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Morphological and functional characterization of a rat vaginal smooth muscle sphincter

International Journal of Impotence Research volume 14, pages 271–282 (2002)Cite this article

Abstract

The aim of the present study was to gain information about adrenergic-, cholinergic- and non-adrenergic, non-cholinergic (NANC)- transmitter systems/mediators in the rat vagina, and to characterize its smooth muscles functionally. Tissue sections from vagina of Sprague Dawley rats were immunolabelled with antibodies against protein gene product 9.5 (PGP), synaptophysin (Syn), tyrosine hydroxylase (TH), vesicular acetylcholine transporter (VAChT), neuropeptide Y (NPY), nitric oxide synthase (NOS), vasoactive intestinal polypeptide (VIP), calcitonin gene-related peptide (CGRP) and pituitary adenylate cyclase-activating polypeptide (PACAP). Circularly cut vaginal smooth muscle preparations from the distal vagina were studied in organ baths. In the paravaginal tissue, a large number of PGP-, NOS-, TH-, VIP-immunoreactive (IR) and few CGRP-IR nerve trunks were observed, giving off branches to the smooth muscle wall. The smooth muscle wall was supplied by a large number of PGP-, Syn-, VAChT-, NPY-, NOS- and TH- IR nerve terminals, whilst only a moderate to few numbers of CGRP-, VIP- and PACAP-IR terminals were identified. Especially the distal part of the vaginal wall, where the circularly running smooth muscle was thickened into a distinct sphincter structure, was very richly innervated, predominantly by PGP- and NOS-IR terminals. Below and within the basal parts of the epithelium in the distal half of the vagina, a large number of PGP- and few NOS- and PACAP-IR varicose terminals were observed. The vaginal arteries were encircled by plexuses of nerve terminals. A large number of these were PGP-, Syn-, VAChT-, NOS-, TH-, NPY- and VIP-IR, and few were CGRP- and PACAP-IR. In isolated preparations of the distal vagina, electrical field stimulation (EFS) caused frequency-dependent contractions, which were reduced by sildenafil, tetrodotoxin (TTX) and phentolamine. In preparations contracted by norepinephrine (NA), EFS produced frequency-dependent relaxations. Pretreatment with the NOS-inhibitor NG-nitro-L-arginine, TTX, or the inhibitor of soluble guanylate cyclase, ODQ, abolished the EFS relaxations. In NE precontracted preparations, cumulative addition of sildenafil caused concentration-dependent relaxation. Carbachol contracted the strips concentration-dependently from baseline. It can be concluded that the distal part of the rat vagina forms a distinct smooth muscle sphincter, which is richly innervated by adrenergic, cholinergic and NANC nerves. The present studies suggest that in the rat the L-arginine/NO-system not only plays an important role in the regulation of vaginal smooth muscle tone, but also affects blood flow, and may have sensory functions.

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References

  1. Levin RJ . The mechanisms of human female sexual arousal Ann Rev Sex Res 1992 3: 1–48

    Article  Google Scholar 

  2. Wagner G, Levin RJ . Vaginal fluid In: Hafez ESE, Evans TN, eds The human vagina. Elsevier/North-Holland Biomedical Press: Amsterdam 1978 pp 121–137

    Google Scholar 

  3. Owman C, Sjöberg NO . Adrenergic innervation of the female genital tract of the dog J Reprod Med 1972 8: 63–66

    CAS  PubMed  Google Scholar 

  4. Adham N, Schenk EA . Autonomic innervation of the rat vagina, cervix and uterus and its cyclic variation Am J Obst Gynecol 1969 104: 508–516

    Article  CAS  Google Scholar 

  5. Blank MA et al. The regional distribution of NPY-, PHM-, and VIP-containing nerves in the human female genital tract Int J Fertil 1986 31: 218–222

    CAS  PubMed  Google Scholar 

  6. Hoyle CH et al. Innervation of vasculature and microvasculature of the human vagina by NOS and neuropeptide-containing nerves J Anat 1996 188: 633–644

    PubMed  PubMed Central  Google Scholar 

  7. Papka RE, Cotton JP, Traurig HH . Comparative distribution of neuropeptide tyrosine-, vasoactive intestinal polypeptide-, substance P-immunoreactive, acetylcholinesterase-positive and noradrenergic nerves in the reproductive tract of the female rat Cell Tissue Res 1985 242: 475–490

