Abstract
Substance P (SP) is a key neurotransmitter involved in the brain stem integration of carotid body chemoreceptor reflexes. In this study, the characteristics and location of SP receptors in the rat brain stem and their regulation by hypoxia were investigated using homogenate radioligand binding and quantitative autoradiography. Specific binding of [125I] Bolton-Hunter SP (BHSP) to brain stem homogenates was saturable (∼0.3 nM) and to a single class of high-affinity sites (Kd , 0.16 nM; maximum density of binding sites, 0.43 fmol/mg wet weight tissue). The order of potency of agonists for inhibition of BHSP binding was SP > [Sar9Met(O2)11]SP ≫ neurokinin A > septide > neurokinin B ≫ [Nle10]-neurokinin A(4–10) = senktide, and for nonpeptide antagonists, RP 67580 > CP-96,345 ≫ RP 68651 = CP-96,344, consistent with binding to NK1 receptors. The effect of single and multiple, 5-min bouts of hypoxia (8.5% O2/91.5% N2) on BHSP binding was investigated using quantitative autoradiography. Binding sites were localized to the lateral, medial and commissural nucleus of the solitary tract (NTS), the hypoglossal nucleus, central gray and the spinal trigeminal tract and nucleus (Sp5 and nSp5, respectively). Five min after a single bout of hypoxia, the density of BHSP binding sites had decreased significantly (P < .05) in the medial NTS (−33%) and lateral NTS (−24%) when compared to normoxic controls. However, the normal receptor complement was restored within 60 min of the hypoxic challenge. In the Sp5, a significant decrease (P < .05) in binding was observed 5 min after hypoxia which was still apparent after 60 min. In contrast, the density of BHSP binding sites in the hypoglossal nucleus decreased slowly and was significantly lower (P < .05) than normoxic controls 60 min after hypoxia. Five min after repetitive hypoxia (3 × 5 min bouts), BHSP binding in the NTS was reduced by more than 40%. Studies in homogenates showed that the affinity of SP for BHSP binding sites was not affected by repetitive hypoxia (Kd s, normoxic, 0.27 nM; hypoxic, 0.24 nM). These data suggest that afferent input from carotid body chemoreceptors may dynamically regulate NK1 receptors in several brain stem nuclei that are intimately involved in stimulating ventilation during hypoxia, and that the time-course of receptor turnover may differ from region to region in the brain stem. The temporary loss of NK1 receptors in the NTS may partly explain why adequate ventilation is often not maintained during hypoxia.
Footnotes
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Send reprint requests to: Dr. Dominic P. Geraghty, Department of Biomedical Science, University of Tasmania at Launceston, P.O. Box 1214, Launceston, Tasmania 7250, Australia.
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↵1 This work was supported by the National Health and Medical Research Council of Australia (D.P.G.) and the National SIDS Council of Australia (D.P.G., C.F.H.).
- Abbreviations:
- BHSP
- [125I]Bolton-Hunter substance P
- CNS
- central nervous system
- cNTS
- commissural nucleus of the solitary tract
- mNTS
- medial nucleus of the solitary tract
- lNTS
- lateral nucleus of the solitary tract
- f
- respiratory frequency
- NK1
- neurokinin 1
- NKA
- neurokinin A
- NKB
- neurokinin B
- CG
- central gray
- PBN
- parabrachial nucleus
- SIDS
- sudden infant death syndrome
- SP
- substance P
- SPLI
- substance P-like immunoreactivity
- Tbc
- body core temperature
- V
- minute ventilation
- Vt
- tidal volume
- Bmax
- maximum density of binding sites
- BSA
- bovine serum albumin
- HGN
- hypoglossal nucleus
- Sp5
- spinal trigeminal tract
- Received December 4, 1996.
- Accepted April 10, 1997.
- The American Society for Pharmacology and Experimental Therapeutics
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