The Double Insult of Neonatal Cystitis Plus Adult Somatic Inflammation Results in Corticotropin Releasing Factor Type II Receptor-Dependent Bladder Hypersensitivity in Female Rats

Published:September 08, 2022DOI:


      • Neonatal bladder inflammation (NBI) and adult Complete Freund's Adjuvant (CFA in hindlimb) interact
      • NBI+CFA increases reflex and dorsal horn neuronal responses to urinary bladder distension
      • The combination of NBI+CFA pretreatments is associated with increased spinal content of CRFR2
      • Responses augmented by NBI+CFA are attenuated by the spinal application of a CRFR2 antagonist
      • Following NBI, hind limb inflammation produced effects similar to re-inflammation of the bladder


      The spinal mechanisms of visceral hypersensitivity are poorly understood, particularly when there is an interaction with somatic systems. Recently we demonstrated that rats which were pretreated with neonatal bladder inflammation (NBI) and subsequently pretreated as adults with bladder re-inflammation had augmented reflex and neuronal responses to urinary bladder distension via a corticotropin-releasing factor receptor type II (CRFR2) mechanism. Another insult producing similar augmented responses is somatic inflammation induced by Complete Freund's Adjuvant (CFA) in the hindlimb. Using neurochemical measures and both reflex and neuronal responses to urinary bladder distension as endpoints, the present study probed the role of CRFR2-related mechanisms in bladder hyperalgesia secondary to NBI and CFA-induced hindlimb inflammation. ELISA measures of the lumbosacral spinal cord demonstrated increased CRFR2 protein following pretreatment with NBI+CFA. Intrathecal CRFR2 antagonists blocked the augmentation of visceromotor responses to distension following pretreatment with both NBI+CFA. Lumbosacral dorsal horn neuronal responses to bladder distension in rats pretreated with NBI+CFA were attenuated by the spinal topical administration of a CRFR2 antagonist. These findings are the first demonstration of a somatovisceral interaction working via CRFR2 receptors and support the therapeutic value of these agents in the treatment of painful bladder disorders, particularly when triggered by somatic events. (Word Count 199).


      Bladder hypersensitivity occurs following neonatal cystitis and an adult insult such as somatic inflammation. This paper demonstrates that CRFR2-related mechanisms are associated with this hypersensitivity. This supports the therapeutic value of these agents in the treatment of painful bladder disorders, particularly when triggered by somatic events.

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        • Abdelhamid RE
        • Kovacs KJ
        • Palsey JD
        • Nunez MG
        • Larson AA.
        Forced swim-induced musculoskeletal hyperalgesia is mediated by CRF2 receptors but not by TRPV1 receptors.
        Neuropharmacol. 2013; 72: 29-37
        • Aldossary SA
        • Alsalen M
        • Kalbouneh H
        • Haddad M
        • Azab B
        • Al-Shboul O
        • Mustafa AG
        • Obiedat S
        • El-Salem K.
        The role of transient receptor potential vanilloid receptor 1 and peroxisome proliferator-activated receptors-alpha in mediating the antinocieptive effects of palmitoylethanolamine in rats.
        Neuroreport. 2019; 30: 32-37
      1. Anonymous. Causes of IC., 2021

        • Bagdas D
        • AlSharari SD
        • Freitas K
        • Tracy M
        • Damaj MI
        The role of alpha5 nicotinic acetylcholine receptors in mouse models of chronic inflammatory and neuropathic pain.
        Biochem Pharmacol. 2015; 97: 590-600
        • Berry SH
        • Elliott MN
        • Suttorp M
        • Bogart LM
        • Stoto MA
        • Eggers P
        • Nyberg L
        • Clemens JQ.
        Prevalence of symptoms of bladder pain syndrome/interstitial cystitis among adult females in the United States.
        J Urol. 2011; 186: 540-544
        • Bogart LM
        • Berry SH
        • Clemens JQ.
        Symptoms of interstitial cystitis, painful bladder syndrome and similar diseases in women: A systematic review.
        J Urol. 2007; 177: 450-456
        • Brumovsky PR
        • Gebhart GF.
        Visceral organ cross-sensitization: An integrated perspective.
        Auton Neurosci. 2010; 153: 106-115
        • Castroman P
        • Ness TJ.
