CRF2 receptor activation prevents colorectal distension induced visceral pain and spinal ERK1/2 phosphorylation in rats.
OBJECTIVE
Activation of corticotropin releasing factor 1 (CRF1) receptors is involved in stress related responses and visceral pain, while activation of CRF2 receptors dampens the endocrine and some behavioural stress responses. We hypothesised that CRF2 receptor activation may influence visceral pain induced by colorectal distension (CRD) in conscious rats, and assessed the possible sites and mechanisms of action.


METHODS
Male Sprague-Dawley rats were exposed to CRDs (60 mm Hg, 10 minutes twice, with a 10 minute rest interval). Visceromotor responses (VMR) were measured by electromyography or visual observation. Spinal (L6-S1) extracellular signal regulated kinase 1/2 (ERK 1/2) activation following in vivo CRD and CRF2 receptor gene expression in the T13-S1 dorsal root ganglia (DRG) and spinal cord were determined. Inferior splanchnic afferent (ISA) activity to CRD (0.4 ml, 20 seconds) was assessed by electrophysiological recording in an in vitro ISA nerve-inferior mesenteric artery (intra-arterial)-colorectal preparation.


RESULTS
In controls, VMR to the second CRD was mean 31 (SEM 4)% higher than that of the first (p<0.05). The selective CRF2 agonist, human urocortin 2 (hUcn 2, at 10 and 20 microg/kg), injected intravenous after the first distension, prevented sensitisation and reduced the second response by 8 (1)% and 30 (5)% (p<0.05) compared with the first response, respectively. RT-PCR detected CRF2 receptor gene expression in the DRG and spinal cord. CRD (60 mm Hg for 10 minutes) induced phosphorylation of ERK 1/2 in neurones of lumbosacral laminae I and IIo and the response was dampened by intravenous hUcn 2. CRD, in vitro, induced robust ISA spike activity that was dose dependently blunted by hUcn 2 (1-3 microg, intra-arterially). The CRF2 receptor antagonist, astressin2-B (200 microg/kg subcutaneously or 20 microg intra-arterially) blocked the hUcn 2 inhibitory effects in vivo and in vitro.


CONCLUSIONS
Peripheral injection of hUcn 2 blunts CRD induced visceral pain, colonic afferent, and spinal L6-S1 ERK 1/2 activity through CRF2 receptor activation in rats.
Million
M
M
CURE/Digestive Diseases Research Center, Department of Medicine, University of California Los Angeles, CA 90073, USA. mmuluget@ucla.edu
Wang
L
L
Wang
Y
Y
Adelson
D W
DW
Yuan
P-Q
PQ
Maillot
C
C
Coutinho
S V
SV
Mcroberts
J A
JA
Bayati
A
A
Mattsson
H
H
Wu
V
V
Wei
J-Y
JY
Rivier
J
J
Vale
W
W
Mayer
E A
EA
Taché
Y
Y
D000818:Animals; D002404:Catheterization; D003106:Colon / Q000473:pathology; D003346:Corticotropin-Releasing Hormone / Q000494:pharmacology; D004576:Electromyography; D005772:Gastrointestinal Transit / Q000187:drug effects / Q000502:physiology; D005786:Gene Expression Regulation / Q000187:drug effects; D007420:Intestine, Large / Q000294:innervation / Q000503:physiopathology*; D008297:Male; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D009043:Motor Activity / Q000187:drug effects / Q000502:physiology; D009619:Nociceptors / Q000502:physiology; D010146:Pain / Q000209:etiology / Q000503:physiopathology / Q000517:prevention & control*; D010766:Phosphorylation; D010812:Physical Stimulation / Q000379:methods; D051381:Rats; D017207:Rats, Sprague-Dawley; D018019:Receptors, Corticotropin-Releasing Hormone / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013116:Spinal Cord / Q000201:enzymology; D054832:Urocortins
C101531:CRF receptor type 2; D018019:Receptors, Corticotropin-Releasing Hormone; C426705:UCN2 protein, human; D054832:Urocortins; D003346:Corticotropin-Releasing Hormone; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3