IP3 production in A10 cells, an established aortic smooth muscle cultured cell line: effects of agonist administration procedure and culture conditions.
1. After normal culture (10% FBS), medium exchange itself evoked marked increases in IP3 levels in A10 cells that masked the net change in IP3 in response to vasopressin (100 nM). 2. Low-serum (0.3% FBS) culture for 10 days abolished the increase in IP3 by medium exchange, and increased expression of cGMP kinase. 3. Stimulation of cells with vasopressin (100 nM) by a pipetting procedure, increased IP3 levels irrespective of culture conditions. 4. This cell line therefore could be used in a contractile form (after low-serum culture) with pipetting procedure to observe IP3 response, especially with regard to interactions with the cGMP signaling system.
Ishibashi
T
T
Department of Pharmacology, Niigata University School of Medicine, Japan. ishitaka@kanazawa-med.ac.jp
Yoshida
Y
Y
Imai
S
S
Nishio
M
M
D000818:Animals; D001011:Aorta / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D003470:Culture Media; D006152:Cyclic GMP / Q000502:physiology; D015544:Inositol 1,4,5-Trisphosphate / Q000096:biosynthesis*; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D051381:Rats; D014667:Vasopressins / Q000494:pharmacology
D003470:Culture Media; D014667:Vasopressins; D015544:Inositol 1,4,5-Trisphosphate; D006152:Cyclic GMP; D002118:Calcium