    Article  CAS  PubMed  Google Scholar 

  8. Lakomy M, Kaleczyc J, Majewski M, Sienkiewicz W . Peptidergic innervation of the bovine vagina and uterus Acta Histochem 1995 97: 53–66

    Article  CAS  PubMed  Google Scholar 

  9. Polak JM, Bloom SR . Localisation and measurement of VIP in the genitourinary system of man and animals Peptides 1984 5: 225–230

    Article  CAS  PubMed  Google Scholar 

  10. Majewski M et al. The distribution and co-localization of immunoreactivity to nitric oxide synthase, vasoactive intestinal polypeptide and substance P within nerve fibres supplying bovine and porcine female genital organs Cell Tissue Res 1995 281: 445–464

    Article  CAS  PubMed  Google Scholar 

  11. Grozdanovic Z, Mayer B, Baumgarten HG, Brüning G . Nitric oxide synthase-containing nerve fibers and neurons in the genital tract of the female mouse Cell Tissue Res 1994 275: 355–360

    Article  CAS  PubMed  Google Scholar 

  12. Jørgensen JC . Neuropeptide Y in mammalian genital tract: localization and biological action Dan Med Bull 1994 41: 294–305

    PubMed  Google Scholar 

  13. Graf AH et al. Helospectin and pituitary adenylate cyclase activating polypeptide in the human vagina Regul Pept 1995 55: 277–286

    Article  CAS  PubMed  Google Scholar 

  14. Steenstup BR et al. Pituitary adenylate cyclase-activating polypeptide: occurrence and relaxant effect in female genital tract Am J Physiol 1995 269: E108–E117

    Google Scholar 

  15. Fahrenkrug J, Hannibal J . Pituitary adenylate cyclase activating polypeptide innervation of the rat female reproductive tract and the associated paracervical ganglia: effect of capsaicin Neuroscience 1996 73: 1049–1060

    Article  CAS  PubMed  Google Scholar 

  16. Boolell M et al. Sildenafil: an orally active type 5 cyclic GMP-specific phosphodiesterase inhibitor for the treatment of penile erectile dysfunction Int J Impot Res 1996 8: 47–52

    CAS  PubMed  Google Scholar 

  17. Andersson KE, Wagner G . Physiology of penile erection Physiol Rev 1995 75: 191–236

    Article  CAS  PubMed  Google Scholar 

  18. Hedlund P et al. Erectile dysfunction in cyclic GMP-dependent kinase I-deficient mice Proc Natl Acad Sci USA 2000 97: 2349–2354

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Norman J . Nitric oxide and the myometrium Pharmacol Ther 1996 70: 91–100

    Article  CAS  PubMed  Google Scholar 

  20. Guslandi M . Nitric oxide: an ubiquitous actor in the gastrointestinal tract Dig Dis 1994 12: 28–36

    Article  CAS  PubMed  Google Scholar 

  21. Ignarro LJ . Nitric oxide: a unique endogenous signaling molecule in vascular biology Biosci Rep 1999 19: 51–71

    Article  CAS  PubMed  Google Scholar 

  22. Andersson KE, Persson K . Nitric oxide synthase and the lower urinary tract: possible implications for physiology and pathophysiology Scand J Urol Nephrol 1995 175: (Suppl) 43–53

    CAS  Google Scholar 

  23. Goldstein I, Berman J . Vasculogenic female sexual dysfunction: vaginal engorgement and clitoral erectile insufficiency syndromes Int J Impot Res 1998 10: S84–S90

    PubMed  Google Scholar 

  24. Koelle GB, Friedenwald JS . A histochemical method for localizing cholinesterase activity Proc Soc Expt Biol Med 1949 70: 617–622

    Article  CAS  Google Scholar 

  25. Holmstedt B . A modification of the thiocholine method for determination of cholinesterase. II. Histochemical application Acta Physiol Scand 1957 40: 331–337

    Article  CAS  PubMed  Google Scholar 

  26. Matthew DA, Nadler JV, Lynch GS, Cotman CW . Development of cholinergic innervation in the hippocampal formation of the rat I. Histochemical demonstration of acetylcholinesterase activity Dev Biol 1974 36: 130–141

