新生大鼠耳蜗Klliker器在体凋亡的研究

The Apoptosis of the Klliker Organ in the Cochlea of Newborn Rat in Vivo

杨军;何圆圆;

1:上海交通大学医学院附属新华医院耳鼻咽喉-头颈外科上海市耳鼻疾病转化医学重点实验室上海交通大学医学院耳科学研究所

2:南京医科大学附属无锡人民医院耳鼻咽喉科

摘要
目的了解不同天龄新生大鼠耳蜗Klliker器的形态变化,研究凋亡相关因子的mRNA及蛋白的表达水平,探讨Klliker器在听觉功能发育过程中凋亡的机制。方法选取出生后不同天龄的Sprague-Dawley大鼠共192只,其中出生后1天(P1)、5天(P5)、12天(P12)各6只大鼠耳蜗冰冻切片后,通过苏木精-伊红染色及免疫荧光染色等方法,观察耳蜗Klliker器的形态结构变化;取P1大鼠6只行耳蜗基底膜免疫荧光观察;提取出生后1、3、5、7、10、12及14天(P1、P3、P5、P7、P10、P12和P14)大鼠耳蜗基底膜mRNA(各6只)及蛋白(各18只),运用real-time PCR及蛋白质印迹的方法,观察出生后各天龄组大鼠耳蜗基底膜Klliker器凋亡过程中bcl-2、caspase-3、caspase-8及caspase-9的表达规律。结果出生后大鼠听力出现之前其耳蜗Klliker器支持细胞的形态自顶回向底回从高柱状向矮柱状变化,同时支持细胞的数量逐渐减少。出生后不同天龄大鼠耳蜗基底膜的caspase-3、caspase-8、caspase-9及bcl-2的mRNA和蛋白的表达水平均呈明显的时间依赖性。结论在大鼠出生后、听力出现前耳蜗发育的整个阶段,Klliker器自底回向顶回逐渐发生退化;caspase-3、caspase-8、caspase-9及bcl-2的mRNA和蛋白的表达表现为时间依赖性。
关键词
大鼠;耳蜗;Klliker器;支持细胞;凋亡
基金项目(Foundation):
国家自然科学基金(81170919,81470689);; 国家科技部973课题(2014CB541705);; 上海市科委重大项目(14DJ1400201,14DZ2260300)资助
作者
杨军;何圆圆;
参考文献

1 Tritsch NX,Yi E,Gale JE,et al.The origin of spontaneous activity in the developing auditory system[J].Nature,2007,450:50.

2 何圆圆,邓杰,杨军.耳蜗发育与三磷酸腺苷的关系[J].中华临床医师杂志,2012,6:112.

3 Liang Y,Huang L,Yang J.Differential expression of ryanodine receptor in the developing rat cochlea[J].Euro J Histochem,2009,52:249.

4 Tritsch NX,Bergles DE.Developmental regulation of spontaneous activity in the Mammalian cochlea[J].J Neurosci,2010,30:1539.

5 Jones TA,Leake PA,Snyder RL,et al.Spontaneous discharge patterns in cochlear spiral ganglion cells before the onset of hearing in cats[J].J Neurophysiol,2007,98:1898.

6 Glueckert R,Wietzorrek G,Kammen-Jolly K,et al.Role of class D L-type Ca2+channels for cochlear morphology[J].Hear Res,2003,178:95.

7 Seal RP,Akil O,Yi E,et al.Sensorineural deafness and seizures in mice lacking vesicular glutamate transporter 3[J].Neuron,2008,57:263.

8 Erazo-Fischer E,Striessnig J,Taschenberger H.The role of physiological afferent nerve activity during in vivo maturation of the calyx of Held synapse[J].J Neurosci,2007,27:1725.

9 McKay SM,Oleskevich S.The role of spontaneous activity in development of the endbulb of Held synapse[J].Hear Res,2007,230:53.

10 Kandler K,Clause A,Noh J.Tonotopic reorganization of developing auditory brainstem circuits[J].Nat Neurosci,2009,12:711.

11 何圆圆,杨军.新生大鼠耳蜗Klliker器支持细胞ATP释放的机制[J].中华耳鼻咽喉头颈外科杂志,2015,50:43.

12 Kamiya K,Takahashi K,Kitamura K,et al.Mitosis and apoptosis in postnatal auditory system of the C3H/He strain[J].Brain Res,2001,901:296.

13 Nicholson DW.Caspase structure,proteolytic substrates,and function during apoptotic cell death[J].Cell Death Differ,1999,6:1028.

14 Adrain C,Martin SJ.The mitochondrial apoptosome:a killer unleashed by the cytochrome seas[J].Trends Biochem Sci,2001,26:390.

15 Maher P,Schubert D.Signaling by reactive oxygen species in the nervous system[J].Cell Mol Life Sci,2000,57:1287.

16 Chen YR,Tan TH.The c-Jun N-terminal kinase pathway and apoptotic signaling(review)[J].Int J Oncol,2000,16:651.

17 Li H,Steyger PS.Systemic aminoglycosides are trafficked via endolymph into cochlear hair cells[J].Sci Rep,2011,1:159.

18 Giorgi C,Romagnoli A,Pinton P,et al.Ca2+signaling,mitochondria and cell death[J].Curr Mol Med,2008,8:119.

19 何圆圆,杨军.新生大鼠Kolliker器支持细胞体外凋亡及增殖的研究[J].临床耳鼻咽喉头颈外科杂志,2015,29:152.

本文信息

PDF(455K)

本文作者相关文章

杨军何圆圆