Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Serotonergic mechanisms on breathing modulation in the rat locus coeruleus

  • 227 Accesses

  • 15 Citations

Abstract

The locus coeruleus (LC) is a noradrenergic nucleus that plays an important role in the ventilatory response to hypercapnia. This nucleus is densely innervated by serotonergic fibers and contains high density of serotonin (5-HT) receptors, including 5-HT1A and 5-HT2. We assessed the possible modulation of respiratory response to hypercapnia by 5-HT, through 5-HT1A and 5-HT2 receptors, in the LC. To this end, we determined the concentrations of 5-HT and its metabolite 5-hydroxyindole-3-acetic acid (5-HIAA) in the LC after hypercapnic exposure. Pulmonary ventilation (V e, plethysmograph) was measured before and after unilateral microinjection (100 nL) of WAY-100635 (5-HT1A antagonist, 5.6 and 56 mM), 8-OHDPAT (5-HT1A/7 agonist, 7 and 15 mM), Ketanserin (5-HT2A antagonist, 3.7 and 37 mM), or (±)-2,5-dimethoxy-4-iodoamphetaminehydrochloride (DOI; 5-HT2A agonist, 6.7 and 67 mM) into the LC, followed by a 60-min period of 7% CO2 exposure. Hypercapnia increased 5-HTIAA levels and 5-HIAA/5-HT ratio within the LC. WAY-100635 and 8-OHDPAT intra-LC decreased the hypercapnic ventilatory response due to a lower tidal volume. Ketanserin increased CO2 drive to breathing and DOI caused the opposite response, both acting on tidal volume. The current results provide evidence of increased 5-HT release during hypercapnia in the LC and that 5-HT presents an inhibitory modulation of the stimulatory role of LC on hypercapnic ventilatory response, acting through postsynaptic 5-HT2A receptors in this nucleus. In addition, hypercapnic responses seem to be also regulated by presynaptic 5-HT1A receptors in the LC.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

References

  1. 1.

    Aguiar CL, Romcy-Pereira RN, Szawka RE, Galvis-Alonso OY, Anselmo-Franci JA, Leite JP (2008) Muscarinic acetylcholine neurotransmission enhances the late-phase of long-term potentiation in the hippocampal-prefrontal cortex pathway of rats in vivo: a possible involvement of monoaminergic systems. Neuroscience 153:1309–1319

  2. 2.

    Astier B, Van Bockstaele EJ, Aston-Jones G, Pieribone VA (1990) Anatomical evidence for multiple pathways leading from the rostral ventrolateral medulla (nucleus paragigantocellularis) to the locus coeruleus in rat. Neurosci Lett 118(2):141–146

  3. 3.

    Aston-Jones G, Rajkowski J, Kubiak P, Valentino RJ, Shipley MT (1996) Role of the locus coeruleus in emotional activation. Prog Brain Res 107:379–402

  4. 4.

    Aston-Jones G, Shipley MT, Chouvet G, Ennis M, van Bockstaele E, Pieribone V, Shiekhattar R, Akaoka H, Drolet G, Astier B et al (1991) Afferent regulation of locus coeruleus neurons: anatomy, physiology and pharmacology. Prog Brain Res 88:47–75

  5. 5.

    Bartlett D, Tenney SM (1970) Control of breathing in experimental anemia. Respir Physiol 10:384–395

  6. 6.

    Berridge CW, Waterhouse BD (2003) The locus coeruleus-noradrenergic system: modulation of behavioral state and state-dependent cognitive processes. Brain Res Brain Res Rev 42(1):33–84

  7. 7.

    Biancardi V, Bícego KC, Almeida MC, Gargaglioni LH (2008) Locus coeruleus noradrenergic neurons and CO(2) drive to breathing. Pflugers Arch 455(6):1119–1128

  8. 8.

    Blandina P, Goldfarb J, Walcott J, Green JP (1991) Serotonergic modulation of the release of endogenous norepinephrine from rat hypothalamic slices. J Pharmacol Exp Ther 256(1):341–347

  9. 9.

    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

  10. 10.

    Brown CM, Mackinnon AC, Mcgrath JC, Spedding M, Kilpatrick AT (1990) α2-Adrenoceptor subtypes and imidazoline-like binding sites in the rat brain. Br J Pharmacol 99:803–809

  11. 11.

