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A F Bennett

Publications and source records attributed to A F Bennett.

At least 37 records · Page 2Linked to original sources

Exercise performance of reptiles.

From the vantage point of thirty years of study, we can sketch the general features of activity capacity and performance ability in reptiles. Extant reptilian groups all share low levels of maintenance metabolism and ectothermy, with their consequent advantages (Pough, 1980) and disadvantages. Among the latter is a limited capacity to expand aerobic metabolism, limited in comparison to the relatively great costs of terrestrial locomotion. Particularly at low body temperatures, reptiles outstrip their aerobic capacities with any exercise more intense than a slow walk. Anaerobic metabolism, particularly anaerobic glycolysis, can be used to fuel bursts of intense activity. As a consequence, however, physiological disruption and exhaustion are entailed. Under field conditions, many reptiles alternate long periods of quiescence or slow movement with very brief bursts of exertion. Other ectotherms with a similar pattern of metabolism have been shown thereby to extend performance beyond that supportable by either aerobic or anaerobic metabolism alone (Weinstein and Full, 1992). Even with careful alternation between these metabolic modes, reptiles remain particularly prone to exhaustion during vigorous activity, as least as judged by our mammalian frame of reference. Their capacities for burst activity and exertion have been shown, at least in some species, to be important determinants of their natural survival.

Anaerobiosis↗

Bundling of microtubules by synapsin 1. Characterization of bundling and interaction of distinct sites in synapsin 1 head and tail domains with different sites in tubulin.

Synapsin 1 is a nerve terminal phosphoprotein whose role seems to encompass the linking of small synaptic vesicles to the cytoskeleton. Synapsin 1 can join small synaptic vesicles to neuronal spectrin, microfilaments and microtubules; it can also bundle microtubules and microfilaments. In this paper, the mode of interaction between synapsin 1 and microtubules has been investigated. Bundling is shown to be highly cooperative: the apparent Hill coefficient is 3.06 +/- 0.3, and bundling is half-maximal at 0.63 +/- 0.02 microM. Bundling occurs either when whole synapsin 1 preparations (containing monomers and oligomers) or when monomeric synapsin 1 is added to microtubules. However, it is not clear that synapsin 1 remains monomeric in the presence of microtubules. Synapsin 1-microtubule mixtures contain two types of filament. One type is characterised by microtubules often with synapsin 1 bound to their surface. The other type is composed of filaments of diameter 15 +/- 5 nm. This filament type is granular and made up in part of 14-nm-diameter particles. These dimensions are consistent with their being made up of polymerised synapsin 1. It is possible that microtubules induce the polymerisation of synapsin 1. Synapsin 1 had independent tubulin binding sites in the N-terminal head domain and in the C-terminal tail domain. Whole synapsin 1 can interact with tubulin after it has been digested to remove the tubulin C terminus (des-C-terminal tubulin). The interaction of des-C-terminal tubulin with synapsin 1 appears to be via the head domain, since 125I-des-C-terminal tubulin only shows specific binding to the head domain on gel blots. By contrast intact tubulin binds to both head and tail domains. Binding to the tail domain can be inhibited by a synthetic peptide representing the microtubule-associated protein 2 (MAP2) binding site of class II beta tubulin. These results suggest a model for microtubule bundling by synapsin 1 in which independent sites in the head and tail domains of synapsin 1 cross-link microtubules by interactions with two distinct sites in tubulin.

Animals↗

Site specificity in the interactions of synapsin 1 with tubulin.

Synapsin 1 is one of a family of phosphoproteins located on small synaptic vesicles (SSV) in the presynaptic terminal, and probably plays a critical role in the process of neuronal exocytosis by providing regulated linkages between SSV and the cytoskeleton. Two forms of synapsin 1 are produced from a single gene by differential mRNA splicing: 1a, 706 amino acid residues, and 1b, 670 residues. Synapsin 1 has two structural domains, a globular N-terminal head domain and an elongated tail domain. Electron microscopy of nerve terminals in situ and reconstitution studies in vitro indicates that synapsin 1 can interact with microtubules, microfilaments and brain spectrin. In vitro, synapsin 1 can bundle microtubules. This could either occur by synapsin 1 being at least bivalent for microtubules, or by univalent synapsin 1 molecules aggregating to form complexes that are more than univalent. To resolve this question, we have taken the approach of preparing defined fragments of synapsin 1 from each structural domain and analysing them for tubulin-binding activity. Our results show that there are tubulin-binding sites in both head and tail domains. We conclude that synapsin 1 monomers should be able to cross-link microtubules.

