Effect of l-triiodothyronine on the mitochondrial α-glycerophosphate dehydrogenase activity,mitochondrial and total protein contents of brain of Singi fish (Heteropneustes fossilis bloch)

A single injection of various doses (0.25, 0.5, 5, 20 and 50 μg/g) of l-triiodothyronine increased the mitochondrial cytochrome-linked α-glycerophosphate dehydrogenase (EC 1.1.99.5) activity and mitochondrial protein content of brain of Singi fish on the 3rd day. l-Triiodothyronine at the dose of 0.1 μg/g did not alter the α-glycerophosphate dehydrogenase activity and mitochondrial protein content of brain. The total protein content of the brain also increased on the 3rd day with 0.5 μg of l-triiodothyronine per g. Increased enzyme activity followed a dose-response relationship of a non-linear fashion. The enhancement of the α-glycerophosphate dehydrogenase activity of fish brain with a dose of 0.5 μg/g was found from the 1st day and it reached to a maximum level from the 3rd to the 5th day. The enzyme activity then sharply declined on the 6th or 7th day. Cycloheximide inhibited the l-triiodothyronine-induced increase in the α-glycerophosphate dehydrogenase activity, mitochondrial and total protein content of fish brain.The present study thus reveals the responsiveness of fish brain to thyroid hormone, and α-glycerophosphate dehydrogenase activity can be taken as a biochemical indices for the expression of thyroid hormone action in fish brain.     

Effect of the epizoic rotifer Brachionus rubens on the population growth of three cladoceran species

Using population densities and growth rates as criteria, we studied interactions between the epizoic rotifer Brachionus rubens and each of three cladoceran species differing in size and reproductive rates — Daphnia carinata, Moina macrocopa and Ceriodaphnia rigaudi. In all mixed — species experiments, B. rubens existed in both the epizoic mode, attached to the cladoceran host, and in the free-swimming mode. Rotifer population growth rates were significantly depressed in the presence of M. macrocopa, presumably as a consequence of exploitative and interference competition. The largest cladoceran, D. carinata probably did not suppress B. rubens, because the epizoic component of the rotifer population escaped from the deleterious effects of mechanical interference. Peak population numbers and initial population growth rates reached by all three cladocerans were lower in the presence of B. rubens, probably because of the adverse effects of the epizoic infestation, which was maximal on D. carinata and least on C. rigaudi. In mixed-species cultures of D. carinata and M. macrocopa, the presence of B. rubens helped D. carinata coexist with M. macrocopa, which otherwise would have suppressed the Daphnia.     

Definition of a Sequence Unique in βII Spectrin Required for Its Axon-Specific Interaction with Fodaxin (A60)

Abstract: Spectrin isotypes segregate in neurons and are differentially distributed between axons and somatodendritic compartments. Their functions in those compartments are likely to be mediated by proteins that interact selectively with one or other isotype. Fodaxin (an axon-specific protein previously termed A60) colocalizes in CNS neurons with axonal spectrin and in vitro binds brain spectrin (a mixture of αI, βI, αII, and βII polypeptides) but not erythrocyte spectrin (αI and βI). Because αII and βII spectrin polypeptides are enriched in axons, we investigated a possible binding of fodaxin to the types of spectrin found in axons. Fodaxin did not bind to isolated brain α chains. Bacterially expressed C-terminal segments 18–19 of βII spectrin bound to fodaxin and inhibited the binding of fodaxin to whole brain spectrin. By contrast, recombinant segments 18–19 of the somatodendritic βIΣ2 spectrin showed no interaction with fodaxin. Within βII, fodaxin binding activity was localized to residues 2,087–2,198, which are unique to βII and link between the end of segment 18 and the pleckstrin homology domain in segment 19. The divergent regions of sequence in segments 19 of βII and βIΣ2 are candidates to mediate the isotype-specific functions of spectrin. Fodaxin is the first protein to be described that discriminates between the unique regions of β spectrin isoforms.     

