ISOLATION AND SOME CHEMICAL PROPERTIES OF OLIGODENDROGLIA FROM CALF BRAIN<sup>1</sup>

Poduslo, Shirley E.; Norton, William T.

doi:10.1111/j.1471-4159.1972.tb01388.x

Cited by 280 publications

(95 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…3A). PL, NL, and GL account for almost all lipids present in OLG and myelin (4,24,25 (18). The cells express myelin basic protein (12) and myelin-associated glycoprotein (17).…”

Section: Methodsmentioning

confidence: 99%

Lipid metabolism of isolated oligodendrocytes maintained in long-term culture mimics events associated with myelinogenesis.

Szuchet¹,

Yim²,

Monsma³

1983

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

made to correlate changes in lipid metabolism with morphologic developments. High incorporation into phospholipids and cholesterol coincided in time with the extensive membrane synthesis required for cell attachment and process extension. Differentiation of these newly formed membranes, as assessed by the incorporation of myelin-characteristic glycolipids, galactocerebrosides, and sulfatides, occurred at a time when an intricate network of processes had already been established. The sequence of metabolic events observed in vitro parallels that observed at the onset of myelinogenesis in vivo. We postulate that mature oligodendrocytes can reenact those early events associated with myelinogenesis.Oligodendrocytes (OLG) Another type of OLG heterogeneity, based on ultrastructural features, has come to light: three prototypes have been described (6,7). It is believed that these types represent stages in the process of maturation of OLG. Although these observations have been made only on murine species and their general validity remains unproven, the fact that myelin changes in composition after its initial deposition, a process referred to as maturation (4), has led to the hypothesis of specialization among the OLG subgroups. It has been suggested that young OLG assemble myelin while mature ones maintain it (8, 9). This formulation presupposes that OLG change from a metabolism suitable for membrane synthesis to one geared to myelin maintenance. Whether these changes are reversible is a question of significance, because it bears on the issue of the capacity of OLG to remyelinate.There is evidence that the onset of myelination is heralded by a cascade of events, morphological, ultrastructural, and biochemical, in OLG. These events reach their maxima midway through myelination and then subside. Once the framework of myelin has been set up, architectural remodeling goes on for months to years depending on the species (4). Thus, perhaps the proper question is not whether there is a change in metabolism; clearly there must be, but rather, is it reversible? Can mature OLG be induced to relive the early episodes of myelination? The availability of procedures to isolate OLG from mature (10)(11)(12)(13)(14) and immature (15, 16) brains and to keep them in long-term culture permits addressing some of these questions.We have improved our procedure for OLG isolation to yield a highly purified (99%) population that can be kept in culture with no loss in purity over time (17) It remains to be shown that this preparation for remyelination can in fact be followed by remyelination. In this paper, we present biochemical evidence to support our con-

show abstract

“…3A). PL, NL, and GL account for almost all lipids present in OLG and myelin (4,24,25 (18). The cells express myelin basic protein (12) and myelin-associated glycoprotein (17).…”

Section: Methodsmentioning

confidence: 99%

Lipid metabolism of isolated oligodendrocytes maintained in long-term culture mimics events associated with myelinogenesis.

Szuchet¹,

Yim²,

Monsma³

1983

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Among glial cells, CNP activity is much higher in oligodendroglia, which produce myelin, than in astroglia (17). Activity is low or absent in non-neural tissues or in cloned cells other than glioma and Schwannoma (13)(14)(15)(16), although CNP can be detected at low specific activity in purified erythrocyte membranes (18).…”

mentioning

confidence: 99%

“…The precise role of CNP in brain function is not known. Within the central nervous system, CNP is generally regarded as specific to myelin and oligodendrocytes (14,17,19). The exact type of glial cell from which the C6 glioma arose is uncertain (33,34) but, although the cells resemble astrocytes morphologically, they express some of the biochemical properties characteristic of oligodendrocytes (15,35).…”

mentioning

confidence: 99%

Norepinephrine induces glial-specific enzyme activity in cultured plasma glioma cells.

