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Gonadotropin-releasing hormone analog buserelin causes neuronal loss in rat gastrointestinal tract

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Abstract

Gonadotropin-releasing hormone (GnRH) analogs are given to women undergoing in vitro fertilization. Case reports describing the development of chronic intestinal pseudo-obstruction and auto-antibodies against GnRH after such treatment suggest a strong association between intestinal dysfunction and GnRH analogs. No experimental model for studying such a relationship is currently at hand. Our main goal was to investigate possible enteric neurodegeneration and titers of GnRH antibodies in response to repeated administration of the GnRH analog buserelin in rat. Rats were treated for 1–4 sessions with daily subcutaneous injections of buserelin or saline for 5 days, followed by 3 weeks of recovery. Buserelin treatment caused significant loss of submucous and myenteric neurons in the fundus, ileum, and colon. The loss of enteric neurons can, at least partly, be explained by increased apoptosis. No GnRH- or GnRH-receptor-immunoreactive (IR) enteric neurons but numerous luteinizing hormone (LH)-receptor-IR neurons were detected. After buserelin treatment, the relative number of enteric LH-receptor-IR neurons decreased, whereas that of nitric-oxide-synthase-IR neurons increased. No intestinal inflammation or increased levels of circulating interleukins/cytokines were noted in response to buserelin treatment. Serum GnRH antibody titers were undetectable or extremely low in all rats. Thus, repeated administrations of buserelin induce neurodegeneration in rat gastrointestinal tract, possibly by way of LH-receptor hyperactivation. The present findings suggest that enteric neurodegenerative effects of GnRH analog treatment in man can be mimicked in rat. However, in contrast to man, no production of GnRH auto-antibodies has been noted in rat.

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References

  • Arciszewski MB, Sand E, Ekblad E (2008) Vasoactive intestinal peptide rescues cultured rat myenteric neurons from lipopolysaccharide induced cell death. Regul Pept 146:218–223

    Article  PubMed  CAS  Google Scholar 

  • Berger H, Sandow J, Heinrich N, Albrecht E, Kertscher U, Oehlke J (1993) Disposition of the 3H-labeled gonadotropin-releasing hormone analog buserelin in rats. Drug Metab Dispos 5:818–822

    Google Scholar 

  • Conn PM, Crowley WF Jr (1994) Gonadotropin-releasing hormone and its analogs. Annu Rev Med 45:391–405

    Article  PubMed  CAS  Google Scholar 

  • Counis R, Laverrière JN, Garrel G, Bleux C, Cohen-Tannoudji J, Lerrant Y, Kottler ML, Magre S (2005) Gonadotropin-releasing hormone and the control of gonadotrope function.Reprod Nutr Dev 45:243–254

    Article  PubMed  CAS  Google Scholar 

  • Dodd PR (2002) Excited to death: different ways to lose your neurons. Biogerontology 3:51–56

    Article  PubMed  CAS  Google Scholar 

  • Ducker TE, Boss JW, Altug SA, Mehrabian H, Dekeratry DR, Clench MH, Mathias JR (1996) Luteinizing hormone and human chorionic gonadotropin fragment the migrating motor complex in rat small intestine. Neurogastroenterol Motil 8:95–100

    Article  PubMed  CAS  Google Scholar 

  • Ekblad E, Bauer AJ (2004) Role of vasoactive intestinal peptide and inflammatory mediators in enteric neuronal plasticity. Neurogastroenterol Motil 1:123–128

    Article  Google Scholar 

  • Ekblad E, Ekman R, Håkanson R, Sundler F (1988) Projections of peptide-containing neurons in rat colon. Neuroscience 27:655–674

    Article  PubMed  CAS  Google Scholar 

  • Ekblad E, Alm P, Sundler F (1994) Distribution, origin and projections of nitric oxide synthase-containing neurons in gut and pancreas. Neuroscience 63:233–248

    Article  PubMed  CAS  Google Scholar 

  • Ekblad E, Sjuve R, Arner A, Sundler F (1998) Enteric neuronal plasticity and a reduced number of interstitial cells of Cajal in hypertrophic rat ileum. Gut 42:836–844

    Article  PubMed  CAS  Google Scholar 

  • Hammar O, Ohlsson B, Veress B, Alm R, Fredrikson GN, Montgomery A (2012a) Depletion of enteric gonadotropin-releasing hormone is found in a few patients suffering from severe gastrointestinal dysmotility. Scand J Gastroenterol 47:1165–1173

    Article  PubMed  CAS  Google Scholar 

  • Hammar O, Veress B, Montgomery A, Ohlsson B (2012b) Expression of luteinizing hormone receptor in the gastrointestinal tract in patients with and without dysmotility. Drug Target Insights 6:13–18

    PubMed  CAS  Google Scholar 

  • He S-H (2004) Key role of mast cells and their major secretory products in inflammatory bowel disease. World J Gastroenterol 10:309–318

