Axonal mapping of neurosecretory Helix bursting cells. Functional aspects of peripheral multibranched axons

• 1.1. The peripheral axon distribution from two bursting neurons was investigated using electrophysiological methods. Both of these cells send out a bundle of axon branches which goes via the visceral nerve to the heart and in the uterine nerve.
• 2.2. The relative number of axon branches in the two nerves was determined through double-shock experiments.
• 3.3. The axon bundle takes the visceral nerve and its uterine branch, supplying the effector systems in parallel.
• 4.4. Slight differences in conduction velocity between the axon branches produce an enlargement of the efferent volley.
• 5.5. The number of active axon branches conveying orthodromic or antidromic spikes is controlled by inhibitory potentials converging onto the initial part of the bundle, so that the two bursting cells amount to a pool of 35 to 40 interconnected nerve cells.
• 6.6. The atypical axonal distribution of the two bursting cells might be related to their neurosecretory properties.

     

The influence of aerial exposure on gas exchange and cardiac activity in the shore crab,Carcinus maenas (L.)

• 1.1. The cardiovascular physiology of adult Carcinus maenas (L.) emerging into air has been investigated at three different air temperatures.
• 2.2. Transition from seawater to air or vice versa triggered transient increases in cardiac and locomotor activity.
• 3.3. However, crabs became inactive 5–10 min after emerging from seawater (15°C) into air at the same temperature (15°C) or at lower temperatures (12–13°C) and heart rate fell.
• 4.4. At higher air temperatures (18–20°C) heart rate rose but to a lesser extent than predicted from aquatic Q₁₀ heart-rate values.
• 5.5. Crabs were again quiescent in aerial conditions.
• 6.6. Mean arterial oxygen tension (Pa_o2) was ~ 74 mmHg in submerged crabs but fell to ~ 38 mmHg in air while mean arterial carbon dioxide tension (Pa_o2) increased from 1 to 4 mmHg resulting in respiratory acidosis.
• 7.7. A model of gill function is proposed to explain the development of internal hypoxia in air.
• 8.8. The results are discussed in relation to the distribution of adult and juvenile C. maenas in situ.

     

Osmotically-induced volume changes in isolated cells of a pond snail

• 1.1. To assess whether the stretch-activated (SA) channels in snail cells could contribute to osmoregulation, information is needed about the behaviour of the cells under anisosmotic conditions.
• 2.2. Cells of Lymnaea stagnalis were therefore examined during acute hyposmotic stress (HOS).
• 3.3. Kidney, heart and neuronal cells (monitored photographically) swelled less than expected for strictly semipermeable cells, but exhibited no regulatory volume decrease.
• 4.4. Long-term viability of the cells was not compromised following acute hyposmotic stress.
• 5.5. Quinidine, which blocks SA channels in Lymnaea, intensified stress-induced swelling most markedly in kidney cells.
• 6.6. The data can, however, be explained without invoking recruitment of SA channels.

     

Identification of neurons involved in the earthworm Amynthas hawayanus reflex activity

• 1.1. The medial (MGF), lateral (LGF) and motor (RMS-2) giant neurons were confirmed as neural components in the earthworm Amynthas hawayanus polysynaptic reflex circuit by simultaneous potential recording and dye injection.
• 2.2. The reflex was initiated from the mechanoreceptors when evoked by mechanical stimulation but electrical stimulation also evoked an antidromic response in the motoneuron.
• 3.3. The primary reflex response propagates decrementally along both giant axons but directly evoked action potentials conduct in an all-or-none fashion.
• 4.4. The secondary reflex response continues to propagate after the primary response disappears.
• 5.5. A rhythmically discharging neuron of uncertain function was also identified.

