Phase-contrast versus off-axis illumination: is a more complex microscope always more powerful?
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In this article, a practical demonstration suitable for any biology college classroom is presented. With the examples of a complex biological specimen (slug's radula) and a simple reference specimen (electron microscopical grid imprint in gelatin), both of which can be easily prepared, the capabilities of two imaging modes commonly used in optical microscopy are demonstrated. The results obtained under phase contrast (a rather sophisticated method, 1953 Nobel Prize to Zernike) and off-axis illumination (a very simple method) are compared. The off-axis illumination setup is capable of delivering noticeably better microscopic images of these two particular specimens, yet it can be easily assembled in a laboratory classroom. The outcome of such a demonstration is expected to be the realization on the part of the students that one needs to carefully choose the apparatus to address a given biological problem, with the "bottom line" being that a more complex one may not necessarily yield better results. An attempt to explain this "paradox" is presented, in the particular case presented here, partly from the physiology of vision perspective (the shape-from-shading problem). The overall aim of the present article is to induce in students critical thinking about the capabilities of a laboratory equipment in general and about data interpretation. (+info)
Habitat associations in gastropod species at East Pacific Rise hydrothermal vents (9 degrees 50'N).
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At deep-sea hydrothermal vents on the East Pacific Rise (9 degrees 50'N), distinct megafaunal assemblages are positioned along strong thermal and chemical gradients. We investigated the distribution of gastropod species to determine whether they associate with specific megafaunal zones and to determine the thermal boundaries of their habitats. Gastropods colonized a series of basalt blocks that were placed into three different zones characterized by vestimentiferan tubeworms, bivalves, and suspension-feeders, respectively. Additional gastropods were collected on selected blocks from higher temperature vestimentiferan habitat and from grab samples of alvinellid polychaetes. On the blocks, gastropod species clustered into a "Cool" group (Clypeosectus delectus, Eulepetopsis vitrea, Gorgoleptis spiralis, and Lepetodrilus ovalis) whose species tended to be most abundant in the suspension-feeder zone, and a "Warm" group (Lepetodrilus cristatus, L. elevatus, L. pustulosus, and Cyathermia naticoides) whose species all were significantly more abundant in the vestimentiferan zone than elsewhere. The temperature ranges of Cool species were generally lower than the ranges of Warm ones, although both groups were present at 3 to 6 degrees C; also present was Bathymargarites symplector, which clustered with neither group. Three additional species, Rhynchopelta concentrica, Neomphalus fretterae, and Nodopelta rigneae, co-occurred with Warm-group species on selected blocks from hotter habitats. Although a few species were found only in alvinellid collections, most species were not exclusive to a specific megafaunal zone. We propose that species in the Cool and Warm groups occupy specific microhabitats that are present in more than one zone. (+info)
Duplication and selection on abalone sperm lysin in an allopatric population.
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While gene duplication is a major source of evolutionary novelty, the importance of this process in reproductive protein evolution has not been widely investigated. Here, we report the first known case of gene duplication of abalone sperm lysin in an allopatric subspecies found in the Eastern Atlantic, Haliotis tuberculata coccinea. Mass spectrometry identified both copies of the lysin protein in testis tissue, and 3-dimensional structural modeling suggests that both proteins remain functional. We also detected positive selection acting on both paralogs after duplication and found evidence of a recent selective sweep. Because H. t. coccinea occurs in geographic isolation from other abalone species, these findings suggest that the evolution of lysin is not driven to create reproductive barriers to unfit hybrid formation with an overlapping species. Instead, sexual selection or sexual conflict acting during abalone fertilization could be responsible for the recent positive selection on this protein. The presence of multiple, rapidly evolving lysin genes in H. tuberculata presents an opportunity to study the early stages of diversification of a protein whose function is well understood. (+info)
Growth in the slow lane: protein metabolism in the Antarctic limpet Nacella concinna (Strebel 1908).
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Growth rates in Antarctic ectotherms are generally considered to be low in comparison to temperate and tropical species. Food consumption plays a major role in determining animal growth rates, but once food is ingested soft tissue growth rates are largely determined by the protein synthesis retention efficiency (PSRE), a measure of the efficiency with which proteins are synthesised and retained as protein growth. The effect of water temperatures on the PSRE of polar organisms has not previously been investigated, and it is possible that reduced PSRE at polar water temperatures may at least partially explain low growth rates in Antarctic organisms. We also currently lack any information on the potential effects of predicted increases in seawater temperatures on protein metabolism in Antarctic ectotherms. We have measured seasonal protein synthesis, degradation and growth rates in free-ranging Antarctic limpets (Nacella concinna), together with protein synthesis rates at temperatures ranging between -1.5 degrees C and 6.0 degrees C. PSRE were not significantly different in summer (15.69+/-4.41%) or winter (20.59+/-4.45%), but values were considerably lower than those previously reported in temperate and tropical species. A meta-analysis of published ectotherm PSRE suggested there was a positive relationship with temperature (y=449.9-114.9x, r(2)=28.8%, P<0.05). In turn, this suggests that temperature may be an important factor in determining ectotherm growth efficiency via an influence on PSRE. Maximal fractional and absolute protein synthesis rates occurred at approximately 1 degrees C in N. concinna, the approximate summer water temperature at the study site, and protein synthesis rates decreased above this temperature. In the absence of adaptation, predicted increases in Antarctic water temperatures would result in reduced, rather than increased, rates of protein synthesis and, in turn, possibly growth. (+info)
Parameter space analysis suggests multi-site plasticity contributes to motor pattern initiation in Tritonia.
