The endoplasmic reticulum-sarcoplasmic reticulum connection. II. Postnatal differentiation of the sarcoplasmic reticulum in skeletal muscle fibers.
The postnatal differentiation of sarcoplasmic reticulum (SR) of rabbit skeletal muscles (the slow-twitch soleus and the fast-twitch adductor muscles) was monitored between Days 1 and 12 by following on Western blots the expression and accumulation of molecular markers specific not only for the muscle endomembrane system, i.e., calsequestrin (CS) and the ryanodine-sensitive Ca2+ release channel, but also for the endoplasmic reticulum (ER) at large, i.e., BiP, calnexin (CN) and calreticulin. Our results demonstrate that SR development, documented by the increase of the SR fractional volume, terminal cisternae proliferation, and reorientation of triads, is accompanied by the accumulation of the SR-specific proteins and also of CN, with no change of the other ER general markers. Moreover, the distribution of two of the markers, BiP and CS, was investigated by immunocytochemistry at both the light and the electron microscope level. At Day 1 CS was found to be concentrated both within the few recognizable triad terminal cisternae and within the lumen of numerous, apparently discrete cisternae and tubules, widely scattered throughout both the contractile and the subplasmalemmal areas of the cytoplasm. These structures remain evident until Day 12, when most triad junctions have acquired proper configuration, composition and orientation. BiP, on the other hand, appears widely distributed within the ER/SR of the fibers. From the early stages of postnatal development it does colocalize with the Ca2+ binding protein in the lumen of the CS-rich structures and appears also within the longitudinal SR and the conventional ER cisternae.
Villa
A
A
Department of Pharmacology, University of Milano, Italy.
Podini
P
P
Nori
A
A
Panzeri
M C
MC
Martini
A
A
Meldolesi
J
J
Volpe
P
P
D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000378:metabolism; D037281:Calnexin; D037282:Calreticulin; D002155:Calsequestrin / Q000378:metabolism; D002454:Cell Differentiation; D004721:Endoplasmic Reticulum / Q000648:ultrastructure*; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D009132:Muscles / Q000648:ultrastructure; D011817:Rabbits; D012261:Ribonucleoproteins / Q000378:metabolism; D012519:Sarcoplasmic Reticulum / Q000648:ultrastructure*; D013997:Time Factors
D002135:Calcium-Binding Proteins; D037282:Calreticulin; D002155:Calsequestrin; D012261:Ribonucleoproteins; D037281:Calnexin