SUMMARY1. Membrane potential and flavoprotein redox state have been measured simultaneously and continuously in brown adipose tissue in order to determine how nerve stimulation and adrenergic agonists control its metabolic activity.2. Both trains of nerve impulses and addition of noradrenaline evoke two temporally distinct cell depolarizations. The first rapid depolarization precedes the increase in flavoprotein reduction.3. With nerve stimulation, at the time of maximum flavoprotein reduction the cell has repolarized or hyperpolarized. The second slow depolarization follows flavoprotein reduction.4. Phentolamine, an a antagonist, selectively blocks the first depolarization, but not the flavoprotein reduction. However the time of maximum flavoprotein reduction is delayed.5. Propranolol, a ,3 antagonist, delays the first repolarization until the end of nerve stimulation and inhibits the transient hyperpolarization, second depolarization and flavoprotein reduction.6. Isoproterenol, a , agonist, or the fatty acid octanoate produce only a transient hyperpolarization and subsequent slow depolarization following flavoprotein reduction.7. Thus brown adipose tissue contains both a-and ,-adrenergic receptors. Stimulation of a receptors produces an early membrane depolarization. Stimulation of ,8 receptors leads to an increase in metabolic activity which then appears to produce slow changes in membrane potential.

Photoreceptor cells of the drone, Apis mellifera male, have a voltage-gated Na+ membrane conductance that can be blocked by tetrodotoxin (TTX) and generates an action potential on abrupt depolarization: an action potential is triggered by the rising phase of a receptor potential evoked by an intense light flash (Autrum and von Zwehl 1964; Baumann 1968). We measured the intracellular voltage response to a small (9%), brief (30 ms) decrease in light intensity from a background, and found that its amplitude was decreased by 1 microM TTX. The response amplitude was maximal when the background intensity depolarized the cell to -38 mV. With intensities depolarizing the cell membrane to -45 to -33 mV the average response amplitude was decreased by TTX from 1.2 mV to 0.5 mV. TTX is also known to decrease the voltage noise during steady illumination (Ferraro et al. 1983) but, despite this, the ratio of peak-to-peak signal to noise was, on average, decreased by TTX. The results suggest that drone photoreceptors use voltage-gated Na+ channels for graded amplification of responses to small, rapid changes in light intensity.

Defects of both diet-induced thermogenesis and cold tolerance have been reported for streptozotocin-diabetic rats. Since brown adipose tissue (BAT) is a major effector of both diet- and cold-induced thermogenesis in the rat, the possible cause of these defects was investigated by comparing BAT metabolism under basal conditions and during activation by nerve stimulation, norepinephrine (NE), or octanoate addition in both streptozotocin-diabetic rats and in controls. The following metabolic indices were measured in rat interscapular BAT (IBAT): 1) tissue composition, 2) heat production rate as measured by direct microcalorimetry, 3) redox state of flavoproteins linked to the acyl-coenzyme A dehydrogenase pathway as measured by reflection spectrometry, 4) redox state of NAD(P) as measured by surface-emitted fluorescence, and 5) fatty acid activation and beta-oxidation activities in IBAT homogenate. In streptozotocin-diabetic rats, IBAT was atrophied (DNA content unmodified, protein and lipid content decreased). The basal and NE-stimulated total heat production rates showed a 75% and 56% decrease, respectively. The specific activity of fatty acid beta-oxidation as measured by flavoprotein redox state or enzymatically was decreased by 52% and 59%, respectively. The basal redox level of NAD(P) was about 3 times higher than in the controls and NE stimulation resulted in oxidation in contrast to the reduction observed in control tissues. These results show that the metabolic capacity of IBAT from streptozotocin-diabetic rats is decreased and further suggest that the reduced capacity for beta-oxidation contributes significantly to the metabolic alteration.

The predominant glial cells of the drone retina (outer pigment cells) respond to an increase in extracellular [K+] (Ko) by a net uptake of K+; thus, they contribute to bringing Ko back toward its baseline value. The authors report herein that there is also a different mechanism by which light stimulation of the retina causes an increase in intracellular free [K+] in the glial cells. In superfused retinal slices, after 5-10 minutes of continuous illumination at physiological intensities, extracellular [K+] often fell back to below its original level in the dark. This fall can be explained by increased activity of the Na/K pump in the photoreceptors and diffusion of K+ down their axons. Despite the absence of raised Ko, K+-selective microelectrodes in glial cells recorded a small increase in intracellular [K+] that was maintained for the duration of the illumination; i.e. a change occurred in the glia that was not mediated by an increase in Ko. The increase in intracellular [K+] is not mediated by illumination of the screening pigment in the glia. Unless the increase is caused by illumination of some other, unknown, pigment in the glia, the results show that some unidentified signal (that is not K+) passes from the photoreceptors to the glia.

The decrease in the metabolic capacity of rat brown adipose tissue during the late postnatal period can be reversed by cold acclimation of the animals. In order to find out whether a parallel decrease in capability for intercellular communication observed during this period is also reversed by cold acclimation, gap junction size and number per unit area of cell surface have been quantified in freeze-fracture replicas; cell diameters have been measured in semi-thin sections. It was found that the specific number of gap junctions remains unchanged during cold acclimation. However, the mean gap junction size increases by 75% and the ratio of gap junctional area per cell volume, an index for intercellular exchange capacity, is doubled. This result illustrates further the parallelism between metabolic capacity and cell communication in brown fat.