Fine‐scale selection by ovipositing females increases egg survival

One of the most important defenses for the eggs of ovipositing female organisms is to avoid being laid in the same habitat as their predators. However, for most organisms, completely avoiding an offspring's predators is not possible. One mechanism that has been largely overlooked is for females to partition an oviposition site into microhabitats that differ in quality for offspring survival. We conducted a series of experiments to examine whether female newts avoid microhabitats utilized by their offspring's primary predator, caddisfly larvae. Female newts avoided laying eggs near predatory caddisflies and shifted egg laying upward in the water column when provided with a vertical dimension. Caddisflies were attracted to chemical stimuli from female newts and their eggs, yet primarily used benthic areas in experimental chambers. Finally, results from a field experiment indicate that the behavioral strategy employed by female newts increases offspring survival. This subset of non‐genetic maternal effects, micro‐oviposition avoidance, is likely an important yet underexplored mechanism by which females increase offspring survival.     

Die Sterilit?t der Bastarde im Lichte des Mendelismus

Ohne Zusammenfassung     

Fluoroplastic Coverslips for Long-Term Nerve Tissue Culture

Fluoroplastics (Aclar and Teflon FEP) were tested for their suitability as coverslip material for nerve tissue cultures. Coverslips prepared from these plastics have a higher transparency over the near-ultraviolet, visible, and infrared regions of the spectrum than glass coverslips, possess exceptional physical and chemical stability, and provide service-ability over a wide temperature range ≥ -320 to 390 F. For nerve tissue culture, circular 22 mm coverslips were cut from 5 mil sheets of Aclar 22A and 33C, and Teflon FEP 100A, cleaned in concentrated HNO₃, rinsed, sterilized in 85% ethyl alcohol, dried, and coated with collagen. Peripheral and central nervous system cultures from fetal rats and newborn mice were set up on these as well as on glass coverslips, and carried in Maximow double coverslip assemblies. Over a 2 mo period of cultivation, neuronal maturation, tissue organization and fiber myelination occurred on plastic coverslips in essentially the same manner as in sister cultures maintained on glass coverslips. Cultures fixed and embedded in Epon for electron microscopy directly on Aclar or Teflon 100A coverslips were easily separated from the coverslip. Some difficulty was encountered with retraction of the collagen coat from Teflon 100A coverslips, however, and Aclar 22A coverslips were warped by some of the reagents used in embedding for electron microscopy. Aclar 33C was not adversely affected by these reagents, and provided the most suitable base for collagenization and culture maintenance. A variety of cells grew satisfactorily on either the bare or the collagenized surface of this plastic, and separated easily from it following embedment in Epon.     

Rapid reversal of a potentially constraining genetic covariance between leaf and flower traits in Silene latifolia

Genetic covariance between two traits generates correlated responses to selection, and may either enhance or constrain adaptation. Silene latifolia exhibits potentially constraining genetic covariance between specific leaf area (SLA) and flower number in males. Flower number is likely to increase via fecundity selection but the correlated increase in SLA increases mortality, and SLA is under selection to decrease in dry habitats. We selected on trait combinations in two selection lines for four generations to test whether genetic covariance could be reduced without significantly altering trait means. In one selection line, the genetic covariance changed sign and eigenstructure changed significantly, while in the other selection line eigenstructure remained similar to the control line. Changes in genetic variance–covariance structure are therefore possible without the introduction of new alleles, and the responses we observed suggest that founder effects and changes in frequency of alleles of major effect may be acting to produce the changes.     

Male competition reverses female preference for male chemical cues

Females must choose among potential mates with different phenotypes in a variety of social contexts. Many male traits are inherent and unchanging, but others are labile to social context. Competition, for example, can cause physiological changes that reflect recent wins and losses that fluctuate throughout time. We may expect females to respond differently to males depending on the outcome of their most recent fight. In Bolitotherus cornutus (forked fungus beetles), males compete for access to females, but copulation requires female cooperation. In this study, we use behavioral trials to determine whether females use chemical cues to differentiate between males and whether the outcome of recent male competition alters female preference. We measured female association time with chemical cues of two size‐matched males both before and after male–male competition. Females in our study preferred to associate with future losers before males interacted, but changed their preference for realized winners following male competitive interactions. Our study provides the first evidence of change in female preference based solely on the outcome of male–male competition.     

