Microglia are the resident macrophages of the central nervous system and are associated with the pathogenesis of many neurodegenerative and brain inflammatory diseases; however, the origin of adult microglia remains controversial. We show that postnatal hematopoietic progenitors do not significantly contribute to microglia homeostasis in the adult brain. In contrast to many macrophage populations, we show that microglia develop in mice that lack colony stimulating factor-1 (CSF-1) but are absent in CSF-1 receptor-deficient mice. In vivo lineage tracing studies established that adult microglia derive from primitive myeloid progenitors that arise before embryonic day 8. These results identify microglia as an ontogenically distinct population in the mononuclear phagocyte system and have implications for the use of embryonically derived microglial progenitors for the treatment of various brain disorders.

/pdf/fate-mapping-analysis-reveals-that-adult-microglia-derive-4q8i4y2udw.pdf

Fate Mapping Analysis Reveals That Adult Microglia Derive from Primitive Macrophages

A novel method has been developed for the preparation of nearly pure separate cultures of astrocytes and oligodendrocytes. The method is based on (a) the absence of viable neurons in cultures prepared from postnatal rat cerebra, (b) the stratification of astrocytes and oligodendrocytes in culture, and (c) the selective detachment of the overlying oligodendrocytes when exposed to sheer forces generated by shaking the cultures on an orbital shaker for 15--18 h at 37 degrees C. Preparations appear greater than 98% pure and contain approximately 1-2 x 10(7) viable cells (20--40 mg of cell protein). Three methods were used to characterize these two culture t ypes. First, electron microscopic examination was used to identify the cells in each preparation (mixed and separated cultures of astrocytes and oligodendrocytes) and to assess the purity of each preparation. Second, two oligodendroglial cell markers, 2',3'-cyclic nucleotide 3'-phosphohydrolase (EC 3.1.4.37) and glycerol phosphate dehydrogenase (EC 1.1.1.8) were monitored. Third, the regulation of cyclic AMP accumulation in each culture type was examined. In addition to these studies, we examined the influence of brain extract and dibutyryl cAMP on the gross morphology and ultrastructure of each preparation. These agents induced astroglial process formation without any apparent morphological effect on oligodendrocytes. Collectively, the results indicate that essentially pure cultures of astrocytes and of oligodendrocytes can be prepared and maintained. These preparations should significantly aid in efforts to examine the biochemistry, physiology, and pharmacology of these two major classes of central nervous system cells.

/pdf/preparation-of-separate-astroglial-and-oligodendroglial-cell-5goxtcftyx.pdf

Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue.

Microglial cells are the resident macrophages in the central nervous system. These cells of mesodermal/mesenchymal origin migrate into all regions of the central nervous system, disseminate through the brain parenchyma, and acquire a specific ramified morphological phenotype termed "resting microglia." Recent studies indicate that even in the normal brain, microglia have highly motile processes by which they scan their territorial domains. By a large number of signaling pathways they can communicate with macroglial cells and neurons and with cells of the immune system. Likewise, microglial cells express receptors classically described for brain-specific communication such as neurotransmitter receptors and those first discovered as immune cell-specific such as for cytokines. Microglial cells are considered the most susceptible sensors of brain pathology. Upon any detection of signs for brain lesions or nervous system dysfunction, microglial cells undergo a complex, multistage activation process that converts them into the "activated microglial cell." This cell form has the capacity to release a large number of substances that can act detrimental or beneficial for the surrounding cells. Activated microglial cells can migrate to the site of injury, proliferate, and phagocytose cells and cellular compartments.

Physiology of Microglia

Transformed stem cells have been isolated from some human cancers. We report that, unlike other brain cancers, the lethal glioblastoma multiforme contains neural precursors endowed with all of the critical features expected from neural stem cells. Similar, yet not identical, to their normal neural stem cell counterpart, these precursors emerge as unipotent (astroglial) in vivo and multipotent (neuronal-astroglial-oligodendroglial) in culture. More importantly, these cells can act as tumor-founding cells down to the clonal level and can establish tumors that closely resemble the main histologic, cytologic, and architectural features of the human disease, even when challenged through serial transplantation. Thus, cells possessing all of the characteristics expected from tumor neural stem cells seem to be involved in the growth and recurrence of adult human glioblastomas multiforme.

/pdf/isolation-and-characterization-of-tumorigenic-stem-like-3p3it406jl.pdf

Isolation and Characterization of Tumorigenic, Stem-like Neural Precursors from Human Glioblastoma

Lang, Florian, Gillian L. Busch, Markus Ritter, Harald Volkl, Siegfried Waldegger, Erich Gulbins, and Dieter Haussinger. Functional Significance of Cell Volume Regulatory Mechanisms. Physiol. Rev. ...

