Studies of RyR function in situ.

The ryanodine receptors (RyRs) are intracellular Ca2+ release channels of the sarcoplasmic reticulum (SR) involved in many cellular responses, including muscle excitation-contraction coupling. Multiple biochemical and biophysical methods are available to study RyR functions. However, most of them are somewhat limited because they can only be used to examine channels which are purified from the SR and no longer in their natural environment. In this review we discuss optical methods for studying RyR functions in situ. We describe several techniques for the investigation of local (microscopic) intracellular Ca2+ signals (a.k.a Ca2+ sparks) by means of confocal microscopy and flash photolysis of caged compounds. We discuss how these studies can and will continue to contribute to our understanding of RyR function in physiological and pathological conditions.