    Cedarbaum JM, Aghajanian GK (1978) Afferent projections to the rat locus coeruleus as determined by a retrograde tracing technique. J Comp Neurol 178(1):1–16

  12. 12.

    Chiang C, Aston-Jones G (1993) 5-Hydroxytryptamine2 agonist augments gamma-aminobutyric acid and excitatory amino acid inputs to noradrenergic locus coeruleus neurons. Neuroscience 54(2):409–420

  13. 13.

    Crespi F, Buda M, Mcrae-Degueurce A, Pujol JF (1980) Alteration in tyrosine hydroxylase activity in the LC after administration of P-chlorophenylalanine. Brain Res 9:501–509

  14. 14.

    Curet O, de Montigny C (1989) Electrophysiological characterization of adrenoceptors in the rat dorsal hippocampus. III. Evidence for the physiological role of terminal alpha 2-adrenergic autoreceptors. Brain Res 499:18–26

  15. 15.

    Dejours P (1981) Principle of comparative respiratory physiology, 2nd edn. Elsevier, New York

  16. 16.

    Dias MB, Nucci TB, Margatho LO, Antunes-Rodrigues J, Gargaglioni LH, Branco LG (2007) Raphe magnus nucleus is involved in ventilatory but not hypothermic response to CO2. J Appl Physiol 103:1780–1788

  17. 17.

    Dobbins EG, Feldman JL (1994) Brainstem network controlling descending drive to phrenic motoneurons in rat. J Comp Neurol 347(1):64–86

  18. 18.

    Done CJG, Sharp T (1992) Evidence that 5-HT2 receptor activation decreases noradrenaline release in rat hippocampus in vivo. Br J Pharmacol 107:240–245

  19. 19.

    Elam M, Yao T, Thoren P, Svensson TH (1981) Hypercapnia and hypoxia: chemoreceptor-mediated control of locus coeruleus neurons and splanchnic, sympathetic nerves. Brain Res 222:373–381

  20. 20.

    Filosa JA, Dean JB, Putnam RW (2002) Role of intracellular and extracellular pH in the chemosensitive response of rat locus coeruleus neurones. J Physiol 541:493–509

  21. 21.

    Foote SL, Bloom FE, Aston-Jones G (1983) Nucleus locus ceruleus: new evidence of anatomical and physiological specificity. Physiol Rev 63:844–914

  22. 22.

    Gargaglioni LH, Steiner AA, Branco LG (2005) Involvement of serotoninergic receptors in the anteroventral preoptic region on hypoxia-induced hypothermia. Brain Res 1044(1):16–24

  23. 23.

    Grzanna R, Molliver ME (1980) The locus coeruleus in the rat: an immunohistochemical delineation. Neuroscience 5:21–40

  24. 24.

    Haddjeri N, de Montigny C, Blier P (1997) Modulation of the firing activity of noradrenergic neurones in the rat locus coeruleus by the 5-hydroxtryptamine system. Br J Pharmacol 120:865–875

  25. 25.

    Haxhiu MA, Yung K, Erokwu B, Cherniack NS (1996) CO2-induced c-fos expression in the CNS catecholaminergic neurons. Respir Physiol 105:35–45

  26. 26.

    Hedlund U, Järvholm B, Lundbäck B (2004) Respiratory symptoms and obstructive lung diseases in iron ore miners: report from the obstructive lung disease in northern Sweden studies. Eur J Epidemiol 19(10):953–958

  27. 27.

    Hilaire G, Viemari JC, Coulon P, Simonneau M, Bévengut M (2004) Modulation of the respiratory rhythm generation by the pontine A5 and A6 groups in rodents. Resp Physiol Neurobiol 143:187–197

  28. 28.

    Hodges MR, Richerson GB (2008) Interaction between defects in ventilatory and thermoregulatory control in mice lacking 5-HT neurons. Respir Physiol Neurobiol 164(3):350–357

  29. 29.

    Hodges MR, Tattersall GJ, Harris MB, McEvoy SD, Richerson DN, Deneris ES, Johnson RL, Chen ZF, Richerson GB (2008) Defects in breathing and 40 thermoregulation in mice with near-complete absence of central serotonin neurons. J Neurosci 28:2495–2505

  30. 30.

