c-Maf基因在小鼠耳蜗内外毛细胞中的表达及意义
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摘要
目的研究c-Maf基因在小鼠耳蜗内、外毛细胞中的表达、分布特点及意义。方法 1)根据形态上的差异分离收集成年小鼠内、外毛细胞,利用Microarray基因芯片技术分别检测内、外毛细胞c-Maf基因RNA水平的表达;2)全耳蜗基底膜铺片,免疫组织化学染色后激光共聚焦显微镜观察c-Maf蛋白在不同发育阶段小鼠耳蜗内、外毛细胞中的分布特点及表达,并使用软件Image Pro Plus进行c-Maf蛋白定量分析。结果 1)耳蜗内、外毛细胞c-Maf基因RNA水平相近;2)c-Maf蛋白主要表达于成年小鼠的耳蜗内外毛细胞胞质,在内毛细胞的核下区表达强于核上区,且在内、外毛细胞的表达强度相近;3)c-Maf蛋白在小鼠出生后0天、7天、12天及成年小鼠耳蜗内、外毛细胞稳定表达。结论 c-Maf蛋白从出生开始在小鼠耳蜗内、外毛细胞上稳定表达,且在成年小鼠内毛细胞的核下区表达较强,表明其可能在毛细胞生长发育以及功能维持中发挥重要的作用。
Objective To investigate the expression, distribution and significance of c-Maf protein in inner and outer hair cells in the mouse cochlea. Methods(1) Inner and outer hair cells in adult mice were collected individually according to their morphological differences, and tested for RNA level c-Maf gene by Gene Chip microarray.(2) Distribution and expression of different developmental stages of the c-Maf protein in inner and outer hair cells on the basilar membrane were examined and recorded by confocal microscopy with immunofluorescence, while quantitative analysis of c-Maf protein was performed using the Image Pro Plus software. Results(1) The level of c-Maf gene RNA was similar in cochlear inner and outer hair cells;(2) c-Maf protein was expressed in the cytoplasm of inner and outer hair cells with similar expression intensity, although expression of the protein in the infranuclear region was stronger than in the supper nuclear region in inner hair cells;(3) c-Maf protein was stably expressed in inner and outer hair cells on Day 0, 7 and 12 after the birth. Conclusions The c-Maf protein is stably expressed in cochlear hair cells after birth and highly concentrated in the subnuclear region in inner hair cells in adult mice. It seems that the c-Maf protein may play an important role in cellular development and function in inner and outer hair cells.
Objective To investigate the expression, distribution and significance of c-Maf protein in inner and outer hair cells in the mouse cochlea. Methods(1) Inner and outer hair cells in adult mice were collected individually according to their morphological differences, and tested for RNA level c-Maf gene by Gene Chip microarray.(2) Distribution and expression of different developmental stages of the c-Maf protein in inner and outer hair cells on the basilar membrane were examined and recorded by confocal microscopy with immunofluorescence, while quantitative analysis of c-Maf protein was performed using the Image Pro Plus software. Results(1) The level of c-Maf gene RNA was similar in cochlear inner and outer hair cells;(2) c-Maf protein was expressed in the cytoplasm of inner and outer hair cells with similar expression intensity, although expression of the protein in the infranuclear region was stronger than in the supper nuclear region in inner hair cells;(3) c-Maf protein was stably expressed in inner and outer hair cells on Day 0, 7 and 12 after the birth. Conclusions The c-Maf protein is stably expressed in cochlear hair cells after birth and highly concentrated in the subnuclear region in inner hair cells in adult mice. It seems that the c-Maf protein may play an important role in cellular development and function in inner and outer hair cells.
引文
1 Géléoc GS,Holt JR.Sound Strategies for Hearing Restoration[J].Science,2014,344(6184):1241062.
2 Liu H,Pecka JL,Zhang Q,et al.Characterization of Transcriptomes of Cochlear Inner and Outer Hair Cells[J].J Neurosci,2014,34(33):11085-11095.