The effect of prey morphology on the feeding behaviour and population growth of the predatory rotifer Asplanchna sieboldi: a case study using five species of Brachionus (Rotifera)

1. We investigated the numerical response, functional response and prey preference of Asplanchna sieboldi to five different prey brachionids. We also analysed the feeding behaviour of the predator in terms of encounters, attacks, capture and prey ingested per unit time. 2. The five prey species (Brachionus havanaensis, B. rubens, B. patulus, B. macracanthus and B. calyciflorus) differed in their body size and spine length. 3. The population growth rates of A. sieboldi ranged from 0.074 ± 0.03 to 0.431 ± 0.02 depending on prey type and density. There was a significant impact of the spine length rather than body size per se on the population growth rates of the predator. 4. The maximum number of prey consumed depended on both body size and spine length. In the functional response analyses, the plateau was reached at a prey density of 4–8 ind. mL^?1. 5. There was a significant impact of prey density on the prey preference of the predator.     

Observations on feed size and capture success in the larval butterfly splitfin (Ameca splendens Miller & Fitzsimons, 1971, Pisces: Goodeidae) reared on zooplankton

In this study, we quantified the feeding behaviour (encounter, attack, capture. and ingestion) of larval A. splendens on micro‐crustacean prey [cladocerans: Alona rectangula, Simocephalus vetulus (separately neonates and adults), Ceriodaphnia dubia, Daphnia pulex (juveniles), Moina macrocopa and ostracods: Heterocypris incongruens]. Although we initially (first 4 weeks) offered rotifers (Brachionus calyciflorus and B. patulus), they were not consumed by the larvae and hence observations with these prey were discontinued. Feeding behaviour was observed during the first 10 weeks. Fifteen observations were made with each prey species (seven diets × four replicates). Experiments were conducted in 50 ml transparent containers with 20 ml fish‐conditioned water into which one fry was introduced. Before introducing the fish, 20 individuals of a given cladoceran prey species or 50 individuals of a rotifer prey species were introduced. Until the fourth week, we used 20 ml of medium and thereafter 30 ml, but the prey density used remained constant (1 ind. ml⁻¹). Observations (10 min per fry per cladoceran replicate) were taken under a stereomicroscope (20×) for the first 2 weeks and later with a lamp and a magnifying lens. The number of encounters (E), attacks (A), captures (C) and ingestions (I) were recorded. During the study period, there was a 60% increase in gape size but only a 30% increase in body length. The number of encounters of larval A. splendens was highest (192) on M. macrocopa and lowest (29) on ostracods and adult S. vetulus (59). The inverse relationship between capture success and prey size was more pronounced during the latter half of the study period. Compared with all the other prey types offered, A. splendens fed maximally on M. macrocopa, which therefore could be a suitable diet for the larval rearing of this fish species.     

Differential regulation of the lateral mobility of plasma membrane phospholipids by the extracellular matrix and cholesterol

In this study, we compared qualitative and quantitative changes in the lateral mobility of phospholipid molecules in the plasma membrane of intact cells under various conditions of specific interaction of integrins in the cell membrane with two extracellular matrix (ECM) components viz. fibronectin (FN) and laminin (LN). We found a strong and specific correlation between the lower lateral mobility of phosphatidylcholine (PC) and higher lateral mobility of phosphatidylethanolamine (PE) when cells were expressing high levels of alpha5beta1 integrin and thus were adherent and motile on FN. The interaction between PC and FN in alpha5 integrin expressing cells was aided by the strong affinity of alpha5 integrin to the FN matrix. Cholesterol was involved in regulating the lateral mobility of PC to a great extent and of PE to a lesser extent without affecting the overall microviscosity of the plasma membrane or the distribution of caveolin-marked domains. The distribution and mobility of PC and PE molecules in the lamellipodial regions differed from that in the rest of the membrane and also in the more motile and in the less motile cells. We propose that these differences in distribution of PC and PE in different regions of cell membrane and their respective lateral mobility are observed due to the specific interaction of PC molecules with FN molecules in the ECM. Our results outline a new role of integrin-matrix interactions in the regulation of membrane phospholipid behavior.     

