Neuroscience
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The senses of hearing and balance in vertebrates are transduced by hair cells in the inner ear. Hair cells from a wide variety of organisms have been described electrophysiologically but this is the first report of the application of these techniques to the genetically tractable zebrafish model system. Auditory and vestibular hair cells isolated from zebrafish lagenae and utricles were patch clamped and both inward and outward currents under voltage clamp, and changes in membrane potential under current clamp were recorded. ⋯ While all cells showed evidence of the presence of fast-inactivating (A-type) K(+) channels, other K(+) channel types, including delayed rectifier, inward rectifier and large conductance Ca(2+)-activated K(+) (BK) channels were less common. Recorded Ca(2+) currents were identified pharmacologically as L-type. Non-linear regenerative voltage responses were evoked in more than half of the cells studied.
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Reproductive success depends on a robust and appropriately timed preovulatory luteinizing hormone (LH) surge, which is induced by the activation of gonadotropin-releasing hormone (GnRH) neurons in response to positive feedback from increasing estrogen levels. Here we document an increase in postsynaptic GluR2-lacking Ca2+ -permeable AMPA-type glutamate receptors (CP-AMPARs) at synapses on GnRH neurons on the day of proestrus in rats, coincident with the increase in estrogen levels. Functional blockade of CP-AMPARs depressed the synaptic responses only on the day of proestrus and concomitantly attenuated the LH surge. Thus, the phasic synaptic incorporation of postsynaptic CP-AMPARs on GnRH neurons is involved in the generation of the LH surge.
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Nerve growth factor (NGF) is an important mediator in the initiation of the inflammatory response and NGF via activation of the p75 neurotrophin receptor (p75(NTR)) and downstream sphingomyelin signaling leads to significant enhancement of the excitability of small-diameter sensory neurons. Because of the interaction between sphingomyelin and cholesterol in creating membrane liquid-ordered domains known as membrane or lipid rafts, we examined whether neuronal NGF-induced sensitization via p75(NTR) was dependent on the integrity of membrane rafts. Here, we demonstrate that the capacity of NGF to enhance the excitability of sensory neurons may result from the interaction of p75(NTR) with its downstream signaling partner(s) in membrane rafts. ⋯ In addition, adding back MβCD with cholesterol restored the NGF-induced sensitization in previously cholesterol-depleted neurons, suggesting that cholesterol and the structural integrity of rafts are key to promoting NGF-mediated sensitization. Using established protocols to isolate detergent-resistant membranes, both p75(NTR) and the neuronal membrane raft marker, flotillin, localize to raft fractions. These results suggest that downstream signaling partners interacting with p75(NTR) in sensory neurons are associated with membrane raft signaling platforms.
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To examine the effect of glucose on the cerebral metabolism of glutamine, rat brain slices were incubated with 5mM [3-(13)C]glutamine without and with 5mM unlabeled glucose. Tissue plus medium extracts were analyzed by using enzymatic and (13)C NMR techniques and fluxes through the enzymatic steps involved were calculated with a mathematical model. ⋯ The results indicate that 77% of the newly appeared glutamine was formed via glutamine synthetase and 23% from endogenous sources; the stimulation of [3-(13)C]glutamine removal by MSO also strongly suggests the existence of a cycle between [3-(13)C]glutamine and [3-(13)C]glutamate. This work also demonstrates that glucose increased fluxes through hexokinase, pyruvate kinase, lactate dehydrogenase, alanine aminotransferase, pyruvate carboxylase, pyruvate dehydrogenase, citrate synthase, flux from α-ketoglutarate to glutamate and flux through glutamine synthetase whereas it inhibited fluxes through aspartate aminotransferase, glutamic acid decarboxylase and GABA aminotransferase.
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Repeated exposure to drugs of abuse is associated with structural plasticity in brain reward pathways. Rats selectively bred for locomotor response to novelty differ on a number of neurobehavioral dimensions relevant to addiction. This unique genetic animal model was used here to examine both pre-existing differences and long-term consequences of repeated cocaine treatment on structural plasticity. ⋯ Changes in spine density occurred specifically near the branch point of terminal dendrites. These findings indicate that structural plasticity associated with prolonged cocaine abstinence varies markedly in two selected strains of rats that vary on numerous traits relevant to addiction. Thus, genetic factors that contribute to individual variation in the behavioral response to cocaine also influence cocaine-induced structural plasticity.