3 Yang Y,Cvekl A.Large Maf Transcription Factors:Cousins of AP-1 Proteins and Important Regulators of Cellular Differentiation[J].Einstein J Biol Med,2007,23(1):2-11.
4 Katsuoka F,Yamamoto M.Small Maf Proteins(Maf F,Maf G,Maf K):History,Structure and Function[J].Gene,2016,586(2):197-205.
5 Yoshida T,Ohkumo T,Ishibashi S,et al.The 5'-AT-rich Half-site of Maf Recognition Element:a Functional Target for b ZIP Transcription Factor Maf[J].Nucleic Acids Res,2005,33(11):3465-3478.
6 Imaki J,Tsuchiya K,Mishima T,et al.Developmental Contribution of C-maf in the Kidney:Distribution and Developmental Study of c-maf m RNA in Normal Mice Kidney and Histological Study of C-maf Knockout Mice Kidney and Liver[J].Biochem Biophys Res Commun,2004,320(4):1323-1327.
7 Cvekl A,Zhang X.Signaling and Gene Regulatory Networks in Mammalian Lens Development[J].Trends Genet,2017,33(10):677-702.
8 Chien CH,Yu HC,Chen SY,et al.Characterization of c-Maf+Foxp3-Regulatory T Cells Induced by Repeated Stimulation of Antigen-Presenting B Cells[J].Sci Rep,2017,7:46348.
9 Wende H,Lechner SG,Cheret C,et al.The Transcription Factor c-Maf Controls Touch Receptor Development and Function[J].Science,2012,335(6074):1373-1376.
10 Wende H,Lechner SG,Birchmeier C.The Transcription Factor c-Maf in Sensory Neuron Development[J].Transcription,2012,3(6):285-289.
11 Sun W,Yang S,Liu K,et al.Hearing Loss and Auditory Plasticity[J].Hear Res,2017,347:1-2.
12张昊昱,张宁,张华等.耳聋基因检测在遗传性耳聋诊断及遗传咨询中的应用[J].中华耳科学杂志2016,14(5):639-643.Zhang HY,Zhang N1,Zhang H et al.The Application of Genetic Testing of Deafness Genes in Hereditary Deafness Diagnosis and Genetic Counseling[J],Chinese Journal of Otology,2016,14(5):639-643.
13 Peng Z,Wang GP,Zeng R,et al.Temporospatial Expression and Cellular Localization of VGLUT3 in the Rat Cochlea[J].Brain Res,2013,1537:100-110.
14 Liu K,Jiang X,Shi C,et al.Cochlear Inner Hair Cell Ribbon Synapse is the Primary Target of Ototoxic Aminoglycoside Stimuli[J].Mol Neurobiol,2013,48(3):647-654.
15 Roux I,Safieddine S,Nouvian R,et al.Otoferlin,Defective in a Human Deafness Form,is Essential for Exocytosis at the Auditory Ribbon Synapse[J].Cell,2006,127(2):277-289.
16杨乐,柳柯,杨仕明等.C57小鼠毛细胞突触带的发育观察[J].中华耳科学杂志2017,15(2):234-238.Yang L,Liu K,Yang SM,et al.Observation of Developmental Changes of Hair Cell Synaptic Ribbons in C57 Mice[J].Chinese Journal of Otology,2017,15(2):234-238.
17 Ring BZ,Cordes SP,Overbeek PA,et al.Regulation of Mouse Lens Fiber Cell Development and Differentiation by the Maf Gene[J].Development,2000,127(2):307-317.
18史珣贝,吴南,郭维维等.腺相关病毒在耳聋基因治疗上的应用[J].中华耳科学杂志2016,14(1):37-41.Shi XB,Wu N,Guo WW et al.Application of Adeno-Associated Virus Gene Vectors in Hereditary Non-Syndromic Sensorineural Hearing Loss[J].Chinese Journal of Otology,2016,14(1):37-41.2018-03-01
2 Liu H,Pecka JL,Zhang Q,et al.Characterization of Transcriptomes of Cochlear Inner and Outer Hair Cells[J].J Neurosci,2014,34(33):11085-11095.
