Mitf-M基因突变对猪耳蜗血管纹发育的影响
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摘要
目的研究Mitf-M基因突变是否会引起血管纹毛细血管、边缘细胞的变化,从而进一步了解Mitf-M基因突变引起Waardenburg综合征的病变机制。方法取出生后7d、10d、30d、40d的野生型和Mitf-M突变型猪共24头,分离解剖耳蜗外侧壁,运用免疫组织化学技术染色,观察野生型和Mitf-M突变型猪耳蜗血管纹毛细血管、边缘细胞在出生后不同时期的免疫荧光的差异。结果 Mitf-M突变型猪与野生型猪在出生后7d、10d及30d,血管纹毛细血管之间没有明显差异,而在40d,Mitf-M突变型猪相对野生型猪的血管纹毛细血管出现明显的减少,并有统计学意义。在7d、40d,Mitf-M突变型猪耳蜗血管纹边缘细胞的细胞骨架均无明显破坏,边缘细胞数量之间没有明显差异。结论 Mitf-M基因突变不引起血管纹边缘细胞骨架的破坏及边缘细胞数量的减少,且Mitf-M突变型猪在出生后早期30d内血管纹毛细血管没有明显变化,在40d后Mitf-M突变型猪较野生型猪的耳蜗血管纹毛细血管出现明显的减少。
Objectives We aim to find out whether Mitf-M gene mutation affects stria capillaries and marginal cells in the swine cochlear stria vascularis to better understand the patheogenic mechanism of Waardenburg syndrome.Methods A total of 24 wild type and mutant swines were selected from P7,P10, P30 and P40 after birth. Cochlear samples were obtained and immunohistochemical staining was used to detect the difference in vascular capillaries and marginal cells. Results There were no significant differences in stria capillary at P7 and P10, but at P40 the mutant swines showed a significant decrease of stria capillaries. Moreover, at P7 and P40, the cytoskeleton of stria stria'marginal cells in mutant swines showed no obvious damage, with no significant decrease in the number of marginal cells. Conclusion Mitf-M mutation only causes a decrease of intermediate cells in swine stria vascularis and no damage to the cytoskeleton and number of marginal cells. While there is no significant change in stria capillaris before P30, there is a significant decrease in stria capillaries at P40 in mutant swines.
Objectives We aim to find out whether Mitf-M gene mutation affects stria capillaries and marginal cells in the swine cochlear stria vascularis to better understand the patheogenic mechanism of Waardenburg syndrome.Methods A total of 24 wild type and mutant swines were selected from P7,P10, P30 and P40 after birth. Cochlear samples were obtained and immunohistochemical staining was used to detect the difference in vascular capillaries and marginal cells. Results There were no significant differences in stria capillary at P7 and P10, but at P40 the mutant swines showed a significant decrease of stria capillaries. Moreover, at P7 and P40, the cytoskeleton of stria stria'marginal cells in mutant swines showed no obvious damage, with no significant decrease in the number of marginal cells. Conclusion Mitf-M mutation only causes a decrease of intermediate cells in swine stria vascularis and no damage to the cytoskeleton and number of marginal cells. While there is no significant change in stria capillaris before P30, there is a significant decrease in stria capillaries at P40 in mutant swines.
引文
1 Lancet,Hearing loss:an important global health concern[J].Lancet,2016.387:p.2351.
2 Price E R,Fisher D E.Sensorineural deafness and pigmentation genes:melanocytes and the Mitf transcriptional network[J].Neuron,2001(1):p.15-8.
3 Bist J,Adhikari P,Sharma A K.Waardenburg syndrome[J].Clin Exp Optom,2011.94(2):p.240-2.
4任丽丽,郭维维,杨仕明.MITF与Waardenburg综合征[J].中华耳科学杂志,2013(01):p.145-150.Ren LL,Guo WW,Yang SM.MITF and Waardenburg syndrome[J].Chinese Journal of Otology,2013.11(1):145-150.
5 Kikuchi,K,Hilding D A.The development of the stria vascularis in the mouse[J].Acta Otolaryngol,1966.62(4):p.277-91.
6 Motohashi H,Hozawa K,Oshima T,et al.Dysgenesis of melanocytes and cochlear dysfunction in mutant microphthalmia(mi)mice[J].Hearing Research,1994(1):p.10-20.
7 Ni C,Zhang D,Beyer L A,et al.Hearing Dysfunction in Heterozygous Mitf(Mi-wh)/+Mice,a Model for Waardenburg Syndrome type 2 and Tietz Syndrome[J].Pigment Cell Melanoma Res,2013.26(1):p.78-87.
