Thyrotropin-releasing hormone-induced cytosolic calcium transients: characterisation of store refilling in bovine anterior pituitary cells.
The intracellular calcium ion concentration ([Ca2+]i) in individual bovine anterior pituitary cells was measured using fura-2 and ratiometric imaging. Addition of thyrotropin-releasing hormone (TRH) in the presence of external calcium ion ([Ca2+]e; 1 mM) caused a rapid transient increase in [Ca2+]i falling to a plateau which remained above pre-stimulation levels in the continued presence of TRH. Decreasing [Ca2+]e to 0.1 microM decreased [Ca2+]i. At 0.1 microM [Ca2+]e, the first TRH addition caused the rapid transient rise in [Ca2+]i but no plateau phase and a second addition of TRH did not cause a second transient rise. However, the second application of TRH in 0.1 microM [Ca2+]e caused a rise in [Ca2+]i if it was preceded by transient exposure of the cells to 2 mM [Ca2+]e. The presence of nitrendipine, 2,5-di-(tert-butyl)-1,4-benzohydroquinone (tBHQ), or TRH during the re-exposure to external calcium blocked this recovery of subsequent responses to TRH in the presence of only 0.1 microM [Ca2+]e. We conclude that refilling of the calcium stores depleted by TRH occurred only after the removal of agonist, used a tBHQ-sensitive uptake mechanism, and was mainly sustained by voltage-gated calcium entry into the cells.
Department of Biochemistry, School of Medical Sciences, University of Bristol, U.K.
S L
SL
Shorte
J G
JG
Schofield
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D016257:Fura-2; D006873:Hydroquinones / Q000494:pharmacology; D007700:Kinetics; D009568:Nitrendipine / Q000494:pharmacology; D010903:Pituitary Gland, Anterior / Q000187:drug effects / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013973:Thyrotropin-Releasing Hormone / Q000494:pharmacology*; D013997:Time Factors
D000975:Antioxidants; D006873:Hydroquinones; D013973:Thyrotropin-Releasing Hormone; D009568:Nitrendipine; C018855:2-tert-butylhydroquinone; D002118:Calcium; D016257:Fura-2