    PubMed  CAS  Google Scholar 

  • Heinrich N, Albrecht E, Sandow J, Kertscher U, Lorenz D, Oehlke J, Berger H (1996) Disposition of 3H-labelled buserelin continuously infused into rats. Eur J Drug Metab Pharmacokinet 21:345–350

    Article  PubMed  CAS  Google Scholar 

  • Huang W, Yao B, Sun L, Pu R, Wang L, Zhang R (2001) Immunohistochemical and in situ hybridization studies of gonadotropin releasing hormone (GnRH) and its receptor in rat digestive tract. Life Sci 68:1727–1734

    Article  PubMed  CAS  Google Scholar 

  • Lin Z, Sandgren K, Ekblad E (2003) Increased expression of vasoactive intestinal peptide in cultured myenteric neurons from rat small intestine. Auton Neurosci 107:9–19

    Article  PubMed  CAS  Google Scholar 

  • Masterson JC, Furuta GT, Lee JJ (2011) Update on clinical and immunological features of eosinophilic gastrointestinal diseases. Curr Opin Gastroenterol 27:515–522

    Article  PubMed  CAS  Google Scholar 

  • Mathias JR, Clench M (1998) Relationship of reproductive hormones and neuromuscular disease of the gastrointestinal tract. Dig Dis Sci 16:3–13

    Article  CAS  Google Scholar 

  • Naor Z (2009) Signaling by G-protein-coupled receptor (GPCR): studies on the GnRH receptor. Front Neuroendocrinol 1:10–29

    Article  Google Scholar 

  • Ohlsson B, Veress B, Janciauskiene S, Montgomery A, Haglund M, Wallmark A (2007) Chronic intestinal pseudo-obstruction due to buserelin-induced formation of anti-GnRH antibodies. Gastroenterology 132:45–51

    Article  PubMed  CAS  Google Scholar 

  • Ohlsson B, Ekblad E, Veress B, Montgomery A, Janciauskine S (2010) Antibodies against gonadotropin-releasing hormone (GnRH) and destruction of enteric neurons in patients suffering from gastrointestinal dysfunction. BMC Gastroenterol 10:48

    Google Scholar 

  • Ohman L, Isaksson S, Lindmark AC, Posserud I, Stotzer PO, Strid H, Sjövall H, Simrén M (2009a) T-cell activation in patients with irritable bowel syndrome.Am J Gastroenterol 104:1205-1212

    Article  Google Scholar 

  • Ohman L, Lindmark AC, Isaksson S, Posserud I, Strid H, Sjövall H, Simrén M (2009b) B-cell activation in patients with irritable bowel syndrome (IBS).Neurogastroenterol Motil 21:644–650

    Article  CAS  Google Scholar 

  • Sand E, Themner Persson A, Ekblad E (2008) Infiltration of mast cells in rat colon is a consequence of ischemia/reperfusion. Dig Dis Sci 53:3158–3169

    Article  PubMed  CAS  Google Scholar 

  • Sand E, Themner-Persson A, Ekblad E (2009) Mast cells reduce survival of myenteric neurons in culture. Neuropharmacology 56:522–530

    Article  PubMed  CAS  Google Scholar 

  • Sand E, Themner-Persson A, Ekblad E (2011) Corticotropin releasing factor—distribution in rat intestine and role in neuroprotection. Regul Pept 166:68–75

    Article  PubMed  CAS  Google Scholar 

  • Sandgren K, Lin Z, Fex Svenningsen Å, Ekblad E (2003) Vasoactive intestinal peptide and nitric oxide promote survival of adult rat myenteric neurons in culture. J Neurosci Res 72:595–602

    Article  PubMed  CAS  Google Scholar 

  • Stridh L, Smith PL, Naylor AS, Wang X, Mallard C (2011) Regulation of toll-like receptor 1 and −2 in neonatal mice brains after hypoxia-ischemia. J Neuroinflammation 8:45

    Article  PubMed  CAS  Google Scholar 

  • Szabo A, Dalmau J, Manley G, Rosenfeld M, Wong E, Henson J, Posner JB, Furneaux HM (1991) HuD, a paraneoplastic encephalomyelitis antigen, contains RNA-binding domains and is homologous to Elav and Sex-lethal. Cell 67:325–333

    Article  PubMed  CAS  Google Scholar 

  • Trindade CR, Camargos AF, Pereira FEL (2008) The effect of buserelin acetate on the uterus of adult rats: morphological aspects. Clin Exp Obstet Gynecol 3:198–201

    Google Scholar 

  • Van der Woude CJ, Stokkers P, Bodegraven A van, Assche G van, Hebzda Z, Paradowski L, D’Haens G, Ghosh S, Feagan B, Rutgeerts P, Dijkstra G, Jong DJ de, Oldenburg B, Farhan M, Richard T, Dean Y, Hommes DW (2010) Phase 1, double-blind, randomized, placebo-controlled, dose-escalation study of NI-0401 (a fully human anti-CD3 monoclonal antibody) in patients with moderate to severe active Crohn’s disease. Inflamm Bowel Dis 16:1708–1716

