Diaphragm disuse reduces Ca2+ uptake capacity of sarcoplasmic reticulum.

Abstract

Chronic phrenic tetrodotoxin (TTX) blockade and phrenic denervation (Dnv) of hamster diaphragm result in decreased maximum specific tension, prolonged contraction time, and improved fatigue resistance (W. Z. Zhan and G. C. Sieck, J. Appl. Physiol. 72: 1445-1453, 1992). An underlying increased relative contribution of type I fibers to total muscle mass appears to be consistent with, but does not completely account for, changes in contractile and fatigue properties. The present study was designed to evaluate a potential role for altered cellular Ca2+ metabolism in the adaptive response of the diaphragm to chronic disuse. An analytic method based on simulation and modeling of long-term 45Ca2+ efflux data was used to estimate Ca2+ contents (nmol Ca2+/g wet wt tissue) and exchange fluxes (nmol Ca2+.min-1.g-1) for extracellular and intracellular compartments in the in vitro hamster hemidiaphragm after prolonged disuse. Three groups were compared: control (Con, n = 5), phrenic TTX blockade (TTX, n = 5), and phrenic denervation (Dnv, n = 5). Experimental muscles were loaded with 45Ca2+ for 1 h, and efflux data were collected for 8 h by using a flow-through tissue chamber. Compartmental analysis of efflux data estimated that the Ca2+ contents and Ca2+ exchange fluxes of the largest and slowest intracellular compartment (putative longitudinal reticulum) were reduced by approximately 50% in TTX and Dnv muscle groups compared with Con. In addition, the kinetic model predicted significant decreases in total intracellular Ca2+ and total diaphragm Ca2+ in TTX and Dnv muscles. We conclude that the data support the hypothesis that the capacity of the sarcoplasmic reticulum for Ca2+ sequestration is reduced in chronic diaphragm disuse. The impact of this effect on diaphragm contractile and fatigue properties is discussed.