Advertisement

Journal of Molecular Neuroscience

, Volume 64, Issue 2, pp 312–320 | Cite as

Antisecretory Factor Modulates GABAA Receptor Activity in Neurons

  • V. Bazzurro
  • E. Gatta
  • Aroldo Cupello
  • S. Lange
  • M. Robello
Article
  • 133 Downloads

Abstract

The antisecretory factor is an endogenous protein found in all mammalian tissues investigated so far. It acts by counteracting intestinal hypersecretion and various forms of inflammation, but the detailed mechanism of antisecretory factor (AF) action is unknown. We tested neuronal GABAA receptors by means of AF-16, a potent AF peptide derived from amino acids 36–51 from the NH2 part of AF. Cultured rat cerebellar granule cells were used, and the effects on the GABA-mediated chloride currents were determined by whole-cell patch clamp. Both the neurotransmitter GABA and AF-16 were added by perfusion of the experimental system. A 3-min AF-16 preincubation was more efficacious than 30 s in significantly elevating the rapidly desensitizing GABA-activated chloride current. No effect was found on the tonic, slowly desensitizing current. The GABA-activated current increase by AF-16 demonstrated a low k of 41 pM with a maximal increase of 37% persisting for some minutes after AF washout, independent from GABA concentration. This indicates an effect on the maximal stimulation (E%Max) excluding an altered affinity between GABA and its receptor. An immunocytochemical fluorescence approach with anti γ2 subunit antibodies demonstrated an increased expression of GABAA receptors. Thus, both the electrophysiological and the immunofluorescence approach indicate an increased appearance of GABAA receptors on the neuronal membrane. The rationale of the experiments was to test the effect of AF on a defined neuronal population of GABAA receptors. The implications of the results on the impact of AF on the enteric nervous system or on brain function are discussed.

Keywords

Antisecretory factor GABAA receptors Cerebellar granule cells Enteric nervous system Central nervous system Patch clamp 

Abbreviations

AF

Antisecretory factor

AF-16

Peptides 36–51 of AF sequence

Notes

Funding Information

This work was supported by MIUR University of Genoa.

Compliance with Ethical Standards

Experimental procedures and care of the animals were according the EU Parliament and Council of September 22 2010 (2010/63/EU). They were approved by the Italian Ministry of Health (protocol number 2207) according to D.M. 116/1992. All efforts were made in order to minimize animal suffering and the number of animals necessary in order to obtain reliable results.