    Article  Google Scholar 

  27. Bredt DS et al. Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase Nature 1991 351: 714–718

    Article  CAS  PubMed  Google Scholar 

  28. Alm P et al. Immunohistochemical localization of peripheral nitric oxide synthase-containing nerves using antibodies raised against synthesized C- and N-terminal fragments of a cloned enzyme from rat brain Acta Physiol Scand 1993 148: 421–429

    Article  CAS  PubMed  Google Scholar 

  29. Herbison AE, Simonian SX, Norris PJ, Emson PC . Relationship of neuronal nitric oxide synthase immunoreactivity to GnRH neurons in the ovariectomized and intact female rat J Neuroendorinol 1996 8: 73–82

    Article  CAS  Google Scholar 

  30. Johnson GD, Araujo GM . A simple method of reducing the fading of immunofluorescence during microscopy J Immunol Methods 1981 43: 349–350

    Article  CAS  PubMed  Google Scholar 

  31. Wessendorf MW, Elde RP . Characterization of an immunofluorescence technique for the demonstration of coexisting neurotransmitters within nerve fibres and terminals J Histochem Cytochem 1985 33: 984–994

    Article  CAS  PubMed  Google Scholar 

  32. Ny L et al. Nitric oxide synthase-containing, peptide-containing, and acetylcholinesterase-positive nerves in the cat lower oesophagus Histochem J 1994 26: 721–733

    Article  CAS  PubMed  Google Scholar 

  33. Lundberg LM, Alm P, Wharton J, Polak JM . Protein gene product 9.5 (PGP 9.5)—a new neuronal marker visualizing the whole uterine innervation and pregnancy-induced and developmental changes in the guinea pig Histochemistry 1988 90: 9–17

    Article  CAS  PubMed  Google Scholar 

  34. Gulbenkian S, Wharton J, Polak JM . The visualization of cardiovascular innervation in the guinea pig using an antiserum to protein gene product 9.5 (PGP 9.5) J Auton Nerv Syst 1987 18: 235–247

    Article  CAS  PubMed  Google Scholar 

  35. Jahn R, Schiebler W, Ouimet C, Greengard P . A 38 000-dalton membrane protein (p38) present in synaptic vesicles Proc Natl Acad Sci USA 1985 82: 4137–4141

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Wiedenmann B, Franke WW . Identification and localization of synaptophysin, an integral membrane glycoprotein of Mr 38 000 characteristic of presynaptic vesicles Cell 1985 41: 1017–1028

    Article  CAS  PubMed  Google Scholar 

  37. Giraldi A et al. Effects of diabetes on neurotransmission in rat vaginal smooth muscle Int J Impot Res 2001 13: 58–66

    Article  CAS  PubMed  Google Scholar 

  38. Arvidsson U, Riedl M, Elde R, Meister B . Vesicular acetylcholine transporter (VAChT) protein: a novel and unique marker for cholinergic neurons in the central and peripheral nervous system J Comp Neurol 1997 378: 454–467

    Article  CAS  PubMed  Google Scholar 

  39. Spraycar M ed.. Stedman's medical dictionary 26thedition Williams & Wilkins: Baltimore, MD 1995 pp 1647

    Google Scholar 

  40. Alm P et al. Nitric oxide synthase-immunoreactive, adrenergic, cholinergic, and peptidergic nerves of the female rat urinary tract: a comparative study J Auton Nerv Syst 1995 56: 105–114

    Article  CAS  PubMed  Google Scholar 

  41. Alm P et al. Nitric oxide synthase-containing neurons in rat parasympathetic, sympathetic and sensory ganglia: a comparative study Histochem J 1995 27: 819–831

    Article  CAS  PubMed  Google Scholar 

  42. Zhang X et al. Nitric oxide synthase-like immunoreactivity in lumbar dorsal root ganglia spinal cord of rat and monkey and effect of peripheral axotomy J Comp Neurol 1993 335: 563–575

    Article  CAS  PubMed  Google Scholar 

  43. Giuliano F et al. Vaginal physiological changes in a model of sexual arousal in anesthetized rats Am J Physiol Regul Integr Comp Physiol 2001 281: R140–R149

