A functional variant in the 5′‐flanking region of the chicken serotonin transporter gene is associated with increased body weight and locomotor activity

In this study, we identified a polymorphism in the 5′‐flanking region of the chicken serotonin transporter (5‐HTT) gene. Sequencing analysis revealed that in comparison with the wild‐type variant (W), a deleted variant (D) is generated by deletion of four nucleotides (5′‐AATT‐3′) and a single nucleotide change (A→T). Using a polyacrylamide gel electrophoresis system, we found that the 360‐bp DNA fragment containing the W variant with the wild‐type sequence 5′‐AATTAATT‐3′ shows intrinsic DNA curvature while the 356‐bp fragment containing the D variant lacking the four base pairs AATT is not curved. Quantitative real‐time RT‐PCR and ELISA demonstrated that the expression of 5‐HTT in D/D chickens was higher than that in W/W and W/D chickens. In addition, transient transfection experiments with chloramphenicol acetyltransferase reporter gene constructs revealed increased 5‐HTT promoter activity mediated by the D variant and a silencer activity of the W variant. Interestingly, females and males with D/D genotype showed significant greater increase in body weight from 6 weeks and 16 weeks of age, respectively, and higher body mass index. Moreover, we found that D/D chickens of both genders were physically more active than W/W and W/D chickens.

     

Systematic Status of Bonefishes (Albula spp.) From the Eastern Pacific Ocean Inferred from Analyses of Allozymes and Mitochondrial DNA

Recent molecular evidence suggests that at least eight species of bonefishes (Albuliformes: Albulidae: Albula) are found worldwide (Colborn et al. 2001). Adults of most of these species, including two that are restricted to the eastern Pacific Ocean (Albula sp. A from the Gulf of California and Albula sp. C from the Gulf of Panama), have not been formally described. Because cryptic species of bonefishes are known to occur sympatrically, population genetics data provide an important tool for taxonomic studies on this group. Isozyme analyses conducted on larval, juvenile and adult bonefish from the Guaymas region of the central Gulf of California confirmed that only a single species (Albula sp. A) was present there. In addition, analysis of a segment of the cytochrome b gene from three adult Albula collected from coastal waters of southern California revealed high sequence homology with Albula sp. A, suggesting that the southern California specimens were conspecific with those from Guaymas. However, the southern distributional limit of Albula sp. A, and whether there are regions where Albula sp. A and C occur sympatrically, are unknown. The historical background of the available name Atopichthys esunculus Garman, 1899 [= Albula esuncula (Garman, 1899)], and problems associated with the nomenclature of the two species of eastern Pacific bonefishes, are summarized. The systematic status of the related shafted bonefish, Dixonina (= Albula) nemoptera Fowler, 1911, is also reviewed.     

Characterization of established,cell lines derived from mutagen-sensitiveDrosophila strains

Summary Six established cell lines have been generated from embryos ofDrosophila melanogaster homozygous for different X-linked mutations. Four of these mutants, confer hypersensitivity to chemical mutagens in larvae. The cell lines derived from the two mutageninsensitive stocks, serve as controls in the analyses of DNA metabolism. One cell line (UCD-Dm-mei-9-2) is uniquely identified by a strong hypersensitivity to ultraviolet radiation. Another (UCD-Dm-mus104-1) expresses an enzyme variant not found in the other lines. The population doubling time for these cultures varies between 24 and 47 h. Labeling indices of 24.4 to 37.5% were found. The duration of the S phase in one of the control cell lines is estimated to be about 9 h. Karyotype stability was monitored for five lines over a period of about 1 y. In general these cultures each, became hypotetraploid with a preferential loss of the Y and fourth chromosomes. DNA synthesis in two of the lines fails to exhibit the pattern of sensitivity to mutagens or caffeine that is observed in the corresponding primary cultures. In primary cultures three classes of cells can be identified by autoradiography. About 50% of the cells label at a moderate rate, 20% do not label within the first 1.5 d of culture, and the remaining cells exhibit a burst of labeling shortly after the cultures are initiated. This research was supported by NIH Grants GM16298 and GM22221 and by DOE Contract AT(04-3)-34 PA 210.     

Preparation of Na,K-ATPase-containing liposomes with predictable transport properties by a procedure relating the Na,K-transport capacity to the ATPase activity

A microprocedure for the preparation of Na,K-ATPase-containing liposomes with a minimal starting material (200 microgram) of purified Na,K-ATPase is presented. Phosphatidylcholine is added gradually to cholate-solubilized Na,K-ATPase of various concentrations and the lipid-induced decrease in enzyme activity is monitored. After removal of the detergent by dialysis, the transport parameters of the resulting Na,K-ATPase-liposomes are established by a microassay. By relating the transport properties to the Na,K-ATPase activity preset before dialysis, a procedure is developed which allows to prepare standardized Na,K-ATPase-liposomes with predictable transport properties.     

Cytologische und physiologische Untersuchungen an Eremothecium ashbyii Guill.

Ohne ZusammenfassungTeilergebnisse aus einer Inauguraldissertation der philosophisch-naturwissenschaftlichen Fakultät der Universität Bern zur Erlangung der Doktorwürde 1961.     

Interaction of hypothalamic Na,K-ATPase inhibitor with isolated human peripheral blood mononuclear cells

A ligand for the digitalis receptor located on the membrane-embedded Na,K-ATPase (NKA; EC 3.6.1.37) has been isolated from bovine hypothalamus (hypothalamic inhibitory factor; HIF) and identified as isomeric ouabain (Tymiaket al, 1993,Proc. Natl. Acad. Sci. 90: 8189–8193). In analogy to cardioactive steroids (CS) derived from plants or from toad, HIF inhibits the Na/K-exchange process and the ATPase activity of isolated Na,K-ATPase although by a different molecular action mechanism. In the present work we show that, as plant-derived ouabain, HIF inhibits⁸⁶Rb-uptake by isolated human lymphocytes with an IC₅₀ of about 20 nM; above this concentration HIF reduces cell viability in contrast to ouabain. The decrease in cell viability by excess HIF is accompanied by discrete morphological alterations (mitochondrial swelling) visible by transmission electron microscopy of ultra-thin sectioned peripheral blood mononuclear cells. Taken together the results show that the hypothalamic NKA inhibitor blocks NKA of isolated human lymphocytes with high potency at nanomolar concentrations without toxicity; concentrations exceeding the ones required to block⁸⁶Rb-uptake reduce cell viability, probably due to leak formation across the NKA molecule. Thus, lymphocytes constitute a potential target for HIF action and by their altered NKA status a possible messenger between the nervous and the immune system.Abbreviations D-PBS Dulbecco's phosphate buffered saline - HBSS Hank's balanced salt Solution - NKA Na,K-ATPase     

Immobilization of yeast ADH by adsorption onto polyaminomethylstyrene

Summary Yeast alcohol dehydrogenase (EC 1.1.1.1) (Y-ADH) was immobilized by adsorption onto polyaminomethylstyrene (PAMS). The adsorption was rapid and yielded a product, the operational and storage stability of which depended on the initial content of enzyme and to a great extent on the composition of the adsorption buffer. The best results were obtained in the presence of 0.05 M phosphate solution, pH 8.8, containing 2 M sodium chloride. The amount of adsorbed ADH correlates with the position of anions in the Hofmeister series. After 1,000 bed volumes of the standard assay mixture have flowed through the column, there is no decrease in activity. It is possible to prepare adsorbates using crude yeast extracts as enzyme source. The stability of the immobilized enzyme is similar to that obtained with crystalline ADH preparations.