        Vigor of visceromotor responses to urinary bladder distension in rats increases withrepeated trials and stimulus intensity.
        Neurosci Letts. 2001; 306: 97-100
        • Coderre TJ
        • Laferriere A.
        The emergence of animal models of chronic pain and logistical and methodological issues concerning their use.
        J Neural Trans. 2020; 127: 393-406
        • Fehrenbacher JC
        • Vasko MR
        • Duarte DB.
        Models of inflammation: Carrageenan- or complete Freund's adjuvant-induced edema and hypersensitivity in the rat.
        Curr Protoc Pharmacol. 2012; 5: 4
        • Hand JR.
        Interstitial cystitis: Report of 223 cases (204 women and 19 men).
        J Urol. 1949; 61: 291-310
        • Huang Y
        • Chen S-R
        • Chen H
        • Pan H-L
        Endogenous transient receptor potential ankyrin 1 and vanilloid1 activity potentials glutamatergic input to spinal lamina I neurons in inflammatory pain.
        J Neurochem. 2019; 149: 381-398
        • Jaggar SI
        • Scott HC
        • Rice AS.
        Inflammation of the rat urinary bladder is associated with a referred thermal hyperalgesia which is nerve growth factor dependent.
        Br J Anaesth. 1999; 83: 442-448
        • Jiang Y-L
        • He X-F
        • Shen Y-F
        • Yin X-H
        • Du J-Y
        • Liang Y
        • Fang J-Q.
        Analgesic roles of perineural intrinsic met-enkephalin and dynorphin in long-lasting inflammatory pain induced by complete Freund's adjuvant in rats.
        Exp Ther Med. 2015; 9: 2344-2348
        • Jones CA
        • Nyberg L.
        Epidemiology of interstitial cystitis.
        Urol. 1997; 49: 2-9
        • Kannampalli P
        • Babygirlja R
        • Zhang J
        • Poe MM
        • Li G
        • Cook JM
        • Shaker R
        • Banerjee B
        • Sengupta JN.
        Neonatal bladder inflammation induces long-term visceral pain and altered responses of spinal neurons in adult rats.
        Neurosci. 2017; 346: 349-364
        • Kiddoo DA
        • Valentino RJ
        • Zderic S
        • Ganesh A
        • Leiser SC
        • Hale L
        • Grigoriadis DE
        Impact of state of arousal and stress neuropeptides on urodynamic function in freely moving rats.
        Am J Physiol. 2006; 290: R1697-R1706
        • Kozaki Y
        • Hori Km Aye-Mon A
        • Okuda H
        • Harada S-I
        • Hayashi K
        • Ozaki N.
        The role of peripheral corticotropin-releasing factor signaling in a rat model of stress-induced gastric hyperalgesia.
        Biochem Biophys Res Comms. 2019; 519: 797-802
        • Koziol JA
        • Clark DC
        • Gittes RF
        • Tan EM.
        The natural history of interstitial cystitis: A survey of 374 patients.
        J Urol. 1993; 149: 465-469
        • Lamb K
        • Zhong F
        • Gebhart GF
        • Bielfeldt K.
        Experimental colitis in mice and sensitization of converging visceral and somatic afferent pathways.
        Am J Physiol. 2006; 290: G451-G457
        • Larson AA
        • Nunez MG
        • Kissel CL
        • Kovacs KJ.
        Intrathecal urocortin I in the spinal cord as a murine model of stress hormone-induced musculoskeletal and tactile hyperalgesia.
        Eur J Neurosci. 2015; 42: 2772-2782
        • Martinez V
        • Ang L
        • Million M
        • Rivier J
        • Tache Y.
        Urocortins and the regulation of gastrointestinal motor function and visceral pain.
        Peptides. 2004; 25: 1733-1744
        • Million M
        • Maillot C
        • Adelson DA
        • Nozu T
        • Gauthier A
        • Rivier J
        • Chrousos GP
        • Bayati A
        • Mattson H
        • Tache Y.
        Peripheral injection of sauvagine prevents repeated colorectal distension-induced visceral pain in female rats.
        Peptides. 2005; 26: 1188-1195
        • Million M
        • Wang L
        • Wang Y
        • Adelson DW
        • Yuan P-Q
        • Maillot C
        • Coutinho SV
        • McRoberts JA
        • Bayati A
        • Mattsson H
        • Wu V
        • Wei J-Y
        • Rivier J
        • Vale W
        • Mayer EA
        • Tache Y
        CRF2 receptor activation prevents colorectal distension induced visceral pain and spinal ERK1/2 phosphorylation in rats.
        Gut. 2006; 55: 172-188
        • Mousa SA
        • Shaqura M
        • Khalefa BI
        • Li L
        • Al-Madol M
        • Treskatsch S
        • Schafer M
        Functional and anatomical characterization of corticotropin-releasing factor receptor subtypes of the rat spinal cord involved in somatic pain relief.
        Mol Neurobiol. 2021; 58: 5459-5472
        • Muley MM
        • Krustev E
        • McDougall JJ.
        Preclinical assessment of inflammatory pain.
        CNS Neurosci Ther. 2016; 22: 88-101
        • Ness TJ
        • Castroman P.
        Evidence for two populations of rat spinal dorsal horn neurons excited by urinary bladder distension.
        Brain Res. 2001; 923: 147-156
        • Ness TJ
        • DeWitte C
        • DeBerry JJ
        • Hart MP
        • Clodfelder-Miller B
        • Gu J
        • Ling J
        • Randich A
        A model in female rats with phenotypic features similar to interstitial cystitis/bladder pain syndrome.
        Front Pain Res. 2021; 2791045
        • Ness TJ
        • DeWitte C
        • Randich A.
        Neonatal cystitis leads to alterations in spinal corticotropin releasing factor receptor-type 2 content and function in adult rats following bladder re-inflammation.
        Brain Res. 2022; 1788147927
        • Ness TJ
        • Gebhart GF.
        Characterization of superficial T13-L2 dorsal horn neurons encoding for colorectal distension in the rat: Comparison with neurons in deep laminae.
        Brain Res. 1989; 486: 301-309
        • Ness TJ
        • Gebhart GF.
        Interactions between visceral and cutaneous nociception in the rat. I. Noxious cutaneous stimuli inhibit visceral nociceptive neurons and reflexes.
        J Neurophysiol. 1991; 66: 20-28
        • Ness TJ
        • Lewis-Sides A
        • Castroman P.
        Characterization of pressor and visceromotor reflex responses to bladder distension in rats: Sources of variability and effect of analgesics.
        J Urol. 2001; 165: 968-974
        • Ness TJ
        • Randich A.
        Neonatal bladder inflammation alters activity of adult rat spinal visceral nociceptive neurons.
        Neurosci Lett. 2010; 472: 210-214
        • Nijsen M
        • Ongenae N
        • Meulemans A
        • Coulie B.
        Divergent role for CRF1 and CRF2 receptors in the modulation of visceral pain.
        Neurogastroenterol Motil. 2005; 17: 423-432
        • Nozu T
        • Takakusaki K
        • Okumura T.
        A balance theory of peripheral corticotropin-releasing factor receptor Type 1 and type 2 signaling to induce colonic contractions and visceral hyperalgesia in rats.
        Endocrinol. 2014; 155: 4655-4664
        • Patel R
        • Calhoun EA
        • Meenan RT
        • O'Keefe Rosetti MC
        • Kimes T
        • Clemens JQ
        Incidence and clinical characteristics of interstitial cystitis in the community.
        Int Urogynecol J Pelvic Floor Dysfunct. 2008; 19: 1093-1096
        • Pattinson D
        • Fitzgerald M.
        The neurobiology of infant pain: Development of excitatory and inhibitory neurotransmission in the spinal cord.
        Reg Anesth Pain Med. 2004; 29: 36-44
        • Peters KM
        • Killinger KA
        • IA Ibrahim
        Childhood symptoms and events in women with interstitial cystitis/painful bladder syndrome.
        Urology. 2009; 73: 258-262
        • Radwani H
        • Roca-Lapirot O
        • Aby F
        • Lopez-Gonzalez MJ
        • Benassouz Errami M
        • Favereaux A
        • Landry M
        • Fossat P
        Group I metabotropic glutamate receptor plasticity after peripheral inflammation alters nociceptive transmission in the dorsal horn of the spinal cord in adult rats.
