1.Brown fat cells exhibit a classical mitochondria1 electron transport system. 2. The respiration of brown fat cells can be stimulated by nor-epinephrine and this hormonally mediated respiration can be mimicked by oleate.3. Both nor-epinephrine and oleate-stimulated respiration can be blocked by rotenone, antimycin, azide and malonate, and the antimycin block can be by-passed by tetramethyl-p-phen y lenediamine .4. a-Glycerol phosphate and succinate were found to stimulate respiration. 5. The potent a-glycerol phosphate oxidase cannot quantitatively contribute to the respiration evoked during catecholamine induced thermogenesis.It is well-known that catecholamines are able to affect a number of systems in the whole animal, such as glycogenolysis by activating the phosphorylase, glycolysis by activating phosphofructokinase and lipolysis by activating the triglyceride lipase. I n recent years the action of catecholamines on brown adipose tissue has received much attention [I -31. Numerous studies carried out in vivo have shown that the sympathetic nervous system, which releases catecholamines on stimulation, controls thermogenesis in brown fat upon cold exposure and probably on arousal from hibernation [3 -81. Several investigators have demonstrated the rich sympathetic innervation of brown fat where the catecholamine content of the tissue was observed to be 0.5-1.0 pg/gram of tissue, a figure equalled only by the heart [9--121. It is known that the thermogenic effect is linked with increased fatty acid mobilisation and oxidation [I3 to i5]. The work presented here has been carried out in order to investigate whether the catecholamine affects all the above systems or rather whether the activity is restricted to one or more of the functions mentioned.The experiments of Heim and Hull The events leading to this large stimulation of 0, utilization and the nature of the oxidative mechanism will be discussed here.
METHODS AND MATERIALS
Isolation of Brown Fat CellsBrown fat cells were isolated by a method similar to that described by Fain et al. [19]. Two important modifications were introduced into the isolation procedure. First, hamsters were starved for 48 to 72 h prior to sacrifice in order to reduce the size of the lipid globules within the cells. Analyses of extractable lipids from rat brown fat show a 40°/, reduction of lipid after 48 h of starvation [21]. Second, micro-hematocrit determinations of the cell volume in the final suspension were made in order to allow adjustment of the suspension concentration prior to experimentation.Adult hamsters, more than 60 days old, weighing 70-105 g, were killed by guillotine to minimize the