    Johnson SM, Haxhiu MA, Richerson GB (2008) GFP expressing locus coeruleus neurons from Prp57 transgenic mice exhibit CO2/H+ responses in primary cell culture. J Appl Physiol 105(4):1301–1311

  31. 31.

    Kaehler ST, Singewald N, Philippu A (1999) Dependence of serotonin release in the locus coeruleus on dorsal raphe neuronal activity. Naunyn Schmiedebergs Arch Pharmacol 359(5):386–393

  32. 32.

    Kanamaru M, Homma I (2007) Compensatory airway dilation and additive ventilatory augmentation mediated by dorsomedial medullary 5-hydroxytryptamine 2 receptor activity and hypercapnia. Am J Physiol Regul Integr Comp Physiol 293(2):R854–R860

  33. 33.

    Kim MA, Lee HS, Lee BY, Waterhouse BD (2004) Reciprocal connections between subdivisions of the dorsal raphe and the nuclear core of the locus coeruleus in the rat. Brain Res 1026(1):56–67

  34. 34.

    Leger L, Descarries L (1978) Serotonin nerve terminals in the locus coeruleus of adult rat: a radioautographic study. Brain Res 145:1–13

  35. 35.

    Li A, Nattie E (2008) Serotonin transporter knockout mice have a reduced ventilatory response to hypercapnia (predominantly in males) but not to hypoxia. J Physiol 586(9):2321–2329

  36. 36.

    Malan A (1973) Ventilation measured by body plethysmography in hibernating mammals and in poikilotherms. Respir Physiol 17(1):32–44

  37. 37.

    Matsumoto M, Yioshioka M, Togashi H, Tochihara M, Ikeda T, Saito H (1995) Modulation of norepinephrine release by serotonergic receptors in the rat hippocampus as measured by in vivo microdialysis. J Pharmacol Exp Ther 272:1044–1051

  38. 38.

    McRae-Degueurce A, Berod A, Mermet A, Keller A, Chouvet G, Joh TH, Pujol JF (1982) Alterations in tyrosine hydroxylase activity elicited by raphe nuclei lesions in the rat locus coeruleus: evidence for the involvement of serotonin afferents. Brain Res 235(2):285–301

  39. 39.

    Mongeau R, De Montigny C, Blier P (1994) Effect of long-term administration of antidepressant drugs on the 5-HT3 receptors that enhance the electrically evoked release of [3H]noradrenaline in the rat hippocampus. Eur J Pharmacol 271(1):121–129

  40. 40.

    Nattie EE, Li A (2008) Muscimol dialysis into the caudal aspect of the nucleus tractus solitarii of conscious rats inhibits chemoreception. Respir Physiol Neurobiol 164(3):394–400

  41. 41.

    Nosjean A, Guyenet PG (1991) Role of ventrolateral medulla in sympatholytic effect of 8-OHDPAT in rats. Am J Physiol 260(3 Pt 2):R600–R609

  42. 42.

    Nucci TB, Branco LGS, Gargaglioni LH (2008) 5-HT1A but not 5HT2 and 5-HT7 receptors in the nucleus raphe magnus. Acta Physiologica Scandinavica 193:403–414

  43. 43.

    Oyamada Y, Ballantyne D, Muckenhoff K, Scheid P (1998) Respiration-modulated membrane potential and chemosensitivity of locus coeruleus neurones in the in vitro brainstem-spinal cord of the neonatal rat. J Physiol 513:381–398

  44. 44.

    Palkovits M (1973) Isolated removal of hypothalamic or other brain nuclei of the rat. Brain Res 59:449–450

  45. 45.

    Paxinos G, Watson C (1998) The rat brain in stereotaxic coordinates. Academic, New York

  46. 46.

    Paxinos G, Watson C (2005) The rat brain in stereotaxic coordinates, 5th edn. Academic, San Diego

  47. 47.

    Pickel VM, Joh TH, Reis DJ (1977) A serotonergic innervation of noradrenergic neurons in nucleus locus coeruleus: demonstration by immunocytochemical localization of the transmitter specific enzymes tyrosine and tryptophan hydroxylase. Brain Res 131(2):197–214

  48. 48.

    Pieribone VA, Aston-Jones G (1991) Adrenergic innervation of the rat nucleus locus coeruleus arises predominantly from the C1 adrenergic cell group in the rostral medulla. Neuroscience 41(2-3):525–542

  49. 49.

