小鼠耳蜗胚胎发育过程中smad4基因的表达

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1、小鼠耳蜗胚胎发育过程中Smad4基因的表达【摘要】目的　探讨小鼠内耳胚胎发育演变过程，检测Smad4基因在小鼠耳蜗中的表达情况。方法　选用耳廓反应灵敏、健康的C57BL/6小鼠作为种鼠交配，用观察阴栓方法获得适龄胎鼠，≤17天取胚胎头，≥18天在显微镜下取耳蜗，胚胎头水平冰冻切片，耳蜗平行于蜗轴冰冻切片，HE染色方法观察小鼠耳蜗发育形态演变过程，免疫组织化学方法检测Smad4蛋白在小鼠胚胎10～20天耳蜗中的表达情况。结果　胚胎10天，听泡发育，胚胎12天听泡下部有蜗管始基形成并开始发育。胚胎18天，蜗管发

2、育2圈，形成了可以辨认的内、外毛细胞，血管纹开始分化。Smad4从胚胎15天才开始表达，最初表达部位为耳蜗底转将发育成蜗轴以及柱状上皮分化为感觉细胞及支持细胞的区域，但在胚胎早期表达较弱，后期在耳蜗内广泛表达，在螺旋器、血管纹、前庭膜均有表达，且表达逐渐增强，而在蜗轴部位的表达逐渐减弱。结论　Smad4在小鼠内耳分化期有阳性表达，且表达具有明显的阶段性和区域性，说明其参与内耳听觉器官的发育过程并且可能在耳蜗的形成过程中起着重要的作用。【关键词】内耳；　发育；　耳蜗；　小鼠；　Smad4Expressiono

3、fSmad4intheembryonicdevelopmentofthemouseinnerearYUYa-ping，YANGShi-ming，HUYin-yan，GUOWei，SUNJian-heYUNing，HANDong-yi，YANGWei-yanDepartmentofOtolaryngologyandHead＆NeckSurgery,InstituteofOtolaryngology,ChinesePLAGeneralHospital,Beijing100853，ChinaCorrespondi

4、ngauthor:YangShi-ming,Email:yangsm301@263.net【Abstract】Objective　ToexplorethedevelopmentalprocessoftheembryonicmouseinnerearandtoinvestigatewhetherSmad4isexpressedintheembryonicmousecochlea.Methods　TheinnerearswereobtainedfromC57BL/6miceatagesrangingfromge

5、stationalday10throughday20.Gestationalagewasdeterminedbythevaginalplugtechnique（E1=dayofthevaginalplug）.Frozensections6μminthicknessforembryoswereprocessedforHEstainingtoobservemorphologicalchangesoftheembryonicmousecochleaandforimmunohistochemistrytode

6、terminetheexpressionofSmad4.Results　Atthetenthdaygestationotocystwasaclosedovoidsac.Thecochlearductstartedtodevelopatthetwelfthdayofgestationasanextensionoftheventralpartoftheotocyst.Attheeighteenthdaytheinnerhaircellsandouterhaircellswereidentified.TheSma

7、d4genewasexpressedincochleafromE15.Atfirst,itwasexpressedinthepartswhichshoulddevelopintothecochlearaxes,sensorycellsandsupportingcells.Thensmad4expressionwasconcentratedinsupportingcells,haircells,spiralganglionsandepitheliaofthestriavascularisduringalate

8、rstage.Conclusion　TheSmad4proteinsexistintheembryonicmousecochleaanditmaybeinvolvedinthecochlearformation.Smad4mightbeanessentialfactorforthedevelopmentofthemousecochlea.【Keyword】　Innerear;　Development;　Cochl