Advertisement

Upregulation of serotonin-receptor-2a and serotonin transporter expression in the pulmonary vasculature of nitrofen-induced congenital diaphragmatic hernia

Published:February 03, 2014DOI:https://doi.org/10.1016/j.jpedsurg.2014.01.015

      Abstract

      Purpose

      Congenital diaphragmatic hernia (CDH) is attributed to severe pulmonary hypoplasia and pulmonary hypertension (PH). PH is characterized by structural changes resulting in vascular remodeling. Serotonin, a potent vasoconstrictor, plays a central role in the development of PH. It exerts its constricting effects on the vessels via Serotonin receptor 2A (5-HT2A) and induces pulmonary smooth muscle cell proliferation via the serotonin transporter (5-HTT). This study was designed to investigate expressions of 5-HT2A and 5-HTT in the pulmonary vasculature of rats with nitrofen-induced CDH.

      Methods

      Rats were exposed to nitrofen or vehicle on D9. Fetuses were sacrificed on D21 and divided into nitrofen and control group (n = 32). Pulmonary RNA was extracted and mRNA level of 5HT2A was determined by qRT-PCR. Protein expression of 5HT2A and 5-HTT was investigated by western blotting. Confocal immunofluorescence double-staining for 5-HT2A, 5-HTT, and alpha smooth muscle actin were performed.

      Results

      Pulmonary 5-HT2A gene expression levels were significantly increased in nitrofen-induced CDH compared to controls. Western blotting and confocal microscopy confirmed increased pulmonary protein expression in CDH lungs compared to controls.

      Conclusion

      Increased gene and protein expression of 5HT2A and 5-HTT in the pulmonary vasculature of nitrofen-induced CDH lungs suggest that 5HT2A and 5-HTT are important mediators of PH in nitrofen-induced CDH.

