胚胎期豚鼠前庭终器细胞凋亡的检测_医学论文.docVIP

胚胎期豚鼠前庭终器细胞凋亡的检测_医学论文 胚胎期豚鼠前庭终器细胞凋亡的检测_医学论文 作者：胡金玮，黄维国，姜鸿彦，王锦玲，刘顺利，邱建华 【关键词】 生长和胚胎发育 关键词: 前庭终器脱噬作用生长和胚胎发育 摘 要:目的 研究豚鼠胚胎正常发育期前庭终器形态变化及细胞凋亡，探讨细胞凋亡在前庭终器发育中的作用. 方法 选用胚胎40和50d正常豚鼠，光镜观察其前庭终器发育形态变化，末端脱氧核苷酸转移酶介导的生物dUTP缺口末端标记技术（TUNEL）测定毛细胞和支持细胞凋亡，并以电镜观察凋亡细胞胞核变化. 结果 胚胎40d，豚鼠前庭壶腹、椭圆囊斑及球囊斑已分化，前庭上皮出现一层毛细胞及多层支持细胞胚胎50d，支持细胞层数减少，毛细胞数目增加，伴有神经纤维出现.TUNEL检测提示大量毛细胞和支持细胞发生凋亡.电镜下前庭上皮细胞发生分化，凋亡的毛细胞和支持细胞胞核染色质浓缩边集.凋亡细胞与新生细胞相邻. 结论 细胞凋亡是豚鼠内耳前庭终发育的必然现象，凋亡的发生对保证内耳正常结构和功能密切相关. 　　Keywords:vestibular end organsapoptosisgrouth and em-bryonic development 　　Abstract:AIM The morphological changes and cell apopto-sis of vestibular organ in normally developing guinea pigs were observed to elucidate the effect of apoptosis in the de-veloping vestibular organ.METHODS Guinea pigs of em-bryonic day（ED）40and50were studied.The morphological changes in the vestibular organ were observed by light micro-scope and apoptosis by TdT-mediated deoxyuridine triphos-phate-biotin nick end labelling（TUNEL）method and trans-mission electro microscopy（TEM）.RESULTS Ampullary cristae or macula of saccule and utricle were well differentiat-ed in ED40.There were a layer of hair cells and several lay-ers of supporting cells.The number of hair cells increased and that of layers of supporting cells decreased，accompanied by the appearance of neural synapses.The differentiated hair cells and supporting cells underwent apoptosis in vestibular sensory epithelium，and were close to newborn cells.CON┐CLUSION Apoptosis is the necessary phenomenon in vestibular end organs of developing guinea pigs.It may be re-lated to the guarantee of the normal structure and the func-tion of inner ear. 　　0 引言 　　脊椎动物内耳发育中，有关细胞增殖、分化的研究已十分详尽，但对于细胞凋亡的报道却为数不多.鸡［1］ 、大鼠［2］ 、小鼠［3］ 及人类［4］ 胚胎中，于耳泡区域发现凋亡细胞，有关细胞凋亡的功能及机制尚无定论.鉴于豚鼠内耳结构近于人耳，是内耳研究领域中重要的实验对象.目前尚未发现有关豚鼠内耳发育过程中细胞凋亡的研究资料.我们利用光镜、电镜和TUNEL技术，观察不同胚胎期豚鼠内耳前庭终器的正常发育及细胞凋亡，旨在探讨细胞凋亡在内耳发育中的意义. 　　1 材料和方法 　　1.1 材料 根据雌鼠出现阴道栓及其体质量和体形变化，选择怀孕40和50d白色豚鼠.戊巴比妥纳（50mg#12539kg-1 ，ip）全身麻醉，40g#12539L-1 多聚甲醛心脏灌注固定.