Altered messenger RNA expression of the neuronal nitric oxide synthase gene in infantile hypertrophic pyloric stenosis

T. Kusafuka1, P. Puri1
1Children's Research Centre, Our Lady's Hospital for Sick Children, Crumlin Dublin, 12, Ireland

Tóm tắt

Nitric oxide (NO) has been described as a mediator of smooth muscle relaxation in the mammalian gastrointestinal tract. The enzyme neuronal nitric oxide synthase (NOS) catalyzes the formation of NO. We examined the expression of the neuronal NOS gene at the messenger RNA (mRNA) level in pyloric smooth-muscle biopsy specimens from six patients with infantile hypertrophic pyloric stenosis (IHPS) using a reverse transcription-polymerase chain reaction (RT-PCR) technique. For controls, smooth-muscle layer specimens of pylorus (n = 3), ileum (n = 2), and colon (n = 2) were used. With 31 cycles of PCR reaction, control specimens revealed detectable signals for neuronal NOS mRNA. In contrast, signals of IHPS specimens were undetectable in five cases and very weak in one. By increasing the PCR to 37 cycles, detectable signals for neuronal NOS mRNA were observed in all IHPS specimens, but they were significantly weaker than those of controls. Since a low level of neuronal NOS mRNA may lead to impaired production of NO, our observations indicate that the excessively contracted, hypertrophied pyloric muscle in IHPS is a result of reduced expression of the neuronal NOS gene at the mRNA level.

Từ khóa


Tài liệu tham khảo

Spicer RD (1982) Infantile hypertrophic pyloric stenosis: a review. Br J Surg 69: 128–135 Bult H, Boeckxstaens GE, Pelckmans PA, Jordaens FH, Van Maercke YM, Herman AG (1990) Nitric oxide as an inhibitory non-adrenergic non-cholinergic neurotransmitter. Nature 345: 346–347 Guslandi M (1994) Nitric oxide: an ubiquitous actor in the gastrointestinal tract. Dig Dis 12: 28–36 Kuo PC, Schroeder RA (1995) The emerging multifaceted roles of nitric oxide. Ann Surg 221: 220–235 Nathan C, Xie QW (1994) Regulation of biosynthesis of nitric oxide. J Biol Chem 269: 13725–13728 Bredt DS, Huang PM, Glatt CE, Lowenstein C, Reed RR, Snyder SH (1991) Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase. Nature 351: 714–718 Bredt DS, Hwang PM, Snyder SH (1990) Localization of nitric oxide synthase indicating a neural role for nitric oxide. Nature 347: 768–770 Marsden PA, Schappert KT, Chen HS, Flowers M, Sundell CL, Wilcox JN, Lamas S, Michel T (1992) Molecular cloning and characterization of human endothelial nitric oxide synthase. FEBS Lett 307: 287–293 Huang PL, Dawson TM, Bredt DS, Snyder SH, Fishman MC (1993) Targeted disruption of the neuronal nitric oxide synthase gene. Cell 75: 1273–1286 Vanderwinden JM, Mailleux P, Schiffmann SN, Vanderhaeghen JJ, De Laet MH (1992) Nitric oxide synthase activity in infantile hypertrophic pyloric stenosis. N Engl J Med 327: 511–515 Kobayashi H, O'Briain DS, Puri P (1995) Immunochemical characterization of neural cell adhesion molecule (NCAM), nitric oxide synthase, and neurofilament protein expression in pyloric muscle of patients with pyloric stenosis. J Pediatr Gastroenterol Nutr 20: 319–325 Nakane M, Schmidt HHHW, Pollock JS, Forstermann U, Murad F (1993) Cloned human brain nitric oxide synthase is highly expressed in skeletal muscle. FEBS Lett 316: 175–180 Hall AV, Antonion H, Wang Y, Cheung AH, Arbus AM, Olson SL, Lu WC, Kau CL, Marsden PA (1994) Structural organization of the human neuronal nitric oxide synthase gene (NOS1). J Biol Chem 269: 33082–33090 Arcari P, Martinelli R, Salvatore F (1984) The complete sequence of a full length cDNA for human liver glyceraldehyde3-phosphate dehydrogenase: evidence for multiple mRNA species. Nucleic Acids Res 12: 9179–9189 Chou Q, Russell M, Birch DE, Raymond J, Bloch W (1992) Prevention of pre-PCR mis-priming and primer dimerization improves low-copy-number amplifications. Nucleic Acids Res 20: 1717–1723 Stark ME, Bauer AJ, Szurszewski JH (1991) Effect of nitric oxide on circular muscle of the canine small intestine. J Physiol (Lond) 444: 743–761 Stark ME, Bauer AJ, Sarr MG, Szurszewski JH (1993) Nitric oxide mediated inhibitory input in human and canine jejunum. Gastroenterology 104: 398–409 Thornbury KD, Ward S M, Dalziel HH, Carl A, Westfall DP, Sanders KM (1991) Nitric oxide and nitrosocysteine mimic nonadrenergic, noncholinergic hyperpolarization in canine proximal colon. Am J Physiol 261: G553-G557 Tottrup A, Svane D, Forman A (1991) Nitric oxide mediating NANC inhibition in opossum lower esophageal sphincter. Am J Physiol 260: G385-G389 Desai KM, Sessa WC, Vane JR (1991) Involvement of nitric oxide in the reflex relaxation of the stomach to accommodate food or fluid. Nature 351: 477–479 Maggi CA, Barbanti G, Turini D, Giuliani S (1991) Effect of NG-mono-methyll-arginine (l-NMMA) and NG-nitrol-argi-nine on nonadrenergic noncholinergic relaxation in the circular muscle of the human ileum. Br J Pharmacol 103: 1970–1972 Shuttleworth CWR, Murphy R, Furness JB (1991) Evidence that nitric oxide participates in non-adrenergic inhibitory transmission to intestinal muscle in the guinea-pig. Neurosci Lett 130: 77–80 Rattan S, Chakder S (1992) Role of nitric oxide as a mediator of internal anal sphincter relaxation. Am J Physiol 262: G107-G112 Ludtke FE, Bertus M, Voth E, Michalski S, Lepsien G (1994) Gastric emptying 16 to 26 years after treatment of infantile hypertrophic pyloric stenosis. J Pediatr Surg 29: 523–526 Vanderwinden JM, Liu H, Menu R, Vanderhaeghen JJ, De Laet MH (1995) The pathology of the infantile hypertrophic pylorus after healing. Presented at the Meeting of the British Association of Paediatric Surgeons, Sheffield, UK, July 1995