McMorris¹

1977

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Addition of 1 #M 1-norepinephrine to cultures of C6TK-rat glioma cells caused a 2-fold increase in specific activity of the glial-specific enzyme 2':3'-cyclic nucleotide 3'-phosphohydrolase (nucleoside-2':3'-cyclic-phosphate 3'-nucleotidohydrolase, EC 3.1.4.16). Specific activity could also be stimulated by analogues of 3':5'-cyclic AMP, and the effect of norepinepbrine could be blocked by P-adrenergic receptor antagonists but not by a-adrenergic antagonists. Norepinephrine or cyclic AMP analogues also increased the specific activity of this enzyme in other clones of glioma and Schwannoma cells and in glioma X neuroblastoma cell hybrids. These results show that the stimulatory effect of norepinephrine on cyclic AMP concentrations in glioma cells leads ultimately to a stimulation of glial-specific cell function. The neurotransmitter l-norepinephrine (NE) induces a dramatic increase in the intracellular concentration of adenosine 3':5'-cyclic monophosphate (cAMP) in brain slices (1) and in glioma cells in tissue culture (2-4). Although glioma cells were the first clonal cell type in which such a response was observed (2-4), the function of this NE-induced cAMP response remains obscure. It has been suggested that this increase in "second messenger" is involved in metabolic coupling between neurons and glia, which are in intimate contact in the central nervous system (2).Exposure of glioma cells to NE or to cAMP analogues affects several functions common to many cell types-including glycogenolysis (5, 6), lactate dehydrogenase (7), ornithine decarboxylase (8), cAMP phosphodiesterase (9, 10), and protein kinase activity (11)-and induces morphological changes in the cells (12), but no glial-specific enzyme or function has been shown to be altered by NE or cAMP.In this paper I report that physiological concentrations of NE cause an increase in the glial-specific enzyme 2':3'-cyclic nucleotide 3'-phosphohydrolase (CNP; nucleoside-2':3'-cyclicphosphate 3'-nucleotidohydrolase, EC 3.1.4.16) in C6TK-rat glioma cells in vitro and that this response is mediated by a #-adrenergic receptor and cAMP.CNP is a membrane-associated enzyme that is present at high specific activity in glial and Schwann cells and in myelin (13-16). Among glial cells, CNP activity is much higher in oligodendroglia, which produce myelin, than in astroglia (17). Activity is low or absent in non-neural tissues or in cloned cells other than glioma and Schwannoma (13-16), although CNP can be detected at low specific activity in purified erythrocyte membranes (18). CNP is therefore considered to be one of the most specific biochemical markers for myelin and oligodendroglia (19). CNP is inactive toward 3':5'-cyclic nucleotides (

show abstract

“…Poduslo and Norton, 1972;Fewster et al, 1973;Chao and Rumsby, 1977;Szuchet et al, 1978;Snyder et al, 1980;McCarthy and de Vellis, 1980;Bhat et al, 1981;Lisak et al, 1981;Gebicke-Harter et al, 1981;Hirayama et al, 1983). As far as rat-brain oligodendrocytes are concerned, the method described by McCarthy and de Vellis (1980) is the only one using physiological conditions as much as possible and yielding enough cells for biochemical investigations.…”

Section: Discussionmentioning

confidence: 99%

“…Isolation and maintenance in culture of this cell type is therefore of great importance for the biochemical investigation of the synthesis of myelin-specific compounds and their assembly in myelin membranes. Several procedures have been described for the isolation of oligodendrocytes; some are only intended for the bulk isolation of these cells (Poduslo and Norton, 1972), whereas other methods have been developed with the aim of obtaining a pure culture of oligodendrocytes (McCarthy andde Vellis, 1980: Hirayama et al, 1983).…”

Section: Introductionmentioning

confidence: 99%

Culture of rat cerebral oligodendrocytes in a serum-free, chemically defined medium

Koper

Lopes‐Cardozo

Romijn

et al. 1984

Journal of Neuroscience Methods

View full text Add to dashboard Cite

Kew'ords: oligodendrocyte--primary culture--serum-free medium--chemically defined medium--brain cells--rat brain Oligodendrocytes were isolated from the cerebra of young rats (5-10 days old) by trypsinization of the tissue followed by cell separation on Percoll gradients. The isolation was carried out in physiological, isotonic media. The cell yield was 2-4 × 106 cells per brain; the plating efficiency was >/70%. Isolated cells were seeded on poly-L-lysine-coated culture dishes and maintained in a serum-free, chemically defined medium for at least 30 days. After 10 days in culture 67+10% of the surviving cells were oligodendrocytes, as judged by immunocytochemical and morphological criteria, whereas most of the other cells reacted positively with antiserum against glial fibrillary acidic protein. The expression of typical oligodendrocyte markers (2':3'-cyclic-nucleotide 3'-phosphodiesterase, galactocerebrosides and myelin basic protein) was greatly enhanced under these serum-free conditions as compared with cultures in serum-containing medium. The antigenic markers (galactocerebrosides, myelin basic protein) were absent in the freshly isolated cells but could be detected after 3 days in culture by immunocytochemistry. The activity of 2':3'-cyclic-nucleotide 3'-phosphodiesterase increased from 75 nmol min 1 mg 1 protein on day 4 to 400 nmol min-1 mg 1 protein on day 14 in culture.

show abstract

ISOLATION AND SOME CHEMICAL PROPERTIES OF OLIGODENDROGLIA FROM CALF BRAIN¹

Cited by 280 publications

References 29 publications

Lipid metabolism of isolated oligodendrocytes maintained in long-term culture mimics events associated with myelinogenesis.

Lipid metabolism of isolated oligodendrocytes maintained in long-term culture mimics events associated with myelinogenesis.

Norepinephrine induces glial-specific enzyme activity in cultured plasma glioma cells.

Culture of rat cerebral oligodendrocytes in a serum-free, chemically defined medium

Contact Info

Product

Resources

About

ISOLATION AND SOME CHEMICAL PROPERTIES OF OLIGODENDROGLIA FROM CALF BRAIN1

Cited by 280 publications

References 29 publications

Lipid metabolism of isolated oligodendrocytes maintained in long-term culture mimics events associated with myelinogenesis.

Lipid metabolism of isolated oligodendrocytes maintained in long-term culture mimics events associated with myelinogenesis.

Norepinephrine induces glial-specific enzyme activity in cultured plasma glioma cells.

Culture of rat cerebral oligodendrocytes in a serum-free, chemically defined medium

Contact Info

Product

Resources

About

ISOLATION AND SOME CHEMICAL PROPERTIES OF OLIGODENDROGLIA FROM CALF BRAIN¹