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Urocortin

 Urocortin and Stresscopin related products

New CRF/Urocortin family members

Human Stresscopin (SCP) and Stresscopin Related Peptide (SRP), two selectively natural ligands for CRFR2, enable to suppress food intake, delay gastric emptying and decrease heat-induced edema. The gene of SCP and SRP  were expressed in central and diverse peripheral tissues. Because CRFR2 is believed to be important in the regulation of the recovery phase of the stress response, SCP and SRP might be important in protecting the organism from damage incurred by prolonged or excessive exposure to stress.  Unlike CRF and Urocortin, SCP and SRP have minimal effects on ACTH release and the resultant elevations in glucocorticoids.

Hsu, S.Y. & Hsueh, A.J.W. Nature Medicine, 7 605-611, 2001


Urocortin III (Human)& Urocortin III (Mouse)

An additional member of CRF family with high affinity for the CRFR2 Lewis, K. et al. Proc. Natl. Acad. Sci. USA 98, 7570-7575, 2001 (June 19)
1. Urocortin III (Human) is identical with Stresscopin (3-40)-NH2 (Human).
2. Urocortin II (Urocortin Related Peptide, URP) (Human) is identical with Stresscopin Related Peptide (6-43)-NH2 (Human).

References:
Lewis, K. et al. Proc. Natl. Acad. Sci. USA  98, 7570-7575, 2001 (June 19);
Hsu, S.Y. & Hsueh, A.J.W. Nature Medicine, 7 605-611, 2001
Review by Jun Yang on June 21, 2001

Urocortin reduces food intake and gastric emptying in lean and ob/ob obese mice
BACKGROUND & AIMS: Gastric emptying plays an important role in regulating food intake. This study was designed to investigate whether intraperitoneally injected urocortin reduces gastric emptying, feeding, and body weight in lean and ob/ob obese mice. METHODS: Food intake and body weight were measured after intraperitoneal injections of one of the following: urocortin, deamidated form of urocortin (urocortin OH), corticotropin-releasing factor (CRF), CRF6-33, cholecystokinin octapeptide (CCK-8), and leptin in 16-hour food-deprived animals. Gastric emptying was assessed 2, 4, or 8 hours after intraperitoneal injection. Repeated injections of urocortin were continued for 5 days in ob/ob mice. RESULTS: Urocortin (0.003-3 nmol) dose-dependently and potently decreased food intake and body weight gain in lean mice. The ranking order of potency was urocortin > urocortin OH >/= CRF > CCK-8 > CRF6-33 > leptin. Gastric emptying was also potently reduced by urocortin with a similar ranking order of potency of urocortin > CRF > urocortin OH > CCK-8. Simultaneous administration of urocortin and CRF receptor antagonist, alpha-helical CRF9-41, blocked the effects of urocortin. Urocortin reduced food intake and body weight gain, as well as the rate of gastric emptying, in ob/ob mice, which was significantly faster than that of lean mice. Five daily injections of urocortin significantly lowered body weight and improved glycemic control in ob/ob mice. CONCLUSIONS: The urocortin-induced decrease in food intake and body weight in lean and ob/ob mice is closely related to gastric emptying and opens new possibilities for the treatment of obesity.

Asakawa A, Inui A, Ueno N, Makino S, Fujino MA, Kasuga M. Gastroenterology 1999 Jun;116(6):1287-92

Urocortin reduces oxygen consumption in lean and ob/ob mice

A vast number of intensive studies have been undertaken to clarify the mechanisms of energy balance. This study was undertaken to investigate the effect of urocortin, an endogenous ligand for corticotropin-releasing factor (CRF) type 2 receptor, on oxygen consumption in lean and genetically obese (ob/ob) mice. Oxygen consumption was measured after intraperitoneal injection in unrestrained mice at an environmental temperature of 22 degrees C of one of the following: urocortin, deamidated form of urocortin (urocortin OH) or CRF. The intraperitoneal injection of urocortin (0.3-3 nmol) dose-dependently decreased oxygen consumption in lean mice. The inhibitory effect induced by urocortin was more potent than that induced by CRF or urocortin OH. The ranking potency was urocortin > urocortin OH > CRF. Urocortin significantly reduced oxygen consumption in ob/ob mice as well as in lean mice. These results suggest that urocortin decreases oxygen consumption, and that the CRF type 2 receptor may influence energy balance in lean and ob/ob mice.

