Neurokinin release in the rat nucleus of the solitary tract via NMDA and AMPA receptors.
Neurokinins (substance P, neurokinin A and neurokinin B) and the neurokinin receptors, the NK1 and NK3 receptors, are largely expressed in the nucleus of the solitary tract (NST) where they are involved in the central regulation of visceral function. Studying the mechanisms that control neurokinin release can provide valuable information concerning the control of autonomic functions subserved by the NST. Glutamate is the principal excitatory neurotransmitter in the NST and the main neurotransmitter of afferent vagal fibers. Neurokinins and glutamate may interact within the NST. In the present study, we have examined the contribution of the N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) subtypes of glutamate receptors on the release of the endogenous neurokinins in the NST. We used internalization of the NK1 or NK3 receptor as an index of endogenous neurokinin release assessed by immunocytochemical visualization of the NK1 or NK3 receptor endocytosis. Experiments were performed in vitro using rat brainstem slices. A first series of experiments were done in order to validate our in vitro preparation. Application of substance P, neurokinin A or neurokinin B induced dose-dependent internalization of NK1 and NK3 receptor. This was blocked by the endocytosis inhibitor, phenylarzine oxide. The NK1 receptor antagonist SR140333 blocked internalization of NK1 receptor induced by the three neurokinins. In addition, the internalization NK1 or NK3 receptor was reversible. These results demonstrate that internalization and recycling mechanisms of NK1 or NK3 receptor were preserved in in vitro brainstem slices. Application of NMDA or AMPA induced internalization of NK1 receptor. This was blocked by the application of SR140333 suggesting that NK1 receptor internalization is due to the binding of endogenous neurokinin released under the effects of NMDA and AMPA. Application of NMDA or AMPA had no effect on NK3 receptor. Application of tetrodotoxin blocked NK1 receptor internalization induced by NMDA, demonstrating that the release of neurokinins is dependent of axon potential propagation. This result excludes the hypothesis of a release on neurokinins via pre-synaptic NMDA receptors located on neurokinin-containing axon terminals. NMDA or AMPA may directly induce neurokinin release in the NST by acting on receptors located on the cell bodies and dendrites of neurokinin-containing neurons. Release of neurokinins may also be the result of a general activation of neuron networks of the NST by NMDA or AMPA. To conclude, our results suggest that glutamate, through activation of post-synaptic NMDA and AMPA receptors, contributes to neurokinin signaling in the NST.
Colin
I
I
ITIS, CNRS, UMR 6150, Batiment N', 31 chemin Joseph Aiguier, 13402 Cedex 20, Marseille, France.
Blondeau
C
C
Baude
A
A
D000818:Animals; D003712:Dendrites / Q000187:drug effects / Q000378:metabolism; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D018698:Glutamic Acid / Q000378:metabolism; D007150:Immunohistochemistry; D016202:N-Methylaspartate / Q000494:pharmacology; D015288:Neurokinin A / Q000378:metabolism; D015287:Neurokinin B / Q000378:metabolism; D017729:Presynaptic Terminals / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D018091:Receptors, AMPA / Q000187:drug effects / Q000378:metabolism*; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects / Q000378:metabolism*; D018040:Receptors, Neurokinin-1 / Q000378:metabolism*; D018042:Receptors, Neurokinin-3 / Q000378:metabolism*; D017552:Solitary Nucleus / Q000187:drug effects / Q000378:metabolism*; D013373:Substance P / Q000378:metabolism; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology; D014630:Vagus Nerve / Q000187:drug effects / Q000378:metabolism; D017833:Visceral Afferents / Q000187:drug effects / Q000378:metabolism*; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid / Q000494:pharmacology
D018691:Excitatory Amino Acid Antagonists; D018091:Receptors, AMPA; D016194:Receptors, N-Methyl-D-Aspartate; D018040:Receptors, Neurokinin-1; D018042:Receptors, Neurokinin-3; D013373:Substance P; D018698:Glutamic Acid; D016202:N-Methylaspartate; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; D015288:Neurokinin A; D015287:Neurokinin B