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丁香实验推荐阅读
Use of Random-Saturation Mutagenesis to Study Receptor-G Protein Coupling

The advent of powerful molecular techniques to study protein structure and function allows the modification of the codmg region of genes to produce proteins with altered structures that can be studied for changes in biochemical properties. The most common way to do structure/function st ...

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Agonist-Induced High-Affinity GTP Hydrolysis as an Index of Receptor-Mediated G Protein Activation in Mammalian Brain Membranes

Heterotrimeric guanine nucleotide-binding regulatory (G) proteins, composed of α-, β-, and γ-subunits, play pivotal roles in many receptor-mediated transmembrane signaling processes. Although detailed regulatory modes of G protein-mediated signal transduction have not y ...

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Identification and Quantitation of G Protein -Subunits

The demonstration that many intracellular signaling processes are mediated by a family of closely related guanine nucleotide binding proteins (G proteins) has led to the development of specific techniques that can be used to identify which of these polypeptide(s) is involved upon rece ...

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Covalent Modification of G Proteins by Affinity Labeling

The basic mechanism of G protein-mediated transmembrane signaling was elucidated in the late 1970s and early 1980s. Subsequently, molecular clomng has identified a large array of closely related receptors (R), G protein subunits (G), and effecters (E). Currently, an important research g ...

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Heterologous Expression of Receptors and Signaling Proteins in Adult Mammalian Sympathetic Neurons by Microinjection

Heterologous expression of receptor proteins provides a means of studying a molecularly defined receptor subtype in isolation from species closely related either by function and/or homology. For example, metabotropic glutamate receptors (mGluRs) comprise a large family (pre ...

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Nuclear Application of Antisense Oligonucleotides by Microinjection and Ballistomagnetic Transfer to Identify G Protein Heterotrimers Activating Phosp

G proteins link heptahelical membrane receptors to their effector systems. The G proteins consist of three subunits, α, β, and γ, of which until now 23 (including splice variants), 6, and 11 different forms are known, respectively (for reviews, see refs. 1 and 2). By sequence homology of G protein α-subuni ...

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Use of Antisense-Generating Plasmids to Probe the Function of Signal Transduction Proteins in Primary Neurons

Investigation of the function of intracellularly located protein components of receptor-effector transduction pathways has been hampered not only by their inaccessibility, but also by the lack of specific tools. The problem is particularly acute when the effect of interest must be m ...

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Autoradiographic Visualization in Brain of Receptor-G Protein Coupling Using [35S]GTPS Binding

Localization of receptors in brain sections using autoradtographlc detection of radioligand binding has been an important technique in the neuroanatomical identification of a large number of neurotransmitter receptors. However, receptor autoradiography provides lit ...

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Neural Stem Cells for Transplantation

Neural stem cell (NSC) biology has generated a great deal of interest in recent years. It can inform on both intrinsic and extrinsic processes that underlie cell diversity within the central nervous system (CNS), and is a powerful method for improving understanding of cell ancestry and potenti ...

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Immortalized Cells

While still in its infancy, developments in the exciting field of cell replacement and gene transfer to the central nervous system (CNS) are providing the basis for potentially new and powerful therapeutic approaches to a wide range of neurological diseases. Neuronal replacement and par ...

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Cells from Testis for Transplantation in the CNS

Neural transplantation as a therapy for neurodegenerative diseases has received much attention over the past two decades. The possibility that disease progress could be arrested and function restored raised hopes that a new treatment, particularly for Parkinson’s disease (PD), was ...

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Intracerebral Gene Transfer Using Viral Vectors

The recent development of recombinant viral vectors that are capable of transducing postmitotic cells may provide a powerful new tool for studying brain function, as well as ameliorative strategies in models of neurological disease. Some of these vectors have recently demonstrated d ...

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Neural Transplantation in the Developing CNS

Since the advent of modern neurotransplantation research about two decades ago, considerable attention has focused on its potential use in the treatment of neurodegenerative disorders, particularly Parkinson’s disease. However, although clinical application is limited to a ...

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Microtransplantation of Nigral Dopamine Neurons: A Step-by-Step Recipe

The cell suspension grafting procedure, as originally described by (1983), has become the standard protocol for the implantation of fetal neuronal cell suspensions to deep brain nuclei. Its major advantages, compared to the earlier studies of transplanting solid pieces of fetal tissue ...

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Morphology and Molecular Properties of Cellular Components of Normal Cerebral Vessels

The blood-brain barrier (BBB) includes anatomical, physicochemical, and biochemical mechanisms that control the exchange of materials between blood and brain and cerebrospinal fluid (CSF). Thus two distinct systems, the BBB and the blood-CSF barrier systems, control cerebral ho ...

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Freeze-Fracture Studies of Cerebral Endothelial Tight Junctions

The tracer experiments of Reese and Karnovsky (1) demonstrated that it was the endothelium that formed a permeability barrier because electron-dense tracers such as horseradish peroxidase did not pass from the vessel lumen through the interendothelial cleft. The structure respon ...

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Studies of Cerebral Vessels by Transmission Electron Microscopy and Morphometry

Transmission electron microscopy (TEM), a once popular research tool, is used less frequently now. However, the isolation of novel proteins in the past decade has led to renewed use of electron microscopy for the subcellular localization of these proteins. This chapter will describe the sta ...

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A Review of Blood-Brain Barrier Transport Techniques

The blood-brain barrier (BBB) is a unique dynamic regulatory interface situated at the border between the blood stream and the brain extracellular (or interstitial) fluid. As the “gatekeeper” to the brain, it determines the ability of drugs to gain entrance to brain extracellular fluid and re ...

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Blood-Brain Barrier Permeability Using Tracers and Immunohistochemistry

The first report of blood-brain barrier (BBB) permeability by Paul Ehrlich (1) involved the use of the exogenous tracer Coerulean-S as described in Chapter 6. Over the years, tracers of different sizes were introduced to study the permeability properties of normal cerebral vessels in physio ...

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In Situ Brain Perfusion Technique

Many techniques have been developed to study transport across the blood-brain barrier (BBB). One that offers particular advantage is the in situ brain perfusion technique. The primary objective of the in situ brain perfusion technique is to take over the circulation to the brain via direct inf ...

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