Role of membranes and energy-producing reactions in cellular processing of insulin in primary cultures of rat hepatocytes

Incubation of primary cultures of rat hepatocytes with the local anesthetics, procaine or lidocaine, had little or no effect on insulin uptake or degradation but caused an inhibition of insulin-stimulated glycogenesis. While exposure of cultures to the amines, monodansylcadaverine or CH3NH2, resulted in significant dose-dependent decreases in glycogenesis, only monodansylcadaverine (an inhibitor of receptor clustering) decreased uptake whereas CH3NH2 (a lysosomotropic agent) caused increases in both insulin uptake and degradation. When cells were treated with agents which inhibit glycolysis (NaF, 2-deoxy-D-glucose) or oxidative metabolism (2,4-dinitrophenol, carbonyl cyanide m-chlorophenyl hydrazone, NaN3, antimycin A), pronounced inhibitions of each of the bioactivities studied (syntheses of glycogen, protein, lipid) were observed, but only the glycolytic inhibitors decreased insulin uptake. These results suggest that insulin is internalized by an endocytotic process involving receptor clustering and requiring metabolic energy derived from glycolysis. The post-receptor biosynthetic processes involved in the expression of the biological activities of insulin (syntheses of glycogen, protein, lipid) require energy produced by oxidative metabolism while the degradation of insulin is carried out by nonlysosomal mechanisms which are not energy-requiring.     

Effect of psychotropic substances on the development of alcoholic motivation in noninbred white rats

The experiments have shown the capacity of antidepressant amitriptylin (0.5 mg/kg, i. p.) and tranquilizer phenazepam (0.1 mg/kg i. p.) to normalize the adaptive behaviour and almost completely prevent the development of alcohol motivation in animals with insufficient adaptive behaviour. It was established that in animals initially rejecting alcohol, chronic treatment with these drugs as well as d-amphetamine promoted alcohol motivation. The results obtained have proved our earlier hypothesis that preclinical search for drugs for the prevention and treatment of early stages of alcoholism is possible only in animals pre-selected according to their inclination to experimental alcoholism.     

Cyclic nucleotide alterations in mixed leukocyte reaction: a preliminary report

Endogenous cyclic adenosine and guanosine monophosphate (cAMP, cGMP) levels were studied in human peripheral blood lymphocytes during mixed leukocyte reactions (MLR). cAMP level was consistently elevated in one-way MLR, with good correlation to 3H-thymidine uptake in these reactions. In contrast, cGMP level was practically unchanged. Irradiation of reacting cell populations resulted in inhibition of cyclic nucleotide phosphodiesterase (PDE) activity. These results suggest that metabolic alterations in cAMP may be associated with immune reactions of cellular recognition.     

A comparison of the respiratory chain in particles from Paracoccus denitrificans and bovine heart mitochondria by EPR spectroscopy

A study is presented on the EPR characteristics of the paramagnetic groups in the respiratory chain present in membrane particles of Paracoccus denitrificans, the respiratory system of which is very similar to that in submitochondrial particles from beef heart. All paramagnetic prosthetic groups of the mitochondrial system are also found in the bacterial plasma membrane. Their properties suggest that the respiratory groups are embedded in very similar protein environments in the two systems.     

Regulation of Apoptosis in Nonapeptidergic Neurons of Rat Hypothalamus by Catecholamines in Experiments in vitro

Effects of noradrenaline (NA) and dopamine (DA) on apoptosis of nonapeptidergic neurons of supraoptic (SON) and paraventricular (PVN) nuclei of hypothalamus of male Wistar rats was studied in experiments in vitro. Incubation of hypothalamic sections in the medium with added NA was shown to induce an increase of the amount of pro-apoptotic protein caspase-9 in the nonapeptidergic neurons of the SON and PVN. A comparison of the level of neuronal NO-synthase with the level of caspase-9 expression in these neurons allows concluding that NA leads to initiation of apoptosis in neurons of the SON with mediation by nitric oxide (NO). In the PVN, the NA-induced initiation of apoptosis does not depend on the NO level. Addition of DA to the incubation medium results in an increase of the caspase-9 amount only in PVN neurons regardless of the NO content. The absence of neuronal death after the NA-induced increase of the caspase-9 level in the cells of SON and PVN seems to be due to increased expression of the anti-apoptotic protein bcl-2. Protection of the PVN neurons from death after addition of DA to the incubation medium is probably independent of the expression level of bcl-2. Thus, in the nonapeptidergic neurons of the SON and PVN, which are related by origin and by performed functions, modulation of the process of apoptosis by elevated concentrations of NA and DA is realized by different mechanisms.