3 Yang Y,Cvekl A.Large Maf Transcription Factors:Cousins of AP-1 Proteins and Important Regulators of Cellular Differentiation[J].Einstein J Biol Med,2007,23(1):2-11.
4 Katsuoka F,Yamamoto M.Small Maf Proteins(Maf F,Maf G,Maf K):History,Structure and Function[J].Gene,2016,586(2):197-205.
5 Yoshida T,Ohkumo T,Ishibashi S,et al.The 5'-AT-rich Half-site of Maf Recognition Element:a Functional Target for b ZIP Transcription Factor Maf[J].Nucleic Acids Res,2005,33(11):3465-3478.
6 Imaki J,Tsuchiya K,Mishima T,et al.Developmental Contribution of C-maf in the Kidney:Distribution and Developmental Study of c-maf m RNA in Normal Mice Kidney and Histological Study of C-maf Knockout Mice Kidney and Liver[J].Biochem Biophys Res Commun,2004,320(4):1323-1327.
7 Cvekl A,Zhang X.Signaling and Gene Regulatory Networks in Mammalian Lens Development[J].Trends Genet,2017,33(10):677-702.
8 Chien CH,Yu HC,Chen SY,et al.Characterization of c-Maf+Foxp3-Regulatory T Cells Induced by Repeated Stimulation of Antigen-Presenting B Cells[J].Sci Rep,2017,7:46348.
9 Wende H,Lechner SG,Cheret C,et al.The Transcription Factor c-Maf Controls Touch Receptor Development and Function[J].Science,2012,335(6074):1373-1376.
10 Wende H,Lechner SG,Birchmeier C.The Transcription Factor c-Maf in Sensory Neuron Development[J].Transcription,2012,3(6):285-289.
11 Sun W,Yang S,Liu K,et al.Hearing Loss and Auditory Plasticity[J].Hear Res,2017,347:1-2.
12张昊昱,张宁,张华等.耳聋基因检测在遗传性耳聋诊断及遗传咨询中的应用[J].中华耳科学杂志2016,14(5):639-643.Zhang HY,Zhang N1,Zhang H et al.The Application of Genetic Testing of Deafness Genes in Hereditary Deafness Diagnosis and Genetic Counseling[J],Chinese Journal of Otology,2016,14(5):639-643.
13 Peng Z,Wang GP,Zeng R,et al.Temporospatial Expression and Cellular Localization of VGLUT3 in the Rat Cochlea[J].Brain Res,2013,1537:100-110.
14 Liu K,Jiang X,Shi C,et al.Cochlear Inner Hair Cell Ribbon Synapse is the Primary Target of Ototoxic Aminoglycoside Stimuli[J].Mol Neurobiol,2013,48(3):647-654.
15 Roux I,Safieddine S,Nouvian R,et al.Otoferlin,Defective in a Human Deafness Form,is Essential for Exocytosis at the Auditory Ribbon Synapse[J].Cell,2006,127(2):277-289.
16杨乐,柳柯,杨仕明等.C57小鼠毛细胞突触带的发育观察[J].中华耳科学杂志2017,15(2):234-238.Yang L,Liu K,Yang SM,et al.Observation of Developmental Changes of Hair Cell Synaptic Ribbons in C57 Mice[J].Chinese Journal of Otology,2017,15(2):234-238.
17 Ring BZ,Cordes SP,Overbeek PA,et al.Regulation of Mouse Lens Fiber Cell Development and Differentiation by the Maf Gene[J].Development,2000,127(2):307-317.
18史珣贝,吴南,郭维维等.腺相关病毒在耳聋基因治疗上的应用[J].中华耳科学杂志2016,14(1):37-41.Shi XB,Wu N,Guo WW et al.Application of Adeno-Associated Virus Gene Vectors in Hereditary Non-Syndromic Sensorineural Hearing Loss[J].Chinese Journal of Otology,2016,14(1):37-41.2018-03-01