     

Hydro- and thermodynamic properties of bovine heart AMP-deaminase

• 1.1. Sedimentation velocity and sedimentation equilibrium studies of bovine heart AMP-deaminase were performed. Molecular weights of the native enzyme and subunit were determined as 161,000 and 43,000 dallons respectively.
• 2.2. The kinetic data indicate that in the presence of 100 mM KCl the enzyme may be active as a dimer.
• 3.3. The influence of temperature on the enzyme kinetics was investigated, from which activation energy (E_a and the heat of enzyme-substrate complex formation (ΔH_s) were calculated.
• 4.4. It is suggested that an equilibrium may exist between a dimeric and tetrameric form of AMP-deaminase in the heart.

     

Kinetic and regulatory properties of phenylalanine ammonia-lyase from Citrus sinensis

• 1.1. The kinetic and regulatory properties of phenylalanine ammonia-lyase from Citrus sinensis fruit tissue were investigated. The substrate specificity of the enzyme was determined as well as the effects of pH and temperature on the catalytic activity.
• 2.2. The enzyme exhibits negative homotropic effects between the substrate binding centra.
• 3.3. Binding of l-phenylalanine to the enzyme is characterized by two K_m-values; K_m^L = 13 μM and K_m^H = 52 μM; with a Hill-interaction coefficient of 0.75.
• 4.4. The enzyme is subject to product inhibition by trans-cinnamate, but the effects of allosteric effectors and inhibitors seem to be of much greater importance in the short-term regulation of phenylpropanoid metabolism in Citrus sinensis.
• 5.5. The enzyme activity was found to be modulated by end-products of diverging metabolic pathways, viz. umbelliferone, scopoletin, naringenin, quercetin, kaempferol, benzoic acid and gallic acid.

     

The effect of γ-radiation on the NADP-malic enzyme from mango fruit,Mangifera indica — II. Kinetic and regulatory properties

• 1.1. Malic enzyme purified from the fruit tissue of Mangifera indica was irradiated in aqueous solution and the effect of γ-irradiation on the catalytic and regulatory properties of the enzyme was investigated.
• 2.2. Significant differences in some of the allosteric properties of the enzyme were found as reflected in the various Hill-coefficients.
• 3.3. Changes in both the kinetic parameters V_max and K_m were observed; suggesting that irradiation leads not only to destruction of the active sites but also to a general denaturation of the enzyme.
• 4.4. The physiological significance of the radiation induced alterations are discussed against the background of ripening and sensescence.

     

The regulation of glucose 6-phosphate dehydrogenase from Dicentrarchus labrax (bass) liver

• 1.1. Kinetic constant values of the reaction catalyzed by bass liver glucose 6-phosphate dehydrogenase show to be modified between 10 and 40°C.
• 2.2. The Arrhenius plot between 10 and 50°C shows two slopes with different activation energies.
• 3.3. These results suggest a regulation of this enzyme by environmental temperature.
• 4.4. Kinetics of ATP inhibition were examined between pH 6.2 and 7.8: patterns and K_i values obtained are affected by the pH variation.
• 5.5. NADH is an effective inhibitor of bass glucose 6-phosphate dehydrogenase but this enzyme does not show NAD-linked activity.
• 6.6. Kinetics of pyridoxal 5′-phosphate inhibition have indicated the presence of a lysine in the catalytic site for NADP⁺.

     

Temperature acclimation in respiratory and cytochrome c oxidase activity in common carp (Cyprinus carpio)

• 1.1. Common carp (Cyprinus carpio) exposed to experimental temperatures of 12, 18, 24, 30 or 36°C for a 4-week period were used to investigate the effect of temperature acclimation on the frequency of opercular movement (FOM), growth and cytochrome c oxidase (CCO) activity in heart, liver and muscle.
• 2.2. An exponential relationship between FOM and temperature after the first week (10₁₀ =1.76) disappeared after the second week.
• 3.3. The initially high FOM at temperatures of 30 or 36°C and the low FOM at 18 or 12°C changed over 4 weeks to approach the FOM of fish at 24°C.
• 4.4. This change in the relationship of FOM to temperature from highly dependent to independent appeared to be thermal compensation.
• 5.5. Heart and liver CCO activities were significantly affected by temperature, with the lowest activity at the approximate optimum temperature for growth, 24°C.
• 6.6. Highest CCO activities for heart and liver occurred at both the highest and lowest temperatures.
• 7.7. Among the three tissues, heart CCO activity was generally the highest and most affected by acclimation temperature.
• 8.8. Muscle tissue had the lowest CCO activity and was unaffected by temperature.
• 9.9. The high CCO activity at a cold acclimation of temperature 12°C was probably due to thermal compensation and the high activity at 36°C may have been a result of thermal stress.

