实验仪器使用技术

This protocol describes an assay for measuring visual recognition memory in larval zebrafish using an automated imaging system. Dubbed the “context recognition assay,” this assay measures the responses of larvae to familiar and unfamiliar visual stimuli displayed on an LCD screen be ...

A long-standing debate on whether fish can feel pain is coming to a close as evidence gathers regarding the existence of nociceptive mechanisms in the peripheral and central nervous system of these vertebrates, as well as nocifensive behavior that can be altered by pharmacological treatme ...

The open tank diving test and the light/dark test are commonly used to assess anxiety-like behavior in zebrafish. The open tank test exerts relatively weak control over behavior, and uses vertical locomotion as its primary measure. The light/dark test exerts relatively strong control over b ...

The open field test is a well-established paradigm for assaying anxiety-related behaviors and has been successfully employed in a broad range of species. We describe an adaptation of this test for the zebrafish, which is increasingly widely used as a model for anxiety-related behavior and st ...

Some teleost fish present scototaxis, a tendency to search for dark places and avoid brightly lit spaces. In zebrafish, it has been proposed that scototaxis (preference for dark environments) is a model of anxiety. In this chapter, we discuss some evidence regarding that proposal, and present a s ...

Mammalian target of rapamycin (mTOR) is a serine/threonine protein kinase known to merge extracellular instructions with information about cellular metabolic resources and control the rate of anabolic and catabolic processes accordingly. In neurons, mTOR has been implicated in ...

Endocytosis and trafficking of Trk receptors are central to their functions in neuronal development. Neurotrophin binding to Trk receptors induces endocytosis of Trk receptors as a ligand–receptor complex in axon terminals. This endocytic ligand–receptor complex, termed the s ...

Diacylglycerol kinase (DGK) is a lipid kinase but not protein kinase. However, the enzyme regulates many protein kinases including PKC, PI4P-Kinase, and mTOR by reducing the amount of diacylglycerol and producing phosphatidic acid. To date, ten mammalian DGKs have been cloned and divided i ...

The protein kinase C (PKC) family contains ten members that share a catalytic core but differ in the composition of their regulatory modules. The conventional PKCs are a subfamily whose regulatory domains bind to Ca2+ and to the lipid second messenger diacylglycerol and thus they are activated ...

Cyclic-AMP dependent protein kinase (PKA) is present in most branches of the living kingdom, and as an example in the nervous system where PKA integrates the cellular effects of various neuromodulators. The recent development of fluorescence resonance energy transfer (FRET) biosens ...

PTEN-induced novel kinase 1 (PINK1) and leucine-rich repeat kinase 2 (LRRK2) are two protein kinases associated with recessive and dominant forms of parkinsonism, respectively. Mutations in PINK1 cause loss of protein function whereas mutations in LRRK2 are less easily defined but, in s ...

Muscle-specific kinase (MuSK) is a RTK that is specifically expressed in skeletal muscle fibers and critical for the formation and maintenance of the neuromuscular junction (NMJ), a peripheral synapse formed between motoneurons and muscle fibers (1, 2). The acetylcholine receptors ( ...

Excitotoxicity via N-methyl-d-aspartate receptors (NMDA-r) is a key mechanism of neurodegeneration following ischemia and other brain pathologies. We here describe the use of a fast, reliable and easily reproducible in vitro model of excitotoxicity induced with NMDA stimulation ...

Protein phosphorylation is one of the key mechanisms controlling cellular signaling networks. Due to the low abundance of phosphorylated proteins and weaker ionization efficiency of phosphopeptides during mass spectrometric analyses, it is highly required to remove abundant ...

Protein kinase profiling can provide a basis for understanding the molecular origins of diseases and, potentially, for developing tools for therapeutic intervention. It is therefore very important to develop advanced experimental procedures for convenient and accurate dete ...

Protein kinases are key modulators of intracellular signal transduction cascades, which determine various events in neuronal cells such as replication and differentiation. For many years, protein kinases were �analyzed mostly by biochemical methods, which could handle the cel ...

As the rate of biomedical discovery is rising exponentially, electronic databases have become particularly effective in organizing and sharing scientific knowledge. Due to a well-characterized genome, robust behavioral responses and physiological similarity to humans, t ...

Egr-1 (also known as zif268, Krox-24, NGFI-A, and ZENK) has been one of the most widely immediate early genes used to map specific neuronal populations activated during behavior in mammals and birds. However, the use of egr-1 mapping technique in nontraditional model systems depends on the evolut ...

The technique of intracerebroventricular (i.c.v.) injection is a useful neuroethological tool for studying the central actions of drugs on behavior of conscious animals because many drugs do not cross the blood–brain barrier, which limits the extent of drug action to the CNS. In this chapter ...

Over the past decades, zebrafish have been presented as a novel and valuable tool for modeling complex human diseases. Epilepsy is a serious brain disorder with multiple genetic and environmental causes. Our poor understanding of its pathogenesis requires novel paradigms and model org ...