神经生物学

A major goal of our research is to develop an implantable device for routine amperometric recordings of l-glutamate and other neurotransmitters in the mammalian central nervous system. Specifically, we wanted to develop a microelectrode that is (1) mass produced such that other laborat ...

The electrode is a vital element of a microelectrode biosensor, which is designed to transform the recognition of a biological molecule into an easily quantifiable electrical signal. The most widely used enzymatic biosensors rely on oxidases for detecting neurotransmitters and met ...

The production of contrast agents sensitive to neuronal signaling events is a rate-limiting step in the development of molecular-level functional magnetic resonance imaging (molecular fMRI) approaches for studying the brain. High-throughput generation and evaluation of po ...

The application of fluorescent receptor ligands has become widespread, incited by two important reasons. “Seeing is believing”—it is possible to visualize in real time in live cells ligand–receptor interactions, and to locate the receptors with subcellular precision allowing one to ...

In this protocol, we describe the procedures we have developed to optimize the performance of voltage-sensitive dyes for recording changes in neuronal electrical activity. We emphasize our experience in finding the best dye conditions for recording backpropagating action poten ...

Fluorescent ion sensors are quite valuable in experimental biology. The development of new sensor molecules requires determination of spectral properties (absorption bands, fluorescence excitation, and emission maxima) in order to characterize the type of optical response to ...

Light is an exquisite reagent for controlling the activity of biological systems, often offering improved temporal and spatial resolution over strictly genetic, biochemical, or pharmacological manipulations. This chapter describes a general approach for developing small m ...

Incorporation of photoisomerizable chromophores into small molecule ligands represents a general approach for reversibly controlling protein function with light. Illumination at different wavelengths produces photostationary states (PSSs) consisting of differ ...

Voltage-gated potassium (K v) channels are membrane proteins that open a selective pore upon membrane depolarization, allowing K+ ions to flow down their electrochemical gradient. In neurons, K v channels play a key role in repolarizing the membrane potential during the falling phase of the a ...

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 ...