The autoradiographic perspective of central benzodiazepine receptors; a short review.

1. We reviewed studies performed to characterize central benzodiazepine binding sites. 2. An overview of the different radioligands used to characterize BZ1 and BZ2 binding sites and a mapping of these central benzodiazepine sites are described. 3. Saturation studies carried out by autoradiogram quantification also are reviewed. 4. The specific use of the autoradiographic technique to carry out studies on ontogeny, development, and phylogeny is discussed, as well as studies performed using this technique on some diseases and experimental conditions, such as drug treatments or chemical and mechanical lesions.

Effects of glycosylation enzymes from membrane fractions, induced by chronic ethanol administration.

To study the effect of chronic ethanol administration on the enzyme activities involved in the glycosylation processes (glycosidases and glycosyltransferases), we used 6-week ethanol-treated female Wistar rats and pair-control rats. Biological material was membrane fractions of microsomes and plasma membrane obtained by subcellular fractionation technique. Ethanol treatment increased with statistical significance the Vmax of N-acetyl-beta-D-glucosaminidase (P less than 0.10), beta-D-glucuronidase and alpha-D-mannosidase (P less than 0.001) activities from microsomal fractions and likewise it increased the Km value of beta-D-glucuronidase (P less than 0.001). In vitro doses of ethanol (0.1-1.7 M) were added to delucidate hypothetical membrane tolerance responses. However in vitro addition of ethanol provoked no differential effects in treated and untreated rats. Glycosyltransferase assays were carried out using [14C]sugar derivatives. We detected several glycosyltransferase activities in both microsome and plasma membrane fractions. Chronic ethanol exposure appeared to affect N-acetyl-neuraminyltransferase activity from both membrane fractions, producing a greatly increased incorporation in the presence of asialofetuin. Glucosyl and mannosyltransferase activities from plasma membrane fractions were also altered by ethanol treatment, producing an increased enzyme activity when reaction was performed in the presence of phosphatidylcholine + dolichol-phosphate liposomes.

Pyrenesulfonyl azide as a fluorescent label for the study of protein-lipid boundaries of acetylcholine receptors in membranes.

Acetylcholine receptor (AcChR) enriched membrane fragments from Torpedo californica electroplax were labeled by in situ photogenerated nitrenes from a hydrophobic fluorescent probe, pyrene-1-sulfonyl azide. Preferential photolabeling of membrane proteins, mainly AcChR, has been achieved and there is a pronounced exposure of the 48,000 and 55,000 molecular weight subunits of AcChR to the lipid environment of the membrane core. Covalent attachment of the photogenerated fluorescence probe does not perturb the alpha-neurotoxins' binding properties of membrane-bound AcChR or the desensitization kinetics induced by prolonged exposures to cholinergic agonists. Non-covalent photoproducts can be conveniently removed from labeled membrane preparations by exchange into lipid vesicles prepared from electroplax membrane lipids. Fluorescence features of model pyrene sulfonyl amide derivatives, such as fine vibrational structure of emission spectra of fluorescence lifetimes, are highly sensitive to the solvent milieu. The covalently bound probe shows similar fluorescence properties in situ. PySA photoproducts have great potential to spectroscopically monitor neurotransmitter induced events on selected AcChR subunits exposed to the hydrophobic environment of membranes.