实验仪器使用技术

Caged molecules are photosensitive molecules with latent biological activity. Upon exposure to light, they are rapidly transformed into bioactive molecules such as neurotransmitters or second messengers. They are thus valuable tools for using light to manipulate biology with e ...

The idea of using light to unleash biologically active compounds from inert precursors (uncaging) was introduced over 30 years ago. Recent efforts prompted the development of photoremovable protecting groups that have increased photochemical efficiencies for one- and two-phot ...

Calcium signaling plays a major role in the function of cells. Measurement of intracellular calcium mobilization is a robust assay that can be performed in a high-throughput manner to study the effect of compounds on potential drug targets. Pharmaceutical companies frequently use calc ...

Analysis of ion channel mutants is a widely used approach for dissecting ion channel function and for characterizing the mechanisms of action of channel-directed modulators. Expression of functional potassium channels in potassium-uptake-deficient yeast together with gene ...

Ion channels function as multi-protein complexes made up of ion-conducting α-subunits and regulatory β-subunits. To detect, identify, and quantitate the regulatory β-subunits in functioning K+ channel complexes, we have chemically derivatized peptide-toxins that specific ...

Potassium channels conduct K+ ions selectively and at very high rates. Central to the function of K+ channels is a structural unit called the selectivity filter. In the selectivity filter, a row of four K+ binding sites are created using mainly the backbone carbonyl oxygen atoms. Due to the involveme ...

An integral part of neural circuit analysis involves understanding cellular-level connectivity. With recent technical advances, this capability has finally been realized. In this chapter, we will discuss various methods of targeting specific cell types in transgenic mice, with an ...

Single-cell electroporation is a recently developed method to introduce polar and charged molecules such as dyes, drugs, peptides, proteins, and nucleic acids into single and identified cells. This feature is advantageous, especially in investigations of the nervous system, becau ...

Peptide toxins derived from venomous animals are widely employed in neuroscience research because of their ability to manipulate specific ionic currents. However, the detailed characterization of particular classes of ion channels and their contributions to given neuronal ci ...

The ability to introduce gain- or loss-of-function constructs into developing organisms is responsible for the majority of advances made in the field of developmental biology. Here we provide step-by-step methods on how to achieve cellular transgenesis in the developing mammalian ne ...

RNA interference (RNAi) is a mechanism that specifically inhibits gene expression via small RNAs such as small interfering RNA (siRNA) and microRNA (miRNA). It not only plays a pivotal role under physiological conditions as cell differentiation and development but it also permits a syste ...

Viral-vector-based gene therapy is a powerful tool that allows experimental studies of species that previously were not amenable to genetic manipulation. Nonhuman primates (NHPs) are an invaluable resource for the study of genetic regulation of disease mechanisms. The main disadv ...

Recombinant AAV vectors currently enjoy an excellent track record in brain applications such as generating preclinical models of neurodegeneration and gene therapy for brain disorders. Indeed, rAAV vectors have been useful in modeling diseases such as Parkinson’s disease (disc ...

The bacteriophage P1-derived Cre/lox recombination system has been extensively used to engineer the genome of cultured cells and experimental animals. Cre recombinase recognizes the loxP site, which is composed of two 13-bp inverted repeats and an 8-bp spacer region, and mediates both i ...

Drug-regulatable site-specific recombinases have become, during the last decade, a central tool for �transgenesis. They allow, depending on the constructs used, the activation or inactivation of target genes in a time-dependent or, if combined with the use of a specific promoter, in a time- a ...

Immunotoxin cell targeting is a transgenic animal technology used to eliminate specific cell types from a complex neural circuitry by using cytotoxic activity of recombinant immunotoxins, which are composed of an antibody variable region fused to bacterial toxin fragments. This te ...

Antipredator behavior is triggered by a combination of internal proximate mechanisms (anatomical receptors and the physiological processes that regulate their function) and external environmental cues that signal the context and timing of when behavior is likely to be effecti ...

Central monoamine systems (e.g., dopamine, serotonin, norepinephrine) are associated with motivation, locomotion, social behavior, emotion, and mood. Biogenic amine transporters regulate neurotransmission by removing neurotransmitters from synapses and extracel ...

Addiction is a complex psychiatric disorder characterised by a spectrum of compulsive drug-seeking behaviours and a persistent tendency to relapse (return to drug taking) even after prolonged periods of abstinence. The most commonly used models for the study of drug reward and depende ...

Alcoholism is a disorder that affects human beings during every stage of the lifespan. Many animal models have been developed to study alcoholism, including those used to assess alcohol preference, the effects of alcohol withdrawal, and the development of tolerance. Knowledge gained fr ...