References

  1. Al-Olama M, Lange S, Lönnroth I, Gatzinsky K, Jennische E (2015) Uptake of the antisecretory peptide AF-16 in rat blood and cerebrospinal fluid and effects on elevated intracranial pressure. Acta Neurochir 157(1):129–137.  https://doi.org/10.1007/s00701-014-2221-7 CrossRefPubMedGoogle Scholar
  2. Ben Ari Y (2002) Excitatory actions of GABA during development: the nature of the nurture. Nat Rev Neurosci 3(9):728–739.  https://doi.org/10.1038/nrn920 CrossRefPubMedGoogle Scholar
  3. Björk S, Bosaeus I, Ek E, Jennische E, Lönnroth I, Johansson E, Lange S (2000) Food induced stimulation of the antisecretory factor can improve symptoms in human inflammatory bowel disease: a study of a concept. Gut 46(6):824–829.  https://doi.org/10.1136/gut.46.6.824 CrossRefGoogle Scholar
  4. Bodrikov V, Pauschert A, Kochlamazashvili G, Stuermer CAO (2017) Reggie-1 and reggie-2 (flotillins) participate in Rab11a-dependent cargo trafficking, spine synapse formation and LTP-related AMPA receptor (GluA1) surface exposure in mouse hippocampal neurons. Exp Neurol 289:31–45.  https://doi.org/10.1016/j.expneurol.2016.12.007 CrossRefPubMedGoogle Scholar
  5. Cupello A (2003) Neuronal transmembrane chloride electrochemical gradient: a key player in GABA-A receptor activation physiological effect. Amino Acids 24(4):335–346.  https://doi.org/10.1007/s00726-002-0350-4 CrossRefPubMedGoogle Scholar
  6. Cupello A, Di Braccio M, Gatta E, Grossi G, Nikas P, Pellistri F, Robello M (2013) GABAA receptors of cerebellar granule cells in culture: interaction with benzodiazepines. Neurochem Res 38(12):2453–2462.  https://doi.org/10.1007/s11064-013-1171-4 CrossRefGoogle Scholar
  7. Davidson TS, Hickey WF (2004a) Antisecretory factor expression is regulated by inflammatory mediators and influences the severity of experimental autoimmune encephalomyelitis. J Leukoc Biol 76(4):835–844.  https://doi.org/10.1189/jlb.0204085 CrossRefPubMedGoogle Scholar
  8. Davidson TS, Hickey WF (2004b) Distribution and immunoregulatory properties of antisecretory factor. Lab Investig 84(3):307–319.  https://doi.org/10.1038/labinvest.3700036 CrossRefPubMedGoogle Scholar
  9. Ekesbo R, Nilsson PM, Sjölund K (2008) Effects of anti-secretory factor (ASF) on irritable bowel syndrome. A double-blind, randomized study. Scand J Prim Health Care 26(2):106–110.  https://doi.org/10.1080/02813430802005894 CrossRefPubMedPubMedCentralGoogle Scholar
  10. Farrant M, Nusser Z (2005) Variations on an inhibitory theme: phasic and tonic activation of GABAA receptors. Nat Rev Neurosci 6(3):215–229.  https://doi.org/10.1038/nrn1625 CrossRefPubMedGoogle Scholar
  11. Furness JB, Callaghan BP, Rivera LR, Cho H-J (2014) The enteric nervous system and gastrointestinal innervation: integrated local and central control. In: Lyte M, Cryan JF (eds) Microbial endocrinology: the microbiota-gut-brain axis in health and disease. Springer, New York, pp 39–71Google Scholar
  12. Galligan JJ (2002) Ligand-gated ion channels in the enteric nervous system. Neurogastroenterol Motil 14(6):611–623.  https://doi.org/10.1046/j.1365-2982.2002.00363.x CrossRefPubMedGoogle Scholar
  13. Glykis J, Mann EO, Mody J (2008) Which GABAA receptor subunits are necessary for tonic inhibition in the hippocampus? J Neurosci 28:1421–1426.  https://doi.org/10.3389/fncir.2013.00136 CrossRefGoogle Scholar
  14. Graber DJ, Harris BT, Hickey WF (2011) Strain-dependent variation in the early transcriptional response to CNS injury using a cortical explant system. J Neuroinfl 8(1):122.  https://doi.org/10.1186/1742-2094-8-122 CrossRefGoogle Scholar
  15. Grøndahl ML, Sørensen H, Unmack MA, Holm A, Skadhauge E (2002) Neuronal involvement in the effect of an antisecretory factor-derived peptide on induced secretion in the porcine small intestine. J Comp Biochem Physiol A Neuroethol Sens Neural Behav Physiol 188(8):589–594.  https://doi.org/10.1007/s00359-002-0330-x CrossRefGoogle Scholar
  16. Gwynne RM, Bornstein JC (2007) Synaptic transmission at functionally identified synapses in the enteric nervous system: roles for both ionotropic and metabotropic receptors. Curr Neuropharmacol 5(1):1–17.  https://doi.org/10.2174/157015907780077141 CrossRefPubMedPubMedCentralGoogle Scholar
  17. Hanner P, Rask-Andersen H, Lange S, Jennische E (2010) Antisecretory factor-inducing therapy improves the clinical outcome in patients with Ménière’s disease. Acta Otolaryngol 130(2):223–227.  https://doi.org/10.3109/00016480903022842 CrossRefPubMedGoogle Scholar
  18. Jennische E, Bergström T, Johansson M, Nyström K, Tarkowski A, Hansson HA, Lange S (2008) The peptide AF-16 abolishes sickness and death at experimental encephalitis by reducing increase of intracranial pressure. Brain Res 1227:189–197.  https://doi.org/10.1016/j.brainres.2008.05.083 CrossRefPubMedGoogle Scholar
  19. Johansson E, Lönnroth I, Lange S, Jonson J, Jennische E, Lönnroth C (1995) Molecular cloning and expression of a pituitary gland protein modulating intestinal fluid secretion. J Biol Chem 270(35):20615–20620.  https://doi.org/10.1074/jbc.270.35.20615 CrossRefPubMedGoogle Scholar
  20. Johansson E, Jennische E, Lange S, Lönnroth I (1997a) Antisecretory factor suppresses intestinal inflammation and hypersecretion. Gut 41(5):642–645.  https://doi.org/10.1136/gut.41.5.642 CrossRefPubMedPubMedCentralGoogle Scholar