      Key words

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Journal of Pediatric Surgery
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Keijzer R.
        • Puri P.
        Congenital diaphragmatic hernia.
        Semin Pediatr Surg. 2010; 19: 180-185
        • Heath D.
        • Smith P.
        • Gosney J.
        • et al.
        The pathology of the early and late stages of primary pulmonary hypertension.
        Br Heart J. 1987; 58: 204-213
        • Belik J.
        • Davidge S.T.
        • Zhang W.
        • et al.
        Airway smooth muscle changes in the nitrofen-induced congenital diaphragmatic hernia rat model.
        Pediatr Res. 2003; 53: 737-743
        • Yamataka T.
        • Puri P.
        Active collagen synthesis by pulmonary arteries in pulmonary hypertension complicated by congenital diaphragmatic hernia.
        J Pediatr Surg. 1997; 32: 682-687
        • Gosemann J.-H.
        • Friedmacher F.
        • Fujiwara N.
        • et al.
        Disruption of the bone morphogenetic protein receptor 2 pathway in nitrofen-induced congenital diaphragmatic hernia.
        Birth Defects Res B. 2013; 98: 304-309
        • Hervé P.
        • Launay J.M.
        • Scrobohaci M.L.
        • et al.
        Increased plasma serotonin in primary pulmonary hypertension.
        Am J Med. 1995; 99: 249-254
        • Eddahibi S.
        • Guignabert C.
        • Barlier-Mur A.-M.
        • et al.
        Cross talk between endothelial and smooth muscle cells in pulmonary hypertension.
        Circulation. 2006; 113: 1857-1864
        • Keegan A.
        • Morecroft I.
        • Smillie D.
        • et al.
        Contribution of the 5-HT(1B) receptor to hypoxia-induced pulmonary hypertension: converging evidence using 5-HT(1B)-receptor knockout mice and the 5-HT(1B/1D)-receptor antagonist GR127935.
        Circ Res. 2001; 89: 1231-1239
        • Guignabert C.
        • Raffestin B.
        • Benferhat R.
        • et al.
        Serotonin transporter inhibition prevents and reverses monocrotaline-induced pulmonary hypertension in rats.
        Circulation. 2005; 111: 2812-2819
        • Drozdov I.
        • Kidd M.
        • Gustafsson B.I.
        • et al.
        Autoregulatory effects of serotonin on proliferation and signaling pathways in lung and small intestine neuroendocrine tumor cell lines.
        Cancer. 2009; 115: 4934-4945
        • Hui A.S.
        Regulation of catecholamines by sustained and intermittent hypoxia in neuroendocrine cells and sympathetic neurons.
        Hypertension. 2003; 42: 1130-1136
        • MacLean M.R.
        Pulmonary hypertension and the serotonin hypothesis: where are we now?.
        Int J Clin Pract. 2007; ([Suppl.]): 27-31
        • MacLean M.R.
        • Dempsie Y.
        Serotonin and pulmonary hypertension—from bench to bedside?.
        Curr Opin Pharmacol. 2009; 9: 281-286
        • Liu Y.
        • Tian H.
        • Yan X.
        • et al.
        Serotonin inhibits apoptosis of pulmonary artery smooth muscle cells through 5-HT2A receptors involved in the pulmonary artery remodeling of pulmonary artery hypertension.
        Exp Lung Res. 2013; 39: 70-79
        • Guignabert C.
        • Izikki M.
        • Tu L.I.
        • et al.
        Transgenic mice overexpressing the 5-hydroxytryptamine transporter gene in smooth muscle develop pulmonary hypertension.
        Circ Res. 2006; 98: 1323-1330
        • Eddahibi S.
        • Hanoun N.
        • Lanfumey L.
        • et al.
        Attenuated hypoxic pulmonary hypertension in mice lacking the 5-hydroxytryptamine transporter gene.
        J Clin Invest. 2000; 105: 1555-1562
        • Greer J.J.
        Current concepts on the pathogenesis and etiology of congenital diaphragmatic hernia.
        Respir Physiol Neurobiol. 2013 Nov 1; 189 (Epub 2013 May 9.): 232-240https://doi.org/10.1016/j.resp.2013.04.015
        • Beurskens N.
        • Klaassens M.
        • Rottier R.
        • et al.
        Linking animal models to human congenital diaphragmatic hernia.
        Birth Defects Res A Clin Mol Teratol. 2007; 79: 565-572
        • Mayer S.
        • Metzger R.
        • Kluth D.
        The embryology of the diaphragm.
        Semin Pediatr Surg. 2011; 20: 161-169
        • Luong C.
        • Rey-Perra J.
        • Vadivel A.
        • et al.
        Antenatal sildenafil treatment attenuates pulmonary hypertension in experimental congenital diaphragmatic hernia.
        Circulation. 2011; 123: 2120-2131
        • MacLean M.R.
        • Hervé P.
        • Eddahibi S.
        • et al.
        5-Hydroxytryptamine and the pulmonary circulation: receptors, transporters and relevance to pulmonary arterial hypertension.
        Br J Pharmacol. 2000; 131: 161-168
        • Rabinovitch M.
        Molecular pathogenesis of pulmonary arterial hypertension.
        J Clin Invest. 2008; 118: 2372
        • Humbert M.
        • Morrell N.W.
        • Archer S.L.
        • et al.
        Cellular and molecular pathobiology of pulmonary arterial hypertension.
        J Am Coll Cardiol. 2004; 43: 13S-24S
        • Eddahibi S.
        • Fabre V.
        • Boni C.
        • et al.
        Induction of serotonin transporter by hypoxia in pulmonary vascular smooth muscle cells relationship with the mitogenic action of serotonin.
        Circ Res. 1999; 84: 329-336
        • MacLean M.R.
        • Deuchar G.A.
        • Hicks M.N.
        • et al.
        Overexpression of the 5-hydroxytryptamine transporter gene effect on pulmonary hemodynamics and hypoxia-induced pulmonary hypertension.
        Circulation. 2004; 109: 2150-2155
        • Marcos E.
        Serotonin-induced smooth muscle hyperplasia in various forms of human pulmonary hypertension.
        Circ Res. 2004; 94: 1263-1270