplasm and glial sheath samples comigrated with a band possessing HSP 70 immunoreactivity. The amount of heat-induced proteins in axoplasm samples was greater after a 2-hour heat shock than after a 1-hour heat shock. These data indicate that MGA axoplasm contains relatively high levels of constitutive HSP 70s and that, after heat shock, MGA axoplasm obtains inducible HSPs of 72 kDa, 84 kDa, and 87 kDa from the glial sheath. These constitutive and inducible HSPs may help MGAs maintain essential structures and functions following acute heat shock.
Sheller
R A
RA
Department of Zoology, University of Texas at Austin, 78712, USA. shellerr@southwestern.edu
Smyers
M E
ME
Grossfeld
R M
RM
Ballinger
M L
ML
Bittner
G D
GD
D000200:Action Potentials; D000818:Animals; D003400:Astacoidea / Q000502:physiology*; D001369:Axons / Q000502:physiology* / Q000648:ultrastructure; D003593:Cytoplasm / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D006358:Hot Temperature; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D009457:Neuroglia / Q000166:cytology / Q000502:physiology*
D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins