1. Volume 140, Issue 2, Pages 627-637.e4 (February 2011)
Corticosterone Mediates Reciprocal Changes in CB 1 and TRPV1 Receptors in Primary Sensory Neurons in the Chronically Stressed Rat 
Shuangsong Hong, Gen Zheng, Xiaoyin Wu, Natasha T. Snider, Chung Owyang, John W. Wiley 
Gastroenterology 
Volume 140, Issue 2, Pages e4 (February 2011)
DOI: /j.gastro
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2. Figure 1
Effect of the corticoid-receptor antagonist RU-486 (2 mg/kg; subcutaneously) on the VMR to CRD. (A) Representative EMG recordings depicting the VMR to CRD at 40 mm Hg in control rats, WA stress rats, and WA stress rats treated with RU-486 during the stress procedure. (B) EMG amplitude expressed as mean change from baseline in control and WA stress rats treated with or without RU-486. (C) Effect of RU-486 on distal colonic motor function in WA stress rats. Data are expressed as mean ± standard error, n = 8 in each group. *P < .05 between control and WA stress rats; +P < .05 between WA stress rats receiving RU-486 or vehicle.
Gastroenterology  , e4DOI: ( /j.gastro )
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3. Figure 2
Treatment with RU-486 prevented the reciprocal changes in TRPV1 and CB1 receptors in DRG neurons innervating the colon in WA stress rats. (A) Representative immunofluorescence images (left) of double-IR–positive neurons for CB1 (red) and TRPV1 (red) in colonic DRG neurons retrograde-labeled with CTB–fluorescein isothiocyanate (green) from control, WA stress, and RU-486–treated WA stress rats. Scale bar: 40 μm. Bar graph (right) depicting the changes in the percentage of double-IR–positive neurons for CB1 and TRPV1 with CTB–fluorescein isothiocyanate. (B) Immunoblots depicting the protein levels of CB1 and TRPV1 in L6–S2 DRGs in WA stress rats in the absence and presence of RU-486. (C) Histogram showing the immunoblot band density of CB1 and TRPV1 protein (n = 5). *P < .05.
Gastroenterology  , e4DOI: ( /j.gastro )
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4. Figure 3
DRG (L6–S2) neurons from WA stress rats showed increased capsaicin-evoked TRPV1 current that was prevented by treatment with RU-486 in situ. (A) Representative inward current traces of neurons responding to the application of 1 μmol/L capsaicin (2 s) in L6–S2 DRG neurons isolated from control, WA stress, and RU-486–treated stress rats. (B) Bar graph depicting the mean peak current density of the capsaicin-evoked currents (n = 8). *P < .05.
Gastroenterology  , e4DOI: ( /j.gastro )
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5. Figure 4
Serial injections of control rats with corticosterone (CORT) in situ induced visceral hyperalgesia that was prevented by co-treatment with RU-486. (A) The level of serum corticosterone was increased significantly in rats after repeated injection with corticosterone. (B) Representative EMG recordings depicting the VMR to CRD at 40 mm Hg in control rats, corticosterone-injected rats, and rats treated with both corticosterone and RU-486. (C) EMG amplitude expressed as mean change from baseline in corticosterone-injected rats in the absence or presence of RU-486 (n = 8). *P < .05.
Gastroenterology  , e4DOI: ( /j.gastro )
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6. Figure 5
Serial injections of corticosterone-induced reciprocal changes in CB1 and TRPV1 expression in L6–S2 DRG neurons. (A) Immunofluorescence staining of double-IR–positive neurons for TRPV1 (red) and CB1 (red) in colonic DRG neurons retrograde-labeled with CTB–fluorescein isothiocyanate (green) from control and serially treated corticosterone (cort)-injected rats. Scale bar: 30 μm. (B) Co-treatment with RU-486 or WIN prevented the changes in TRPV1 and CB1 protein expression in the corticosterone-injected control rats. (C) Statistical analysis of the immunoblot band intensity of TRPV1 and CB1 receptors (n = 5). *P < .05.
Gastroenterology  , e4DOI: ( /j.gastro )
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7. Figure 6
Changes in endocannabinoid degradation enzymes and anandamide content in L6–S2 DRGs. (A) Western blot analysis showed decreased expression levels of COX-2 and FAAH protein in L6–S2 DRGs in both chronic WA stress rats and corticosterone-injected control rats compared with untreated controls. (B) Histogram depicting the relative changes of COX-2 and FAAH protein levels in chronic WA stress rats and corticosterone-injected control rats (n = 6). (C) The bar graph illustrated the increase in anandamide content in L6–S2 DRGs in rats after corticosterone injection compared with controls receiving vehicle only. L6–S2 DRGs from 10 rats in each group were combined for sample extraction. *P < .05.
Gastroenterology  , e4DOI: ( /j.gastro )
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8. Figure 7
Effects of corticosterone in the absence and presence of RU-486 or WIN on the levels of CB1 and TRPV1 in L6–S2 DRGs isolated from control rats in vitro. (A) Representative Western blots for TRPV1 and CB1 in L6–S2 DRGs isolated from control rats treated with corticosterone in the absence and presence of RU-486 (500 nmol/L) or WIN (1 μmol/L). (B) Exposure of control DRG explants to corticosterone significantly decreased the expression level of CB1 and increased the level of TRPV1-receptor protein levels (P < .05; n = 4). Co-treatment with RU-486 or WIN blocked corticosterone-induced changes in CB1 and TRPV1. *P < .05.
Gastroenterology  , e4DOI: ( /j.gastro )
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9. Supplementary Figure 1
Treatment with RU-486 prevented the reciprocal changes in TRPV1 and CB1 receptors in whole L6–S2 DRG neurons in WA stress rats. Representative immunofluorescence images of double-IR–positive neurons for CB1 (green) and TRPV1 (red) in DRGs from control, WA stress, and RU-486–treated WA stress rats were shown. Scale bar: 50 μm. Bar graph depicting the changes in the percentage of double-IR–positive neurons for CB1 and TRPV1. Data are expressed as mean ± standard error, n = 5 in each group. *P < .05.
Gastroenterology  , e4DOI: ( /j.gastro )
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10. Supplementary Figure 2
Serial injections of corticosterone (cort)-induced reciprocal changes in TRPV1 and CB1 expression in whole L6–S2 DRG neurons. (A) Immunofluorescence staining of TRPV1 (red; a and b) and CB1 (green; c and d) in L6–S2 DRGs from the corticosterone-injected rats (b and d) and the controls (a and c). Scale bar: 35 μm. (B) Bar graph depicting the changes in staining intensities of TRPV1 and CB1 after repeated CORT injection (n = 5). *P < .05.
Gastroenterology  , e4DOI: ( /j.gastro )
Copyright © 2011 AGA Institute Terms and Conditions