Ligand Binding to A1 Adenosine Receptors is Influenced by Protonation

Abstract

Studies on the pH dependence of ligand binding to A₁ adenosine receptors revealed that protonation of a histidine residue in the binding pocket is accompanied by high affinity agonist binding.     

Oncogenes and inflammation rewire host energy metabolism in the tumor microenvironment

Here, we developed a model system to evaluate the metabolic effects of oncogene(s) on the host microenvironment. A matched set of “normal” and oncogenically transformed epithelial cell lines were co-cultured with human fibroblasts, to determine the “bystander” effects of oncogenes on stromal cells. ROS production and glucose uptake were measured by FACS analysis. In addition, expression of a panel of metabolic protein biomarkers (Caveolin-1, MCT1, and MCT4) was analyzed in parallel. Interestingly, oncogene activation in cancer cells was sufficient to induce the metabolic reprogramming of cancer-associated fibroblasts toward glycolysis, via oxidative stress. Evidence for “metabolic symbiosis” between oxidative cancer cells and glycolytic fibroblasts was provided by MCT1/4 immunostaining. As such, oncogenes drive the establishment of a stromal-epithelial “lactate-shuttle”, to fuel the anabolic growth of cancer cells. Similar results were obtained with two divergent oncogenes (RAS and NFκB), indicating that ROS production and inflammation metabolically converge on the tumor stroma, driving glycolysis and upregulation of MCT4. These findings make stromal MCT4 an attractive target for new drug discovery, as MCT4 is a shared endpoint for the metabolic effects of many oncogenic stimuli. Thus, diverse oncogenes stimulate a common metabolic response in the tumor stroma. Conversely, we also show that fibroblasts protect cancer cells against oncogenic stress and senescence by reducing ROS production in tumor cells. Ras-transformed cells were also able to metabolically reprogram normal adjacent epithelia, indicating that cancer cells can use either fibroblasts or epithelial cells as “partners” for metabolic symbiosis. The antioxidant N-acetyl-cysteine (NAC) selectively halted mitochondrial biogenesis in Ras-transformed cells, but not in normal epithelia. NAC also blocked stromal induction of MCT4, indicating that NAC effectively functions as an “MCT4 inhibitor”. Taken together, our data provide new strategies for achieving more effective anticancer therapy. We conclude that oncogenes enable cancer cells to behave as selfish “metabolic parasites”, like foreign organisms (bacteria, fungi, viruses). Thus, we should consider treating cancer like an infectious disease, with new classes of metabolically targeted “antibiotics” to selectively starve cancer cells. Our results provide new support for the “seed and soil” hypothesis, which was first proposed in 1889 by the English surgeon, Stephen Paget.     

Force Measurement During Contraction to Assess Muscle Function in Zebrafish Larvae

Zebrafish larvae provide models of muscle development, muscle disease and muscle-related chemical toxicity, but related studies often lack functional measures of muscle health. In this video article, we demonstrate a method to measure force generation during contraction of zebrafish larval trunk muscle. Force measurements are accomplished by placing an anesthetized larva into a chamber filled with a salt solution. The anterior end of the larva is tied to a force transducer and the posterior end of the larva is tied to a length controller. An isometric twitch contraction is elicited by electric field stimulation and the force response is recorded for analysis. Force generation during contraction provides a measure of overall muscle health and specifically provides a measure of muscle function. Although we describe this technique for use with wild-type larvae, this method can be used with genetically modified larvae or with larvae treated with drugs or toxicants, to characterize muscle disease models and evaluate treatments, or to study muscle development, injury, or chemical toxicity.