Asakawa A, Inui A, Ueno N, Makino S, Fujimiya M, Fujino MA, Kasuga M. Int J Mol Med 2001 May;7(5):539-41
Rat brain tissue was stained with Urocortin Antibody (catalog No.: H-019-06)

Rat brain tissue was stained with Urocortin Antibody (catalog No.: H-019-06)

 Urocortin and Stresscopin related products

Human Urocortin II (Urocortin Related Peptide, URP) is identical with Stresscopin Related Peptide (SRP) (6-43)-NH2 (Human).
Human Urocortin III is identical with human Stresscopin (3-40)-NH2

References:
Lewis, K. et al. Proc. Natl. Acad. Sci. USA 98, 7570-7575, 2001 (June 19)
Hsu, S.Y. & Hsueh, A.J.W. Nature Medicine, 7 605-611, 2001
Review by Jun Yang on June 21, 2001



Amino Acid Sequence of Prepro-Urocortin II (Mouse)

 
1   M T R W A L V V F V V L M L D R I L F V   20
21   P G T P I P T F Q L L P Q N S L E T T P   40
41   S S V T S E S S S G T T T G P S A S w S   60
61   N S K A S P Y L D T R V I L S L D V P I   80
81   G L L R I L L E Q A R Y K A A R N Q A A   100
101   T N A Q I L A H V G R R                   112
 


 
 

 

Human Urocortin II, a Selective Agonist for the Type 2 Corticotropin-Releasing Factor Receptor, Decreases Feeding and Drinking in the Rat
Corticotropin-releasing factor (CRF) has been hypothesized to modulate consummatory behavior through the Type 2 CRF (CRF(2)) receptor. However, behavioral functions subserved by the CRF(2) receptor remain poorly understood. Recently, human urocortin II (hUcn II), a selective CRF(2) receptor agonist, was identified. To study the effects of this neuropeptide on ingestive behavior, we examined the effects of centrally infused hUcn II (i.c.v. 0, 0.01, 0.1, 1.0, 10.0 &mgr;g) on the microstructure of nose-poke responding for food and water in nondeprived, male rats. Malaise-inducing properties of the peptide were monitored using conditioned taste aversion (CTA) testing. To identify potential sites of action, central induction of Fos protein expression was examined. hUcn II dose dependently reduced the quantity and duration of responding for food and water at doses lower (0.01-1.0 &mgr;g) than that forming a CTA (10 &mgr;g). Effects were most evident during hours 4 to 6 of the dark cycle. Meal pattern analysis showed that hUcn II potently (0.1 &mgr;g) increased the satiating value of food. Rats ate and drank smaller and shorter meals without changing meal frequency. Rats also ate more slowly. hUcn II induced Fos in regions involved in visceral sensory processing and autonomic/neuroendocrine regulation and resembling those activated by appetite suppressants. hUcn II is a promising neuropeptide for investigating the role of the CRF(2) receptor in ingestive behavior.

Inoue K,et al. J Pharmacol Exp Ther 2003 Apr 1;305(1):385-393

Human urocortin II, a new CRF-related peptide, displays selective CRF(2)-mediated action on gastric transit in rats
Human urocortin (hUcn) II is a new member of the corticotropin-releasing factor (CRF) family that selectively binds to the CRF(2) receptor. We investigated the CRF receptors involved in mediating the effects of hUcn II and human/rat CRF (h/rCRF) on gut transit. Gastric emptying, 4 h after a solid meal, and distal colonic transit (bead expulsion time) were monitored simultaneously in conscious rats. CRF antagonists were given subcutaneously 30 min before intravenous injection of peptides or partial restraint (for 90 min). hUcn II (3 or 10 microg/kg i.v.) inhibited gastric emptying (by 45% and 55%, respectively) and did not influence distal colonic transit. The CRF(2) peptide antagonist astressin(2)-B blocked hUcn II action. h/rCRF, rat Ucn, and restraint delayed gastric emptying while accelerating distal colonic transit. The gastric response to intravenous h/rCRF and restraint was blocked by the CRF(2) antagonist but not by the CRF(1) antagonist CP-154,526, whereas the colonic response was blocked only by CP-154,526. None of the CRF antagonists influenced postprandial gut transit. These data show that intravenous h/rCRF and restraint stress-induced delayed gastric emptying involve CRF(2) whereas stimulation of distal colonic transit involves CRF(1). The distinct profile of hUcn II, only on gastric transit, is linked to its CRF(2) selectivity.