     

Na+/H+ exchange in the kidneys of eels (Anguilla anguilla) adapted to sea water or to freshwater environments: Studies with brush border membrane vesicles

• 1.1. In brush border membrane vesicles isolated from eel kidneys, adapted either to sea water or freshwater environments, a Na⁺/H⁺ antiporter is present.
• 2.2. Using a calibration plot it is possible to evaluate the amount of protons that this antiporter can accumulate inside the vesicular space.
• 3.3. The activity of the antiporter seems to be affected by the salinity of the water; it is higher in animals adapted to seawater.
• 4.4. This adaptation seems to occur by a J_max regulation of the antiporter.

     

Detection of 180 kDa proteins in electroplax sodium channel preparations

• 1.1. Co-isolating proteins (M_r 170,000–220,000) from sodium channel preparations made from the electric organ of the electric eel (Electrophorus electricus) were detected on Western blots using monoclonal a antibodies.
• 2.2. Similar protein patterns were seen on immunoblots containing immunoprecipitated protein from eel muscle and brain tissues but not heart.
• 3.3. These co-isolating proteins could be separated from the mature TTX-sensitive channel protein (M_r 280,000) using a lentil lectin-Sepharose column.
• 4.4. The 180 kDa proteins do not appear to be channel-related and can be detected as contaminants in electroplax sodium channel preparations using the monoclonal antibodies described here.

     

Heart rate and its cholinergic control in the sole (Solea vulgaris), acclimatized to different temperatures

• 1.1. The effects of thermal acclimatization at 10 and 24°C on heart rate were investigated on unrestrained soles (Solea vulgaris).
• 2.2. The sensitivity of heart rate to temperature changes induced by temperature acclimatization was higher in cold-acclimatized than in warm-acclimatized soles.
• 3.3. Heart rate of cold-acclimatized fish to temperature changes was not affected by blocking the vagal tone with atropine.
• 4.4. After atropine treatment the ability of heart rate to show thermal compensation decreased in warm-acclimatized soles.
• 5.5. It is suggested that the vagus nerve can function differently at different temperatures.

     

NADPH/NADP ratio could regulate the glyoxylate cycle in Tetrahymena pyriformis

• 1.1. The glyoxylic acid cycle pathway could be regulated through the modulation of the isocitrate dehydrogenase-NADP activity. This enzyme is inhibited by NADPH.
• 2.2. The effect on the glyoxylate cycle flux of variations in the rate of the NADPH-consuming pathways has been studied.
• 3.3. Increase in the rate of NADPH-consuming activity by addition of H₂O₂ produces inhibition of the glyoxylate cycle and decrease in the NADPH/NADP ratio.
• 4.4. These results suggest that the glyoxylate flux in Tetrahymena could be modulated by regulation of NADP-dependent isocitrate dehydrogenase by the NADPH/NADP ratio.

     

Characteristics of Crassostrea virginica crystalline style chitin digestion

• 1.1. Fundamental chitin digestion characteristics of Crassostrea virginica crystalline style were investigated.
• 2.2. Optimum temperature and pH were 34°C and 4.8. respectively.
• 3.3. The colloidal regenerated chitin (0.56mol/0.5 ml: GlcNAc equivalents) was saturating under all enzyme levels encountered.
• 4.4. There was no evidence of end product inhibition, even after 100 hr incubation.
• 5.5. Calculated K_m for the chitinase complex was 1.19mM when determined using a 30 min assay, but was only 0.70 mM when determined using a 4.6 hr assay.
• 6.6. Both K_m values are lower than reported for similar assays in other molluscs and for most bacteria.
• 7.7. Effect of substrate preparation on the kinetics are discussed.
• 8.8. Eight peaks of chitinase activity were resolved by DEAE-Fractogel ion exchange chromatography.

