One of the most powerful tools in the identification of various neuronal types and then connectivities is immunocytochemical localization of specific neuronal markers, e g., synthetic enzymes for neurotransmitters at precise cellular and subcellular levels. Several methods ex ...

Five amino acids have received considerable attention as putative neurotransmitters in mammalian brain; namely, γ-ammobutyric acid (GABA), glycine, glutamate, aspartate, and taurine Fonnum, 1978).

Some amino acids may function as neurotransmitters and are presumably released at nerve terminals to interact with specific receptors postsynaptically. Biochemical studies of high-affinity uptake, a potential marker of nerve terminals and of receptor binding, have been employ ...

Astrocytes communicate with the vascular endothelium via direct cell–cell contacts as well as a variety of secreted growth factors and extracellular matrix (ECM) proteins. Integrins are heterodimeric cell surface receptors for ECM protein ligands, and many integrin subunits are e ...

Astrocytes are the predominant nonneuronal cell type in the central nervous system. Although they are electrically nonexcitable, they have been found to play an active role in modulation of neuronal function and plasticity through Ca2+ excitability. Thus, Ca2+ signaling in astrocytes ...

Genetic tools are enabling the molecular dissection of the functions and mechanisms of many biological processes. Transgenic manipulations provide powerful tools with which to test hypotheses regarding functions of specific cell types and molecules in vivo in combination with d ...

The goal of this chapter is to highlight methods used to demonstrate in vivo changes in astrocyte expression at the blood–brain barrier (BBB). Loss of BBB integrity is seen in many acute and chronic disease conditions. However, despite the importance of the BBB to homeostasis and correct functio ...

Direct conversion of glia into neurons by cellular reprogramming represents a novel approach toward a cell-based therapy of neurodegenerative processes. Here we describe a protocol that allows for the direct and efficient in vitro reprogramming of mouse astroglia from the early post ...

Brain stroke is a devastating cerebrovascular disease and ranks as the third most common cause of death and disability in the US. Altered blood–brain barrier (BBB) signaling and permeability characteristics during stroke can increase the risk for life-threatening hemorrhagic tran ...

The chapter provides an introduction and brief overview of currently available in vitro blood–brain barrier models, pointing out the major advantages and disadvantages of the respective models and potential applications. Bovine brain microvessel endothelial cell isolation, ...

In vitro blood–brain barrier (BBB) models using primary cultured brain endothelial cells are important for establishing cellular and molecular mechanisms of BBB function. Co-culturing with BBB-associated cells especially astrocytes to mimic more closely the in vivo condition ...

Multiple sclerosis (MS) is characterized by CNS demyelination and oligodendrocyte depletion, axonal loss, and reactive astrogliosis. Myelin loss causes conduction block, while remyelination is associated with recovery of conduction and return of function. Reactive astroc ...

Oligodendrocyte migration is required for the myelination of axons during development and also following demyelinating lesions of the central nervous system. Oligodendrocytes arise from oligodendrocyte precursor cells (OPCs) which are present within the brain and spinal co ...

Schwann cells are one of the cellular candidates used in repair strategies following trauma and demyelination of the spinal cord. One of the major obstacles in the use of Schwann cells is their limited migratory ability within the astrocytic environment of the CNS and boundary formation betw ...

Microglia are the principal immune effector cells of the central nervous system (CNS). Under normal conditions, they occupy a quiescent surveillance phenotype, but following stimulation by microorganisms or inflammatory cytokines, microglia transform into highly activat ...

Astrocytes perform critical functions necessary for neuronal survival. Thus, examining the influence of astrocyte function on neuronal cell death during disease, including hypoxia/ischemia, has become an important avenue of investigation. In this chapter we detail the method ...

Astrocytes secrete factors that promote neuron survival, synapse formation, and plasticity. Understanding how these factors perform these roles requires a robust in vitro system that can effectively assess the impact of individual glial factors on neuronal properties. A classic ...

Astrogliosis, whereby astrocytes in the central nervous system (CNS) become reactive in response to tissue damage, is a prominent process leading to the formation of the glial scar that inhibits axon regeneration after CNS injury. Upon becoming reactive, astrocytes undergo various mo ...

Protocols are described for measurement in primary cultures of astrocytes of unidirectional fluxes of glutamate (influx and efflux), glutamate metabolism to glutamine or CO2, glucose influx, glycolysis, pyruvate dehydrogenation, oxidative metabolism of glucose, pyruvate ...

A typical feature of astrocytes is their high level of connexin expression. These membrane proteins constitute the molecular basis of two types of channels: gap junction channels that allow direct cytoplasm-to-cytoplasm communication and hemichannels that provide a pathway for ex ...