Million M, et al. Am J Physiol Gastrointest Liver Physiol 2002 Jan;282(1):G34-40

Differential actions of peripheral corticotropin-releasing factor (CRF), urocortin II, and urocortin III on gastric emptying and colonic transit in mice: role of CRF receptor subtypes 1 and 2
Peripheral CRF inhibits gastric emptying and stimulates colonic motor function in rats. We investigated the role of CRF(1) and CRF(2) receptors in i.p. CRF-induced alterations of gut transit in conscious mice using selective CRF(1) and CRF(2) ligands injected i.p. Gastric emptying 2 h after ingestion of a solid chow meal and colonic transit (time to expel a bead inserted into the distal colon) were determined simultaneously. Rat/human (r/h)CRF, which has CRF(1) > CRF(2) binding affinity, decreased distal colonic transit time at lower doses (6-12 microg/kg) than those inhibiting gastric emptying (20-60 microg/kg). Ovine CRF, a preferential CRF(1) receptor agonist (6-60 microg/kg), reduced significantly the colonic transit time without altering gastric emptying, whereas the selective CRF(2) receptor agonists mouse urocortin II (20-60 microg/kg) and urocortin III (120 microg/kg) inhibited significantly gastric emptying without modifying colonic transit. The CRF(1)/CRF(2) receptor antagonist, astressin (30-120 microg/kg), dose dependently prevented r/hCRF (20 microg/kg)-induced inhibition of gastric emptying and reduction of colonic transit time. The selective CRF(1) receptor antagonists, NBI-27914 (C(18)H(20)Cl(4)N(4)C(7)H(8)O(3)S) and CP-154,526 (butyl-[2,5-dimethyl-7-(2,4,6-trimethylphenyl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl]e thylamine) (5-30 mg/kg), dose dependently blocked r/hCRF action on the colon without influencing the gastric response, whereas the CRF(2) receptor antagonist, antisauvagine-30 (30-100 microg/kg), dose dependently abolished r/hCRF-induced delayed gastric emptying and had no effect on colonic response. These data show that i.p. r/hCRF-induced opposite actions on upper and lower gut transit in conscious mice are mediated by different CRF receptor subtypes: the activation of CRF(1) receptors stimulates colonic propulsive activity, whereas activation of CRF(2) receptors inhibits gastric emptying.

Martinez V,et al. J Pharmacol Exp Ther 2002 May;301(2):611-7

 Urocortin and Stresscopin related products



Protocol for Urocortin II (Mouse)  Immunohistochemistry
Tissue Sample Mouse Brain
Fixative 10% Formalin
Embedding Paraffin
Negative control No primary antibody
Pretreatment Target Retrieval 25 min (Steam)
Blocking 2% Normal Goat Serum
Primary Antibody Anti-Urocortin II (Mouse) serum (catalog No.: H-019-24, Lot No.R601-4)
Optimal Dilution 1: 50
Secondary Antibody Goat anti-Rabbit IgG, Biotinylated (1:400)
Amplification  ABC (Vector)
Detection system HRP
substrate DAB (Sigma)
Counterstained Hematoxylin
Protocol for Stresscopin Related peptide (SRP)  Immunohistochemistry
Tissue Sample Mouse Brain
Fixative 10% Formalin
Embedding Paraffin
Negative control No primary antibody
Pretreatment Target Retrieval 25 min (Steam)
Blocking 2% Normal Goat Serum
Primary Antibody Anti-Stresscopin Related Peptide (SRP) (Human) serum (catalog No.: H-019-27, Lot No.R605-2)
Optimal Dilution 1: 50
Secondary Antibody Goat anti-Rabbit IgG, Biotinylated (1:400)
Amplification  ABC (Vector)
Detection system HRP
substrate DAB (Sigma)
Counterstained Hematoxylin


Protocol for Urocortin II Immunohistochemistry

Tissue Sample Mouse brain
Fixative 10% formalin
Embedding paraffin
Negative Control No primary antibody
Pretreatment Target retrieval 25 min (Steam)
Blocking 2% Normal Goat Serum
Primary Antibody Anti-Urocortin II (Mouse),  Antibody (Catalog No.:H-019-24)
Optimal Dilution 1:50
Secondary Antibody Goat Anti-Rabbit IgG, Biotinylated (1:400)
Amplification ABC (Vector)
Detection System HRP
Substrate DAB (Sigma)
Counterstained Hematoxylin


Human Urocortin III Immunohistochemistry Protocol
Human Urocortin III Immunohistochemistry Protocol
   

 


Binding Properties and Functional Activities of Selective CRG Receptor Ligand

 
 
  CRF-R2 CRF-R1
  EC50, nM(cAMP) ki, nM (binding) EC50, nM(cAMP) ki, nM (binding)
Ucn II (Mouse) 0.14 0.66 < 100 < 100
URPI (Human) 0.42 0.50 < 100 < 100
Ucn (Rat) 0.17 0.62 0.29 0.32
 

 

 Urocortin and Stresscopin related products

T-019-24;019-24;G-019-24;H-019-24;FG-019-24-A;FR-019-24;019-30;FG-019-30-A;FR-019-30;T-019-31;019-31;T-G-019-24


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