Amino Acid Sequence↗

Selective Ca2(+)-dependent interaction of calmodulin with the head domain of synapsin 1.

The calcium-dependent regulatory protein calmodulin is a critical element in the machinery regulating exocytosis at nerve terminals. Okabe & Sobue [(1987) FEBS Lett. 213, 184-188] showed that calmodulin interacts with one of the proteins intimately connected with the neuronal exocytotic process, i.e. synapsin 1. We have investigated the site at which calmodulin interacts with synapsin 1. We find that it is possible to generate chemically cross-linked Ca2(+)-dependent complexes between synapsin 1 and calmodulin in vitro, and have used covalent cross-linking in conjunction with calmodulin affinity chromatography to identify fragments of synapsin 1 that interact with calmodulin. Ca2(+)-dependent calmodulin binding is restricted to the 'head' domain (residues 1-453 in bovine synapsin 1). Within this domain the binding site is located in a unique 11 kDa Staphylococcus aureus V8 proteinase generated fragment. This fragment does not contain the site for cyclic-AMP-dependent phosphorylation and therefore does not represent the N-terminus of the protein.

Animals↗

The evolution of activity capacity.

The capacities of animals for activity (burst speed, maximal exertion, endurance) are examined in relation to their selective importance in extant populations and the pattern of their evolution in major animal taxa. Activity capacities have been demonstrated to be both heritable and highly variable in natural populations and hence susceptible to natural selection. Some field studies have demonstrated significant positive associations between activity capacities, particularly burst speed, and survivorship; other studies have not. The potential for such selection therefore clearly exists, although it may not operate in all populations. Comparative studies of major taxa have linked endurance capacities to maximal rates of oxygen consumption; speed and exertion are correlated with capacities for anaerobic metabolism, either the catabolism of phosphagens or the production of lactic acid or octopine, depending on taxon. In vertebrates, the primitive metabolic pattern involved the use of aerobic metabolism to support moderate swimming performance, supplemented by bursts of activity fuelled through lactic acid production. Because of much greater locomotor costs, the transition of vertebrates onto land entailed a decrease in endurance, which was greatly expanded again only after the evolution of the higher rates of aerobic metabolism characteristic of the birds and mammals. These greater aerobic capacities may have been selected for thermoregulatory reasons and/or for increased activity capacity itself.

Animals↗

Rapid evolution in response to high-temperature selection.

Temperature is an important environmental factor affecting all organisms, and there is ample evidence from comparative physiology that species and even conspecific populations can adapt genetically to different temperature regimes. But the effect of these adaptations on fitness and the rapidity of their evolution is unknown, as is the extent to which they depend on pre-existing genetic variation rather than new mutations. We have begun a study of the evolutionary adaptation of Escherichia coli to different temperature regimes, taking advantage of the large population sizes and short generation times in experiments on this bacterial species. We report significant improvement in temperature-specific fitness of lines maintained at 42 degrees C for 200 generations (about one month). These changes in fitness are due to selection on de novo mutations and show that some biological systems can evolve rapidly in response to changes in environmental factors such as temperature.

Biological Evolution↗

Thermal dependence of locomotor capacity.

The thermal dependence of locomotor performance capacity, particularly speed and endurance, in vertebrate ectotherms is examined. Most studies have found an optimal speed for performance at relatively high body temperatures, close to upper lethal limits. These performance capacities decrease markedly at low body temperatures and may be compensated by increments in aggressive or evasive behaviors. Relative ranking of performance is maintained among individuals across body temperatures. Acclimation of performance capacities is generally incomplete or entirely absent: most animals compensate locomotor performance rather poorly to cold exposure. Locomotor performance in different groups has been shown to possess the attributes (e.g., variability, repeatability, heritability, and differential survivorship) necessary for evolutionary adaptation, but interpretation of comparative data is complicated by phylogenetic differences among species studied. Controlled studies show partial but incomplete adaptation to environmental temperature.

