The signaling mechanisms of long distance intercellular calcium waves (far waves)in cultured human uterine myocytes
Tóm tắt
Cultured human myocytes exhibit intercellular calcium waves that travel farther than 100 μm (‘far waves’). This work investigates the mechanism of far wave propagation. Culture lines were initiated from myometrial biopsies of term pregnant women. Calcium green-1 was used as a fluorescence probe for intracellular free calcium. Serial imaging was performed at a frame rate of 0.83 frames/s. Intercellular calcium waves were mechanically initiated by atraumatically applying small drops of mineral oil onto the surface of the monolayer. Each intercellular calcium wave was scored using a standardized grid, and points were assigned depending upon the distance the wave traveled and the fluorescent intensity observed within each region. Experiments were performed in the presence of inhibitors of gap junctions and connexin hemichannels (octanol), ATP (apyrase and MDL 12330 A), prostaglandins (indomethacin, high concentrations of lanthanum), the prostaglandin transporter, PGT (DIDS), and transmembrane calcium flux (low concentrations of lanthanum). Octanol, apyrase and MDL 12330 A failed to modify the far waves, indicating gap junctions, connexin hemichannels and ATP do not participate in the paracrine mechanism. Indomethacin at 30, 100 and 300 μM, in a dose dependent manner, reduced the far wave score to 0, suggesting a prostaglandin was critically involved in the mechanism. DIDS reduced the far wave score, but did not fully inhibit wave propagation, suggesting the presence of PGT-dependent and -independent components to the mechanism. Lanthanum at 0.1 mM had no effect, but at 1 mM, reduced the far wave score. These results are consistent with PGF2α and/or PGE2 being the signal molecule for the PGT-dependent component. Taken together, these data indicate that long distance intercellular calcium waves in cultured human myocytes utilizes a paracrine signaling mechanism, but with more than one extracellular signaling compound.
Tài liệu tham khảo
Brodt-Eppley J and Myatt L (1999) Obstet Gynecol 93: 89-93.
Bruzzone S, Guida L, Zocchi E, Franco L and De Flora A (2001) Faseb J 15: 10-12.
Chan BS, Satriano JA, Pucci M and Schuster VL (1998) J Biol Chem 273: 6689-6697.
Colliard-Rouiller C and Durand J (1997) Respir Physiol 107: 263-273.
Faraci FM, Sobey CG, Chrissobolis S, Lund DD, Heistad DD and Weintraub NL (2001) Am J Physiol Regul Integr Comp Physiol 281: R246-R253.
Fu X, Favini R, Kindahl K and Ulmsten U (2000) Am J Obstet Gynecol 182: 582-588.
Gibb W (1998) Ann Med 30: 235-241.
Grynkiewicz G, Poenie M and Tsein RY (1985) J Biol Chem 260: 3440-3450.
Guthrie PB, Knappenberger J, Segal M, Bennett MV, Charles AC and Kater SB (1999) J Neurosci 19: 520-528.
Ishizawa M (1991) Prostaglandins 42: 579-586.
Kanai N, Lu R, Satriano JA, Bao Y, Wolko. AW and Schuster VL (1995) Science 268: 866-869.
Landen CN Jr, Zhang P and Young RC (2001) Am J Obstet Gynecol 184: 1100-1103.
Li H, Liu TF, Lazrak A, Peracchia C, Goldberg GS, Lampe PD and Johnson RG (1996) J Cell Biol 134: 1019-1030.
Paemeleire K, Martin PE, Coleman SL, Fogarty KE, Carrington WA, Leybaert L, Tuft RA, Evans WH and Sanderson MJ (2000) Mol Biol Cell 11: 1815-1827.
Panter KR, Hannah ME, Amankwah KS, Ohlsson A, Jefferies AL and Farine D (1999) Br J Obstet Gynaecol 106: 467-473.
Sanderson MJ, Charles AC and Dirksen ER (1990) Cell Regul 1: 585-596.
Schuster VL (1998) Annu Rev Physiol 60: 221-242.
Sparey C, Robson SC, Bailey J, Lyall F and Europe-Finner GN (1999) J Clin Endocrinol Metab 84: 1705-1710.
Tamai I, Nezu J, Uchino H, Sai Y, Oku A, Shimane M and Tsuji A (2000) Biochem Biophys Res Commun 273: 251-260.
Tegeder I, Pfeilschifter J and Geisslinger G (2001) Faseb J 15: 2057-2072.
Young RC (2000) J Soc Gynecol Investig 7: 146-152.
Young RC and Hession RO (1996) J Muscle Res Cell Motil 17: 349-355.
Young RC and Hession RO (1997) Obstet Gynecol 90: 928-932.
Young RC and Zhang P (2001) Am J Obstet Gynecol 184: 1228-1234.