Determination of indoles and catechols in rat brain and pineal using liquid chromatography with fluorometric and amperometric detection

Tryptophan, serotonin, 5-hydroxyindoleacetic acid, and homovanillic acid were determined in rat brain by the direct injection of a centrifuged tissue homogenate into a liquid chromatographic—fluorometric/amperometric system. The above indoles, along with melatonin, were also determined in single rat pineal glands. The utility of the system in determining several additional catechols and indoles in brain was examined.     

The hypothalamo-choroidal tract

Summary Light and electron microscopic examination of choroid plexuses from lateral ventricles of water-deprived and subcutaneously or intravenously vasopressin administered rats reveal morphologic changes typical for vasopressin responsive fluid transporting epithelia during hormonal stimulation. Ultrastructural changes noted in both dehydrated and vasopressin treated animals included: the frequent occurrence of choroidal dark cells, dilatation of the lateral and basal intercellular spaces, increased vacuolization of the apical cytoplasm, and a change in microvillar conformation from the normal clavate type to those with a filiform shape. On the basis of the ultrastructural changes observed it is proposed that the choroid plexus be regarded as a target tissue for vasopressin. These findings indicated that a vasopressinmediated transchoroidal cerebrospinal fluid absorption capability exists.Supported by U.S.P.H.S. Grant HD-08867This work submitted as partial requirement for the Master of Science degree in the Department of Anatomy, Colorado State University     

The multiple forms of brain acetylcholinesterase

Summary Micro-polyacrylamide gradient electrophoresis followed by active staining is applied for the demonstration of the multiple forms of acetylcholinesterase. Among other advantages the very small samples that enable the analysis of well-defined brain material as well as the almost histochemical conditions of incubation enable its successful use in topochemical investigations of the multiple form pattern of brain acetylcholinesterase.The acetylcholinesterase of bovine nc. caudatus could be separated into 4 multiple forms and the pattern was analysed microdensitometrically. These forms differ in their molecular weight as well as well as in their degree of membrane binding. Increasing ionic strength (NaCl) is followed by changes in the pattern. This result is discussed as caused by aggregation of enzyme subunits.The research reported in this paper was supported by the Ministerium für Wissenschaft und Technik der DDR     

Polysaccharide component in the stigmatic exudate from Lilium longiflorum

The polysaccharide component of the stigmatic exudate from Lilium longiflorum has the composition, arabinose (26%), rhamnose (6%), galactose (57%) and glucuronic acid (11%). The highly branched polysaccharide bears a striking resemblance to the acidic polysaccharide exudate from Araucaria bidwillii in belonging to the galactan group and in carrying outer chains terminated by arabinofuranose, rhamnopyranose, galactopyranose and glucuronic acid residues. Both polysaccharides contain the sequence O-rhamnopyranosyl-(1→4)-glucopyranosyluronic acid-(1→6)-galactopyranose in some of the outer chains.     

The influence of gamma radiation on arachidonic acid release and prostacyclin synthesis

Cultured pulmonary artery endothelial cells produce PGI₂ as their primary prostaglandin. Conditions which inhibit cell division have been shown to accelerate the synthesis of this compound. Exposure of endothelial cells to γ raidation results in an irreversible cessation of growth and enhanced production of PGI₂. The level of PGI₂ measured after radiation exposure exceeds that observed in cultures rendered quiescent by serum reduction. This indicates a role for γ radiation in the elevation of PGI₂ levels which is distinct from its effect on cell division. Result presented indicate that exposure to γ radiation does not, in and of itself, elevate PG levels but capacitates cells for enhanced production when presented with appropriate stimuli. Increased PGI₂ synthesis appears to be a result of an observed increase in arachidonic acid release and an activation of cyclooxygenase.     

The Cytoskeleton and Nuclear Disassembly during Germinal Vesicle Breakdown in Starfish Oocytes

In response to maturation-inducing hormone, prophase-arrested oocytes of the starfish Pisaster ochraceus resume meiosis and undergo nuclear disassembly during a process referred to as germinal vesicle breakdown (GVBD). Time-lapse video recordings of maturing oocytes reveal that the nucleus lengthens along the animal-vegetal axis of the oocyte directly prior to GVBD. Neither taxol (10 μM) nor microtubule-depolymerizing agents [colcemid (50 μM), colchicine (250 μM), or nocodazole (1 μM)] prevent the pre-GVBD changes in nuclear shape from occurring, although correlative microscopical studies demonstrate that microtubules are nucleated (taxol) or depolymerized (colcemid, colchicine, nocodazole) at the concentrations listed above. The microtubule-altering drugs also do not affect the time at which GVBD begins or ends. A 10 μM solution of the microfilament-disrupting drug cytochalasin B (CB), on the other hand, essentially eliminates the pre-GVBD elongation of the nucleus. CB also slightly delays the onset of GVBD and significantly lengthens the time required to complete GVBD. Such studies suggest that: (i) drug-sensitive microtubules are not required for GVBD to proceed in a normal fasion; (ii) the pre-GVBD changes in nuclear shape involve microfilament-mediated events; and (iii) cytochalasin-induced depolymerization of microfilaments retards the normal timing of GVBD.     

