Issue Archive

Comparison of the synchronous changes of vascular tension and intracellular Ca²⁺ signal in third-order branches of mesenteric arteries under the different objective amplification of confocal microscope

MU Yun-Ping, YAN Fu-Rong, LIN Da-Cen, ZHENG Si-Yi, LIN Mo-Jun^*

Department of Physiology and Pathophysiology, The Key Laboratory of Fujian Province Universities on Ion Channel and Signal Transduction in Cardiovascular Diseases, Fujian Medical University, Fuzhou 350108, China

Abstract

This study was aimed to establish an optimized method to observe the synchronous changes of vascular tension and intracellular Ca²⁺ signal in the third-order branches of mesenteric arteries (sMA, diameter: 100–300 μm). The vascular tension and intracellular Ca²⁺ signal changes in response to potassium chloride (KCl), endothelin-1 (ET-1) and Gd³⁺ were detected using confocal wire myograph system and confocal laser scanning microscopy imaging technique, respectively. The experimental results were analyzed to explore the optimal experimental conditions. The results showed that KCl caused contraction in sMA significantly, and the intracellular Ca²⁺ level of vascular smooth muscle cells (VSMCs) was also increased under 20× and 40× objective lens. Compared with those under the 40× objective lens, the Ca²⁺ signal change was larger and the fluorescence value was more stable under the 20× objective lens, whereas the Ca²⁺ signal change was not obvious under the 10× objective lens. ET-1 (1–10 nmol/L) caused concentration dependent contraction in sMA significantly, and the intracellular Ca²⁺ signal of VSMCs was also enhanced in a concentration dependent manner. Additionally, Gd³⁺ significantly reduced the contraction of sMA and the intracellular Ca²⁺ signal of VSMCs caused by ET-1. The results suggest that the intracellular Ca²⁺ signal of VSMCs changes with vascular contraction or relaxation caused by the agonists or antagonists of Ca²⁺ channels. We successfully recorded both changes synchronously using confocal wire myograph system and confocal laser scanning microscopy imaging technique at the same time. Based on the analysis of the experimental results, we concluded that 20× objective lens provides the best experimental condition. Compared to combination of vascular tone detection method and real- time cellular fluorescence imaging technique, the present synchronous method is convenient and helpful to reduce experimental error.

Key words: small artery; tension ; calcium signal; calcium channels ; synchronous

Received: 2017-03-27　　Accepted: 2017-11-29

Corresponding author: 林默君　　E-mail: mjlin@mail.fjmu.edu.cn

DOI: 10.13294/j.aps.2017.0087

Citing This Article：

MU Yun-Ping, YAN Fu-Rong, LIN Da-Cen, ZHENG Si-Yi, LIN Mo-Jun. Comparison of the synchronous changes of vascular tension and intracellular Ca²⁺ signal in third-order branches of mesenteric arteries under the different objective amplification of confocal microscope. Acta Physiol Sin 2017; 69 (6): 785-793 (in Chinese with English abstract).