斑马鱼听觉诱发电位检测系统的建立和应用

Establishment and Application of Zebrafish Auditory Evoked Potential(AEP) Detection System

潘豪来;王际平;于栋帧;杨光;时海波;王坚;殷善开;

1:上海交通大学附属第六人民医院耳鼻咽喉头颈外科

2:加拿大达尔豪斯大学人类交流障碍学院

摘要
目的建立斑马鱼听觉诱发电位(auditory evoked potential,AEP)检测系统。方法设计并制作斑马鱼听觉诱发电位检测水箱;应用该系统检测和记录35条体长为1046mm斑马鱼的AEP,将斑马鱼分为5组:体长1246mm斑马鱼的AEP,将斑马鱼分为5组:体长1215mm,6条;体长1715mm,6条;体长1720mm,4条;体长2220mm,4条;体长2226mm,4条;体长3226mm,4条;体长3237mm,9条;体长4237mm,9条;体长4246mm,12条。对照组为金鱼,体长3846mm,12条。对照组为金鱼,体长3854mm,8条。通过麻醉实验、鱼鳔实验、噪声暴露实验并以金鱼为对照验证斑马鱼AEP检测系统的可靠性。结果实验证实斑马鱼AEP检测系统获得的响应为听觉电生理反应;所有体长斑马鱼的听力频率范围为100Hz54mm,8条。通过麻醉实验、鱼鳔实验、噪声暴露实验并以金鱼为对照验证斑马鱼AEP检测系统的可靠性。结果实验证实斑马鱼AEP检测系统获得的响应为听觉电生理反应;所有体长斑马鱼的听力频率范围为100Hz12kHz,在发育过程中无听觉频率扩展的现象;成鱼最佳听力范围为600Hz12kHz,在发育过程中无听觉频率扩展的现象;成鱼最佳听力范围为600Hz1kHz,5组斑马鱼AEP阈值分别为141.7±1.32、124.8±1.31、121.8±1.49、117.8±1.09和124.4±1.87dB w;斑马鱼AEP阈值随体长增加而变化,具有随着发育成熟AEP阈值降低,随着衰老阈值升高的特征。结论本研究建立了稳定的斑马鱼听觉诱发电位检测系统,该系统可用于斑马鱼AEP的检测。
关键词
斑马鱼;听觉诱发电位;检测系统
基金项目(Foundation):
上海市科委重点项目子课题(4DJ1400202);上海市科委国际合作项目(14430720900)联合资助
作者
潘豪来;王际平;于栋帧;杨光;时海波;王坚;殷善开;
参考文献

1 Steiner AB,Kim T,Cabot V,et al.Dynamic gene expression by putative hair-cell progenitors during regeneration in the zebrafish lateral line[J].Proc Natl Acad Sci USA,2014,111:E1393.

2 Higgs DM,Rollo AK,Souza MJ,et al.Development of form and function in peripheral auditory structures of the zebrafish(Danio rerio)[J].J Acoust Soc Am,2003,113:1145.

3 Zeddies DG,Fay RR.Development of the acoustically evoked behavioral response in zebrafish to pure tones[J].J Exp Biol,2005,208:1363.

4 Higgs DM,Souza MJ,Wilkins HR,et al.Age-and sizerelated changes in the inner ear and hearing ability of the adult zebrafish(Danio rerio)[J].J Assoc Res Otolaryngol,2002,3:174.

5 Kenyon TN,Ladich F,Yan HY.A comparative study of hearing ability in fishes:the auditory brainstem response approach[J].J Comp Physiol A,1998,182:307.

6 Yan HY,Fine ML,Horn NS,et al.Variability in the role of the gasbladder in fish audition[J].J Comp Physiol A,2000,186:435.

7 Smith ME,Kane AS,Popper AN.Noise-induced stress response and hearing loss in goldfish(Carassius auratus)[J].J Exp Biol,2004,207:427.

8 Lechner W,Wysocki LE,Ladich F.Ontogenetic development of auditory sensitivity and sound production in the squeaker catfish synodontis schoutedeni[J].BMC Biol,2010,8:10.

9 Nicolson T.The genetics of hearing and balance in zebrafish[J].Annu Rev Genet,2005,39:9.

10 Vlasits AL,Simon JA,Raible DW,et al.Screen of FDAapproved drug library reveals compounds that protect hair cells from aminoglycosides and cisplatin[J].Hear Res,2012,294:153.

11 Jiang L,Romero-Carvajal A,Haug JS,et al.Gene-expression analysis of hair cell regeneration in the zebrafish lateral line[J].Proc Natl Acad Sci USA,2014,111:E1383.

12 Monroe JD,Manning DP,Uribe PM,et al.Hearing sensitivity differs between zebrafish lines used in auditory research[J].Hear Res,2016,341:220.

13 Trapani JG,Nicolson T.Physiological recordings from zebrafish lateral-line hair cells and afferent neurons[J].Methods Cell Biol,2010,100:219.