Evolutionary relationships among the permease proteins of the bacterial phosphoenolpyruvate: Sugar phosphotransferase system. Construction of phylogenetic trees and possible relatedness to proteins of eukaryotic mitochondria

Summary The amino acid sequences of 15 sugar permeases of the bacterial phosphoenolpyruvatedependent phosphotransferase system (PTS) were divided into four homologous segments, and these segments were analyzed to give phylogenetic trees. The permease segments fell into four clusters: the lactose-cellobiose cluster, the fructose-mannitol cluster, the glucose-N-acetylglucosamine cluster, and the sucrose--glucoside cluster. Sequences of the glucitol and mannose permeases (clusters 5 and 6, respectively) were too dissimilar to establish homology with the other permeases, but short regions of statistically significant sequence similarities were noted. The functional and structural relationships of these permease segments are discussed.Some of the homologous PTS permeases were found to exhibit sufficient sequence similarity to subunits 4 and 5 of the eukaryotic mitochondrial NADH dehydrogenase complex to suggest homology. Moreover, subunits 4 and 5 of this complex appeared to be homologous to each other, suggesting that these PTS and mitochondrial proteins comprise a superfamily. The integral membrane subunits of the evolutionarily divergent mannose PTS permease, the P and M subunits, exhibited limited sequence similarity to subunit 6 of the mitochondrial F₁F₀-ATPase and subunit 5b of cytochrome oxidase, respectively. These results suggest that PTS sugar permeases and mitochondrial proton-translocating proteins may be related, although the possibility of convergent evolution cannot be ruled out.     

Evidence forCis- andTrans-acting element coevolution of the 2-μm circle genome inSaccharomyces cerevisiae

Summary We compared the DNA sequence of the yeas 2-μm plasmidcis-actingSTB andtrans-actingREP1 partition loci of laboratory haploid and industrial amphiploid strains. Several industrial strains had a uniqueSTB sequence (type 1) sharing only 70% homology with laboratorySTB (type 2). Type 1 plasmids had a REP1 protein with 6–10% amino acid substitutions when compared to REP1 of type 2 plasmids. All 2-μm variants that shared a similarSTB consensus sequence exhibited a high degree ofREP1 nucleotide and amino acid sequence conservation. These observations suggest molecular coevolution oftrans-acting elements with cognate target DNA structure. Based on DNA sequencing and Southern hybridization analyses, we classified 2-μm variants into two main evolutionary lineages that differ atSTB as well asREP1 loci. The role of molecular coevolution in yeast intra- and interspecies plasmid evolution was discussed.     

Sexual differences in cricket calling song recognition

Summary Phonotactic behavior was studied in male crickets,Teleogryllus oceanicus. Tethered flying males were presented with electronically synthesized calling song models in a two-choice phonotaxis assay, and their song preferences were determined and compared with previous findings for females.Males are poorer at discriminating between songs than females; they do not display choice behavior as frequently as females, and the choices they do make are not as consistent as those of females (Figs. 3, 4). T. oceanicus calling song is composed of rhythmically different chirp and trill sections. The selectivity of males for these two components differs from that of females. Females prefer chirp to trill, but the opposite is true for males (Fig. 5B-F). Males are similar to females in that they prefer either a conspecific song model or its separate components to a heterospecific model (Fig. 5A, G, H).Behavioral and neural implications of these findings are discussed.     

Chemotaxis ofAeromonas hydrophila to the surface mucus of fish

Isolates ofAeromonas hydrophila from various sources show different chemotactic responses to mucus from the surface of freshwater fish. Some isolates were nonchemotactic to fish surface mucus. Isolates ofA. hydrophila from fish lesions had a significantly higher chemotactic index than isolates ofA. hydrophila from water. Maximum chemotactic responses occurred more often to diluted fish mucus than to undiluted samples. Fish which were experimentally stressed did not produce mucus that was more or less chemotactic than that of unstressed fish. Fish with red-sore lesions produced surface mucus which was not chemotactic toA. hydrophila. Differences between fish, for any isolate, were also not significant. The chemotactic substance(s) in fish mucus has a molecular weight of approximately 100,000 and did not appear to be labile when heated to 56°C.