     

Purification and properties of hydroxypyruvate reductase from Lemna minor L

• 1.1. Hydroxypyruvate reductase has been purified 193-fold from Lemna minor L. by affinity chromatography on Blue Sepharose.
• 2.2. The enzyme has activity over a broad pH range (optimum pH 6), a K_m hydroxypyruvate of 59 μ M and K_m NADH of 12μM.
• 3.3. Crude extracts of Lemna exhibit substrate inhibition of activity above 1 mM hydroxypyruvate, a property which is lost on purification.
• 4.4. Oxaloacetate inhibits purified preparations of the enzyme and a possible role for such regulation in vivo is discussed.

     

Two kinds of peripheral afferent neurons in the earthworm reflex arc

• 1.1. Two kinds of neurons were identified in the body-wall longitudinal muscle layer of the earthworm, Amynthas hawayanus, by the simultaneous potential recording and Lucifer Yellow-CH injection method with a single microelectrode.
• 2.2. Both kinds of neurons have their somata, neuntes and longitudinal processes imbedded in the longitudinal muscle layer. Those with two circular processes extending into the third segmental nerve trunk are tentatively named “intra-nerve-trunk” neurons and those with four circular processes extending into four setae shafts are tentatively named “intramural” neurons.
• 3.3. Both kinds of neurons responded to electrical and mechanical stimuli applied in an afferent direction to them.
• 4.4. The “intra-nerve-trunk” neuron decreased its response amplitudes to these stimuli after the third nerve trunk was sectioned in correlation to the response amplitude decrease recorded from the nerve trunk after it was sectioned.
• 5.5. The response amplitude decrease due to denervation implies a nonlinear structure of the earthworm reflex circuits.
• 6.6. The “intramural” neurons are believed to be primary sensory neurons connected to the mechanoreceptors in the setae.

     

Respiration in the eastern water dragon,Physignathus lesueurii (agamidae)

• 1.1. Some aspects of the gas exchange system of a diving lizard, Physignathus lesuewii were studied.
• 2.2. Breathing patterns were analysed.
• 3.3. Breathing rate increases logarithmically with temperature and Q₁₀ = 1.8. LogBR = −0.237 + 0.0256 T.
• 4.4. Gas tensions in lung air and arterial and venous blood were measured. Arterial pH declines with increasing temperature.
• 5.5. Temperature has a marked effect on oxygen affinity of the blood (ΔH = −10.1 kcal mol⁻). A Bohr effect was also noted.
• 6.6. CO₂ equilibrium curves were drawn.
• 7.7. The results are considered with a view to anticipating the efficiency of the gas exchange system of this species under conditions of variable temperature and during diving.

     

Effects of water stress on hemolymph volume,osmotic potential and chemical composition in Megetra cancellata

• 1.1. Rates of water loss in Megetra cancellata were very high compared to those reported for other xeric arthropods.
• 2.2. Hemolymph weight in hydrated animals was 43.0% of the total body weight while it was 24.7% in desiccated animals that had lost 16.1% of their body weight as water.
• 3.3. Hemolymph osmotic potential increased from 417 to 447 mOsm/kg in desiccated beetles, but osmotic regulation was evident.
• 4.4. Total hemolymph protein mass and concentration decreased in desiccated beetles while amino acid concentrations remained constant (at about 70 mM).
• 5.5. Na⁺ and −PO₄ concentrations increased in desiccated beetles.
• 6.6. Cl⁻ and K⁺ concentrations in desiccated beetles were equal to those in undesiccated beetles.