Involvement of ecto-phosphoryl transfer in contractions of the smooth muscle of the guinea pig vas deferens to adenosine 5'- triphosphate.

Details

• Personal Author:
  Fedan, Jeffery S. ; Lamport-Vrana SJ ; Vrana KE
• Description:
  The involvement of ecto-phosphoryl transfer transduction in the ATP induced contraction of the vas deferens was examined. Vas deferens obtained from English-short-hair-guinea-pigs were treated with 10(- 2) molar (M) periodate-oxidized-ATP (P-ATP), an inhibitor of the second phase of the vas deferens contraction to ATP, or 10(-4)M arylazido-aminopropionyl-ATP (ANAPP3), an inhibitor of the first phase of the contraction. Control preparations were treated with saline. Intact and homogenized vas deferens were then incubated with 3x10(-3)M phosphorus (32P) radiolabeled ATP or sulfur (35S) radiolabeled adenosine-5'-O-(3-thiotriphosphate) (ATP-gamma-S) for 5 to 60 seconds. The tissues were analyzed using sodium-dodecyl- sulfate polyacrylamide-gel electrophoresis (SDS/PAGE). Controls were incubated with the radiolabel only. Vas deferens incubated with 10(-2)M radiolabeled P-ATP for 5 minutes were also subjected to SDS/PAGE. Intact vas deferens incubated with radiolabeled ATP-gamma- S or radiolabeled ATP incorporated 35S or 32P, respectively, into a 19 to 21 kilodalton (kD) protein, at an intensity which corresponded to the time course of the contraction. Thus, 35S labeling occurred within 5 seconds, peaked within 10 seconds, and decreased significantly thereafter. The incorporation of 32P peaked within 5 seconds and decreased significantly thereafter. The incorporation of 35S was not observed in homogenized tissue. Treatment with ANAPP3 caused a significant decrease in 35S incorporation at 60 seconds. Treatment with P-ATP induced significant decreases in 35S incorporation at 60 seconds in intact tissue and 5 seconds in homogenized tissue. Treatment with P-ATP had no effect on 32P incorporation. In intact vas deferens, radiolabeled P-ATP was incorporated into several proteins, not including the 19 to 21kD protein. The presence of excess ATP enhanced 35S incorporation and inhibited P-ATP labeling. Neither 35S nor P-ATP labeling were affected by either norepinephrine or histamine. The authors conclude that an ecto-phosphoryl transfer transduction is involved in the second phase of contraction to ATP. [Description provided by NIOSH]
• Subjects:
  Animals Biochemistry Laboratories Musculoskeletal System Occupational Health Safety
• Keywords:
  Laboratory-animals; Laboratory-techniques; Animals; Animal-studies; Biochemical-analysis; Protein-biochemistry; Chemical-kinetics; Adenosines; Tissue-culture; Muscle-contraction; Muscle-tissue
• ISSN:
  0022-3565
• Document Type:
  Text
• Genre:
  Journal Article
• Place as Subject:
  OSHA Region 3 ; West Virginia
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Division:
  DRDS - Division of Respiratory Disease Studies
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  339-348
• Volume:
  258
• Issue:
  1
• NIOSHTIC Number:
  nn:00239131
• Citation:
  J Pharmacol Exp Ther 1991 Jul; 258(1):339-348
• Federal Fiscal Year:
  1991
• Peer Reviewed:
  True
• Source Full Name:
  Journal of Pharmacology and Experimental Therapeutics
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:7dc847c09aedf140bc731323e8446b4703641ae07d6ad9e7b7238120dab30b8f7dcaa9b938d34d035f87873a0fe7fec262d5d13dcaf290925445e82b6de3b57d
• Download URL:
  https://stacks.cdc.gov/view/cdc/247914/cdc_247914_DS1.pdf
• File Type:
  [PDF - 168.02 KB ]