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Neurochemical Research

, Volume 12, Issue 10, pp 839–850 | Cite as

Induction of the high-affinity nerve growth factor receptor on embryonic chicken sensory nerve cells by elevated potassium

  • David J. Ennulat
  • Robert W. Stach
Original Articles

Abstract

Culture medium with elevated K+ has been shown to enhance the survival of neurons isolated from several different regions of the nervous system. Nerve growth factor binds to binding sites on sensory and sympathetic neurons through two sites, one of high-affinity (Kd1∼3×10−11 M) and the other of low-affinity (Kd2∼2×10−9 M). Equilibrium binding data generated on dissociated cells derived from E9 chicken embryo dorsal root ganglia, has shown that there is a two-fold increase in the number of high affinity (type I) receptors, with no effect on the affinity, when cells are incubated for 2 hours in buffer containing 59 mM K+. There does not appear to be a significant change in the affinity or the number of low-affinity binding sites. This two-fold increase in type I receptors is dependent on temperature, Ca2+, and active protein synthesis. There does not appear to be an intracellular pool of the type I receptor sufficient to account for this increase. The induction is not observed on sensory nerve cells cultured in 59 mM K+ for 24 hours, either in the presence or absence of nerve growth factor. Additionally, the induction in the number of type I receptors requires that both nerve growth factor and K+ be present simultaneously. Taken in total, this data suggests that there may be a critical period in which the sensory neurons require nerve growth factor exposure to respond. Evidence is presented which indicates that nerve growth factor responsive cells are able to elicit neurites after an acute exposure to nerve growth factor of as little as 4 hours. Finally, there is an approximate two-fold decrease in the concentration of nerve growth factor needed to elicit maximal fiber outgrowth, consistent with the two-fold increase in the number of type I receptors.

Key Words

Nerve growth factor receptor induction 

Abbreviations

NGF

nerve growth factor

7S NGF

the high molecular weight form of NGF

βNGF

the β-subunit of 7S NGF

125I-βNGF

125I-labeled βNGF

mβNGF−rAb

polyclonal rabbit IgG raised against mouse βNGF

DRG

dorsal root ganglia

Kd

the equilibrium dissociation constant

N

the maximal number of binding sites for the ligand βNGF

βNGFR

the biologically relevant receptor through which the neurite outgrowth and neuron survival are mediated

GBS

Gey's balanced salts

HKGBS

high K+ GBS

PBG

phosphate buffered GBS

HKPBG

high K+ PBG

CFHKPBG

Ca+2 free high K+ PBG

PBG-cyt c

PBG containing 2 mg/ml cytochrome c

HKPBG-cyt c

HKPBG containing 2 mg/ml cytochrome c

AbU

antibody unit

BU

biological unit PBS, phosphate buffered saline

HKPBS

high K+ PBS

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Copyright information

© Plenum Publishing Corporation 1987

Authors and Affiliations

  • David J. Ennulat
    • 1
  • Robert W. Stach
    • 1
  1. 1.Departments of Biochemistry & Molecular Biology and Anatomy & Cell BiologyState University of New York, Health Science CenterSyracuse

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