Nerve Growth Factor Modulates Synaptic Transmission between Sympathetic Neurons and Cardiac Myocytes

Regulation of heart rate by the sympathetic nervous system involves the release of norepinephrine (NE) from nerve terminals onto heart tissue, resulting in an elevation in beat rate. Nerve growth factor (NGF) is a neurotrophin produced by the heart that supports the survival and differentiation of sympathetic neurons. Here we report that NGF also functions as a modulator of sympathetic synaptic transmission. We determined the effect of NGF on the strength of synaptic transmission in co-cultures of neonatal rat cardiac myocytes and sympathetic neurons from the superior cervical ganglion (SCG). Synaptic transmission was assayed functionally, as an increase in the beat rate of a cardiac myocyte during stimulation of a connected neuron. Application of NGF produced a pronounced, reversible enhancement of synaptic strength. We found that TrkA, the receptor tyrosine kinase that mediates many NGF responses, is expressed primarily by neurons in these cultures, suggesting a presynaptic mechanism for the effects of NGF. A presynaptic model is further supported by the finding that NGF did not alter the response of myocytes to application of NE. In addition to the acute modulatory effects of NGF, we found that the concentration of NGF in the growth medium affects the level of synaptic transmission in cultures of sympathetic neurons and cardiac myocytes. These results indicate that in addition to its role as a survival factor, NGF plays both acute and long-term roles in the regulation of developing sympathetic synapses in the cardiac system.

[1]  G. Yancopoulos,et al.  The low affinity NGF receptor, p75, can collaborate with each of the Trks to potentiate functional responses to the neurotrophins , 1994, Neuron.

[2]  M. Schwab,et al.  Selective induction of tyrosine hydroxylase and dopamine β-hydroxylase by nerve growth factor: comparison between adrenal medulla and sympathetic ganglia of adult and newborn rats , 1977, Brain Research.

[3]  F. Mudry,et al.  Conditions increasing the adrenergic properties of dissociated chick superior cervical ganglion neurons grown in long-term culture. , 1986, Developmental biology.

[4]  T. Yamashita,et al.  Sympathetic nerve disturbance and fine structure of adrenergic nerve terminals in nasal mucosa after experimentally-induced nasal hypersensitivity. , 1993, Acta oto-laryngologica. Supplementum.

[5]  L. Conforti,et al.  Influence of sympathetic innervation on the membrane electrical properties of neonatal rat cardiomyocytes in culture. , 1991, Journal of developmental physiology.

[6]  D. Purves.  Body and Brain: A Trophic Theory of Neural Connections , 1988 .

[7]  A. Davies,et al.  Regulation of nerve growth factor receptor gene expression in sympathetic neurons during development , 1995, The Journal of cell biology.

[8]  Nancy Y. Ip,et al.  Potentiation of developing neuromuscular synapses by the neurotrophins NT-3 and BDNF , 1993, Nature.

[9]  R. Dobrowsky,et al.  Neurotrophins Induce Sphingomyelin Hydrolysis , 1995, The Journal of Biological Chemistry.

[10]  M. Schwab,et al.  Induction of tyrosine hydroxylase by nerve growth factor and by elevated K+ concentrations in cultures of dissociated sympathetic neurons , 1982, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[11]  R. Campenot,et al.  Development of sympathetic neurons in compartmentalized cultures. Il Local control of neurite growth by nerve growth factor. , 1982, Developmental biology.

[12]  M. Rydén,et al.  Differential Modulation of Neuron Survival during Development by Nerve Growth Factor Binding to the p75 Neurotrophin Receptor* , 1997, The Journal of Biological Chemistry.

[13]  R. Dobrowsky,et al.  Activation of the sphingomyelin cycle through the low-affinity neurotrophin receptor. , 1994, Science.

[14]  G G Turrigiano,et al.  Brain-Derived Neurotrophic Factor Mediates the Activity-Dependent Regulation of Inhibition in Neocortical Cultures , 1997, The Journal of Neuroscience.

[15]  I. Black,et al.  Brain-derived neurotrophic factor rapidly enhances synaptic transmission in hippocampal neurons via postsynaptic tyrosine kinase receptors. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[16]  D. Anderson,et al.  Sympathetic neuroblasts undergo a developmental switch in trophic dependence. , 1993, Development.

[17]  H. Thoenen,et al.  Physiology of nerve growth factor. , 1980, Physiological reviews.

[18]  H. G. Kim,et al.  Neurotrophin 3 potentiates neuronal activity and inhibits gamma-aminobutyratergic synaptic transmission in cortical neurons. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[19]  H. C. Hartzell,et al.  Mechanisms of slow postsynaptic potentials , 1981, Nature.

[20]  M. Chao,et al.  Expression and structure of the human NGF receptor , 1986, Cell.

