Purification and preliminary characterization of a glial growth factor from the bovine pituitary.

Abstract. We have immortalized rat central nervous system (CNS) cells of primary cultures of rat optic nerve with murine leukemia virus w2,SV-40-6, which is defective in assembly and contains the SV-40 large T antigen and neomycin resistance genes, to produce a cell line that we named A7. After drug selection, >90% of the growing cells expressed nuclear SV-40 large T cells and a fraction of these contained the astrocyte-specific marker, glial fibrillary acidic protein.The majority of these cells also expressed surface marker A4 (specific for neural tube derivatives), Ran 2, p185 (the 185-kD phosphoprotein product of the neu oncogene), and fibronectin, but did not express the astrocyte enzymes glutamine synthetase and monoamine oxidase B. Surface markers characteristic of glial progenitors (A2B5) and oligodendrocytes (galactocerebroside) were not detected. After two rounds of cell cloning, subclone A7.6-3 expressed Ran 2, fibronectin, and the neural cell adhesion molecule (N-CAM) but not glial fibrillary acidic protein and A4. The A7 cell line and subclones also displayed certain functions of type 1 astrocytes: the conditioned medium of these cells had a potent mitogenic activity for glial progenitor cells which could be neutralized by anti-platelet-derived growth factor antibodies and monolayers of these cells supported the growth of embryonic hypothalamic neurons. We conclude that a retrovirus containing SV-40 large T antigen can immortalize rat CNS cells and that such immortalized glial cells retain at least two important functions of type 1 astrocytes: the ability to secrete platelet-derived growth factor and to support the growth of embryonic CNS neurons. Moreover, such stable immortalized clonal cell lines can be used to study gene regulation in glial cells.

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“…None of our clones were found to secrete glial growth factor activity as tested on Schwann cells (Brockes, 1980).…”

Section: Mitogenic Activity Of the Supernatantmentioning

confidence: 87%

“…CM from cell line A7-6.3 was tested for glial growth factor activity by Dr. J. Brockes at Kings College, London, England, as previously described (Brockes et al, 1980).…”

Section: Glial Growth Factor Activitymentioning

confidence: 99%

Antigenic and functional characterization of a rat central nervous system-derived cell line immortalized by a retroviral vector.

Geller

Dubois-Dalcq

1988

The Journal of Cell Biology

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“…Schwann cells were grown on poly-D-lysine (1 mg/ml in distilled H20) coated tissue culture dishes, in DME with 1.0 g/l glucose (DME H-16; UCSF Cell Culture Facility) supplemented with 10% fetal calf serum, partially purified glial growth factor (12 i~g/ml), forskolin (2 p.M), glutamine (2 mM), and penicillin/streptomycin (100 U/ml), following a procedure kindly provided by Dr. Greg Lemke. The partially purified glial growth factor used in these experiments was prepared from a bovine pituitary extract by differential ammonium sulfate precipitation and carboxymethyl cellulose chromatography (Brockes et al, 1980).…”

Section: Cell Culturementioning

confidence: 99%

Distinct molecular interactions mediate neuronal process outgrowth on non-neuronal cell surfaces and extracellular matrices.

Tomaselli¹,

Reichardt²,

Bixby³

1986

The Journal of Cell Biology

242

183

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Abstract.We have compared neurite outgrowth on extracellular matrix (ECM) constituents to outgrowth on glial and muscle cell surfaces. Embryonic chick ciliary ganglion (CG) neurons regenerate neurites rapidly on surfaces coated with laminin (LN), fibronectin (FN), conditioned media (CM) from several non-neuronal cell types that secrete LN, and on intact extracellular matrices. Neurite outgrowth on all of these substrates is blocked by two monoclonal antibodies, CSAT and JG22, that prevent the adhesion of many cells, including neurons, to the ECM constituents LN, FN, and collagen. Neurite outgrowth is inhibited even on mixed LN/poly-D-lysine substrates where neuronal attachment is independent of LN. Therefore, neuronal process outgrowth on extracellular matrices requires the function of neuronal cell surface molecules recognized by these antibodies.The surfaces of cultured astrocytes, Schwann cells, and skeletal myotubes also promote rapid process outgrowth from CG neurons. Neurite outgrowth on these surfaces, though, is not prevented by CSAT or JG22antibodies. In addition, antibodies to a LN/proteoglycan complex that block neurite outgrowth on several LN-containing CM factors and on an ECM extract failed to inhibit cell surface-stimulated neurite outgrowth. After extraction with a nonionic detergent, Schwann cells and myotubes continue to support rapid neurite outgrowth. However, the activity associated with the detergent insoluble residue is blocked by CSAT and JG22 antibodies. Detergent extraction of astrocytes, in contrast, removes all neurite-promoting activity.These results provide evidence for at least two types of neuronal interactions with cells that promote neurite outgrowth. One involves adhesive proteins present in the ECM and ECM receptors on neurons. The second is mediated through detergent-extractable macromolecules present on non-neuronal cell surfaces and different, uncharacterized receptor(s) on neurons. Schwann cells and skeletal myotubes appear to promote neurite outgrowth by both mechanisms.

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“…Neuregulins [also known as glial growth factors (GGF), neu differentiation factor, heregulins, and ARIA] are a group of peptide growth factors expressed by embryonic neurons (Marchionni et al, 1993) and mesenchymal cells (Meyer and Birchmeier, 1994). The forms of neuregulins that correspond to the GGF were identified originally as an activity from pituitary glands that stimulated proliferation of astrocytes and Schwann cells (Brockes et al, 1980;Lemke and Brockes, 1984;Lemke, 1990). Subsequent purification of the peptides and cloning of the neuregulin gene demonstrated that alternative splicing of the neuregulin gene generates multiple isoforms, including ones that are predicted to be intracellular, transmembrane, or secreted (Marchionni et al, 1993;Wen et al, 1994).…”

mentioning

confidence: 99%

Axonal Interactions Regulate Schwann Cell Apoptosis in Developing Peripheral Nerve: Neuregulin Receptors and the Role of Neuregulins

et al. 1996

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Programmed cell death during development resulting from the lack of appropriate survival factors has been demonstrated in both neurons and oligodendrocytes and occurs mostly in the form of apoptosis. We now demonstrate that Schwann cells in the rat sciatic nerve undergo apoptosis during early postnatal development and that the amount of apoptosis is markedly increased by axotomy. The apoptotic Schwann cells express the low-affinity nerve growth factor receptor but not myelin-related proteins, indicating that they are in the premyelinating state. Apoptosis resulting from normal development or from axotomy can be inhibited markedly by exogenous neuregulin. Consistent with this, the neuregulin receptor components erbB2 and erbB3 are expressed and phosphorylated in developing sciatic nerve. These data suggest that Schwann cell number in developing peripheral nerve is regulated by apoptosis through competition for axonally derived neuregulin.

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Purification and preliminary characterization of a glial growth factor from the bovine pituitary.

Cited by 216 publications

References 19 publications

Antigenic and functional characterization of a rat central nervous system-derived cell line immortalized by a retroviral vector.

Antigenic and functional characterization of a rat central nervous system-derived cell line immortalized by a retroviral vector.

Distinct molecular interactions mediate neuronal process outgrowth on non-neuronal cell surfaces and extracellular matrices.

Axonal Interactions Regulate Schwann Cell Apoptosis in Developing Peripheral Nerve: Neuregulin Receptors and the Role of Neuregulins

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