8 Liu H,Li Y,Chen L,et al.Organ of Corti and Stria Vascularis:Is There an Interdependence for Survival?[J].PLoS One,2016.11(12):p.e0168953.
9 Yi HJ,Guo W,Wu N,et al.The Temporal Bone Microdissection of Miniature Pigs as a Useful Large Animal Model for Otologic Research[J].Acta Otolaryngol,2014.134(1):p.26-33.
10 Chen L,Guo W,Ren L,et al.A de Novo Silencer Causes Elimination of MITF-M Expression and Profound Hearing Loss in Pigs[J].BMC Biol,2016.14(1):p.52.
11 Neng L,Zhang F,Kachelmeier A,et al.Endothelial Cell,Pericyte,and Perivascular Resident Macrophage-Type Melanocyte Interactions Regulate Cochlear Intrastrial Fluid-Blood Barrier Permeability[J].Springer-Verlag,2013(2):p.175-185.
12 Coppens A G,Salmon I,Heizmann C W,et al.Postnatal Maturation of the Dog Stria Vascularis--an Immunohistochemical Study[J].Anat Rec A Discov Mol Cell Evol Biol,2003.270(1):p.82-92.
13塞娜,张桐,吴军等.小鼠耳蜗血管纹血-迷路屏障免疫组织化学研究[J].中华耳科学杂志,2018(02):p.205-211.Sai N,Zhang T,Wu J,et al.Immunohistochemical Study of Cochlear Blood-labyrinth Barrier in mice[J].Chinese Journal of Otology,2018(02):p.205-211.
14 Wangemann P,Itza E M,Albrecht B,et al.Loss of KCNJ10 Protein Expression Abolishes Endocochlear Potential and Causes Deafness in Pendred Syndrome Mouse Model[J].BMC Med,2004.2:p.30.
15 Ingham N J,Carlisle F,Pearson S,et al.S1PR2 Variants Associated with Auditory Function in Humans and Endocochlear Potential Decline in Mouse[J].Sci Rep,2016.6:p.28964.
16 Ohlemiller K K,Rice M E,Lett J M,et al.Absence of Strial Melanin Coincides with Age-associated Marginal Cell Loss and Endocochlear Potential Decline[J].Hear Res,2009.249(1-2):p.1-14.
17 Steingrimsson,E,Copeland N G,Jenkins N A.Melanocytes and the Microphthalmia Transcription Factor Network[J].Annu Rev Genet,2004.38:p.365-411.
18 Levy C,Khaled M,Fisher D E.MITF:Master Regulator of Melanocyte Development and Melanoma Oncogene[J].Trends Mol Med,2006.12(9):p.406-14.
19陈伟,刘日渊,张亮等.荣昌猪电子耳蜗植入方法建立及听功能初步观察[J].中华耳科学杂志,2016(01):p.15-20.Chen W;Liu RY;Zhang L,et al.Establish the Standard Method of Rongchang Pig Cochlear Implant and Preliminary Observation of Hearing Function[J].Chinese Journal of Otology,2016(01):p.15-20.
20任丽丽,杨仕明.白化荣昌猪耳聋的分子病理机制研究[D].2013,解放军医学院.Ren LL,Yang SM.Probing the Molecular Pathological Mechanism Underlying the Deafness in Albino Rongchang Swine[D].Chinese Medical School PLA,2013.
21 Gao X,Tao Y,Lamas V,et al.Treatment of Autosomal Dominant Hearing Loss by in Vivo Delivery of Genome Editing Agents[J].Nature,2018.553(7687):p.217-221.
22 Pan B,Askew C,Galvin A,et al.Gene Therapy Restores Auditory and Vestibular Function in a Mouse Model of Usher Syndrome Type 1c[J].Nature biotechnology,2017.35(3):p.264-272.
23 Hai T,Guo W,Yao J,et al.Creation of Miniature Pig Model of Human Waardenburg Syndrome Type 2A by ENU Mutagenesis[J].Hum Genet,2017.136(11-12):p.1463-1475.
2 Price E R,Fisher D E.Sensorineural deafness and pigmentation genes:melanocytes and the Mitf transcriptional network[J].Neuron,2001(1):p.15-8.
3 Bist J,Adhikari P,Sharma A K.Waardenburg syndrome[J].Clin Exp Optom,2011.94(2):p.240-2.
4任丽丽,郭维维,杨仕明.MITF与Waardenburg综合征[J].中华耳科学杂志,2013(01):p.145-150.Ren LL,Guo WW,Yang SM.MITF and Waardenburg syndrome[J].Chinese Journal of Otology,2013.11(1):145-150.
