Complete complementary DNA of rat tyrosine aminotransferase messenger RNA. Deduction of the primary structure of the enzyme

The primary structure of rat tyrosine aminotransferase (L-tyrosine:2-oxoglutarate aminotransferase; EC 2.6.1.5), a liver-specific enzyme involved in gluconeogenesis, has been deduced from the nucleotide sequence of a cloned full-length cDNA. The mRNA is 2362 nucleotides long (excluding the poly(A) tail) and codes for a polypeptide of 454 amino acids with a molecular weight of 50634. Unambiguous identification was obtained by comparison of this sequence with the amino acid sequences of several peptides obtained from the purified enzyme.     

Single-channel analysis of a potassium inward rectifier in myocytes of newborn rat heart

Summary Unitary K⁺ currents in single cells isolated from ventricular muscle of newborn rat hearts were measured in response to different potentials and [K] _o . TheI/V curves were linear for potentials more negative than the zero-current voltage: especially in high [K] _o (150nm KCl), no clear outward currents could be detected indicating a drastic rectification in the inward direction. The channel is mainly selective to K⁺ but Na⁺ ions are also carried (P _Na/P _K=0.056). The channel conductance is proportional to the square root of [K] _o but Na⁺ ions seem to have a facilitatory effect on _K, the single-channel conductance. The channel activity, measured asP _o, i.e. the probability to find the channel in open state, decreased as the membrane was hyperpolarized. This behavior was tentatively explained by an inactivation process as the membrane becomes more negative. The rate constants of the transitions between the different states were calculated according to a C–O–C model. A control of the gating process by permeant ion K⁺ was postulated, based on the increase of one of the rate constants from the closed to the open state with [K] _o . Finally, the macroscopicI/V curves calculated fromP _o and i, the unit current, were found to be characteristic of a ion-blocked inward rectifier.     

Opposite effects of two ligands for peripheral type benzodiazepine binding sites, PK 11195 and RO5-4864, in a conflict situation in the rat

The effects of two drugs acting at the peripheral type benzodiazepine binding sites, PK 11195 and RO5-4864, were examined in shock-induced suppression of drinking in rats. These two compounds have opposite effects : RO5-4864 (3.1-1205 mg/kg i.p.) enhanced whereas PK 11195 (25-50 mg/kg i.p.) decreased the punished responding, and PK 11195 (6.25 mg/kg, a dose which did not alter the punished responding) blocked the proconflict action of RO5-4864 (6.25 and 12.5 mg/kg). The effects of RO5-4864 and PK 11195 were not antagonized by RO15-1788, a selective antagonist of the central benzodiazepine site. In addition, PK 11195 (6.25 mg/kg) did not reverse the proconflict effect of two beta-carbolines : beta-CEE and FG 7142. AS picrotoxin did not change the punished responding, these data imply that the effects of RO5-4864 and PK 11195 on the one hand and those of chlordiazepoxide and beta-carbolines on the other hand are differentially mediated and suggest that the peripheral type benzodiazepine binding sites are involved in this conflict model.     

Effect of endurance training and chronic isoproterenol treatment on skeletal muscle sensitivity to norepinephrine

The purpose of this study was to determine the influence of metabolic stresses, such as endurance training and chronic isoproterenol treatment, on skeletal muscle sensitivity to norepinephrine. Using an isolated perfused rat hindlimb preparation, dose-response curves for skeletal muscle oxygen consumption (VO2) and vascular resistance were obtained with control, endurance trained, and isoproterenol treated rats. No significant difference was found between control and experimental groups for non-stimulated VO2. In response to NE infusion, trained rats showed a significantly greater increase in VO2 compared to control rats while the response of the isoproterenol treated rats was of the same magnitude as the one for their respective control rats. At the highest dose of NE infused, the vasopressor response was significantly lower in trained rats compared to control rats. At none of the doses was there a significant difference in the vasopressor response between control and isoproterenol treated rats. These results suggest that repeated exposures to high levels of catecholamines, as produced during endurance training, leads to an increased sensitivity of skeletal muscle to the effect of norepinephrine.     

Barley straw decomposition with varied levels of microbial grazing by Folsomia fimetaria (L.) (Collembola,Isotomidae)

Summary Folsomia fimetaria (L.) were added (0, 5, 10, 20 animals) to 0.100 g barley straw which had been inoculated 10 days (244 h) earlier with a natural soil microflora. Respiration (CO₂ evolution) was monitored continuously. Mass loss, fungal standing crop (total and FDA-active), bacterial and protozoan biomass were estimated 42 days (1,000 h) after microbial inoculation. The degree of surface cover by hyphae was surveyed at regular intervals. No significant differences (P>0.05) were found in respiration, mass loss or microbial biomass, but the density of surface hyphae were reduced by addition of Collembola. Fungal production was low, less than 5% of the estimated microbial production, and could not account for all collembolan growth during incubation. F. fimetaria appeared to consume mainly bacteria and protozoa, and had little impact on carbon mineralization.     

