Plasticity of the transverse tubules following denervation and subsequent reinnervation in rat slow and fast muscle fibres.
We have studied the effects of short term denervation followed by reinnervation on the ultrastructure of the membrane systems and on the content of and distribution of key proteins involved in calcium regulation of fast-twitch (FT) extensor digitorum longus (EDL) and slow-twitch (ST) soleus (SOL) muscle fibres. Ischiadic nerve freezing resulted in total lack of neuromuscular transmission for 3 days followed by a slow recovery, but no decline in twitch force elicited by direct stimulation. The latter measurements indicate no significant atrophy within this time frame. The membrane systems of skeletal muscle fibres were visualized using Ca92+)-K3Fe(CN)6-OsO4 techniques and observed using a high voltage electron microscope. [3H]nitrendipine binding was used to detect levels of dihydropyridine receptor (DHPR) expression. The Ca2+ pumping free sarcoplasmic reticulum domains were not affected by the denervation, but the Ca2+ release domains were dramatically increased, particularly in the FT-EDL muscle fibres. The increase is evidenced by a doubling up of the areas of contacts between SR and transverse (t-) tubules, so that in place of the normal triadic arrangement, pentadic and heptadic junctions, formed by multiple interacting layers of ST and t-tubules are seen. Frequency of pentads and heptads increases and declines in parallel to the denervation and reinnervation but with a delay. Immunofluorecence and electron microscopy observations show presence of DHPR and ryanodine receptor clusters at pentads and heptads junctions. A significant (P < 0.01) positive correlation between the level of [3H]nitrendipine binding component and the frequency pentads and heptads was observed in both the FT-EDL and ST-SOL muscle fibres indicating that overexpression of DHPRs accompanies the build up extra junctional contacts. The results indicate that denervation reversibly affects the domains of the membrane systems involved in excitation-contraction coupling.
Takekura
Hiroaki
H
Department of Physiological Sciences, National Institute of Fitness and Sports, Kanoya, Kagoshima 891-2393, Japan. takekura@nifs-k.ac.jp
Tamaki
Hiroyuki
H
Nishizawa
Tomie
T
Kasuga
Norikatsu
N
D000818:Animals; D002118:Calcium / Q000378:metabolism; D020746:Calcium Channels, L-Type / Q000378:metabolism; D020013:Calcium Signaling; D005260:Female; D007425:Intracellular Membranes / Q000648:ultrastructure*; D008854:Microscopy, Electron; D009119:Muscle Contraction / Q000502:physiology; D009121:Muscle Denervation; D018656:Muscle Fibers, Fast-Twitch / Q000648:ultrastructure*; D018657:Muscle Fibers, Slow-Twitch / Q000648:ultrastructure*; D018482:Muscle, Skeletal / Q000294:innervation / Q000648:ultrastructure*; D009409:Nerve Crush; D051381:Rats; D012519:Sarcoplasmic Reticulum / Q000648:ultrastructure*
D020746:Calcium Channels, L-Type; D002118:Calcium