Purification of the Chick Eye Ciliary Neuronotrophic Factor

Purification of the Chick Eye Ciliary Neuronotrophic Factor Abstract: Dissociated 8‐day chick embryo ciliary ganglionic neurons will not survive for even 24 h in culture without the addition of specific supplements. One such supplement is a protein termed the ciliary neuronotrophic factor (CNTF) which is present at very high concentrations within intraocular tissues that contain the same muscle cells innervated by ciliary ganglionic neurons in vivo. We describe here the purification of chick eye CNTF by a 2½‐day procedure involving the processing of intraocular tissue extract sequentially through DE52 ion‐exchange chromatography, membrane ultrafiltration‐concentration, sucrose density gradient ultracentrifugation, and preparative sodium dodecyl sulfate‐polyacrylamide gradient electrophoresis. An aqueous extract of the tissue from 300 eyes will yield about 10–20 μg of biologically active, electrophoretically pure CNTF with a specific activity of 7.5 × 106 trophic units/mg protein. Purified CNTF has an Mr of 20,400 daltons and an isoelectric point of about 5, as determined by analytical gel electrophoresis. In addition to supporting the survival of ciliary ganglion neurons, purified CNTF also supports the 24‐h survival of cultured neurons from certain chick and rodent sensory and sympathetic ganglia. CNTF differs from mouse submaxillary nerve growth factor (NGF) in molecular weight, isoelectric point, inability to be inactivated by antibodies to NGF, ability to support the in vitro survival of the ciliary ganglion neurons, and inability to support that of 8‐day chick embryo dorsal root ganglionic neurons. Thus, CNTF represents the first purified neuronotrophic factor which addresses parasympathetic cholinergic neurons. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Neurochemistry Wiley

Purification of the Chick Eye Ciliary Neuronotrophic Factor

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Publisher
Wiley
Copyright
Copyright © 1984 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0022-3042
eISSN
1471-4159
DOI
10.1111/j.1471-4159.1984.tb05410.x
Publisher site
See Article on Publisher Site

Abstract

Abstract: Dissociated 8‐day chick embryo ciliary ganglionic neurons will not survive for even 24 h in culture without the addition of specific supplements. One such supplement is a protein termed the ciliary neuronotrophic factor (CNTF) which is present at very high concentrations within intraocular tissues that contain the same muscle cells innervated by ciliary ganglionic neurons in vivo. We describe here the purification of chick eye CNTF by a 2½‐day procedure involving the processing of intraocular tissue extract sequentially through DE52 ion‐exchange chromatography, membrane ultrafiltration‐concentration, sucrose density gradient ultracentrifugation, and preparative sodium dodecyl sulfate‐polyacrylamide gradient electrophoresis. An aqueous extract of the tissue from 300 eyes will yield about 10–20 μg of biologically active, electrophoretically pure CNTF with a specific activity of 7.5 × 106 trophic units/mg protein. Purified CNTF has an Mr of 20,400 daltons and an isoelectric point of about 5, as determined by analytical gel electrophoresis. In addition to supporting the survival of ciliary ganglion neurons, purified CNTF also supports the 24‐h survival of cultured neurons from certain chick and rodent sensory and sympathetic ganglia. CNTF differs from mouse submaxillary nerve growth factor (NGF) in molecular weight, isoelectric point, inability to be inactivated by antibodies to NGF, ability to support the in vitro survival of the ciliary ganglion neurons, and inability to support that of 8‐day chick embryo dorsal root ganglionic neurons. Thus, CNTF represents the first purified neuronotrophic factor which addresses parasympathetic cholinergic neurons.

Journal

Journal of NeurochemistryWiley

Published: Nov 1, 1984

References

  • The nerve growth factor: biochemistry, synthesis and mechanism of action
    Greene, Greene; Shooter, Shooter
  • A series of normal stages in the development of the chick embryo
    Hamburger, Hamburger; Hamilton, Hamilton
  • Nerve growth factor: biological significance, measurement, and distribution
    Harper, Harper; Thoenen, Thoenen
  • Cholinergic neuronotrophic factors: Fractionation properties of an extract from selected chick embryonic eye tissues
    Manthorpe, Manthorpe; Skaper, Skaper; Adler, Adler; Landa, Landa; Varon, Varon
  • Neurite‐promoting factor in conditioned medium from RN22 Schwannoma cultures: bioassay, fractionation, and properties
    Manthorpe, Manthorpe; Varon, Varon; Adler, Adler
  • Isoelectric focusing of the chick eye ciliary neuronotrophic factor
    Manthorpe, Manthorpe; Barbin, Barbin; Varon, Varon
  • Cholinergic neuronotrophic factors. Concurrent activities on certain nerve growth factor‐responsive neurons
    Manthorpe, Manthorpe; Skaper, Skaper; Barbin, Barbin; Varon, Varon
  • Characterization of chick gizzard extract that promotes neurite outgrowth in cultured ciliary neurons
    Miki, Miki; Hayashi, Hayashi; Higashida, Higashida
  • Nerve growth factors and control of nerve growth
    Varon, Varon; Adler, Adler

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