Characterization of cross-bridge elasticity and kinetics of cross-bridge cycling during force development in single smooth muscle cells

Force development in smooth muscle, as in skeletal muscle, is believed to reflect recruitment of force-generating myosin cross-bridges. However, little is known about the events underlying cross-bridge recruitment as the muscle cell approaches peak isometric force and then enters a period of tension maintenance. In the present studies on single smooth muscle cells isolated from the toad (Bufo marinus) stomach muscularis, active muscle stiffness, calculated from the force response to small sinusoidal length changes (0.5% cell length, 250 Hz), was utilized to estimate the relative number of attached cross-bridges. By comparing stiffness during initial force development to stiffness during force redevelopment immediately after a quick release imposed at peak force, we propose that the instantaneous active stiffness of the cell reflects both a linearly elastic cross-bridge element having 1.5 times the compliance of the cross-bridge in frog skeletal muscle and a series elastic component having an exponential length-force relationship. At the onset of force development, the ratio of stiffness to force was 2.5 times greater than at peak isometric force. These data suggest that, upon activation, cross-bridges attach in at least two states (i.e., low-force-producing and high-force-producing) and redistribute to a steady state distribution at peak isometric force. The possibility that the cross-bridge cycling rate was modulated with time was also investigated by analyzing the time course of tension recovery to small, rapid step length changes (0.5% cell length in 2.5 ms) imposed during initial force development, at peak force, and after 15 s of tension maintenance. The rate of tension recovery slowed continuously throughout force development following activation and slowed further as force was maintained. Our results suggest that the kinetics of force production in smooth muscle may involve a redistribution of cross-bridge populations between two attached states and that the average cycling rate of these cross-bridges becomes slower with time during contraction.     

Auxin action: the search for the receptor

Abstract. The molecular specificity of the substances which have auxin activity implies the existence of specific receptors. There have been many efforts to identify and isolate these receptors on the assumption that they should bind auxins with affinities coordinate to their activities in bioassays. However, the known complexity of auxin uptake and metabolism make this assumption seriously deficient. Although several such binding sites have, in fact, been identified, proof of a connection between these sites and auxin action has been lacking. Definite proof would include a requirement that the site be reconstituted, together with the appropriate macro-molecular machinery, to construct a model of an auxin response. At the moment, our ignorance of the biochemistry and molecular biology of auxin growth responses makes such a proof difficult. However, two avenues of research promise to accelerate the rate of progress. The increasingly potent tools of molecular biology should soon allow the dissection of auxin-regulated gene expression, while improved knowledge of plasma membrane proton pumps and the mechanism of cell wall biosynthesis should produce, in parallel, an understanding of the auxin regulation of acid growth.     

FLAGELLAR MOTILITY IS NOT INVOLVED IN THE INCORPORATION OF THE SPERM INTO THE EGG AT FERTILIZATION*

Cinemicrography of sea urchin fertilization reveals that the fertilizing sperm is one of the first sperm to attach to the egg. Just before the cortical reaction the fertilizing sperm ceases motility and then is incorporated into the egg without flagellar beating. The rate of incorporation is 5–11 μm/sec and is constant. Lytechinus pictus sperm rendered immotile by azide treatment can bind to and fertilize eggs but binding, and therefore fertilization, is blocked by azide treatment of Strongylocentrotus purpuratus gametes.     

Carbohydrate Composition of Variant-Specific Surface Antigen Glycoproteins from Trypanosoma brucei

SYNOPSIS. The carbohydrate of variant-specific surface antigen glycoproteins from bloodstream forms of 13 cloned variants of Trypanosoma brucei was analyzed by gas-liquid chromatography. The glycoproteins contained from 6 to 17% carbohydrate by weight, and all contained the same 4 sugars: mannose, galactose, glucose, and glucosamine (probably as N-acetyl-glucosamine). The glycoprotein from variant 048, strain 427 contained (±20%) 11 mannose, 4 galactose, 4 glucose, and 5 glucosamine residues/mole of glycoprotein (molecular weight 65,000). Glucose was an integral component of the glycoproteins, not dissociable by sodium dodecyl sulphate, 8 M urea, or 1 M acetic acid. Some of the glucose was dissociated by trichloroacetic acid. Most of the glycoproteins formed precipitin bands with concanavalin A in Ouchterlony double diffusion, but none formed such bands with wheat germ agglutinin or Ricinus communis lectin (molecular weight 120,000).     