[21]  R. Rush,et al.  Elevated concentrations of nerve growth factor in heart and mesenteric arteries of spontaneously hypertensive rats , 1993, Brain Research.

[22]  M. Barbacid,et al.  K252a is a selective inhibitor of the tyrosine protein kinase activity of the trk family of oncogenes and neurotrophin receptors. , 1992, Oncogene.

[23]  H. Thoenen,et al.  Nerve growth factor changes the relative levels of neuropeptides in developing sensory and sympathetic ganglia of the chick embryo. , 1985, Developmental biology.

[24]  D. Lowenstein,et al.  Neurotrophins and Their Receptors—Current Concepts and Implications for Neurologic Disease , 1993, Experimental Neurology.

[25]  Marlies Knipper,et al.  Positive Feedback between Acetylcholine and the Neurotrophins Nerve Growth Factor and Brain‐derived Neurotrophic Factor in the Rat Hippocampus , 1994, The European journal of neuroscience.

[26]  L. Reichardt,et al.  Expression of the beta-nerve growth factor gene correlates with the density of sympathetic innervation in effector organs. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[27]  Dale Purves,et al.  Trophic regulation of nerve cell morphology and innervation in the autonomic nervous system , 1988, Nature.

[28]  William J. Rutter,et al.  Directed expression of NGF to pancreatic β cells in transgenic mice leads to selective hyperinnervation of the islets , 1989, Cell.

[29]  N. Matsuki,et al.  Inhibition of GABAA Synaptic Responses by Brain-Derived Neurotrophic Factor (BDNF) in Rat Hippocampus , 1997, The Journal of Neuroscience.

[30]  D. Kaplan,et al.  The trk proto-oncogene product: a signal transducing receptor for nerve growth factor. , 1991, Science.

[31]  Takayoshi Kobayashi,et al.  Fine structure of the synaptic endings between sympathetic axons and skeletal muscle cells and of the varicosities in the bundles of neurites in tissue culture , 1984, Experimental Neurology.

[32]  I. Black,et al.  NT-3 stimulates sympathetic neuroblast proliferation by promoting precursor survival , 1993, Neuron.

[33]  J. Mallet,et al.  Regulation of neurotransmitter metabolic enzymes and tyrosine hydroxylase mRNA level by nerve growth factor in cultured sympathetic neurones. , 1988, Development.

[34]  J. Fawcett,et al.  Regressive events in neurogenesis. , 1984, Science.

[35]  Y. Barde,et al.  The Neurotrophin Receptor p75 Binds Neurotrophin-3 on Sympathetic Neurons with High Affinity and Specificity , 1997, The Journal of Neuroscience.

[36]  R. Campenot,et al.  Concentration-dependent regulation of neuronal gene expression by nerve growth factor , 1992, The Journal of cell biology.

[37]  R. Campenot,et al.  Development of sympathetic neurons in compartmentalized cultures. II. Local control of neurite survival by nerve growth factor. , 1982, Developmental biology.

[38]  Yasuhisa Endo,et al.  Brain‐Derived Neurotrophic Factor Increases the Stimulation‐Evoked Release of Glutamate and the Levels of Exocytosis‐Associated Proteins in Cultured Cortical Neurons from Embryonic Rats , 1997, Journal of neurochemistry.

[39]  C. Murakata,et al.  K-252 compounds, novel and potent inhibitors of protein kinase C and cyclic nucleotide-dependent protein kinases. , 1987, Biochemical and biophysical research communications.

[40]  O. Lindvall,et al.  Neurotrophins and their Receptors , 1992 .

[41]  H. Thoenen,et al.  Nerve growth factor-mediated induction of tyrosine hydroxylase in rat superior cervical ganglia in vitro. , 1978, The Journal of biological chemistry.

[42]  R. Levi‐montalcini,et al.  Nerve growth factor. , 1975, Science.

[43]  M. Washburn,et al.  Bodily Changes in Pain, Hunger, Fear, and Rage. , 1917 .

[44]  H. Thoenen,et al.  Timing and site of nerve growth factor synthesis in developing skin in relation to innervation and expression of the receptor , 1987, Nature.

[45]  M. Barbacid,et al.  The trk proto-oncogene encodes a receptor for nerve growth factor , 1991, Cell.

[46]  D. Clegg,et al.  Regulation of nerve growth factor mRNA levels in developing rat heart ventricle is not altered by sympathectomy. , 1989, Developmental biology.

[47]  R. Levi‐montalcini,et al.  Destruction of the sympathetic ganglia in mammals by an antiserum to a nerve-growth protein* , 1960, Proceedings of the National Academy of Sciences.

[48]  D. Potter,et al.  Synaptic functions in rat sympathetic neurons in microcultures. I. Secretion of norepinephrine and acetylcholine , 1986, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[49]  R. Levi‐montalcini,et al.  The nerve growth factor 35 years later. , 1987, Science.

