Mitochondrial Ca(2+) uptake is crucial for the regulation of the rate of oxidative phosphorylation, the modulation of spatio-temporal cytosolic Ca(2+) signals and apoptosis. Although the phenomenon of mitochondrial Ca(2+) sequestration, its characteristics and physiological consequences have been convincingly reported, the actual protein(s) involved in this process are unknown. Here, we show that the uncoupling proteins 2 and 3 (UCP2 and UCP3) are essential for mitochondrial Ca(2+) uptake. Using overexpression, knockdown (small interfering RNA) and mutagenesis experiments, we demonstrate that UCP2 and UCP3 are elementary for mitochondrial Ca(2+) sequestration in response to cell stimulation under physiological conditions - observations supported by isolated liver mitochondria of Ucp2(-/-) mice lacking ruthenium red-sensitive Ca(2+) uptake. Our results reveal a novel molecular function for UCP2 and UCP3, and may provide the molecular mechanism for their reported effects. Moreover, the identification of proteins fundemental for mitochondrial Ca(2+) uptake expands our knowledge of the physiological role for mitochondrial Ca(2+) sequestration.
In extrahepatic tissues lipoprotein lipase (LPL) hydrolyzes triglycerides thereby generating FFA for tissue uptake and metabolism. To study the effects of increased FFA uptake in muscle tissue, transgenic mouse lines were generated with a human LPL minigene driven by the promoter of the muscle creatine kinase gene. In these mice human LPL was expressed in skeletal muscle and cardiac muscle, but not in other tissues. In proportion to the level of LPL overexpression, decreased plasma triglyceride levels, elevated FFA uptake by muscle tissue, weight loss, and premature death were observed in three independent transgenic mouse lines. The animals developed a severe myopathy characterized by muscle fiber degeneration, fiber atrophy, glycogen storage, and extensive proliferation of mitochondria and peroxisomes. This degree of proliferation suggests that FFA play an important role in the biogenesis of these organelles. Our experiments indicate that LPL is rate limiting for the supply of muscle tissue with triglyceride-derived FFA. Improper regulation of muscle LPL can lead to major pathological changes and may be important in the pathogenesis of some human myopathies. Muscle-specific LPL transgenic mouse lines will serve as a useful animal model for the investigation of myopathies and the biogenesis of mitochondria and peroxisomes. (J. Clin. Invest. 1995. 96:976-986.)
The objective of the present study was to investigate the involvement of key players in reverse cholesterol/ 24(S)OH-cholesterol transport in primary porcine brain capillary endothelial cells (pBCEC) that constitute the BBB. We identified that, in addition to scavenger receptor class B, type I (SR-BI), pBCEC express ABCA1 and
Lipoprotein lipase (LPL) is the rate-limiting enzyme for the import of triglyceride-derived fatty acids by muscle, for utilization, and adipose tissue (AT), for storage. Relative ratios of LPL expression in these two tissues have therefore been suggested to determine body mass composition as well as play a role in the initiation and͞or development of obesity. To test this, LPL knockout mice were mated to transgenics expressing LPL under the control of a musclespecific promoter (MCK) to generate induced mutants with either relative (L2-MCK) or absolute AT LPL deficiency (L0-MCK). L0-MCK mice had normal weight gain and body mass composition. However, AT chemical composition indicated that LPL deficiency was compensated for by large increases in endogenous AT fatty acid synthesis. Histological analysis confirmed that such up-regulation of de novo fatty acid synthesis in L0-MCK mice could produce normal amounts of AT as early as 20 h after birth. To assess the role of AT LPL during times of profound weight gain, L0-MCK and L2-MCK genotypes were compared on the obese ob͞ob background. ob͞ob mice rendered deficient in AT LPL (L0-MCKob͞ob) also demonstrated increased endogenous fatty acid synthesis but had diminished weight and fat mass. These findings reveal marked alterations in AT metabolism that occur during LPL deficiency and provide strong evidence for a role of AT LPL in one type of genetic obesity.Lipoprotein lipase (LPL), located on the capillary endothelium of extrahepatic tissues, catalizes the rate-limiting step in the hydrolysis of triglycerides (TGs) from circulating chylomicrons and very low density lipoprotein (for reviews, see refs. 1 and 2). Quantitatively, most LPL is found in adipose tissue (AT) and muscle, where the liberated free fatty acids are taken up and either stored or oxidized, respectively (3). It has been hypothesized that relative levels of LPL activity in AT and muscle determine how fat calories are partitioned toward storage or utilization, and that imbalances in tissue expression can therefore lead to obesity or weight loss (4, 5). Until the advent of induced mutant mice, experimental systems to directly test this hypothesis have not been available.We have previously generated LPL knockout mice (6) as well as transgenic mice expressing human LPL exclusively in muscle (7). LPL-deficient mice are normal at birth, but develop lethal hypertriglyceridemia within the first day of life, at which point they have markedly reduced intracellular lipid stores. Transgenic mice that express high levels of human LPL in muscle showed increased muscle free fatty acid concentrations and increased numbers of fatty acid-metabolizing organelles (mitochondria and peroxisomes). Together these observations suggest that tissue LPL expression is a major determinant of fatty acid entry into cells. Unfortunately, the previous studies could not answer questions about the longterm metabolic consequences for the host of altered tissue expression of LPL. Neonatal death in the knockout mice precluded examinat...
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