Relationships between EUF-N fractions,N uptake and quality of sugar beet in deep loess soils of Southern Lower Saxony

Summary In field experiments conducted in 1981 on deep loess soils a significant correlation was obtained between EUF-N content in the topsoil and N in sugar beet roots as well as in whole plants. A very close correlation was found to exist between EUF-N values in June 1981 and -amino N in sugar beet roots at harvest. Also other quality criteria such as the contents of sugar, K and Na in roots and yield parameters correlated with EUF-N contents in topsoils.The inorganic and organic EUF-N fractions changed during the vegetation period due to both N uptake by the crop and mineralization. Therefore it appears to be expedient to use both fractions when calculating N fertilizer requirements.The correlation between EUF-N (topsoil samples drawn in the year preceding sugar beet) and sugar yield increases obtained by N fertilization that was found in Austria¹² was confirmed in our experiments with soils from Southern Lower Saxony.     

Performance ofin vitro propagatedAlnus glutinosa (L.) Gaertn. clones inoculated withFrankiae

Summary ThreeAlnus glutinosa (L.) Gaertn. clones, obtained byin vitro propagation techniques, were inoculated with four strains ofFrankia. The ability of these clones to nodulate and fix nitrogen was previously reported; this study deals with the performance of 12 different combinations of pairs of symbionts.Shoot fresh weight, shoot height and collar diameter were measured 60 and 82 days after inoculation. Shoot fresh weight seems to be more sensitive and reliable than the other parameters. Nitrogenase activity, measured by the acetylene reduction assay, was assayed 78 days after inoculation and was consistent with the biomass measurements.Better growth was observed when type N strains were used. Significant growth differences were observed between clones AG-2 and AG-8 on the one hand and clone AG-4 on the other. Thus, the use of genetically defined host plants and microsymbionts permitted the demonstration of significant performance variation even among cloned plants from the same provenance (AG-4 and AG-8).The duration of the experiment influenced the results with differences becoming less significant with time. This might be caused by an external limiting factor such as the pot size, competition for light,etc. But it could also be indicative of differences in nodulation speed among the treatments.     

Control of seed coat thickness and permeability in soybean : a possible adaptation to stress

Although the seed coat, through its thickness and permeability, often regulates seed germination, very little is known about the control of its development. Using soybean (Glycine max [L.] Merrill) explants, podbearing cuttings in which defined solutions can be substituted for the roots, we have demonstrated that cytokinin and mineral nutrients moving through the xylem can control soybean seed coat development. Lack of cytokinin and minerals in the culture solution, causes a thicker, less permeable seed coat to develop. The seeds with thickened coats will imbibe water rapidly if scarified; furthermore, these scratched seeds also germinate and produce normal plants. Inasmuch as stress (e.g. drought) decreases mineral assimilation and cytokinin production by the roots, the resulting delay in germination could be an adaptive response to stress.     

Calcium-binding and its effect on circular dichroism of plant calmodulin

The Ca²⁺-binding properties of calmodulin purified from zucchini (Cucurbita pepo L.) has been determined. A value of 3.3 mol Ca²⁺ per mol of zucchini calmodulin was measured at pH 7.5 by equilibrium chromatography. The far-and near-UV circular-dichroic spectra of the Ca²⁺-and Mg²⁺-saturated as well as from the metal-free forms of zucchini calmodulin reveal that upon Ca²⁺-binding the -helix content increases. A comparison with the spectra of vertebrate calmodulin indicates that both calmodulin have a similar secondary structure, similar Ca²⁺-induced conformational changes and the same number of Ca²⁺-binding sites.Abbreviations CAPP 10-(3-aminopropyl)-2-chloro-phenothiazine - EGTA ethylene glycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid - EDTA ethylenediaminetetraacetic acid Dedicated to Prof. Dr. Karl Decker on the occasion of his 60th birthday     

Glutamate synthase in greening callus of Bouvardia ternifolia Schlecht

The distribution of the two glutamate-synthase (GOGAT) activities known to exist in higher plants (NADH dependent, EC 2.6.1.53; and ferredoxin dependent, EC 1.4.7.1) was studied in non-chlorophyllous and chlorophyllous cultured tissue as well as in young leaves of Bouvardia ternifolia. The NADH-GOGAT was present in all three tissues. Using a sucrose gradient we found it in both the soluble and the plastid fraction of non-chlorophyllous and chlorophyllous tissue, but exclusively in the chloroplast fraction of the leaves. Ferredoxin-GOGAT was found only in green tissues and was confined to the chloroplasts. Ferredoxin-GOGAT activity increased in parallel with the chlorophyll content of the callus during the greening process in Murashige-Skoog medium (nitrate and ammonium as the nitrogen sources), while NADH-GOGAT was not affected by the greening process in this medium. Furthermore, both activities were differentially affected by either nitrate or ammonium as the sole nitrogen source in the medium during this process. It is suggested that each GOGAT activity is a different entity or is differently regulated.Abbreviations GOGAT glutamate synthase - MS Murashige-Skoog (1962) medium - PMSF phenylmethylsulfonyl fluoride