title: MicroRNA-mediated GABA Aα-1 receptor subunit down-regulation in adult spinal cord following neonatal cystitis-induced chronic visceral pain in rats.
abstract: The nociceptive transmission under pathological chronic pain conditions involves transcriptional and/or translational alteration in spinal neurotransmitters, receptor expressions, and modification of neuronal functions. Studies indicate the involvement of microRNA (miRNA) - mediated transcriptional deregulation in the pathophysiology of acute and chronic pain. In the present study, we tested the hypothesis that long-term cross-organ colonic hypersensitivity in neonatal zymosan-induced cystitis is due to miRNA-mediated posttranscriptional suppression of the developing spinal GABAergic system. Cystitis was produced by intravesicular injection of zymosan (1% in saline) into the bladder during postnatal (P) days P14 through P16 and spinal dorsal horns (L6-S1) were collected either on P60 (unchallenged groups) or on P30 after a zymosan re-challenge on P29 (re-challenged groups). miRNA arrays and real-time reverse transcription-polymerase chain reaction (RT-PCR) revealed significant, but differential, up-regulation of mature miR-181a in the L6-S1 spinal dorsal horns from zymosan-treated rats compared with saline-treated controls in both the unchallenged and re-challenged groups. The target gene analysis demonstrated multiple complementary binding sites in miR-181a for GABA(A) receptor subunit GABA(Aα-1) gene with a miRSVR score of -1.83. An increase in miR-181a concomitantly resulted in significant down-regulation of GABA(Aα-1) receptor subunit gene and protein expression in adult spinal cords from rats with neonatal cystitis. Intrathecal administration of the GABA(A) receptor agonist muscimol failed to attenuate the viscero-motor response (VMR) to colon distension in rats with neonatal cystitis, whereas in adult zymosan-treated rats the drug produced significant decrease in VMR. These results support an integral role for miRNA-mediated transcriptional deregulation of the GABAergic system in neonatal cystitis-induced chronic pelvic pain.
authors.affiliation: Division of Gastroenterology and Hepatology, Medical College of Wisconsin, Milwaukee, WI 53226, USA Division of Pediatric Gastroenterology, Hepatology and Nutrition, Medical College of Wisconsin, Milwaukee, WI, USA Cancer Genomics Facility, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, USA.
authors.forename: Jyoti N
authors.initials: JN
authors.lastname: Sengupta
authors.forename: Soumiya
authors.initials: S
authors.lastname: Pochiraju
authors.forename: Pradeep
authors.initials: P
authors.lastname: Kannampalli
authors.forename: Mitchell
authors.initials: M
authors.lastname: Bruckert
authors.forename: Sankar
authors.initials: S
authors.lastname: Addya
authors.forename: Priyanka
authors.initials: P
authors.lastname: Yadav
authors.forename: Adrian
authors.initials: A
authors.lastname: Miranda
authors.forename: Reza
authors.initials: R
authors.lastname: Shaker
authors.forename: Banani
authors.initials: B
authors.lastname: Banerjee
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D000367:Age Factors; D000818:Animals; D018236:Carcinoma, Embryonal; D045744:Cell Line, Tumor; D059350:Chronic Pain / Q000209:etiology / Q000235:genetics / Q000503:physiopathology*; D003556:Cystitis / Q000139:chemically induced / Q000150:complications / Q000503:physiopathology*; D004195:Disease Models, Animal; D015536:Down-Regulation / Q000235:genetics; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D017699:Pelvic Pain / Q000209:etiology / Q000235:genetics / Q000503:physiopathology; D020671:Posterior Horn Cells / Q000502:physiology; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D011963:Receptors, GABA-A / Q000235:genetics* / Q000378:metabolism; D013116:Spinal Cord / Q000502:physiology*; D059265:Visceral Pain / Q000209:etiology / Q000235:genetics / Q000503:physiopathology*; D015054:Zymosan / Q000494:pharmacology
chemical_list: D020413:3' Untranslated Regions; C546242:Gabra1 protein, rat; C581717:MIRN181 microRNA, rat; D035683:MicroRNAs; D011963:Receptors, GABA-A; D015054:Zymosan