    Article  CAS  PubMed  Google Scholar 

  44. Henning RJ, Sawmiller DR . Vasoactive intestinal polypeptide: cardiovascular effects Cardiovasc Res 2001 49: 27–37

    Article  CAS  PubMed  Google Scholar 

  45. Ottesen B et al. Vasoactive intestinal polypeptide (VIP) provokes vaginal lubrication in normal women Peptides 1987 8: 797–800

    Article  CAS  PubMed  Google Scholar 

  46. Fahrenkrug J, Hannibal J . Pituitary adenylate cyclase activating polypeptide immunoreactivity in capsaicin-sensitive nerve fibres supplying the rat urinary tract Neuroscience 1998 83: 1261–1272

    Article  CAS  PubMed  Google Scholar 

  47. Min K et al. Sildenafil augments pelvic nerve-mediated female genital sexual arousal in the anesthetized rabbit Int J Impot Res 2000 12: S32–S39

    Article  PubMed  Google Scholar 

  48. Hedlund P, Larsson B, Alm P, Andersson KE . Nitric oxide synthase-containing nerves and ganglia in the dog prostate: a comparison with other transmitters Histochem J 1996 28: 635–642

    Article  CAS  PubMed  Google Scholar 

  49. Persson K et al. Co-existence of nitrergic, peptidergic and acetylcholine esterase-positive nerves in the pig lower urinary tract J Auton Nerv Syst 1995 52: 225–236

    Article  CAS  PubMed  Google Scholar 

  50. Kilbinger H, Wolf D . Increase by NO synthase inhibitors of acetylcholine release from guinea-pig myenteric plexus Naunyn Schmiedebergs Arch Pharmacol 1994 349: 543–545

    Article  CAS  PubMed  Google Scholar 

  51. Rae MG, Khoyi MA, Keef KD . Modulation of cholinergic neuromuscular transmission by nitric oxide in canine circular smooth muscle Am J Physiol 1998 275: G1324–G1322

    CAS  PubMed  Google Scholar 

  52. Berman J et al. Effect of vasoactive agents in modulating vaginal smooth muscle contractility In: Proceedings of the Function Female Sexual Forum Meeting. Boston 1999 59

  53. Forsberg JG, Kalland T . Embryology of the genital tract in humans and in rodents In: Herbst AL, Bern HA, eds Developmental effects of diethylstibestrol (DES) in pregnancy Thieme-Stratton Inc: New York 1981 pp 4–25

    Google Scholar 

  54. Cellek S, Moncada S . Nitrergic neurotransmission mediates the non-adrenergic non-cholinergic responses in the clitoral corpus cavernosum of the rabbit Br J Pharmacol 1998 125: 1627–1629

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Hedlund P, Larsson B, Alm P, Andersson KE . Distribution and function of nitric oxide-containing nerves in canine corpus cavernosum and spongiosum Acta Physiol Scand 1995 155: 445–455

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by the Swedish Medical Research Council (grant numbers 11205 and 6837) and by Pfizer, DK. The authors are grateful to Lillemor Thuresson, Brita Sunden-Andersson and Agneta Kristensen, for technical assistance and Dr Anders Nylen, Department of Clinical Pharmacology, Lund for help with the oestrus cycle studies. The generous supplies of goat NPY and mouse monoclonal PACAP antisera from Professor Thue Schwartz and Dr Jens Hannibal, Departments of Clinical Chemistry, Rigshospitalet and Bispebjerg Hospital, Copenhagen, Denmark are greatly appreciated.

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Authors and Affiliations

  1. Division of Sexual Physiology, Department of Medical Physiology, University of Copenhagen, Rigshospitalet, Copenhagen, Denmark

    A Giraldi & G Wagner

  2. Department of Clinical Pharmacology, Lund University Hospital, Lund, Sweden

    V Werkström, L Myllymäki & K E Andersson

  3. Department of Pathology, Lund University Hospital, Lund, Sweden

    P Alm

  4. Department of Clinical Pharmacology, Rigshospitalet, Copenhagen, Denmark

    A Giraldi

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  1. A Giraldi
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Correspondence to A Giraldi.

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Giraldi, A., Alm, P., Werkström, V. et al. Morphological and functional characterization of a rat vaginal smooth muscle sphincter. Int J Impot Res 14, 271–282 (2002). https://doi.org/10.1038/sj.ijir.3900886

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