Ca2+-sparks constitute elementary building blocks for global Ca2+-signals in myocytes of retinal arterioles.
Spontaneous Ca2+-events were imaged in myocytes within intact retinal arterioles (diameter <40 microm) freshly isolated from rat eyes. Ca2+-sparks were often observed to spread across the width of these small cells, and could summate to produce prolonged Ca2+-oscillations and contraction. Application of cyclopiazonic acid (20 microM) transiently increased spark frequency and oscillation amplitude, but inhibited both sparks and oscillations within 60s. Both ryanodine (100 microM) and tetracaine (100 microM) reduced the frequency of sparks and oscillations, while tetracaine also reduced oscillation amplitude. None of these interventions affected spark amplitude. Nifedipine, which blocks store filling independently of any action on L-type Ca2+-channels in these cells, reduced the frequency and amplitude of both sparks and oscillations. Removal of external [Ca2+] (1mM EGTA) also reduced the frequency of sparks and oscillations but these reductions were slower in onset than those in the presence of tetracaine or cyclopiazonic acid. Cyclopiazonic acid, nifedipine and low external [Ca2+] all reduced SR loading, as indicated by the amplitude of caffeine evoked Ca2+-transients. This study demonstrates for the first time that spontaneous Ca2+-events in small arterioles of the eye result from activation of ryanodine receptors in the SR and suggests that this activation is not tightly coupled to Ca2+-influx. The data also supports a model in which Ca2+-sparks act as building blocks for more prolonged, global Ca2+-signals.
Tumelty
James
J
Cell and Metabolic Signalling Group, School of Medicine and Dentistry, The Queen's University of Belfast, Medical Biology Centre, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, United Kingdom.
Scholfield
Norman
N
Stewart
Michael
M
Curtis
Tim
T
McGeown
Graham
G
D000818:Animals; D001160:Arterioles / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects; D007211:Indoles / Q000494:pharmacology; D008297:Male; D032342:Muscle Cells / Q000187:drug effects / Q000378:metabolism*; D009543:Nifedipine / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012161:Retinal Artery / Q000166:cytology*; D012433:Ryanodine / Q000494:pharmacology; D019837:Ryanodine Receptor Calcium Release Channel / Q000378:metabolism; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000378:metabolism; D013748:Tetracaine / Q000494:pharmacology; D013997:Time Factors
D007211:Indoles; D019837:Ryanodine Receptor Calcium Release Channel; D013748:Tetracaine; D012433:Ryanodine; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D009543:Nifedipine; D002118:Calcium; C000543:cyclopiazonic acid