5 Kikuchi,K,Hilding D A.The development of the stria vascularis in the mouse[J].Acta Otolaryngol,1966.62(4):p.277-91.
6 Motohashi H,Hozawa K,Oshima T,et al.Dysgenesis of melanocytes and cochlear dysfunction in mutant microphthalmia(mi)mice[J].Hearing Research,1994(1):p.10-20.
7 Ni C,Zhang D,Beyer L A,et al.Hearing Dysfunction in Heterozygous Mitf(Mi-wh)/+Mice,a Model for Waardenburg Syndrome type 2 and Tietz Syndrome[J].Pigment Cell Melanoma Res,2013.26(1):p.78-87.
8 Liu H,Li Y,Chen L,et al.Organ of Corti and Stria Vascularis:Is There an Interdependence for Survival?[J].PLoS One,2016.11(12):p.e0168953.
9 Yi HJ,Guo W,Wu N,et al.The Temporal Bone Microdissection of Miniature Pigs as a Useful Large Animal Model for Otologic Research[J].Acta Otolaryngol,2014.134(1):p.26-33.
10 Chen L,Guo W,Ren L,et al.A de Novo Silencer Causes Elimination of MITF-M Expression and Profound Hearing Loss in Pigs[J].BMC Biol,2016.14(1):p.52.
11 Neng L,Zhang F,Kachelmeier A,et al.Endothelial Cell,Pericyte,and Perivascular Resident Macrophage-Type Melanocyte Interactions Regulate Cochlear Intrastrial Fluid-Blood Barrier Permeability[J].Springer-Verlag,2013(2):p.175-185.
12 Coppens A G,Salmon I,Heizmann C W,et al.Postnatal Maturation of the Dog Stria Vascularis--an Immunohistochemical Study[J].Anat Rec A Discov Mol Cell Evol Biol,2003.270(1):p.82-92.
13塞娜,张桐,吴军等.小鼠耳蜗血管纹血-迷路屏障免疫组织化学研究[J].中华耳科学杂志,2018(02):p.205-211.Sai N,Zhang T,Wu J,et al.Immunohistochemical Study of Cochlear Blood-labyrinth Barrier in mice[J].Chinese Journal of Otology,2018(02):p.205-211.
14 Wangemann P,Itza E M,Albrecht B,et al.Loss of KCNJ10 Protein Expression Abolishes Endocochlear Potential and Causes Deafness in Pendred Syndrome Mouse Model[J].BMC Med,2004.2:p.30.
15 Ingham N J,Carlisle F,Pearson S,et al.S1PR2 Variants Associated with Auditory Function in Humans and Endocochlear Potential Decline in Mouse[J].Sci Rep,2016.6:p.28964.
16 Ohlemiller K K,Rice M E,Lett J M,et al.Absence of Strial Melanin Coincides with Age-associated Marginal Cell Loss and Endocochlear Potential Decline[J].Hear Res,2009.249(1-2):p.1-14.
17 Steingrimsson,E,Copeland N G,Jenkins N A.Melanocytes and the Microphthalmia Transcription Factor Network[J].Annu Rev Genet,2004.38:p.365-411.
18 Levy C,Khaled M,Fisher D E.MITF:Master Regulator of Melanocyte Development and Melanoma Oncogene[J].Trends Mol Med,2006.12(9):p.406-14.
19陈伟,刘日渊,张亮等.荣昌猪电子耳蜗植入方法建立及听功能初步观察[J].中华耳科学杂志,2016(01):p.15-20.Chen W;Liu RY;Zhang L,et al.Establish the Standard Method of Rongchang Pig Cochlear Implant and Preliminary Observation of Hearing Function[J].Chinese Journal of Otology,2016(01):p.15-20.
20任丽丽,杨仕明.白化荣昌猪耳聋的分子病理机制研究[D].2013,解放军医学院.Ren LL,Yang SM.Probing the Molecular Pathological Mechanism Underlying the Deafness in Albino Rongchang Swine[D].Chinese Medical School PLA,2013.
21 Gao X,Tao Y,Lamas V,et al.Treatment of Autosomal Dominant Hearing Loss by in Vivo Delivery of Genome Editing Agents[J].Nature,2018.553(7687):p.217-221.
22 Pan B,Askew C,Galvin A,et al.Gene Therapy Restores Auditory and Vestibular Function in a Mouse Model of Usher Syndrome Type 1c[J].Nature biotechnology,2017.35(3):p.264-272.
23 Hai T,Guo W,Yao J,et al.Creation of Miniature Pig Model of Human Waardenburg Syndrome Type 2A by ENU Mutagenesis[J].Hum Genet,2017.136(11-12):p.1463-1475.