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 ...
The extremely elongated processes of neurons—especially axons—present an unusual challenge to the metabolic machinery of the cell. A consequence of this geometry is that the metabolic and specialized (e.g., transmission) needs of axon terminals are dependent on the perikaryon (the p ...
In this chapter on methods for identifying and mapping receptors, most of the procedures to be described will involve NGF receptors. The current status of knowledge about NGF receptors will be reviewed briefly before describing some of the techniques that yielded this information.
The introduction of recombinant technology has dramatically influenced progress in the studies of many proteins and systems. In the areas of neurochemistry and neurobiology, perhaps the most notable effect has been on work with nerve growth factor (NGF), and its related protein family me ...
The technique of neuronal transplantation provides a powerful tool to deliver many pharmacologically active agents into the brain to address some fundamental questions regarding the basic principles of brain function. For example, grafting of embryonic tissues or cells into the ce ...
The neurotrophins are expressed in many cell types including fibroblasts, Schwann cells, salivary gland secretory cells, glia, and neurons. Neurotrophin gene expression in these cells is regulated developmentally, and once switched on it is regulated by diverse extracellular sig ...
During development of the vertebrate nervous system, neurons are produced in excess and, at a restricted time period, a significant portion degenerate, a phenomenon referred to as naturally occurring cell death (Hamburger, 1975). The neurons that survive this phase are those that form fun ...
Many proteins have “neurotrophic” activities in in vitro or in vivo test systems. Originally these were discovered as certain neurotrophic activities with unknown molecular structures. Nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), and ciliary neurotrop ...
Neurotrophic proteins have been broadly defined as those that promote the survival or differentiation of embryonic neurons. The first neurotrophic protein and the protein whose properties are responsible for this definition is nerve growth factor (NGF) (Cohn, 1960). In vivo, NGF parti ...
An injury to neural tissue that disrupts axonal continuity causes disconnection of pre- and postsynaptic elements of a neuronal circuit. The axon also transports to the cell body neurotrophic factors from the innervation territory, and axotomy, by interrupting this trophic supply, may ...
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