上海交通大学学报（医学版）

上海交通大学学报（医学版） >

2022 , Vol. 42 >Issue 10: 1361 - 1374

DOI: https://doi.org/10.3969/j.issn.1674-8115.2022.10.001

创新团队成果专栏

B细胞淋巴瘤/白血病11B在小鼠耳蜗螺旋器发育中的表达研究

  • 姜正麟 ,
  • 高云鸽 ,
  • 吴皓
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  • 1.上海交通大学医学院附属第九人民医院耳鼻咽喉头颈外科，上海 200011
    2.上海交通大学医学院耳科学研究所，上海 200125
    3.上海市耳鼻疾病转化医学重点实验室，上海 200125
姜正麟（1993—），男，硕士生；电子信箱：jiangzhenglin@sjtu.edu.cn。
吴 皓，电子信箱：wuhao@shsmu.edu.cn。

收稿日期: 2022-05-07

  录用日期: 2022-08-09

  网络出版日期: 2022-12-02

基金资助

国家自然科学基金(81730028);上海市科学技术委员会科技创新行动计划(21JC1404000);上海市耳鼻疾病转化医学重点实验室(14DZ2260300)

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Study on the expression of B-cell leukemia/lymphoma 11B in the organ of Corti during mice cochlea development

  • Zhenglin JIANG ,
  • Yunge GAO ,
  • Hao WU
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  • 1.Department of Otolaryngology-Head and Neck Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
    2.Ear Institute, Shanghai Jiao Tong University School of Medicine, Shanghai 200125, China
    3.Shanghai Key Laboratory of Translational Medicine on Ear and Nose Diseases, Shanghai 200125, China
WU Hao, E-mail: wuhao@shsmu.edu.cn.

Received date: 2022-05-07

  Accepted date: 2022-08-09

  Online published: 2022-12-02

Supported by

National Natural Science Foundation of China(81730028);Science and Technology Innovation Action Plan of Shanghai Municipal Science and Technology Commission(21JC1404000);Shanghai Key Laboratory of Translational Medicine on Ear and Nose Diseases(14DZ2260300)

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摘要

目的·探索B细胞淋巴瘤/白血病11B（B-cell leukemia/lymphoma 11B，BCL11B）在小鼠耳蜗螺旋器（organ of Corti）发育过程中的表达。方法·选择出生前后不同时期/年龄［胚胎期14.5 d（E14.5）、E15.5、E16.5、E18.5，出生后0日龄（P0）、P2、P4、P7］、随机性别的野生型C57BL/6J小鼠24只，每个年龄3只。将其处死后，取出小鼠双侧耳蜗并固定，对左侧耳蜗解剖获得完整基底膜，右侧耳蜗经蔗糖脱水后进行包埋并制作冰冻切片。采用BCL11B抗体与肌球蛋白6（MYOSIN 6）抗体对小鼠耳蜗基底膜解剖样品、冰冻切片耳蜗样品进行免疫组化分析。采用共聚焦荧光显微镜对上述染色后的不同年龄点的组织样品进行观察，并对BCL11B阳性表达的毛细胞的计数及百分比进行统计。结果·成功制备了8个年龄点的小鼠耳蜗基底膜解剖样品及冰冻切片耳蜗样品。共聚焦荧光显微镜下显示，最先于E15.5胎鼠耳蜗螺旋器底中圈发现BCL11B的表达，其表达特异性出现在MYOSIN6阳性的外侧3排外毛细胞中；随着耳蜗发育，BCL11B在外毛细胞内的表达逐渐增强，至P2时达到顶峰，于P4时逐渐减弱，直至P7时已无法检测到。在BCL11B于外毛细胞表达的整个时期内，螺旋器中的其他细胞均未有BCL11B的表达。分别对BCL11B阳性表达毛细胞计数及百分比进行统计分析，结果显示E16.5、P0和P4这3个年龄点间的差异均无统计学意义（均P>0.05）。结论·通过免疫组化共染BCL11B与MYOSIN6证实，BCL11B是一个在耳蜗螺旋器中外毛细胞发育早期的特异性表达蛋白。

关键词： 螺旋器; 表达模式; 外毛细胞; B细胞淋巴瘤/白血病11B

本文引用格式

姜正麟 , 高云鸽 , 吴皓 . B细胞淋巴瘤/白血病11B在小鼠耳蜗螺旋器发育中的表达研究[J]. 上海交通大学学报（医学版）, 2022 , 42(10) : 1361 -1374 . DOI: 10.3969/j.issn.1674-8115.2022.10.001

Abstract

Objective ·To explore the expression of B-cell leukemia/lymphoma 11B (BCL11B) in the organ of Corti during mice cochlea development. Methods ·A total of 24 wild-type C57BL/6J mice of random sex at different ages before and after birth [embryonic 14.5 days (E14.5), E15.5, E16.5 and E18.5, and postnatal 0 day (P0), P2, P4 and P7] were selected, with 3 at each age point. After the mice were killed, the bilateral cochleae of the mice were sampled and fixed. The left cochleae were dissected to obtain complete basement membranes, and right cochleae were dehydrated by sucrose solution following cross section. BCL11B antibody was combined with MYOSIN 6 antibody were used for immunohistochemical analysis of whole mount samples of mouse cochlea basement membrane and cross section samples. Confocal fluorescence microscope was used for observing the tissue samples at different age points after staining, and counting the numbers to calculate the percentage of BCL11B-expressing hair cells. Results ·Whole mount samples of mouse cochlear basement membrane and cross section samples at eight age points were successfully prepared. Confocal fluorescence microscope showed that BCL11B expression was first detected specifically in outer three rolls of MYOSIN6 positive hair cells in E15.5 middle-basal turn of mouse cochlea; with the development of mouse cochlea, BCL11B expression gradually strengthened until P2, at which time point BCL11B reached top expression level and started to weaken in P4, with almost no BCL11B expression detected in P7. During the whole period of BCL11B expression in outer hair cells, no BCL11B expression was detected in other cell types in the organ of Corti. The results of the number and percentage of BCL11B-expressing hair cells showed that there was no significant difference among the three age points of E16.5, P0, and P4 (both P>0.05). Conclusion ·It is confirmed that BCL11B is a specifically expressed protein in the early development of outer hair cells in the organ of Corti by co-immunostaining of BCL11B and MYOSIN6.

Key words： organ of Corti; expression pattern; outer hair cell; B-cell leukemia/lymphoma 11B (BCL11B)

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