A chemical method for intracellular loading of the calcium indicator aequorin in mammalian skeletal muscle.
The bioluminescent calcium indicator aequorin was loaded into bundles of skeletal muscle fibers from the rat extensor digitorum longus by macroinjection, a technique previously applied only to cardiac muscle. After loading, the amplitude and time course of the twitch returned to control values, indicating lack of damage to the fibers. Individual light signals (i.e., calcium transients) were recorded during each twitch or tetanus without the need for signal averaging. The calcium transients obtained were qualitatively and quantitatively similar to those reported previously with microinjection of aequorin. Our data suggest that macroinjection may be the method of choice for loading aequorin into mammalian skeletal muscle.
Perreault
C L
CL
Charles A. Dana Research Institute, Boston, Massachusetts.
Gonzalez-Serratos
H
H
Litwin
S E
SE
Morgan
J P
JP
D000331:Aequorin / Q000008:administration & dosage*; D000818:Animals; D002118:Calcium / Q000502:physiology; D003080:Cold Temperature / Q000009:adverse effects; D046508:Culture Techniques; D007267:Injections / Q000379:methods*; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects; D009132:Muscles; D051381:Rats; D011919:Rats, Inbred Strains
D000331:Aequorin; D002118:Calcium