Recovery Patterns of Moina Macrocopa Exposed Previously to Different Concentrations of Cadmium and Methyl Parathion: Life-Table Demography and Population Growth Studies

In most toxicity studies using Cladocera, bioassays are routinely done to determine median lethal concentration (LC₅₀) or the responses to sublethal exposure. However, information on the patterns of recovery of cladocerans exposed to different concentrations of toxicants is scarce. This is important because cladocerans exposed to toxicants for a short duration may later recuperate under favourable conditions. Using the life table demographic and population growth, the present study was conducted to evaluate the recovery patterns of Monia macrocopa exposed to five different concentrations (0, 25, 50, 75 and 100% of 24 h LC₅₀ for CdCl₂ or methyl parathion) and then returned to toxicant-free medium containing alga (Chlorella vulgaris) at low (0.25 × 10⁶cells ml^–1), medium (0.5 × 10⁶cells ml^–1) or high (1 × 10⁶cells ml^–1) levels. We measured selected life history variables such as average lifespan, life expectancy at birth, gross and net reproductive rates, generation time and the rate of population increase. Results indicated that regardless of food concentration, surviving individuals of M. macrocopa exposed to a median lethal concentration did not recover. The effect of food level was significant at 25 and 50% of the median lethal concentration for cadmium or methyl parathion. Age-specific fecundity curves showed that exposure to either toxicant for a duration as short as 24 h at one-fourth of the LC₅₀ showed reduced output of offspring, especially at a lower food level. At and above exposures of 0.037 mgl^–1 of methyl parathion, no reproduction occurred. The highest gross and net reproductive rates (127 and 55 offspring female^–1) were obtained in controls at the high (1 × 10⁶ cells ml^–1) algal food level. The rate of population increase obtained from life table data was around 0.7 per day in controls but decreased when exposed to toxicant concentrations. The rates of population increase per day derived from population growth data varied from 0.22 to 0.33 per day for the controls, depending on the food levels.     

Ab initio studies on the tri- and diphosphate fragments of adenosine triphosphate

Pyrophosphate prototypes such as methyl triphosphate and methyl diphosphate molecules in their different protonation states have been investigated at high levels of quantum chemical calculations. The optimized geometries, the thermochemistry of the hydrolysis and the molecular orbitals contributing to the high energy of these compounds have been analyzed. These investigations provide insights into the "high energy" character of ATP molecule. Further, the dependence of vibrational frequencies on the number of phosphate groups and the charged states has also been presented. These results can aid the interpretation of spectra obtained by experiments on complexes containing pyrophosphate prototypes.     

Nebulin interacts with CapZ and regulates thin filament architecture within the Z-disc

The barbed ends of actin filaments in striated muscle are anchored within the Z-disc and capped by CapZ; this protein blocks actin polymerization and depolymerization in vitro. The mature lengths of the thin filaments are likely specified by the giant "molecular ruler" nebulin, which spans the length of the thin filament. Here, we report that CapZ specifically interacts with the C terminus of nebulin (modules 160-164) in blot overlay, solid-phase binding, tryptophan fluorescence, and SPOTs membrane assays. Binding of nebulin modules 160-164 to CapZ does not affect the ability of CapZ to cap actin filaments in vitro, consistent with our observation that neither of the two C-terminal actin binding regions of CapZ is necessary for its interaction with nebulin. Knockdown of nebulin in chick skeletal myotubes using small interfering RNA results in a reduction of assembled CapZ, and, strikingly, a loss of the uniform alignment of the barbed ends of the actin filaments. These data suggest that nebulin restricts the position of thin filament barbed ends to the Z-disc via a direct interaction with CapZ. We propose a novel molecular model of Z-disc architecture in which nebulin interacts with CapZ from a thin filament of an adjacent sarcomere, thus providing a structural link between sarcomeres.