Physiology

Finding novel compounds that affect neuronal or muscular function is of great interest, as they can serve as potential pharmacological agents for a variety of neurological disorders. For instance, conopeptides have been developed into powerful drugs like the painkiller PrialtTM. Most conopeptides, however, have yet to be characterized, revealing the need for a rapid and straightforward screening method. We have designed a novel bioassay, which allows for unbiased screening of biological activity of compounds in vivo against numerous molecular targets on a wide variety of neurons and muscles in a rapid and straightforward manner. For this, we paired nanoinjection of compounds with electrophysiological recordings from the Giant Fiber System of Drosophila melanogaster, which mediates the escape response of the fly.

Elasmobranch fishes use electroreception to detect electric fields in the environment, particularly minute bioelectric fields produced by potential prey. A single elasmobranch family (Potamotrygonidae) is composed of obligate freshwater stingrays endemic to the Amazon River. A freshwater existence has imposed morphological adaptions on their electrosensory system due to life in a high impedance medium. Because their electrosensory morphology differs from their marine relatives, freshwater stingrays may demonstrate corresponding differences in behavioral sensitivity. The objective of this study was to quantify behavioral sensitivity of the obligate freshwater stingray Potamotrygon motoro to prey-simulating voltage. The voltage produced by common teleost prey of P. motoro were measured and replicated for behavioral trials. The best response was 10.62 cm, and the smallest voltage gradient detected was 0.005 mVcm-1. This sensitivity is reduced compared to marine species. The conductivity of the medium, more so than ampullary morphology, may dictate sensitivity of the elasmobranch electrosensory system.

The electrosensory and visual adaptations of elasmobranchs to the environment have been more studied than most other senses, however, work on these senses is mostly limited to descriptive analyses of sensitivity, morphology, and behavior. The goal of this work was to explore electrosensory and visual capabilities in a more ecological context. To gain an understanding of the content of bioelectric signals, the magnitude and frequency of these stimuli were recorded from a broad survey of elasmobranch prey items... Color vision adaptations also correlated to the photic environment of each species; cownose rays inhabit turbid, green-dominated waters and had two cone visual pigments that maximize contrast of objects against the green background... Yellow stingrays were trichromatic and likely possess the ability to discriminate colors in their clear, reef and seagrass habitats, which are spectrally rich. Both species showed evidence of ultraviolet sensitivity, which may aid in predator and conspecific detection as an enhanced communication channel. Future studies should investigate the integration of sensory input and sensory involvement in intraspecific communication to gain more insight into ecological adaptations.

Experimentally naive rats show variance in their locomotor reactivity to novelty, some displaying higher (HR) while others displaying lower (LR) reactivity, associated with vulnerability to stress. LRHR phenotype is proposed as an antecedent to the development of stress hyper responsiveness. Results presented here show emergence of antidepressive-like behavior following peripubertal-juvenile exposure to chronic variable physical (CVP) and chronic variable social stress (CVS) in HR rats, and depressive-like behavior following CVP in the LRs. The antidepressive-like behavior in HR rats was accompanied by increased levels of acetylated Histone3 (acH3) and acetylated Histone4 (acH4) at the hippocampal brain-derived neurotrophic factor (BDNF) P2 and P4 promoters respectively. This effect may mediate increased mossy fibre (MF) terminal field size, particularly the suprapyramidal mossy fibre projection volume (SP-MF), in the HR animals following both stress regimens. These findings show that chronic variable stress during adolescence induces individual differences in molecular, neuromorphological and behavioral parameters between LRs and HRs, which provides further evidence that individual differences in stress responsiveness is an important factor in resistance or vulnerability to stress-induced depression and/or anxiety.

