南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (7): 1102-1109.doi: 10.12122/j.issn.1673-4254.2023.07.06

• • 上一篇    下一篇

大鼠小脑浦肯野神经元发育过程中NaV亚基表达与电生理特性的发育成熟具有相关性

傅鸣宇，纪晓宏，钟 磊，吴 琼，李海福，王宁黔   

  1. 南方医科大学基础医学院生理教研室，南方医院医疗质量管理科，广东 广州 510515；解放军第74集团军医院疾控科，广东 广州 510300；广东江门中医药职业学院基础医学院，广东 江门 529000；南方医科大学生物医学工程学院数学物理系，广东 广州 510515
  • 出版日期:2023-07-20 发布日期:2023-07-19

Expression changes of NaV channel subunits correlate with developmental maturation of electrophysiological characteristics of rat cerebellar Purkinje neurons

FU Mingyu, JI Xiaohong, ZHONG Lei, WU Qiong, LI Haifu, WANG Ningqian   

  1. Department of Physiology, School of Basic Medical Sciences, Southern Medical University; Department of Quality Management, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China; Disease Control Department, 74th Army Group Hospital of PLA, Guangzhou 510300, China; Department of Basic Medicine, Guangdong Jiangmen Chinese Medicine College, Jiangmen 529000, China; Department of Mathematical Physics, School of Biomedical Engineering, Southern Medical University, Guangzhou 510515, China
  • Online:2023-07-20 Published:2023-07-19

RichHTML

16

PDF

185

摘要： 目的 通过免疫组化和电生理记录来探讨大鼠小脑浦肯野神经元发育过程中电压门控钠通道亚基的表达差异特性及其与电生理特性发育成熟的相关关系。方法 将大鼠根据发育阶段分为4组：出生后5~7 d（婴幼儿组）、12~14 d（幼年组）、21~24 d（青少年组）、42~60 d（成年组）。运用免疫组化染色方法检测NaV1.1、1.2、1.3、1.6在浦肯野神经元发育过程中的表达变化，并分析不同年龄段的平均光密度值量化发育过程中各亚基的差异变化特性。同时，运用离体脑片全细胞膜片钳技术记录自发性电活动研究发育过程中浦肯野神经元电生理基本特性变化。结果 NaV各亚基在大鼠浦肯野神经元发育过程中表达差异明显。NaV1.1亚基在整个发育阶段呈高表达，且随发育时间增加而增加（P<0.05）。NaV1.2在整个发育阶段无表达（P>0.05）。NaV1.3随发育表达量降低（P<0.05），青少年期后无表达。NaV1.6婴幼年期无表达，随发育时间增加，表达量增多（P<0.05）。神经元发育早期主要表达NaV1.1和 NaV1.3，而随发育增加，NaV1.3消失，NaV1.6增加。青少年期后主要表达NaV1.1和NaV1.6。NaV蛋白总量随发育逐渐增加（P<0.05），青少年期后趋于成熟。浦肯野神经元自发放频率和兴奋性随发育逐渐增强，到青少年期达到成熟水平。NaV亚基的发育表达变化与放电频率呈正相关（r=0.9942，P<0.05），与阈值变化呈负相关（r=-0.9891，P<0.05）。结论 NaV亚基在浦肯野神经元表达发育变化与神经元高频电生理特性的发育成熟具有相关性，提示NaV亚基的表达成熟是电生理特性不断成熟的基础。

关键词: 电压门控型钠通道；浦肯野神经元发育；免疫组化；自发电活动；相关性

Abstract: Objective To investigate the variations in the expression of voltage-gated sodium (Nav) channel subunits during development of rat cerebellar Purkinje neurons and their correlation with maturation of electrophysiological characteristics of the neurons. Methods We observed the changes in the expression levels of NaV1.1, 1.2, 1.3 and 1.6 during the development of Purkinje neurons using immunohistochemistry in neonatal (5-7 days after birth), juvenile (12-14 days), adolescent (21-24 days), and adult (42-60 days) SD rats. Using whole-cell patch-clamp technique, we recorded the spontaneous electrical activity of the neurons in ex vivo brain slices of rats of different ages to analyze the changes of electrophysiological characteristics of these neurons during development. Results The expression of NaV subunits in rat cerebellar Purkinje neurons showed significant variations during development. NaV1.1 subunit was highly expressed throughout the developmental stages and increased progressively with age (P<0.05). NaV1.2 expression was not detected in the neurons in any of the developmental stages (P>0.05). The expression level of NaV1.3 decreased with development and became undetectable after adolescence (P<0.05). NaV1.6 expression was not detected during infancy, but increased with further development (P<0.05). NaV1.1 and NaV1.3 were mainly expressed in the early stages of development. With the maturation of the rats, NaV1.3 expression disappeared and NaV1.6 expression increased in the neurons. NaV1.1 and NaV1.6 were mainly expressed after adolescence. The total NaV protein level increased gradually with development (P<0.05) and tended to stabilize after adolescence. The spontaneous frequency and excitability of the Purkinje neurons increased gradually with development and reached the mature levels in adolescence. The developmental expression of NaV subunits was positively correlated with discharge frequency (r=0.9942, P<0.05) and negatively correlated with the excitatory threshold of the neurons (r=0.9891, P<0.05). Conclusion The changes in the expression levels of NaV subunits are correlated with the maturation of high frequency electrophysiological properties of the neurons, suggesting thatmature NaV subunit expressions is the basis of maturation of electrophysiological characteristics of the neurons.

Key words: voltage gated sodium channel; Purkinje neuron development; immunohistochemistry; spontaneous electrical activity; correlation analysis