Acclimatization↗

Quantitative genetics of maximal oxygen consumption in a garter snake.

Broad-sense heritabilities and genetic correlations of maximal oxygen consumption (VO2max), blood hemoglobin levels, and ventricle mass were estimated in a natural population of snakes. Traits were measured for six or fewer presumed full-sibling offspring from each of 45 wild-caught gravid garter snakes (Thamnophis sirtalis). VO2max was highly reproducible between replicate trial days (r = 0.88). In an attempt to reduce maternal effects, correlations of each character with body mass, snout-vent length, age at testing, litter size, dam mass, and dam snout-vent length were removed by computing residuals from multiple-regression equations. These residuals were used in subsequent genetic analyses. Approximate coefficients of variation of residuals were 16% for VO2max, 19% for hemoglobin level, and 13% for ventricle mass. Broad-sense heritabilities were highly significant for all characters [P less than 0.0001; VO2max heritability (h2) = 0.88; hemoglobin level h2 = 0.63; ventricle mass h2 = 0.41], suggesting that they could respond genetically to selection. Phenotypic correlations (rP) among residual characters were significant only between VO2max and ventricle mass (rP = +0.27). VO2max and ventricle mass exhibited a significant (broad-sense) genetic correlation of +0.64; this might facilitate the correlated evolution of these two traits in response to natural or artificial selection. Ventricle mass and hemoglobin level showed a significant environmental correlation of +0.43. Treadmill endurance crawling time (Evolution 42: 335-350, 1988) showed a weak but significantly positive rP with VO2max (rP = +0.17).

Aging↗

Individual correlation of morphology, muscle mechanics, and locomotion in a salamander.

Measurements were made on locomotor performance (burst run and swim speed, run and swim endurance), morphology (body, tail, and hindlimb length, body mass), and skeletal muscle mechanics (isometric: twitch and tetanic tension, rates of force development and relaxation; isotonic: maximal velocity of shortening and power output) in a size range of individual salamanders (Ambystoma tigrinum nebulosum) at 10 and 20 degrees C. The size dependence of each factor was determined, and the interindividual correlations among factors were measured after removal of size effects. Locomotor performance is positively related to body size: larger animals are faster and have higher endurance. Isometric tensions (standardized for muscle cross-sectional area) and isotonic properties are mass independent; rates of isometric force development and relaxation are negatively related to body mass. Locomotor performance capacities are not intercorrelated among individuals. Isometric force and rates of force development are highly intercorrelated as are maximal shortening velocity and power output. Several statistical techniques failed to uncover correlations among sets of variables (performance, morphology, muscle mechanics): neither fast individuals nor individuals with high endurance necessarily have relatively fast muscles or long limbs or tails after the effects of body size have been removed.

Ambystoma↗

Evidence that the cytoplasmic aldehyde dehydrogenase-catalysed oxidation of aldehydes involves a different active-site group from that which catalyses the hydrolysis of 4-nitrophenyl acetate.

Acylation of the aldehyde dehydrogenase.NADH complex by acetic anhydride leads to the production of acetaldehyde and NAD+. By monitoring changes in nucleotide fluorescence, the rate constant for acylation of the active site of the *enzyme.NADH complex was found to be 11 +/- 3 s-1. The rate of acylation by acetic anhydride at the group that binds aldehydes on the oxidative pathway is clearly rapid enough to maintain significant steady-state concentrations of the required active-site-acylated *enzyme.NADH intermediate despite the rapid hydrolysis of this *enzyme.acyl.NADH intermediate (5-10 s-1) [Blackwell, Motion, MacGibbon, Hardman & Buckley (1987) Biochem. J. 242, 803-808]. Hence reversal of the normal oxidative pathway can occur. However, although acylation of the aldehyde dehydrogenase.NADH complex by 4-nitrophenyl acetate also occurs rapidly with a rate constant of 10.9 +/- 0.6 s-1, even under the most extreme trapping conditions only very small amounts of acetaldehyde are detected [Loomes & Kitson (1986) Biochem. J. 235, 617-619]. Furthermore enzyme-catalysed hydrolysis of 4-nitrophenyl acetate is limited by the rate of deacylation of a group on the enzyme (0.4 s-1), which is an order of magnitude less than deacylation of the group at the active site (5-10 s-1). It is concluded that the enzyme-catalysed 4-nitrophenyl ester hydrolysis involves a group on the enzyme that is different from the active-site group that binds aldehydes on the normal oxidative pathway.

