神经生物学

The dynamic expression pattern analysis provides the primary information of gene function. Differences of the RNA and/or protein location will provide valuable information for gene expression regulation. Generally, in situ hybridization (ISH) and immunohistochemistry (I ...

Antisense morpholino oligonucleotides (MOs) have become a valuable method to knock down protein levels, to block mRNA splicing, and to interfere with miRNA function. MOs are widely used to alter gene expression during development of Xenopus and zebra fish, where they are typically inject ...

Microinjection has a long and distinguished history in Xenopus and has been used to introduce a surprisingly diverse array of agents into embryos by both intra- and intercellular means. In addition to nuclei, investigators have variously injected peptides, antibodies, biologically ...

Representatives of the phylum Mollusca have long been important models in neurobiological research. Recently, the routine application of immunocytochemistry in combination with confocal laser scanning microscopy has allowed fast generation of highly detailed reconstr ...

Injection of fluorochromes such as Alexa Fluor� 568 into single cells in brain slices reveals a network of dye-coupled cells to be associated with the central complex. Subsequent immunolabeling shows these cells to be repo positive/glutamine synthetase positive/horseradish pero ...

The Drosophila visual system is an excellent model system to study the switch from proliferating to differentiating neural stem cells. In the developing larval optic lobe, symmetrically dividing neuroepithelial cells transform to asymmetrically dividing neuroblasts in a high ...

Mosaic analysis with a repressible cell marker (MARCM) generates positively labeled, wild-type or mutant mitotic clones by unequally distributing a repressor of a cell lineage marker, originally tubP-driven GAL80 repressing the GAL4/UAS system. Variations of the technique incl ...

Visualization of single neurons within their complex environment is a pivotal step towards uncovering the mechanisms that control neural circuit development and function. This chapter provides detailed technical information on how to use Drosophila variants of the mouse Brain ...

Proper functioning of the brain relies on an enormous diversity of neural cells generated by neural stem cell-like neuroblasts (NBs). Each of the about 100 NBs in each side of brain generates a nearly invariant and unique cell lineage, consisting of specific neural cell types that develop in defin ...

Immunostaining is used to visualize the spatiotemporal expression pattern of developmental control genes that regulate the genesis and specification of the embryonic and larval brain of Drosophila. Immunostaining uses specific antibodies to mark expressed proteins and all ...

In Drosophila, the brain arises from about 100 neural stem cells (called neuroblasts) per hemisphere which originate from the neuroectoderm. Products of developmental control genes are expressed in spatially restricted domains in the neuroectoderm and provide positional cues t ...

Extracellular microelectrodes have been used for many years to apply focal electrical stimulation to individual cells (for example, see Pratt and Eisenberger and Huxley and Taylor ). Strickholm (1961) was the first to use a single extracellular electrode for both voltage control and rec ...

The technique described in this chapter provides electrophysiologists using patch-clamp with a convenient method to link electrophysiological data to a molecular analysis of the mRNAs expressed in a single cell. This molecular analysis can be used either to correlate cell responses ...

The patch-clamp technique allows the measurement of current through a wide variety of channels under reasonably realistic conditions, while controlling (“voltage clamping”) one component of the driving force for current, the electrical potential. The other component of the drivi ...

When membrane patches are isolated from cells, one gains the ability to regulate precisely the composition of the solution bathing both surfaces of the membrane and to change the composition rapidly. However, in detaching the membrane from the underlying cytoskeleton, one irreversibly ...

It is approaching 20 years since the introduction of the single-channel patch-clamp recording technique (Neher and Sakmann, 1976), and over the last two decades its refinements and diverse applications have served to maintain it as the dominant technique in membrane physiology (Neher, 1 ...

The extracellular patch voltage clamp technique has allowed the currents through single ionic channels to be studied from a wide variety of cells. In its early form (Neher and Sakmann, 1976), the resolution of this technique was limited by the relatively low (∽50 MΩ) resistances that isolated the i ...

It is now over 20 years since the seminal studies by John Gurdon and colleagues established that Xenopus laevis oocytes, when injected with messenger RNA (mRNA), were able after a period of incubation to translate the mRNA and appropriately synthesize the relevant protein (Gurdon et al., 1971; Gu ...

The technique of patch-clamp recording in brain slices is applicable to a large variety of cell types in slices from nearly all areas of the central nervous system (CNS) in animals at many different stages of development (Blanton et al., 1989; Edwards et al., 1989; Konnerth, 1990). To date, the technique has ...

The finding that nerve growth factor (NGF) influences the survival and maintenance of only selective neuronal populations has led to the identification of many related polypeptide factors, constituting a family of neurotrophic factors. Since NGF is required for neural crest-deriv ...