A silver-reducing component in rat striated muscle. II. Isolated sarcoplasmic reticulum vesicles.
In previous work on rat striated muscle cells a silver-reducing component was found selectively localized at the terminal cistern/transverse tubule system (Tandler and Pellegrino de Iraldi 1989). To further investigate that problem we performed the Hg-Ag argentaffin reaction on a sarcoplasmic reticulum fraction from rat skeletal muscle. Circular profiles corresponding to vesicular structures were found outlined by silver grains. The number of silver "stained" vesicles were less than the total number vesicles stained by conventional procedures. The correlation between argentaffinities in the intact muscle fiber and their subcellular organelles indicated that the Hg-Ag reactive vesicles must be those derived from the terminal cisternae of the sarcoplasmic reticulum. The silver-reducing constituent aggregates in the presence of 1 mM CaCl2 or 0.5 M K cacodylate. The state of aggregation induced by Ca2+ was not affected by incubation with 0.5% Triton X-100 or by 2 mM EDTA, thus suggesting a localization at or near the membrane of the terminal cistern vesicle facing the junctional gap. In Laemmli SDS-acrylamide gels the Hg-Ag reaction stained all proteins in a manner similar to Coomasie blue. It is suggested that the selective histochemical staining is the result of differential reactivities due to steric requirements of the chemical reaction.
Tandler
C J
CJ
Instituto de Biología Celular, Facultad de Medicina, Universidad de Buenos Aires, República Argentina.
Gonzalez
D A
DA
Remorini
P G
PG
Pellegrino de Iraldi
A
A
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D006651:Histocytochemistry; D008628:Mercury / Q000378:metabolism; D008854:Microscopy, Electron; D009124:Muscle Proteins / Q000032:analysis*; D009132:Muscles / Q000032:analysis* / Q000648:ultrastructure; D051381:Rats; D012519:Sarcoplasmic Reticulum / Q000032:analysis* / Q000648:ultrastructure; D012834:Silver / Q000378:metabolism; D013194:Staining and Labeling; D013572:Synaptic Vesicles / Q000032:analysis
D009124:Muscle Proteins; D012834:Silver; D008628:Mercury