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- Title
Store-operated Ca[sup 2+] entry is exaggerated in fresh preglomerular vascular smooth muscle cells of SHR.
- Authors
Fellner, Susan K; Arendshorst, William J
- Abstract
Background. Regulation of preglomerular vasomotor tone vessels ultimately control glomerular filtration rate, sodium reabsorption and systemic blood pressure. To gain insight into the complex renal hemodynamic factors that may result in hypertension, we studied calcium signaling pathways. Methods. Fresh, single, preglomerular vascular smooth muscle cells (VSMC) were isolated from 5- to 6-week-old SHR and WKY utilizing a magnetized microsphere/sieving technique. Cytosolic Ca[sup 2+] ([Ca[sup 2+]][sub i]) was measured with fura-2 ratiometric fluorescence. To examine store-operated calcium entry (SOC), VSMC were activated in calcium-free buffer containing nifedipine. To deplete the sarcoplasmic reticulum (SR) of Ca[sup 2+], vasopressin-1 receptor agonist [V[sub 1]R; inositol trisphosphate (IP[sub 3])-mediated mobilization], ryanodine (non-IP[sub 3] induced mobilization), and cyclopiazonic acid (CPA; Ca[sup 2+]-ATPase inhibition) were utilized. Addition of external calcium followed by quenching of the fura/Ca[sup 2+] signal with Mn[sup 2+] permitted assessment of divalent cation entry via SOC. Results. V[sub 1]R caused greater mobilization in SHR than WKY (P < 0.01) as well as greater calcium entry (P < 0.001). Ryanodine and CPA both caused SR calcium depletion that was not statistically different between strains, but absolute calcium entry through SOC was more than double in SHR following either maneuver (P < 0.001). 2-Amino-ethoxybiphenyl borane (2-APB), an inhibitor not only of IP[sub 3] receptors, but also of SOC, blocked calcium entry in the ryanodine and CPA experiments independent of IP[sub 3]. As well, Gd[sup 3+], a selective inhibitor of SOC, inhibited the Ca[sup 2+] response. We also studied L-channel calcium entry stimulated by V[sub 1]R. The total calcium response was greater in SHR as was the absolute inhibition by nifedipine. As a percent of the total response, participation of L-type channels sensitive to nifedipine was about 45 % in both strains of...
- Subjects
MUSCLE cells; VASCULAR smooth muscle; LABORATORY rats; CALCIUM
- Publication
Kidney International, 2002, Vol 61, Issue 6, p2132
- ISSN
0085-2538
- Publication type
Article
- DOI
10.1046/j.1523-1755.2002.00383.x