神经生物学

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 ...

Astrocytes produce numerous mediators under conditions of inflammation in the central nervous system. One such mediator is nitric oxide (NO) derived from nitric oxide synthase-2 (NOS-2), the high output, inducible NOS isoform. Expression of NOS-2 and production of NO can be stimulated in a ...

Astrocytes are intimately involved in immunological and inflammatory events occurring in the central nervous system (CNS), due to their ability to secrete and respond to a large number of immunoregulatory cytokines/chemokines such as IL-1β, IL-6, IL-8, IL-10, IL-17, IL-27, TNF-α, TGF-β, IFN- ...

Glia synthesize, package, and secrete several species of matrix proteases, including the gelatinases (pro-)MMP-2 and (pro-)MMP-9. In appropriate settings (e.g., experimental ischemia), these MMPs can be assayed from cerebral tissues or from astrocytes and microglia in culture by enz ...

Protocols are presented describing a unique in vitro injury model and how to culture and mature mouse, rat, and human astrocytes for its use. This injury model produces widespread injury and astrocyte reactivity that enable quantitative measurements of morphological, biochemical, a ...

Astrocytes are highly polarized cells. This is manifest not only during development and in the adult brain, but also following injury. In response to a wound, astrocytes extend processes that participate in formation of a glial scar, which walls off lesions in the brain or spinal cord. Similarly, a ...

We describe here a method to obtain primary cell cultures from the cerebral cortex and the hypothalamus of bovine fetuses. We report how tissue origin, developmental stages, and culture medium conditions influence cell differentiation and the prevalence of glial cells vs. neurons. We com ...

Detailed methods for the preparation of optic nerve head (ONH) astrocytes from human donor eyes and retinal astrocytes from rat eyes are described. Included is the immunopanning method used for ONH astrocyte isolation as well as cell characterization. The isolation of purified retinal a ...

Astrocytes are possibly the most numerous cells of the vertebrate central nervous system, yet a detailed characterization of their functions is still missing. One potential reason for the obscurity of astrocytic function is that they represent a diverse population of cells that all share ...

Astrocytes play fundamental roles in the establishment and maintenance of tissue homeostasis in the central nervous system (CNS). To examine these different functions in vitro, it is important to be able to generate pure astrocyte cultures. While many “enriched” astrocyte cultures ha ...