Vitamin D insufficiency/deficiency is widespread in asthma, and epidemiological studies point to an association between low serum 25-hydroxyvitamin D level and poor asthma control and increased severity. In humans. Vitamin D is principally derived from sunlight induced cutaneous conversion of 7-dehydrocholesterol to vitamin D and oral supplementation. We sought to determine if established and chronic-persistent adult asthma patients from a South-Florida pulmonary patient population, with abundant sunshine availability and oral vitamin D supplementation exhibit vitamin D insufficiency/deficiency. A trend to vitamin D insufficiency was observed in approximately 65% of both adult asthma patients and apparently healthy (non-asthmatic) volunteers. . The transcription factors required for Th9 conversion, PU.1 and IRF-4, were down-regulated by vitamin D. The generation of Th9 cells was inhibited equally by vitamin D and dexamethasone when used alone, but the effect was additive when both steroids were used in combination. Our studies using non-specifically stimulated cells were extended by analyzing the effect of vitamin D on allergen specific stimulation. The response of CD4+ T cells obtained from the blood of house dust mite positive asthmatics was studied. House dust mite allergen elicited a classical Th2 phenotype response (IL-4, IL-5, IL-9, and IL-13 cytokine profile) and vitamin D effectively inhibited those key Th2 cytokines. We conclude that vitamin D appears to be of significant clinical benefit in our cohort of patients, i.e., established chronic adult human asthma, by down-regulating key immune cells including Th9, Th17, and Th2 involved in this disorder.

Nitrogen (N) has traditionally been regarded as the primary limiting nutrient to algal growth in marine coastal waters, but recent studies suggest that phosphorus (P) can be limiting in carbonate-rich environments. To better understand the importance of P. alkaline phosphatase activity (APA) was measured in reef macroalgae in seven counties of south Florida ; several significant trends emerged : 1) APA decreased geographically from the highest values in Dada>Monroe>Palm Beach>St. Lucie>Broward>Martin>Lee counties 2) APA varied temporally with increasing nutrient-rich runoff in the wet season 3) APA varied due to taxonomic division Phaeophyta>Rhodophyta>Chlorophyta 4) Nutrient enrichment experiments demonstrated that increased N-enrichment enhanced P-limitation while increased P decreased P-limitation. These results suggest that high APA observed in carbonate-rich waters of Dade County and low APA in Broward County resulted from high nutrient inputs associated with anthropogenic nutrient pollution.

The olfactory system is the most highly developed system for molecular sensing in vertebrates. Despite their reputation for being particularly olfactory driven, little is known about how this sense functions in elasmobranch fishes. The goal of this dissertation was to examine the morphology and physiology of elasmobranchs to compare their olfactory system with teleost fishes and more derived vertebrates. To test the hypotheses that elasmobranchs possess greater olfactory sensitivities than teleosts and that lamellar surface area is correlated to sensitivity, I compared the surface area of the olfactory lamellae and the olfactory sensitivities of five phylogenetically diverse elasmobranch species. The olfactory thresholds reported here (10-9 to 10-6 M) were comparable to those previously reported for teleosts and did not correlate with lamellar surface area. Since aquatic species are subject to similar environmental amino acid levels, they appear to have converged upon similar amino acid sensitivities. To test the hypothesis that elasmobranchs are able to detect bile salt odorants despite lacking ciliated olfactory receptor neurons (ORNs), the type of ORN that mediates bile salt detection in the teleosts, I quantified the olfactory specificity and sensitivity of two elasmobranch species to four, teleost-produced C24 bile salts. Both species responded to all four bile salts, but demonstrated smaller relative responses and less sensitivity compared to teleosts and agnathans. This may indicate that elasmobranchs don't rely on bile salts to detect teleost prey. Also, the olfactory system of elasmobranchs contains molecular olfactory receptors for bile salts independent of those that detect amino acids, similar to teleosts.

