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A High-Throughput and Inexpensive Assay for Anxiety-Related Behaviors in the Zebrafish, Based on Place Preference and Latency to Feed

In this protocol, we describe a high-throughput and inexpensive assay for anxiety-related behaviors in zebrafish. The behavioral observations occur while fish are housed individually in 1-L tanks commonly found in commercial zebrafish rearing systems. Behavioral indices incl ...

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Ca2+/Calmodulin-Dependent Protein Kinase II (CaMKII)

The Ca2+/calmodulin (CaM)-dependent protein kinase II (CaMKII) is a major regulator of synaptic �plasticity. CaMKII function depends on complex regulation of its activity and localization by Ca2+/CaM and several auto-phosphorylation reactions. Auto-phosphorylation at T286 ...

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Electrophysiological Technique for Analysis of Synaptic Function of PKN1 in Hippocampus

PKNs are serine/threonine protein kinases that have conserved catalytic domains homologous to those of protein kinase C (PKC) family members and regulatory regions containing antiparallel coiled-coil (ACC) domains and C2-like domains. PKN1 in particular is abundant in the brain, a ...

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CASK: A Specialized Neuronal Kinase

CASK, a scaffolding protein present in neuronal synapses and other cell junctions, contains a CaM-kinase domain at its N terminus. Due to lack of adequate biochemical data and based on bioinformatics, this domain was classified as a pseudokinase. But new evidence suggests that CASK is a unique ki ...

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A Method for Sorting Zebrafish on the Exploratory-Boldness Behavioral Axis

A major axis of inter-individual behavioral variation is correlated behavioral traits, known variously as behavioral syndromes, personality, or temperament. The zebrafish is a good model organism with which to explore the molecular genetic mechanisms of personality. The first st ...

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Methods to Quantify Basal and Stress-Induced Cortisol Response in Larval Zebrafish

The zebrafish model is being increasingly used in stress and anxiety research. The homologue of the hypothalamic-pituitary-adrenal (HPA) axis has been found in fish and is referred to as the hypothalamic-pituitary-�interrenal (HPI) axis. HPA (mammalian) and HPI (fish) axes are homolog ...

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Automated Conditioning in Larval Zebrafish

A primary goal in neuroscience is to understand how the brain gives rise to innate and learned behavior. One way to expand the contributions of zebrafish in this respect is to use a computerized system that can train larval zebrafish in various behavior assays. This will allow more efficient genetic ...

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Living Human Brain Slices: Network Analysis Using Voltage-Sensitive Dyes

The study of diseases of the nervous system relies heavily on the use of animal models which try to replicate the human condition via various methods. The underlying assumption is that the root causes of the impairment are similar in the animal model and the human patient, thus validating the findings ...

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Defined Medium Organotypic Cultures of Spinal Cord Put Pain in a Dish

Nerve injury and/or disease are known to provoke the release of various neurotrophic and inflammatory agents from spinal microglia and astrocytes. Prolonged exposure of spinal neurons to these ‘nociceptive mediators’ invokes pathophysiological changes that lead to the ‘centr ...

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The Study of Respiratory Chemoreflexes Using a Novel Dual-Perfused Rodent Preparation

Artificially perfused brainstem preparations have several advantages over both in vivo and superfused en bloc preparations for studying respiratory circuits. They allow the analysis of neuronal function without confounding influences of anesthesia and major peripheral f ...

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Neuronal Networks in the In Vitro Isolated Guinea Pig Brain

Neuronal networks can be properly analyzed in experimental preparations that preserve the largest �connectivity within and between brain structures. The isolated whole guinea pig brain maintained in vitro by arterial perfusion is ideal for this purpose. The methods and techniques ...

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Spontaneous Neural Network Oscillations in Hippocampus, Cortex, and Locus Coeruleus of Newborn Rat and Piglet Brain Slices

Rises of cytosolic Ca2+ (Cai) associated with early network oscillations (ENOs) are important for brain maturation. Thus, developing neural networks are often studied by combining Cai imaging with electrophysiological recording of extracellular activity and/or intracellu ...

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Intact In Vitro Preparations of the Neonatal Rodent Cortex: Analysis of Cellular Properties and Network Activity

In vitro preparations of one whole hippocampus or one entire neocortical hemisphere (so-called intact preparations) combine the technical possibilities of conventional in vitro slice preparations with the advantage of a preserved intrinsic connectivity of a defined brain reg ...

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Autonomic Nervous System In Vitro: Studying Tonically Active Neurons Controlling Vagal Outflow in Rodent Brainstem Slices

Central mammalian neurons often show spontaneous discharge of single action potentials at regular rate. Several types of such ‘tonic’ neurons are involved in sensing or control of the intake and processing of food. Brain slices containing tonically active hypothalamic neurons are one ...

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Rodent Isolated Spinal Cord Preparations to Examine Motor Output

Understanding mammalian neural networks such as the central pattern generator (CPG) for locomotion requires the use of multiple experimental models. In vitro approaches provide easy access to the spinal cord, thus allowing more experimental tools to be used. In this chapter, some estab ...

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Anatomically Calibrated Isolated Respiratory Networks from Newborn Rodents

En bloc and transversal slice preparations from perinatal rodents are established in vitro models for studying control of breathing by neurons and neighboring glial cells in the lower brainstem. These neural “respiratory networks in the dish” show features complementary with those in ...

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Physiological Properties of Human Fetal Cortex In Vitro

Emergence of neuronal membrane excitability (e.g., voltage-gated ion conductances) and spontaneous electrical activity (e.g., spontaneous firing of action potentials) is an imperative for the normal development and maturation of brain circuits. Understanding the interplay ...

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Histological Characterization of Physiologically Determined Fast-Spiking Interneurons in Slices of Primate Dorsolateral Prefrontal Cortex

GABA (γ-aminobutyric acid)-ergic interneurons are a structurally and functionally diverse group of cells that, together, regulate the activity of neuronal networks giving rise to the brain oscillations necessary for information processing. The phenotypes of GABAergic neuro ...

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Neural Circuits Underlying Circadian Oscillations in Mammals: Clocks in a Dish

In mammals, the part of the nervous system responsible for most circadian behavior can be localized to a bilaterally paired structure in the hypothalamus known as the suprachiasmatic nucleus (SCN). Understanding this mammalian circadian system will require a detailed multilevel an ...

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Establishing In Vivo-Like Activity in Rat Cerebellar Cells Maintained In Vitro

Purkinje cells of the cerebellar cortex and neurons in the deep cerebellar nuclei (DCN) were among the first central neurons to be studied extensively through the use of in vitro preparations. Yet, the degree to which the patterns of action potential (“spike”) output recorded in these cells in vitro ...

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