Secondary production,longevity and resource consumption rates of freshwater shrimps in two tropical streams with contrasting geomorphology and food web structure

1. Freshwater shrimps often dominate the biomass of tropical island streams and are known to have strong effects on stream ecosystem structure and function, but little effort has been dedicated toward quantifying basic energetic and life history attributes such as growth, production and longevity. Such information is critical for understanding both the role of shrimps in ecosystem dynamics and the gravity of threats to shrimp populations posed by human activities such as shrimp harvesting, dam construction and water withdrawal. 2. We quantified growth rates and secondary production of dominant freshwater shrimps for 3 years in two Puerto Rican headwater streams that differ in food web structure because of the presence or absence of predatory fishes that are excluded from reaches above waterfalls. Using growth data, we constructed a minimum longevity model to explore the likely minimum life spans of the two dominant taxa (Atya spp. and Xiphocaris elongata). Finally, we used a bioenergetics model to quantify annual consumption rates of major basal resources by the two taxa. 3. Daily growth rates ranged from ?0.001 to 0.011 day^?1, were inversely related to body size, and were higher for small individuals of X. elongata than Atya spp. Mean annual shrimp biomass and secondary production were an order of magnitude higher in the stream that lacked predatory fishes (biomass: 4.34 g AFDM m^?2; production: 0.89 g AFDM m^?2 year^?1) than in the stream with predatory fishes (biomass: 0.12 g AFDM m^?2; production: 0.02 g AFDM m^?2 year^?1). Production : biomass ratios ranged from 0.01 to 0.38. 4. Our longevity model predicted a minimum life span of 8 years for Atya spp. and 5 years for X. elongata in the stream lacking predatory fishes. In contrast, due to a larger average size of X. elongata in the stream with predatory fishes, our model predicted a minimum life span of 11 years. Actual life spans of these taxa are likely to be much longer based on long‐term observations of marked individuals. 5. Estimated consumption rates from the bioenergetics model indicated that Atya spp. and X. elongata are important processors of organic matter resources in streams where they occur at high densities. Atya spp. and X. elongata appeared capable of consuming a large proportion of algal and insect production and the proportion of direct leaf litter inputs consumed was also appreciable (c. 40–60%). However, the consumption of suspended fine particulate organic matter (SFPOM) by Atya spp. is probably only a minor proportion of total SFPOM flux in these streams. 6. Our study suggests that geomorphic features such as waterfalls may play an important role in controlling the distribution and production of freshwater shrimps through their effects on predatory fish movement. Spatial differences in shrimp densities result in landscape‐scale variation in the significance to ecosystem processes of these long‐lived organisms, particularly as processors of major organic matter resources.     

Characterization of six microsatellite loci in the Bermuda skink (Eumeces longirostris)

Conservation strategies for the critically endangered Bermuda skink will require knowledge of genetic variation and population structure. We describe the isolation of six polymorphic microsatellites for this species suitable for use in population genetics studies or for monitoring success of potential breeding programmes. Tissue sampling for DNA extraction involved no blood loss or tail autonomy. Three of these loci cross‐amplify and are polymorphic in a related North American skink (Eumeces fasciatus) and should also be informative for this species.     

Heat-stress effects on reproduction and seed set in Linum usitatissimum L. (flax)

Heat stress can detrimentally affect the reproductive capacity of many plants. The effect of a 7 or 14 d heat stress on flowering, seed set, pollen viability and germinability of flax (Linum usistatissimum L.) was assessed under growth chamber conditions. An incremental (2 °C/h), cyclical (daytime high 40 °C and night‐time low 18 °C) heat stress was applied 12 d after the initiation of flowering. Although flower formation in flax was not affected by heat stress, boll formation and seed set were reduced with onset of the heat stress. On removal of heat stress the stressed plants showed a compensatory response, flowering and producing bolls at a greater rate than the control plants. Heat stress significantly prolonged flowering by 17 d. Boll weight and seed weight were reduced with heat stress and the number of malformed, sterile seed increased three‐fold after 14 d of heat stress. Pollen viability and appearance were negatively affected after 6 and 10 d of heat stress, respectively. Pollen germinability decreased by the sixth day of heat stress, with no pollen germinating by the tenth day. Effects of heat stress on pollen viability and germinability alone, which did not occur until after the sixth day of the stress, could not account for the decreased boll formation due to heat stress in flax. These observations suggest that a combined effect of heat stress on both pollen and ovules contributes to decreased boll formation and seed set in flax.