Perfect cellular platform for electrophysiology:acutely isolated vagal ganglion neurons from adult canines
10.3867/j.issn.1000-3002.2020.07.002
- VernacularTitle:理想的电生理学实验平台:急性分离的成年比格犬迷走神经节神经元
- Author:
Zhao QIAN
1
;
Guo-Fen QIAO
;
Bai-Yan LI
Author Information
1. 哈尔滨医科大学药学院药理学教研室,黑龙江 哈尔滨 150081;哈尔滨医科大学黑龙江省生物医药工程重点实验室-省部共建国家重点实验室培育基地,黑龙江 哈尔滨 150081;哈尔滨医科大学心血管药物研究教育部重点实验室,黑龙江 哈尔滨 150081;哈尔滨医科大学附属第一医院药学部,黑龙江 哈尔滨 150081
- Keywords:
nodose ganglion;
electrophysiology;
patch-clamp technique;
canine
- From:
Chinese Journal of Pharmacology and Toxicology
2020;34(7):490-501
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To establish a platform of electrophysiology using vagal ganglion neurons (VGNs) isolated from adult canines. METHODS The VGNs were enzymatically isolated from adult canines of either gender and cultured under experimental conditions. Action potential (AP), repetitive firing, voltage-gated outward K+ currents (IK) and hyperpolarization-mediated inward currents (Ih) were recorded under current-and voltage-clamp configurations before and after treatment. RESULTS Analysis of AP waveform showed that ① inaddition to traditionally classified myelinated A- and unmyelinated C-types, myelinated Ah-types could also be identified in females rather than in males; ② step current depolarization evoked a stimulus intensity-dependent repetitive discharge, and to reach a similar firing frequency, the lowest stimulus intensity was required for A-types, a similar or slightly higher stimulus intensity was needed for Ah-types, and the highest stimulus intensity was required for C-types;③tetro?dotoxin significantly reduced the rate of depolarization and positively shifted the AP firing threshold of Ah-types, and iberiotoxin dramatically increased the neuroexcitability of Ah-types;④all tested neurons functionally expressed IK and Ih, and the current density for both channels on average was A-types>Ah-types>C-types; ⑤ although the distribution of afferent types of VGNs differed between males and females, the known difference in discharge profiles of A- and C-types isolated from male and female rats was not studied here. CONCLUSION The VGNs can be successfully isolated from adult canines, AP, IK and Ih can be recorded. The tight seal can be held for at least 30 min, which may be enough for pharmacological investigation.
