Thiamine and Cholinergic Transmission in the Electric Organ of Torpedo

The electric organ of Torpedo marmorata was found to contain as much as 120 +/- 24 nmol of thiamine per g of fresh tissue. The vitamin was distributed as nonesterified thiamine (32%), thiamine monophosphate (22%), thiamine diphosphate (8%), and an important proportion of thiamine triphosphate (38%). A high level of thiamine triphosphate was found in synaptosomes isolated from the electric organ. In contrast, the synaptic vesicles did not show any enrichment in thiamine, whereas they contained a marked peak of acetylcholine (ACh) and ATP. Thus thiamine seems to be very abundant in cholinergic nerve terminals; its localization is apparently extravesicular, either in the axoplasm or in association with plasma membrane. When calcium was reduced and magnesium increased in the external medium, the efficiency of transmission was diminished, owing to inhibition of ACh release; in a parallel manner the degree of thiamine phosphorylation was found to increase--this condition is known to modify the repartition of ACh between vesicular and extravesicular compartments. Electrical stimulation, which causes periodic variations of the level of ACh and ATP, also caused significant changes in thiamine esters. In addition, related changes of the vitamin and the transmitter were observed under other conditions, suggesting a functional link between the metabolism of thiamine and that of ACh in cholinergic nerve terminals.     

Biodegradation of [C]Tri-p-Cresyl Phosphate in a Laboratory Activated-Sludge System

Biodegradation of [C]tri-p-cresyl phosphate was studied in a laboratory model sewage treatment system to develop information on the nature of its transformation products. In 24-h experiments, 70 to 80% of tri-p-cresyl phosphate added at the 1-mug/ml level was degraded. The remaining parent compound was associated with the sludge solids. The major metabolite extracted with ethyl ether from the aqueous phase was identified as p-hydroxybenzoic acid by gas chromatography-mass spectrometry. Two unstable ether-extractable metabolites were not identified. The half-life of [C]tri-p-cresyl phosphate was estimated to be 7.5 h.     

Thin-layer chromatographic collector-eluter.

A combination thin-layer chromatographic collector and elution apparatus is described. The device is designed to minimize manipulations and maximize accuracy and can be used for both analytical and small-scale preparative separations.     

Voltage-dependent conductance induced by hemocyanin in black lipid films

When hemocyanin is added to a black lipid film, the conductance increases in discrete steps. For negative potentials the single step conductance is constant, but for positive potentials the step conductance appears to decrease as the potential increases. At high positive potentials the conductance fluctuates between several levels. These data suggest that, in lipid membranes, hemocyanin conducts ions through discrete channels. The voltage-dependent conductance observed at high levels of conductance seems to be a consequence of the properties of the conductance of the single channel.     

The nature of the voltage-dependent conductance of the hemocyanin channel.

The electrical responses of individual hemocyanain channels in oxidized cholesterol membranes demonstrate that the voltage-dependent conductance of many-chanel membranes arises from two different mechanisms. These are the voltage-dependent redistribution of channels among several discrete single-channel conductance states themselves. The relaxation time for the discrete conductance changes is of the order of seconds nd the relaxation time of the continuous conductance changes is of the order 10(-4) seconds. As salt concentration in the bathing medium is increased, the single-channel conductance first increases lineary and then saturates. The characteristics of the saturation curves suggest that the continuous conductance changes occur at the edges of the channel and that the mean time an ion spends in the channel is 4 nanoseconds...     

Distribution of interstitial retinol-binding protein (IRBP) in the vertebrates

Immunoblots of interphotoreceptor matrix preparations from 20 species belonging to six vertebrate classes were probed with antibodies against bovine interstitial retinol-binding protein (b-IRBP). Each preparation displayed an immunoreactive protein band. In the Osteichthyes, the apparent Mr of this band was 67,600 +/- 2,700 (mean +/- SD, n = 8). In two of the Osteichthyes, the band was resolved into a closely spaced doublet. Including previously published data for five mammals and one amphibian, species from the other classes (Chondrichthyes, one species; Amphibia, four species; Reptilia, one species; Aves, one species; Mammalia, nine species) had IRBPs with Mr that averaged 2.0 times that of the Osteichthyes, namely 134,200 +/- 8,600 (mean +/- SD, n = 17). Frog IRBP was very similar to mammalian IRBP in terms of its immunohistochemical distribution (determined with rabbit anti-frog IRBP antibodies), its molecular weight (sodium dodecyl sulfate polyacrylamide gel electrophoresis and gel-filtration chromatography), retinol- and concanavalin A-binding ability, and because it was synthesized and secreted in vitro by the isolated retina but not by the pigmented layers of eye. Goldfish IRBP apparently binds exogenous (3H)-retinol but does not bind concanavalin A and has about half the Mr of frog IRBP. The occurrence of IRBP-like proteins cross-reacting with anti b-IRBP antibodies in the interphotoreceptor matrix of all six major vertebrate classes is consistent with the hypothesis that IRBP is an important element in the vertebrate visual cycle.