Acetaldehyde↗

Thermal adaptations in lizard muscle function.

This study was undertaken to investigate thermal adaptations in muscle contractile properties in closely-related lizards with different preferred body temperatures (PBT). The species examined all belong to the Sphenomorphus group of Australian skinks (Scincidae: Lygosominae). Preferred body temperatures are conservative at the generic level as follows: Ctenotus, 35 degrees C; Sphenomorphus, 30 degrees C; Eremiascincus, 25 degrees C. Contractile properties of the fast glycolytic portion of the iliofibularis muscle were measured. Translational adaptations are evident in several isometric factors, including tetanic tension (Po), twitch tension (Pt), twitch time to peak tension (TPT), and twitch half-relaxation time (1/2 RT). Capacity adaptations are not evident in rates of tetanic tension development (dPo/dt) or in maximal velocities of isotonic shortening (Vmax). Rotational adaptations are not evident in any contractile properties. Thermal limits on upper response temperatures are about 5 degrees C warmer in Ctenotus than in the more cryophilic species, indicative of resistance adaptation in muscle performance. Despite these adaptive shifts, there is little indication that muscle functional capacities are optimized or equalized at PBT in these lizards.

Acclimatization↗

The thermal dependence of locomotor performance and muscle contractile function in the salamander Ambystoma tigrinum nebulosum.

The thermal dependence of locomotor performance and in vitro muscle mechanical properties were studied after acclimation at 10 degrees and 20 degrees C in the salamander Ambystoma tigrinum nebulosum Hallowell. Performance measurements included burst and endurance capacities on land and in water. No significant differences in locomotor performance or muscle contractile properties were found between acclimation groups. Locomotor performance had only a slight thermal dependence, with Q10 values of 0.99-1.36 for running and swimming burst capacities (i.e. maximum speed and leg/tail cycling frequency). Running and swimming endurance capacities had thermal ratios of 1.58-1.66. Thermal dependence of muscle contractile rates was higher than that of locomotor performance: rates of force development for both isometric twitch and tetanus and maximal shortening velocity had Q10 values of 1.89-2.01. Maximal power output was also thermally dependent (Q10 = 2.33) and occurred at 38% of maximal (tetanic) force output. Force-generating capacities in isometric twitch and tetanus were relatively temperature-independent.

Acclimatization↗

Thermal dependence of contractile properties of skeletal muscle from the lizard Sceloporus occidentalis with comments on methods for fitting and comparing force-velocity curves.

The isometric and isotonic contractile properties of fast-twitch glycolytic fibres of the iliofibularis muscle (FG-IF) in the lizard Sceloporus occidentalis were measured in vitro at 5 degrees C intervals form 10 to 40 degrees C. The mean isometric parameters at 35 degrees C, the preferred body temperature of this species, were as follows: maximum isometric force (Po), 187 +/- 8 (S.E.M.) kNm-2; ratio of twitch force to tetanic force (PTW/Po), 0.46 +/- 0.02; time to peak twitch tension (tPTW), 7.0 +/- 0.3 ms; and time from peak twitch force to 50% relaxation (t50% ), 8.2 +/- 0.3 ms. From 20 to 35 degrees C Po was almost constant (within 8% of the value at 35 degrees C). At 10 and 15 degrees C C. Po dropped to approximately 80% of the value at 35 degrees C. Po was very unstable at 40 degrees C. PTW/Po was almost constant at all temperatures. The time-related isometric parameters were positively modified by temperature at all temperatures measured (Q10 greater than 1.9). The force-velocity curves of the FG-IF deviated from the simple hyperbolic relation of A. V. Hill's characteristic equation. We present two alternative equations for fitting these data. These equations resulted in residual sums of squares from nonlinear least-squares analysis that were at least seven-fold lower than those from Hill's equation. The equation that best describes our data is a hyperbola modified by the addition of a linear component: V = B(1 - P/Po)/(A + P/Po) + C(1 - P/Po). To describe the curvature of this or any other force-velocity relationship, we propose the power ratio, Wmax/VmaxPo (where Wmax is the maximum power calculated from the force-velocity relationship and Vmax is the predicted maximum velocity of shortening at zero force). Vmax of the FG-IF was 21.9LoS-1 at 35 degrees C (where Lo is muscle length). This parameter was directly related to temperature between 10 and 35 degrees C with Q10 greater than 1.8. The shape of the force-velocity curve is not influenced by temperature (Wmax/VmaxPo = 0.11).