        Mol Pain. 2017; 131744806917737934
        • Randich A
        • Shaffer AD
        • Ball CL
        • Mebane H.
        Serotonergic and noradrenergic facilitation of the visceromotor reflex evoked by urinary bladder distension in rats with inflamed bladders.
        Neurosci Lett. 2008; 442: 253-256
        • Randich A
        • Uzzell TW
        • DeBerry JJ
        • Ness TJ.
        Neonatal urinary bladder inflammation produces adult bladder hypersensitivity.
        J Pain. 2006; 7: 469-479
        • Rivier J
        • Gulyas J
        • Kirby D
        • Low W
        • Perrin MH
        • Kunitake K
        • Gruccio M
        • Vaughan J
        • Reubi JC
        • Waser B
        • Koerber SC
        • Martinez V
        • Wang L
        • Tache Y
        • Vale W
        Potent and long-acting corticotropin releasing factor (CRF) receptor 2 selective peptide competitive antagonists.
        J Med Chem. 2002; 45: 4737-4747
        • Rivier J
        • Gulyas J
        • Kirby D
        • Low W
        • Perrin MH
        • Kunitake K
        • DiSchwaller F
        • Fitzgerald M.
        The consequences of pain in early life: Injury-induced plasticity in developing pain pathways.
        Eur J Neurosci. 2014; 39: 344-352
        • Sengupta JN
        • Pochiraju S
        • Kannampalli P
        • Bruckert M
        • Addys S
        • Yadav P
        • Miranda A
        • Shaker R
        • Banerjee B.
        MicroRNA-mediated GABAAa-1 receptor subunit downregulation in adult spinal cord following neonatal cystitis-induced chronic visceral pain in rats.
        Pain. 2013; 154: 59-70
        • Shaffer AD
        • Ness TJ
        • Robbins MT
        • Randich A.
        Early in life bladder inflammation alters opioid peptide content in the spinal cord and bladder of adult female rats.
        J Urol. 2013; 189: 352-358
        • Shaffer AD
        • Ness TJ
        • Randich A.
        Early-in-life bladder inflammation alters U50,488H but not morphine-induced inhibition of visceromotor responses to urinary bladder distension.
        Neurosci Lett. 2013; 534: 150-154
        • Uta D
        • Kato G
        • Ddoi A
        • Andoh T
        • Kume T
        • Yoshimura M
        • Koga K
        Animal models of chronic pain increase spontaneous glutamatergic transmission in adult rat spinal dorsal horn in vitro and in vivo.
        Biochem Biophys Res Comm. 2019; 512: 352-359
        • Uta D
        • Yoshimura M
        • Koga K.
        Chronic pain models amplify transient receptor potential vanilloid 1 (TRPV1) receptor responses in adult rat spinal dorsal horn.
        Neuropharm. 2019; 160107753
        • Vincent K
        • Wang SF
        • Laferriere A
        • Kumar N
        • Coderre TJ.
        Spinal intracellular metabotropic glutamate receptor 5 (mGluR5) contributes to pain and c-fos expression in a rat model of inflammatory pain.
        Pain. 2017; 158: 705-716
        • Warren JW
        • Brown V
        • Jacobs S
        • Horne L
        • Langenberg P
        • Greenberg P.
        Urinary tract infection and inflammation at onset of interstitial cystitis/painful bladder syndrome.
        Urology. 2008; 71: 1085-1090
        • Yu L
        • Yang F
        • Luo H
        • Liu F-Y
        • Han J-S
        • Xing G-G
        • Wan Y.
        The role of TRPV1 in different subtypes of dorsal root ganglion neurons in rat chronic inflammatory nocicep0tioin induced by complete Freund's adjuvant.
        Mol Pain. 2008; 4: 61
        • Zhu H
        • Yu Y
        • Zheng L
        • Wang L
        • Li C
        • Yu J
        • Wei J
        • Wang C
        • Zhang J
        • Xu S
        • Wei X
        • Cui W
        • Wang Q
        • Chen X
        Chronic inflammatory pain upregulates expression of P2Y2 receptor in small-diameter sensory neurons.
        Metab Brain Dis. 2015; 30: 1349-1358
        • Zimmermann M.
        Ethical guidelines for investigations of experimental pain in conscious animals.
        Pain. 1983; 16: 109-110