摘要
目的:探究在神经示踪结合膜片钳全细胞记录的实验中,示踪染料DiI是否会对小鼠背根神经节(dorsal root ganglion,DRG)神经元兴奋性产生影响。方法:采用SPF级雄性C57BL/6J小鼠,经5%葡聚糖硫酸钠(dextran sulfate sodium,DSS)诱导溃疡性结肠炎(ulcerative colitis,UC)模型,在T12~L2皮节多点注射DiI逆向示踪躯体感觉神经元,分离全组织DRG并进行全细胞膜片钳记录。结果:HE染色结果显示模型诱导成功。膜片钳记录未经标记的小鼠DRG神经元,模型组小鼠膜电位兴奋性高于正常组小鼠。两组小鼠示踪后,正常示踪组和模型示踪组DiI+神经元兴奋性均低于本组DiI-神经元;与两未示踪组的神经元相比较,两示踪组的DiI-神经元兴奋性分别升高。结论:本实验结果明确了示踪剂DiI会降低DRG神经元的兴奋性,提示相关实验设计中应注意示踪剂的应用对神经元兴奋性的影响,以保证实验结果的可靠性。
Objective:To investigate whether the tracer dye DiI affects the excitability of dorsal root ganglion(DRG)neurons in mice in an experiment applying neural tracing combined with patch clamp technique.Methods:Using SPF-grade male C57BL/6J mice,the ulcerative colitis(UC)model was induced by 5%dextran sulfate sodium(DSS).DiI was injected at multiple points in T12-L2 dermatomes to retrogradely label somatosensory neurons.Whole mount DRG were isolated and subjected to whole-cell patch clamp recording.Results:In the HE staining,the structure of colonic mucous glands of model group were destroyed,indicating successful induction of the model.Patch clamp recording of unlabeled animal DRG neurons showed that the excitability of the cell membrane from the mice in DSS group was higher than that from the Con group.In the groups treated with DiI,excitability of DiI+neurons in both the Con+DiI and DSS+DiI groups was lower than that of DiIneurons in the same group.The excitability of DiI+neurons and DiI-neurons in the DSS+DiI group was lower than that of the corresponding neurons in the Con+DiI group,respectively.Compared with DRG neurons in two untracked groups,the excitability of DiI-neurons in the two tracking groups was increased,respectively.Conclusion:Our results demonstrated that the tracing dye DiI mainly reduces the excitability of dorsal root ganglion neurons,suggesting that the influence of tracer should be avoided in the design of relevant experiments for the reason that it may mask the changes in neuronal excitability caused by animal models.
作者
雒炜
刘永斌
刘坤
刘允
高昕妍
乔海法
LUO Wei;LIU Yong-bin;LIU Kun;LIU Yun;GAO Xin-yan;QIAO Hai-fa(College of Acupunture and Massage,Shaanxi University of Chinese Medicine,Xianyang 712046,China;Department of Physiology,Institute of Acupuncture and Moxibustion,China Academy of Chinese Medical Science,Bejing 100700,China)
出处
《中国疼痛医学杂志》
CAS
CSCD
北大核心
2024年第5期331-340,共10页
Chinese Journal of Pain Medicine
基金
国家自然科学基金面上项目(81973963、82074557)
中央级公益性科研院所基本科研业务费专项资金(ZZ-YQ2023002)。
关键词
神经示踪
背根神经节
兴奋性
DII
膜片钳
neural tracing
dorsal root ganglion
excitability
DiI
patch clamp
