神经生物学

Tract tracing is a fundamental technique in neuroanatomy for examining fiber connections in the nervous system. After the introduction of horseradish peroxidase 40 years ago, many tracing substances have been used for neuroanatomical studies on various nervous systems. Here, we des ...

Biotinylated dextran amine (BDA) is a highly efficient and powerful marker for bidirectional tracing of nerve pathways in a wide variety of species at the light and electron microscopic level. The BDA tract-tracing method can readily be combined with other anterograde or retrograde trac ...

One of the best neurohistologic methods to reveal the cytoarchitecture of the brain and detailed morphology of neurons with unsurpassed clarity has been the Golgi staining. It is based on the principle of metallic impregnation of neurons, allowing visualization in their entirety inclu ...

The preparation of tissue for histological study is a multi-step process in which potential loss of quality and the introduction of artifacts can occur during each step. Knowledge of the process and the potential pitfalls at each step will serve the investigator well. Here I describe the most bas ...

Immunofluorescence or IF is a technique allowing the visualization of a specific protein or antigen in cells or tissues by binding a specific antibody chemically conjugated with a fluorescence dye. Immunofluorescent staining is widely used in life science research, particularly for ...

Time-lapse imaging techniques are widely used to monitor dendritic spine dynamics, a measurement of synaptic plasticity. However, it is challenging to follow the dynamics of spines over an extended period in vivo during development or in deep brain structures that are beyond the reach of tra ...

Quantifying dendrite morphology is a method for determining the effect of biochemical pathways and extracellular agents on neuronal development and differentiation. Quantification can be performed using Sholl analysis, dendrite counting, and length quantification. The ...

Patterned distributions of signalling molecules play fundamental roles during embryonic development. Several attempts have been made to reproduce these patterns in vitro. In order to study substrate-bound or membrane proteins, microcontact printing (μCP) is a suitable method ...

Stripe assays are frequently used for studying binary growth decisions of cells and axons towards surface-bound molecules in vitro. In particular in the fields of neurodevelopment and axon guidance, stripe assays have become a routine tool. Several variants of the stripe assay have been de ...

Growth cone collapse is an easy and efficient test for detecting and characterizing axon guidance activities secreted or expressed by cells. It can also be used to dissect signaling pathways by axon growth inhibitors and to isolate therapeutic compounds that promote axon regeneration. H ...

Luciferase reporter systems are widely employed to provide a quantitative readout of gene expression for studies of transcriptional regulation, translation efficiency, and cell signaling. The most common application of luciferase involves transient transfections into c ...

Protein–DNA interactions are critical to maintain genome stability, DNA replication, chromosome �segregation and to regulate gene expression. Chromatin immunoprecipitation (ChIP) is a powerful technique to study these interactions within living neurons and nervous tis ...

The lacZ gene product, β-galactosidase, has classically been used as a reporter of gene expression. β-Galactosidase activity can be detected using a chromogenic substrate, X-gal, which leaves an intense blue precipitate when cleaved by the enzyme. Insertion of the lacZ coding DNA targeted ...

Immunofluorescence (IF), a form of immunohistochemistry (IHC) with specific applications, is commonly used for both basic research and clinical studies, including diagnostics, and involves visualizing the cellular distribution of target molecules (e.g., proteins, DNA, and sm ...

In situ hybridization (ISH) is a type of hybridization that uses a labeled complementary DNA or RNA strand (i.e., probe) to localize a specific DNA or RNA sequence in a portion or section of tissue (In Situ) or in the entire tissue (whole mount ISH). Localization of endogenous transcripts is a desirable app ...

In utero electroporation has been extensively used to study a variety of developmental questions in the developing brain. This protocol aims to provide the basic knowledge for a beginner to get familiar with the technique. Basically, by electroporating a DNA construct into a subpopulati ...

In ovo electroporation is a popular technique to study gene function during development. This technique enables precise temporal and spatial genetic manipulation with the added advantages of being quick and inexpensive. In this chapter the transient transfection of a construct into ...

The developing spinal cord is a well-established model system widely used to study the signaling pathways and genetic programs that control neuronal/glial differentiation and neural circuit assembly. This is largely due to the relatively simple organization (compared to other CNS ...

Lentiviral vectors have become very useful tools for transgene delivery. Based on their ability to transduce both dividing and nondividing cells and to produce long-term transgene expression, lentiviruses have found numerous applications in the biomedical sciences, including ...

Biolistic transfection and diolistic labeling are techniques in which subcellular-sized particles, coated with DNA and lipophilic dyes, respectively, are propelled into cells. The gene-gun approach is particularly applicable for use on ex vivo organized tissue such as brain slic ...