Functional Significance of Cell Volume Regulatory Mechanisms

Introduction, Marston Manthorpe Outgrowth Assays and Cortical Slice Cultures, Valerie Castellani and Jurgen Bolz Microexplant Cultures of the Cerebellum, Bernard Rogister and Gustave Moonen Aggregating Neural Cell Cultures, Paul Honegger and Florianne Monnet-Tschudi Primary Cell Cultures of Peripheral and Central Neurons and Glia, Mary I Johnson and Richard P Bunge Chick Spinal Somatic Motoneurons in Culture, Bernhard H J Juurlink Neural Cells in Microcultures: In Vitro Biological Assays for Neuroactive Agents, Marston Manthorpe Construction and Use of Compartmented Cultures, Robert B Campenot Astrocyte and Oligodendrocyte Cultures, Ruth Cole and Jean de Vellis Generation of Mouse Astroglia and Microglia Cultures from Mouse Neopallium, Sergey Fedoroff and Arleen Richardson Generation of Oligodendroblasts from Primary Cultures of Rat Neopallium, Bernhard H J Juurlink, Shawn K Thorburne, and Richard M Devon Culture of Glial Cells from Human Brain Biopsies, Voon Wee Yong and Jack P Antel Colony Cultures, Sergey Fedoroff and Arleen Richardson Herpes Simplex Virus Vectors for Gene Therapy of the Nervous System , Angela P Dyer and Frank Tufaro Immunostaining and Identification of Antigens, Colin J Barnstable Elimination of Cell Types from Mixed Neural Cell Cultures, Richard M Devon Quantification of Cells in Culture, Arleen Richardson and Sergey Fedoroff Procedures for Subculturing Cells, Arleen Richardson and Sergey Fedoroff Preparation of Tissue-Cultured Material for Electron Microscopical Observation, Richard M Devon Tips in Tissue Culture, Sergey Fedoroff and Arleen Richardson Use of Dissecting Instruments in Tissue Culture, Arleen Richardson Abbreviations and Definitions Index

Protocols for Neural Cell Culture

Kinetics for uptake and release of glutamate were measured in normal, i.e., nontransformed, astrocytes in cultures obtained from the dissociated, cortexenriched superficial parts of the brain hemispheres of newborn DBA mice. The uptake kinetics indicated a minor, unsaturable component together with an intense uptake following Michaelis-Menten kinetics. TheK

 m
 (50 μM) was reasonably comparable to the corresponding values in brain slices and in other glial preparations. TheV
max (58.8 nmol min−1 mg−1 protein) was, however, much higher than that observed in glial cell lines or peripheral satellite cells, and also considerably higher than that generally reported for brain slices. The release of glutamate was much smaller than the uptake, and only little affected by an increase of the external glutamate concentration, suggesting a net accumulation of glutamate rather than a homoexchange. Such an intense accumulation of glutamate into normal astrocytes may play a major role in brain metabolism and may help keep the extracellular glutamate cohcentration below excitatory levels.

Kinetic characteristics of the glutamate uptake into normal astrocytes in cultures

Let's read! We will often find out this sentence everywhere. When still being a kid, mom used to order us to always read, so did the teacher. Some books are fully read in a week and we need the obligation to support reading. What about now? Do you still love reading? Is reading only for you who have obligation? Absolutely not! We here offer you a new book enPDFd advances in cellular neurobiology to read.

Advances in Cellular Neurobiology

The relationship between astrocytes forming in the presence of dibutyryl cyclic AMP (dBcAMP) in culture and reactive astrocytes responding to a cerebral cortex stab wound was investigated using computerized image analysis (Zeiss IBAS 1) and immunocytochemical staining. The diameters of the nuclei of astrocytes in primary cultures of newborn mouse neopallial cells were compared to those of the nuclei of normal and reactive astrocytes in histological sections of mouse cerebral cortex. We found that the nuclei of astrocytes that formed in the presence of dBcAMP in cultures are significantly larger than those of spontaneously occurring small stellate astrocytes in culture and of normal astrocytes of the cerebral cortex in vivo but corresponded more closely to the nuclei of reactive astrocytes in the area surrounding a stab wound in the cerebral cortex. Large stellate cells formed in the presence of dBcAMP had vimentin and an increase in GFP-containing intermediate filaments. Formation of reactive astrocytes in vivo is also associated with an increase in both vimentin and GFP-containing intermediate filaments. These observations indicate a closer relationship of astrocytes formed in the presence of dBcAMP in cultures to the reactive astrocytes in the cerebral cortex than to normal astrocytes. We propose, therefore, that the large stellate astrocytes that form in the presence of dBcAMP be referred to as reactive astrocytes in culture.

Sergey Fedoroff

Papers

Protocols for Neural Cell Culture

Kinetic characteristics of the glutamate uptake into normal astrocytes in cultures

Advances in Cellular Neurobiology

Astrocyte cell lineage. V. Similarity of astrocytes that form in the presence of dBcAMP in cultures to reactive astrocytes in vivo.

Cell, tissue, and organ cultures in neurobiology.