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Introduction Androgen deficiency commonly occurs in middle-aged and older men ( Clinical studies have shown that low levels of endogenous testosterone are also associated with cardiovascular diseases ( Testosterone plays an important role in the regulation of carbohydrate, fat and protein metabolism ( In terms of energy production, cardiac cells contain two spatially distinct mitochondrial subpopulations: the subsarcolemmal subpopulation (SSM), located just below the sarcolemma and the interfibrillar subpopulation (IFM) which is located between the myofibrils ( The present study aims to assess (1) cardiac contractility, (2) mitochondrial function, (3) oxidative phosphorylation, (4) morphology and mitochondrial membrane potential, and (5) permeability transition pore opening in the two subpopulations of cardiac mitochondria, thus providing new information on the effect of testosterone deficiency on cardiac contractility and mitochondrial function