Although most batoids (skates and rays) are benthic, only the skates (Rajidae) have been described as performing benthic locomotion, termed 'punting'. While keeping the rest of the body motionless, the skate's specialized pelvic fins are planted into the substrate and then retracted caudally, which thrusts the body forward. This may be advantageous for locating and feeding on prey, avoiding predators, and reducing energetic costs. By integrating kinematic, musculoskeletal, material properties, and compositional analyses across a range of morphologically and phylogenetically diverse batoids, this dissertation (i) demonstrates that punting is not confined to the skates, and (ii) provides reliable anatomical and mechanical predictors of punting ability. Batoids in this study performed true punting (employing only pelvic fins), or augmented punting (employing pectoral and pelvic fins). Despite the additional thrust from the pectoral fins, augmented punters failed to exceed the punting c apabilities of the true punters. True punters' pelvic fins had greater surface area and more specialized and robust musculature compared to the augmented punters' fins. The flexural stiffness of the main skeletal element used in punting, the propterygium, correlated with punting ability (3.37 x 10-5 - 1.80 x 10-4 Nm2). Variation was due to differences in mineral content (24.4-48-9% dry mass), and thus, material stiffness (140-2533 MPa), and second moment of area. The propterygium's radius-to-thickness ratio (mean = 5.52 +-0.441 SE) indicated that the propterygium would support true and augmented punters, but not non-punters, in an aquatic environment. All propterygia would fail on land. Geometric and linear morphometric analyses of 61 batoid pelvic girdles demonstrated that pelvic girdle shape can predict punting and swimming ability and taxonomic attribution to Order.

Taurine is the second most abundant amino acid in the CNS after glutamate and its functions have been found largely related to intracellular calcium ([Ca2+]i) modulation, osmoregulation, membrane stabilization, reproduction and immunity. The action of taurine has also been implicated in neurotransmission and neuromodulation though its specific sites of action are not fully understood. Isolated retinal neurons from the larval tiger salamanders (Ambystoma tigrinum) were used as a model to study the neuromodulatory role of taurine in the CNS and to gain insights into its potential sites of action. A combination of techniques was used, including whole-cell patch clamp recording to study taurine's regulation of voltage-gated potassium (K+) and Ca2+ channels and Fluo-4AM Ca2+-imaging to study taurine's regulation of glutamate-induced [Ca2+] I,. Taurine was shown to suppress of glutamate-induced [Ca2+] l, in a dose dependent manner. This suppression was mostly sensitive to the glycine rece ptor antagonist Strychnine but insensitive to any GABA receptor antagonist. The remaining strychnine-insensitive effect was inhibited with the protein kinase A (PKA) inhibitor, PKI, suggesting that there was an additional metabotropic pathway. Moreover, using the protein kinase C (PKC) inhibitor, GF109203X, there was an enhancement in strychnine-insensitive taurine's regulation. Taurine inhibits voltage-gated Ca2+ channels in the retinal neurons and has a dual effect on voltage-gated K+ channels. Taurine causes an increase in K+ current amplitude which is further enhanced with PKI and blocked with GF109203X, suggesting that it is through a PKC-dependent pathway negatively controlled by PKA-dependent pathway.

The enzyme Methionine sulfoxide reductase A (MsrA) repairs oxidized proteins, and may act as a scavenger of reactive oxygen species (ROS), making it a potential therapeutic target for age-related neurodegenerative diseases. The anoxia-tolerant turtle offers a unique model to observe the effects of oxidative stress on a system that maintains neuronal function following anoxia and reoxygenation, and that ages without senescence. MsrA is present in both the mitochondria and cytosol, with protein levels increasing respectively 3- and 4-fold over 4 hours of anoxia, and remaining 2-fold higher than basal upon reoxygenation. MsrA was knocked down in neuronally-enriched cell cultures via RNAi transfection. Propidium iodide staining showed no significant cell death during anoxia, but this increased 7-fold upon reoxygenation, suggesting a role for MsrA in ROS suppression during reperfusion. This is the first report in any system of MsrA transcript and protein levels being regulated by oxygen levels.