    Google Scholar 

  • Voss U, Sand E, Hellström PM, Ekblad E (2012) Glucagon-like peptides 1 and 2 and vasoactive intestinal peptide are neuroprotective on cultured and mast cell co-cultured rat myenteric neurons. BMC Gastroenterol 12:30

    Google Scholar 

  • Westwood FR (2008) The female rat reproductive cycle: a practical histological guide to staging. Toxicol Pathol 36:375–384

    Article  PubMed  Google Scholar 

  • Yao KM, Samson ML, Reeves R, White K (1993) Gene elav of Drosophila melanogaster: a prototype for neuronal-specific RNA binding protein gene family that is conserved in flies and humans. J Neurobiol 24:723–739

    Article  PubMed  CAS  Google Scholar 

  • Yu B, Ruman J, Christman G (2011) The role of peripheral gonadotropin-releasing hormone receptors in female reproduction. Fertil Steril 95:465–473

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Anna Themner-Persson for valuable technical assistance.

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Authors and Affiliations

  1. Department of Experimental Medical Sciences, Neurogastroenterology Unit, Lund University, BMC B11, 22184, Lund, Sweden

    Elin Sand, Ulrikke Voss & Eva Ekblad

  2. Department of Clinical Sciences, Division of Gastroenterology, Skåne University Hospital, Malmö, Sweden

    Elin Sand, Oskar Hammar & Bodil Ohlsson

  3. Department of Clinical Sciences, Experimental Cardiovascular Research Unit, Skåne University Hospital, Malmö, Sweden

    Ragnar Alm & Gunilla Nordin Fredrikson

  4. Lund University, Lund, Sweden

    Oskar Hammar, Ragnar Alm, Gunilla Nordin Fredrikson & Bodil Ohlsson

Authors
  1. Elin Sand
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  2. Ulrikke Voss
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  3. Oskar Hammar
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  4. Ragnar Alm
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  5. Gunilla Nordin Fredrikson
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  6. Bodil Ohlsson
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  7. Eva Ekblad
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Corresponding author

Correspondence to Elin Sand.

Additional information

This work were supported by the Swedish Society for Medical Research, the Royal Physiographic Society, the Bengt Ihres and A. Påhlssons Foundations, the Gustaf V:th and Queen Victorias Frimurare Foundation and the Lund Medical Faculty.

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Supplementary Fig. 1

Cryo-sections of hypothalamus (a), ileum (b), pituitary (c), and colon (d) from control rat immunostained with antibodies against GnRH (a, b) or GnRH receptor (c, d). The hypothalamus harbors nerve fibers intensely immunoreactive (IR) for GnRH (a), whereas no GnRH-IR enteric nerve cellbodies or fibers can be detected in the gastrointestinal tract (b). FITC labeling of vascular lamina elastica interna in the submucosa is unspecific (b). A pituicyte intensely IR for GnRH receptors (c). Enteric nerve cell bodies or fibers devoid of GnRH receptor immunostaining (d). Bars 10 μm (a, c), 100 μm (b, d). (JPEG 37 kb)

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Supplementary Fig. 2

Top Numbers of mast cells per millimeter section in the fundus (a), ileum (b), and colon (c) from saline-treated (control) and buserelin-treated (B1-B4) rats were quantified in longitudinal toluidine-blue-stained sections. The submucosa (sm), circular (cm), and longitudinal (lm) muscle layers and serosa (s) were evaluated separately. The fundus in rats treated with one session of buserelin (B1) had higher numbers of mast cells in the circular muscle layer and serosa compared with controls. Bottom Relative numbers of activated-caspase-3-immunoreactive (IR) neurons in submucous (SG in d) and myenteric (MG in e) ganglia, expressed in percent of HuC/D-IR neurons in the colon from rats treated with saline (control) or buserelin (B1-B4). Rats treated with two sessions of buserelin (B2) had an increased number of apoptotic activated-caspase-3-IR submucous neurons compared with controls. Results are presented as medians and spreads expressed as the 25th and 75th percentile. Statistical analyses were performed by the Kruskal-Wallis test followed by Dunn’s multiple comparison test (all comparisons against control), n=3–11, with statistical significance being indicated by *P<0.05, **P<0.01. (JPEG 37 kb)

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Sand, E., Voss, U., Hammar, O. et al. Gonadotropin-releasing hormone analog buserelin causes neuronal loss in rat gastrointestinal tract. Cell Tissue Res 351, 521–534 (2013). https://doi.org/10.1007/s00441-012-1534-1

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  • DOI: https://doi.org/10.1007/s00441-012-1534-1

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