摘要
目的比较不同切片液对小鼠下丘脑视前区(mPOA)脑片神经元电生理特性的影响。方法选取6~8周龄C57雌性小鼠,异氟烷麻醉后心脏灌注(通混合气950 mL/L O_(2),50 mL/L CO_(2))4℃氯化胆碱切片液或蔗糖切片液后取脑,振动切片机制备含mPOA脑区的冠状脑片,全细胞膜片钳记录mPOA电生理特性。结果采用蔗糖切片液制作的脑片mPOA神经元的内在兴奋性要优于氯化胆碱切片液,具体表现为细胞静息膜电位更超极化,动作电位幅度、不同电流刺激平均放电幅度、放电个数均高,半宽、基强度较小。而氯化胆碱切片液制作的脑片突触活性要更好,表现为兴奋性微小突触后电流频率高于蔗糖切片液制作脑片的细胞。结论蔗糖切片液和氯化胆碱切片液在保证细胞活性的情况下,蔗糖切片液更适合记录mPOA神经元的内在兴奋性,而氯化胆碱切片液更适合突触特性的记录。
Objective To compare the effects of different cutting solutions on the electrophysiological characteristics of neurons in medial preoptic area of hypothalamus(mPOA) in mice. Methods C57 female mice aged 6-8 weeks were anesthetized with isoflurane and their brains were obtained by cardiac perfusion(950 mL/L O_(2), 50 mL/L CO_(2)) with choline chloride or sucrose cutting solution at 4 ℃. Coronal brain slices containing mPOA were prepared by a vibratome. The electrophysiological characteristics of mPOA neurons were recorded by whole-cell patch clamp. Results The intrinsic excitability of mPOA neurons in brain slices prepared with sucrose cutting solution was better than that with choline chloride cutting solution, specifically manifested as more hyperpolarized resting membrane potential of cells, higher amplitude of action potential, average discharge amplitude and the number of discharges under different current stimuli, and smaller half wide and rheobase. However, the synaptic activity of brain slices prepared with choline chloride cutting solution was better, manifested as higher frequency of miniature excitatory postsynaptic currents than those of brain slices prepared with sucrose cutting solution. Conclusion Sucrose cutting solution is more suitable for recording the intrinsic excitability of mPOA neurons, while choline chloride cutting solution is more suitable for recording the synaptic characteristics of mPOA neurons.
作者
马乖乖
毛宏晖
奚楷文
刘明月
肖昊翔
袁滋铎
武胜昔
王文挺
殷松娜
MA Guaiguai;MAO Honghui;XI Kaiwen;LIU Mingyue;XIAO Haoxiang;YUAN Ziduo;WU Shengxi;WANG Wenting;YIN Songna(Department of Neurobiology,School of Basic Medical Sciences,Yan'an University,Yan'an 716000,China;Department of Neurobiology,School of Basic Medical Sciences,Air Force Medical University,Xi'an 710032,China)
出处
《空军军医大学学报》
CAS
2023年第2期111-116,共6页
Journal of Air Force Medical University
基金
国家自然科学基金(82071536)
陕西省重点研发计划项目(2020ZDLSF01-09)
陕西省自然科学基础研究基金(S2020-JC-QY-1437)。
关键词
氯化胆碱
蔗糖
下丘脑内侧视前区
脑片
choline chloride
sucrose
medial preoptic area of hypothalamus
brain slice