[50]  M. Poo,et al.  Synaptic Modulation by Neurotrophic Factors: Differential and Synergistic Effects of Brain-Derived Neurotrophic Factor and Ciliary Neurotrophic Factor , 1996, The Journal of Neuroscience.

[51]  M. Parish,et al.  NGF modulates sympathetic innervation of lymphoid tissues , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[52]  R. Rush,et al.  Sympathetic neurons in neonatal rats require endogenous neurotrophin-3 for survival , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[53]  H. Thoenen,et al.  Selective induction by nerve growth factor of tyrosine hydroxylase and dopamine- -hydroxylase in the rat superior cervical ganglia. , 1971, Proceedings of the National Academy of Sciences of the United States of America.

[54]  R. Levi‐montalcini,et al.  Essential role of the nerve growth factor in the survival and maintenance of dissociated sensory and sympathetic embryonic nerve cells in vitro. , 1963, Developmental biology.

[55]  David J. Anderson,et al.  p75LNGFR regulates Trk signal transduction and NGF-induced neuronal differentiation in MAH cells , 1994, Neuron.

[56]  J. J. Steinberg,et al.  Overexpression of NGF within the heart of transgenic mice causes hyperinnervation, cardiac enlargement, and hyperplasia of ectopic cells. , 1995, Developmental biology.

[57]  E. Schuman,et al.  Long-lasting neurotrophin-induced enhancement of synaptic transmission in the adult hippocampus , 1995, Science.

[58]  B. Lu,et al.  Regulation of synaptic responses to high-frequency stimulation and LTP by neurotrophins in the hippocampus , 1996, Nature.

[59]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[60]  B. Davis,et al.  Overexpression of nerve growth factor in epidermis of transgenic mice causes hypertrophy of the peripheral nervous system , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[61]  A. Davies,et al.  Sympathetic neuron survival and TrkA expression in NT3‐deficient mouse embryos , 1997, The EMBO journal.

[62]  H. Thoenen,et al.  Developmental changes of nerve growth factor levels in sympathetic ganglia and their target organs. , 1988, Developmental biology.

[63]  David J. Anderson,et al.  Neurotrophins regulate sequential changes in neurotrophin receptor expression by sympathetic neuroblasts , 1994, Neuron.

[64]  P. Patterson,et al.  ROLE OF NERVE GROWTH FACTOR IN THE DEVELOPMENT OF RAT SYMPATHETIC NEURONS IN VITRO , 1977 .

[65]  E. Shooter,et al.  Gene transfer and molecular cloning of the rat nerve growth factor receptor , 1987, Nature.

[66]  S. Landis.  Rat sympathetic neurons and cardiac myocytes developing in microcultures: correlation of the fine structure of endings with neurotransmitter function in single neurons. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[67]  P. Distefano,et al.  Expression and possible function of nerve growth factor receptors on Schwann cells , 1988, Trends in Neurosciences.

[68]  D. Anderson,et al.  Expression of trk in MAH cells lacking the p75 low-affinity nerve growth factor receptor is sufficient to permit nerve growth factor-induced differentiation to postmitotic neurons. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[69]  D. Potter,et al.  Chemical transmission between rat sympathetic neurons and cardiac myocytes developing in microcultures: evidence for cholinergic, adrenergic, and dual-function neurons. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[70]  T Bonhoeffer,et al.  Hippocampal long-term potentiation is impaired in mice lacking brain-derived neurotrophic factor. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[71]  R. Zigmond,et al.  Regulation of Neuropeptide Expression in Sympathetic Neurons , 1997, Annals of the New York Academy of Sciences.

[72]  W D Snider,et al.  Nerve growth factor enhances dendritic arborization of sympathetic ganglion cells in developing mammals , 1988, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[73]  S. Landis,et al.  Morphological studies of synapses and varicosities in dissociated cell cultures of sympathetic neurons , 1983, Journal of neurocytology.

[74]  B. Lu,et al.  Neurotrophins promote maturation of developing neuromuscular synapses , 1995, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[75]  G. Barrett,et al.  The p75 nerve growth factor receptor mediates survival or death depending on the stage of sensory neuron development. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[76]  G. Yancopoulos,et al.  K-252a and staurosporine selectively block autophosphorylation of neurotrophin receptors and neurotrophin-mediated responses. , 1992, Molecular biology of the cell.

[77]  E. Hawrot,et al.  Long-term culture of dissociated sympathetic neurons. , 1979, Methods in enzymology.

[78]  H. Thoenen,et al.  Nerve growth factor in sympathetic ganglia and corresponding target organs of the rat: correlation with density of sympathetic innervation. , 1983, Proceedings of the National Academy of Sciences of the United States of America.