Troponin T expression in trout red muscle correlates with muscle activation

Abstract

SUMMARY Red or aerobic muscle from the anterior of rainbow trout Oncorhynchus mykiss activates (generates force) more quickly than that from the posterior. TnT is a component of the troponin complex that modulates muscle activation once Ca2+ is bound. Since trout express at least two forms of TnT in their red muscle (S1 and S2), the differential expression of these two forms was predicted to explain variations in contractile properties. TnT isoforms from trout muscle were identified through hydroxy-apatite chromatography of purified myofibrillar proteins followed by SDS-PAGE. Western blots employing a mammalian anti-troponin T monoclonal antibody were used to identify TnT isoforms. The relative expression of the two isoforms of TnT was then examined at seven longitudinal positions from each of three fish using SDS-PAGE and densitometry on the silver-stained TnT bands. A significant shift in expression was detected from anterior to posterior in all three fish with TnT S1 becoming more dominant in the posterior red muscle. As predicted, a shift in TnT expression was associated with the decrease in activation rate along the length of the fish. This study was then extended to include a different species of salmonid, brook trout Salvelinus fontinalis, to explore the generality of TnT modulation of muscle activation. Muscle contractile properties were determined from anterior and posterior muscle, and relative expression of S1 and S2 was determined. Unlike rainbow trout, there is no consistent longitudinal pattern of muscle activation in brook trout:some fish have kinetically faster muscle in the anterior, some in the posterior. Similarly, there is no consistent pattern of TnT expression. Individual analysis of muscle activation and TnT expression in brook trout provides insight into the role of TnT in modulating muscle activation in slow fish muscle.