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This research examines the mechanisms that initiate rhythmic activity in the episodic central pattern generator (CPG) underlying escape swimming in the gastropod mollusk Tritonia diomedea. Activation of the network is triggered by extrinsic excitatory input but also accompanied by intrinsic neuromodulation and the recruitment of additional excitation into the circuit. To examine how these factors influence circuit activation, a detailed simulation of the unmodulated CPG network was constructed from an extensive set of physiological measurements. In this model, extrinsic input alone is insufficient to initiate rhythmic activity, confirming that additional processes are involved in circuit activation. However, incorporating known neuromodulatory and polysynaptic effects into the model still failed to enable rhythmic activity, suggesting that additional circuit features are also required. To delineate the additional activation requirements, a large-scale parameter-space analysis was conducted (~2 x 10(6) configurations). The results suggest that initiation of the swim motor pattern requires substantial reconfiguration at multiple sites within the network, especially to recruit ventral swim interneuron-B (VSI) activity and increase coupling between the dorsal swim interneurons (DSIs) and cerebral neuron 2 (C2) coupling. Within the parameter space examined, we observed a tendency for rhythmic activity to be spontaneous and self-sustaining. This suggests that initiation of episodic rhythmic activity may involve temporarily restructuring a nonrhythmic network into a persistent oscillator. In particular, the time course of neuromodulatory effects may control both activation and termination of rhythmic bursting. (+info)
Serotonin-like immunoreactivity in the central and peripheral nervous systems of the interstitial acochlidean Asperspina sp. (Opisthobranchia).
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Species of Acochlidea are common members of the marine interstitial environment and defined in part by their minuscule size and highly divergent morphology relative to other benthic opisthobranchs. Despite these differences, acochlideans such as species of Asperspina display many plesiomorphic characteristics, including an unfused condition of their neural ganglia. To gain insight into the distribution of specific neural subsets within acochlidean ganglia, a species of Asperspina was studied by using anti-serotonin immunohistochemistry and epifluorescence and confocal laser scanning microscopy. Results reveal similarities between Asperspina and larger opisthobranchs in the general distribution of serotonergic perikarya in the central nervous system. Specifically, the arrangement of perikarya into regional clusters within the cerebral and pedal ganglia and the absence of immunoreactive perikarya in the pleural ganglia are similar to the model species of Aplysia californica, Pleurobranchaea californica, and Tritonia diomedea. Moreover, serotonergic innervation of the rhinophores in all opisthobranchs, including Asperspina sp., originates from the cerebral ganglion instead of directly from the rhinophoral ganglion. Serotonergic innervation of the body wall, including the epithelium, muscles, and pedal sole, appears to arise exclusively from pedal and accessory ganglia. These observations indicate a general conservation of serotonin-like immunoreactivity in the central and peripheral nervous systems of acochlidean and other benthic opisthobranchs. (+info)
Photosynthesis drives oxygen levels in macrophyte-associated gastropod egg masses.
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Many aquatic animals deposit fertilized eggs in adherent clutches or gelatinous masses. Egg aggregation carries certain risks, including the potential for inadequate oxygen supply to embryos. Physical and biological conditions alter such risks. We examined the effects of light levels and associated photosynthetic organisms on the distribution of oxygen inside gelatinous egg masses of four temperate gastropod species. Egg masses of two species, the opisthobranchs Melanochlamys diomedea and Haminoea callidegenita, contained significant populations of diatoms but generally were not associated with macrophytes. Egg masses of the other two species, the opisthobranch Haminoea vesicula and the prosobranch Lacuna sp., occurred commonly on subtidal macrophytes and appeared not to contain significant populations of diatoms. In the laboratory, we used microelectrodes to measure oxygen levels inside masses exposed to alternating dark and light conditions; light level had an enormous influence on oxygen profiles in egg masses of all four species. Masses of H. vesicula and Lacuna sp., when experimentally separated from their macrophytes, showed only slight increases in oxygen upon light exposure, indicating that the main source of oxygen in situ was the macrophyte rather than associated microalgae. Our findings indicate that photosynthesis by macrophytes can drive large changes in internal oxygen profiles. (+info)
Influence of macrofaunal assemblages and environmental heterogeneity on microphytobenthic production in experimental systems.
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Despite the complexity of natural systems, heterogeneity caused by the fragmentation of habitats has seldom been considered when investigating ecosystem processes. Empirical approaches that have included the influence of heterogeneity tend to be biased towards terrestrial habitats; yet marine systems offer opportunities by virtue of their relative ease of manipulation, rapid response times and the well-understood effects of macrofauna on sediment processes. Here, the influence of heterogeneity on microphytobenthic production in synthetic estuarine assemblages is examined. Heterogeneity was created by enriching patches of sediment with detrital algae (Enteromorpha intestinalis) to provide a source of allochthonous organic matter. A gradient of species density for four numerically dominant intertidal macrofauna (Hediste diversicolor, Hydrobia ulvae, Corophium volutator, Macoma balthica) was constructed, and microphytobenthic biomass at the sediment surface was measured. Statistical analysis using generalized least squares regression indicated that heterogeneity within our system was a significant driving factor that interacted with macrofaunal density and species identity. Microphytobenthic biomass was highest in enriched patches, suggesting that nutrients were obtained locally from the sediment-water interface and not from the water column. Our findings demonstrate that organic enrichment can cause the development of heterogeneity which influences infaunal bioturbation and consequent nutrient generation, a driver of microphytobenthic production. (+info)