Animals↗

Thermal dependence of sprint performance of the lizard Sceloporus occidentalis.

Sprint velocity of the lizard Sceloporus occidentalis was maximal at preferred body temperature (Tb, 35 degrees C). Mean running velocity (VR) and stride frequency (f) at this temperature were 3.23 +/- 0.7 (mean +/- S.E.M.) ms-1 and 15.6 +/- 0.3 s-1, respectively. VR and f did not change significantly when Tb was raised to 40 degrees C. At Tb values between 25 and 35 degrees C the thermal dependencies of VR (Q10 = 1.23) and f (Q10 = 1.12) were quite low. At Tb values below 25 degrees C the thermal dependence of these factors increased markedly. Stride length (LS) was independent of Tb from 15 to 40 degrees C. Lizards with a Tb of 10 degrees C were largely incapacitated, and VR, f and LS were all greatly reduced. Comparisons with measurements of the contractile properties of skeletal muscle of this species suggest that stride frequency is limited by the twitch contraction time at temperatures below 23 degrees C. At higher temperatures, sprint performance is nearly independent of the thermal effects on the muscles.

Animals↗

Thermal dependence of isotonic contractile properties of skeletal muscle and sprint performance of the lizard Dipsosaurus dorsalis.

Contractile properties of the fast-twitch glycolytic (FG) portion of the iliofibularis muscle and sprint running performance were studied at approximately 5 degrees C intervals from 15-44 degrees C in the lizard Dipsosaurus dorsalis. Maximal running velocity (VR) and stride frequency (f) were both greatest when body temperature (Tb) was 40 degrees C, the field-active Tb in Dipsosaurus. At 40 degrees C VR was 4.3 +/- 0.2 m/s and f was 13.5 +/- 0.5 s-1. Between 25 and 40 degrees C, the thermal dependencies of VR and f were approximately constant (Q10's of 1.31 and 1.36 got VR and f, respectively). Below 25 degrees C performance declined more markedly with decreasing temperature. At 20 degrees C strides were qualitatively normal, but VR was only half of the value at 25 degrees C. At 15 degrees C the lizards were substantially incapacitated, and VR was 10% of the value at 20 degrees C. Stride length was approximately 0.33 m and changed very little with Tb from 20-44 degrees C. The time dependent contractile properties of FG muscle were affected more by temperature than was sprint performance. The maximal velocity of shortening at zero load (VO) was 18.7 0/s at 40 degrees C and had a Q10 of 1.7 from 25-40 degrees C. Maximal power output (Wmax) determined from the force-velocity curve was 464 W/kg at 40 degrees C. Below 40 degrees C max varied with temperature with a Q10 of 2-3. The shape of the force-velocity curve changed little with temperature (Wmax/POVO = 0.11). Between 25 and 40 degrees C a relatively temperature-independent process must modulate the effects of temperature on the contractile properties of the muscles that supply the power for burst locomotion. Storage and recovery of elastic energy appears to be a likely candidate for such a process. Below 25 degrees C, however, the contraction time is prolonged to such an extent that the f attainable is limited by the minimum time taken to contract and relax the muscles.

Animals↗