    Poliacek I, Halasova E, Jakus J, Murin P, Barani H, Stransky A, Bolser DC (2007) Brainstem regions involved in the expiration reflex. A c-fos study in anesthetized cats. Brain Res 1184:168–177

  50. 50.

    Poncet L, Denoroy L, Dalmaz Y, Pequignot JM (1997) Activity of tryptophan hydroxylase and content of indolamines in discrete brain regions after a long-term hypoxic exposure in the rat. Brain Res 765(1):122–128

  51. 51.

    Pudovkina OL, Cremers TI, Westerink BH (2002) The interaction between the locus coeruleus and dorsal raphe nucleus studied with dual-probe microdialysis. Eur J Pharmacol 445(1-2):37–42

  52. 52.

    Reader TA, Briere R, Grondin L, Ferron A (1986) Efects of p-chlorophenylalanine on cortical monamines and on electrical activity of noradrenergic neurons. Neurochem Res 11:1025–1035

  53. 53.

    Shannon NJ, Gunnet JW, Moore KE (1986) A comparison of biochemical indices of 5-hydroxytryptaminergic neuronal activity following electrical stimulation of the dorsal raphe nucleus. J Neurochem 47:958–965

  54. 54.

    Stunden CE, Filosa JA, Garcia AJ, Dean JB, Putnam RW (2001) Development of in vivo ventilatory and single chemosensitive neuron responses to hypercapnia in rats. Respir Physiol 127(2–3):135–155

  55. 55.

    Stupfel M (1974) Carbon dioxide and temperature regulation of homeothermic mammals. In: Nahas G, Schaefer KE (eds) Carbon dioxide and metabolic regulations. Springer, New York, pp 163–186

  56. 56.

    Svensson TH, Bunney BS, Aghajanian GK (1975) Inhibition of both NA and 5-HT neurons in brain by the α-adrenergic agonist clonidine. Brain Res 92:291–306

  57. 57.

    Szabo ST, Blier P (2001) Effect of the selective noradrenergic reuptake inhibitor reboxetine on the firing activity of noradrenaline and serotonin neurons. Eur J Neurosci 13(11):2077–2087

  58. 58.

    Szabo ST, Blier P (2002) Effects of serotonin (5-hydroxytryptamine, 5-HT) reuptake inhibition plus 5-HT(2A) receptor antagonism on the firing activity of norepinephrine neurons. J Pharmacol Exp Ther 302(3):983–991

  59. 59.

    Szawka RE, Rodovalho GV, Helena CV, Franci CR, Anselmo-Franci JA (2007) Prolactin secretory surge during estrus coincides with increased dopamine activity in the hypothalamus and preoptic area and is not altered by ovariectomy on proestrus. Brain Res Bull 73:127–134

  60. 60.

    Teppema LJ, Veening JG, Kranenburg A, Dahan A, Berkenbosch A, Olievier C (1997) Expression of c-fos in the rat brainstem after exposure to hypoxia and to normoxic and hyperoxic hypercapnia. J Comp Neurol 388:169–190

  61. 61.

    Viemari JC, Bévengut M, Burnet H, Coulon P, Pequignot JM, Tiveron MC, Hilaire G (2004) Phox2a gene, A6 neurons, and noradrenaline are essential for development of normal respiratory rhythm in mice. J Neurosci 24:928–937

Download references

Acknowledgments

This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and Instituto Nacional de Ciência e Tecnologia (INCT) em Fisiologia Comparada. We thank Aretuza C. Carregari, Euclides Roberto Secato, and Ruither O.G. Carolino for excellent technical assistance and Dr. Patricia M. de Paula that kindly provided Idazoxan. Vanessa de Souza Moreno was the recipient of a FAPESP undergraduate scholarship (2005/56128-9).

Author information

Correspondence to Luciane H. Gargaglioni.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

de Souza Moreno, V., Bícego, K.C., Szawka, R.E. et al. Serotonergic mechanisms on breathing modulation in the rat locus coeruleus. Pflugers Arch - Eur J Physiol 459, 357–368 (2010). https://doi.org/10.1007/s00424-009-0741-4

Download citation

Keywords

  • Chemosensitivity
  • A6 region
  • Hypercapnia
  • Ventilation
  • Pons
  • Serotonin
  • 5-HT1A
  • 5HT2A