  21. Johansson E, Lange S, Lönnroth I (1997b) Identification of an active site in the antisecretory factor protein. Biochim Biophys Acta 1362(2-3):177–182.  https://doi.org/10.1016/S0925-4439(97)00066-5 CrossRefPubMedGoogle Scholar
  22. Johansson E, Jonson I, Bosaeus M, Jennische E (2008) Identification of flotillin-1 as an interacting protein for antisecretory factor. Regul Pept 146(1-3):303–309.  https://doi.org/10.1016/j.regpep.2007.11.005 CrossRefPubMedGoogle Scholar
  23. Kim M, Wasling P, Xiao M-Y, Jennische E, Lange S, Hanse E (2005) Antisecretory factor modulates GABAergic transmission in the rat hippocampus. Regul Pept 129(1-3):109–118.  https://doi.org/10.1016/j.regpep.2005.01.018 CrossRefPubMedGoogle Scholar
  24. Krantis A (2000) GABA in the mammalian enteric nervous system. New Physiol Sci 15:284–290Google Scholar
  25. Lange S, Jennische E, Johansson E, Lönnroth I (1999) The antisecretory factor: synthesis and intracellular localisation in porcine tissues. Cell Tissue Res 296(3):607–617.  https://doi.org/10.1007/s004410051322 CrossRefPubMedGoogle Scholar
  26. Lange S, Lönnroth I (2001) The antisecretory factor: synthesis, anatomical and cellular distribution, and biological action in experimental and clinical studies. Int Rev Cytol 210:39–75.  https://doi.org/10.1016/S0074-7696(01)10003-3 CrossRefPubMedGoogle Scholar
  27. Lange S, Bosaeus I, Jennische E, Johansson E, Lundgren BK, Lönnroth I (2003) Food-induced antisecretory factor activity is correlated with small bowel length in patients with intestinal resections. APMIS 111(10):985–988.  https://doi.org/10.1034/j.1600-0463.2003.1111011.x CrossRefPubMedGoogle Scholar
  28. Laurenius A, Wängberg B, Lange S, Jennische E, Lundgren BK, Bosaeus I (2003) Antisecretory factor counteracts secretory diarrhoea of endocrine origin. Clin Nutr 22(6):549–552.  https://doi.org/10.1016/S0261-5614(03)00057-8 CrossRefPubMedGoogle Scholar
  29. Leong SC, Narayan S, Lesser TH (2013) Antisecretory factor-inducing therapy improves patient-reported functional levels in Ménière’s disease. Ann Otol Rhinol Laryngol 122(10):619–624PubMedGoogle Scholar
  30. Lönnroth I, Lange S (1984) Purification and characterization of a hormone-like factor which inhibits cholera secretion. FEBS Lett 177(1):104–108.  https://doi.org/10.1016/0014-5793(84)80990-4 CrossRefPubMedGoogle Scholar
  31. Lönnroth I, Lange S (1986) Purification and characterization of the antisecretory factor: a protein in the central nervous system and in the gut which inhibits intestinal hypersecretion induced by cholera toxin. Biochim Biophys Acta 883(1):138–144.  https://doi.org/10.1016/0304-4165(86)90144-3 CrossRefPubMedGoogle Scholar
  32. Lönnroth I, Lange S, Skadhauge E (1988) The antisecretory factors: inducible proteins which modulate secretion in the small intestine. Comp Biochem Physiol 90A:611–617CrossRefGoogle Scholar
  33. Mann EO, Mody I (2010) Control of hippocampal gamma oscillation frequency by tonic inhibition and excitation of interneurons. Nat Neurosci 13(2):205–212.  https://doi.org/10.1038/nn.2464 CrossRefPubMedGoogle Scholar
  34. Rapallino MV, Cupello A, Lange S, Lönnroth I (2003) Antisecretory factor peptide derivatives specifically inhibit [3H]-γ-amino-butyric acid/36Cl out→in permeation across the isolated rabbit Deiters’ neuronal membrane. Acta Physiol Scand 179(4):367–371.  https://doi.org/10.1111/j.1365-201X.2003.01173.x CrossRefPubMedGoogle Scholar
  35. Reis HJ, Vanden Berghe P, Romano-Silva MA, Smith TK (2006) GABA-induced calcium signaling in cultured enteric neurons is reinforced by activation of cholinergic pathways. Neuroscience 139(2):485–494.  https://doi.org/10.1016/j.neuroscience.2005.12.023 CrossRefPubMedGoogle Scholar
  36. Robello M, Amico C, Cupello A (1993) Regulation of GABAA receptor in cerebellar granule cells in culture: differential involvement of kinase activities. Neuroscience 53(1):131–138.  https://doi.org/10.1016/0306-4522(93)90291-M CrossRefPubMedGoogle Scholar
  37. Saliba RS, Kretschmannova K, Moss SJ (2012) Activity-dependent phosphorylation of GABAA receptors regulates receptor insertion and tonic current. EMBO J 31(13):2937–2951.  https://doi.org/10.1038/emboj.2012.109 CrossRefPubMedPubMedCentralGoogle Scholar
  38. Strandberg J, Lindquist C, Lange S, Asztely F, Hanse E (2014) The endogenous peptide antisecretory factor promotes tonic GABAergic signaling in CA1 stratum radiatum interneurons. Front Cell Neurosci 8:1–9.  https://doi.org/10.3389/fncel.2014.00013 CrossRefGoogle Scholar
  39. Svensson K, Lange S, Lönnroth I, Widström AM, Hanson LÅ (2004) Induction of anti-secretory factor in human milk may prevent mastitis. Acta Paediatr 93(9):1228–1231CrossRefPubMedGoogle Scholar
  40. Zaman S, Aamir K, Lange S, Jennische E, Silfverdal SA, Hanson LÅ (2014) Antisecretory factor effectively and safely stops childhood diarrhoea: a placebo-controlled, randomised study. Acta Pediatr 103(6):659–664.  https://doi.org/10.1111/apa.12581 CrossRefGoogle Scholar
  41. Watanabe M, Fukuda A (2015) Development and regulation of chloride homeostasis in the central nervous system. Front Cell Neurosci 9:1–14.  https://doi.org/10.3389/fncel.2015.00371 CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of PhysicsUniversity of GenoaGenoaItaly
  2. 2.Department of Medical Biology and Cell Biology, Institute of BiomedicineUniversity of GothenburgGothenburgSweden

Personalised recommendations