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国家自然科学基金(30872836)

作品数:5 被引量:4H指数:1
相关作者:张劲松赵江月何雪菲赵琪赵芳坤更多>>
相关机构:中国医科大学中国医科大学附属第四医院大连医科大学更多>>
发文基金:国家自然科学基金辽宁省自然科学基金更多>>
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骨形成蛋白受体ⅠA在晶状体内条件敲除后对胚眼晶状体发育的影响
2014年
目的通过研究骨形成蛋白受体ⅠA(BMPRⅠA)基因在晶体内条件敲除小鼠中的作用,阐述胚胎发育过程中BMPRⅠA在胚眼晶状体发育中产生的影响。方法采用BMPRⅠA基因在晶体内条件敲除的小鼠alk3fl/fl;lecre和alk3fl/+;lecre,母鼠alk3fl/fl配公鼠lecre alk3fl/+或者母鼠alk3fl/+配公鼠lecre alk3fl/fl产生实验组lecre alk3fl/fl(3只),对照组为lecre alk3fl/+(3只)。提取鼠尾DNA,然后进行基因型鉴定,取胚胎固定,石蜡包埋,HE染色。接下来进行免疫荧光检查,一抗是sc-9305 bmp7,二抗是IF-0314 aG0IgG/FITC,荧光镜下观察。结果 BMPRⅠA晶体内条件敲除的小鼠与对照组比较,晶体的形态学变化差异从E13.5 d开始可以分辨,在E15.5 d已经有显著差异,但实验组和对照组均有晶体的发育。在BMPRⅠA晶体内条件敲除的小鼠中,骨形成蛋白7(BMP7)在晶体内的表达显著减少,尤其在胚胎发育的中后期(E13.5 d^E15.5 d);而对照组中BMP7的表达不受影响。结论 BMPRⅠA在晶体的发育过程中起重要的调节作用,其在晶体内条件敲除可以导致晶体发育不良,但不会导致晶体的缺失。BMP7在胚胎发育过程中表达于晶体中,而且其表达受BMPRⅠA的影响。
赵琪赵江月张劲松
关键词:骨形成蛋白7晶状体胚胎发育
Mutually inductive interactions between the lens and retina require ALK3 functions during mouse embryonic development被引量:1
2012年
AIM: To investigate the mutually inductive interactions that occur between the lens and retinal tissue during the development of the vertebrate eye. METHODS: Cre-positive mice were mated with Cre-negative mice to generate 50% Cre-positive (conditional knockout, CKO) and 50% Cre-negative offspring (wild type, WT). The embryos were fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned to a thickness of 4 mu m. The sections were processed for hematoxylin and eosin staining. The primary antibody used for immunofluorescence detection was sc-9305 bone morphogenetic proteins (bmp7) (Santa Cruz, US). The secondary antibody was IF-0314 aG0IgG/FITC (Santa Cruz) in combination with the primary antibody. Bright-field and fluorescent images were taken. RESULTS: Changes in the lens and retina were associated with specific alterations to the expression of type IA BMP receptor [BMPR-IA (ALK3)], which have already been implicated in eye growth. BMPR-IA was required for lens and retinal growth, but was not essential for the formation of lens. We observed that the expression of Bmp7 in the embryonic retina was reduced in the ALK3 lens of CKO mice. This phenomenon became increasingly visible in accordance with embryo development. This apparent alteration was present at stage E15.5. CONCLUSION: ALK3 is essential for lens and retinal growth. Mutually inductive interactions between the lens and retina are present in the developing mouse eye.
Qi Zhao,Zhi-Yu Zhang
关键词:BMP7LENSRETINA
Influence of bone morphogenetic protein type IA receptor conditional knockout in lens on expression of bone morphogenetic protein 4 in lens被引量:2
2015年
AIM: To investigate the influence of bone morphogenetic protein type IA receptor [BMPR-IA(ALK3)] conditional knockout in lens on expression of bone morphogenetic protein 4(BMP4) in lens during the development of the vertebrate eye.METHODS: Cre-positive mice were mated with Crenegative mice to generate 50% Cre-positive(conditional knockout, CKO) 4 embryos, 8 eyes and 50% Cre-negative offspring(wild type, WT) 4 embryos, 8 eyes. The embryos were fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned to a thickness of 4 μm.Removal of paraffin wax and dehydrating for sections,and then the procedure of in situ hybridization was processed, BMP4 MK1784-m(BOSTER) was used, and observed the expression of BMP4 in the lens in experimental group and control group. We selected SPSS11.0 software for statistical analysis, P<0.05 showed that the difference was statistically significant.· RESULTS: Four embryos of each genotype were examined, totally we had 8 embryos, 16 eyes. We got the uniform outcomes in all the embryos. We found ALK3 was required during lens growing, but was not essential for the formation of lens. We observed that the expression of BMP4 in the lens was significantly reduced in all 8 ALK3 CKO lens, BMP4 expression was normal in all the 8 WT lens, P <0.01. This phenomenon became increasingly visible in accordance with embryo development. The most apparent alteration was present at stage E15.5.CONCLUSION: ALK3 is essential for lens growth. The influence of ALK3 on the expression of BMP4 is present during the development of mice lens.
Qi ZhaoJiang-Yue ZhaoJin-Song Zhang
关键词:LENS
角膜发育的分子调控机制研究进展被引量:1
2011年
角膜的发育异常可引起先天性角膜内皮营养不良、Peters异常、Axenfeld—Rieger综合征及眼表鳞状上皮瘤(OSSN)等先天性眼病。角膜的发育经历了从诱导、发生到逐渐成熟的过程,在此期间的细胞增生、凋亡、分化等一系列过程受到多种信号转导通路和转录因子的精确调控。近年来通过胚胎的组织和基因操作技术,人们发现了一系列信号转导通路,如骨形态发生蛋白(BMP)、成纤维细胞生长因子(FGF)、Notch等,和多种转录因子,如Pax6、Krfippel样因子6(KLF6)、转化生长因子8(TGF-β)、TGF—α等均参与了角膜的发育调控过程,它们分别或协同作用于角膜发育的不同阶段,发挥各自的重要功能。主要介绍几种已知在角膜胚胎发育过程中起重要作用的信号转导通路和细胞因子。
何雪菲赵江月张劲松
关键词:角膜胚胎发育信号转导
紫外线诱导的人晶状体上皮细胞氧化损伤与小窝蛋白1 mRNA表达量下降有关
2011年
目的紫外线(UV)照射导致的氧化损伤是晶状体上皮细胞凋亡的原因之一,而小窝蛋白(Caveolin)参与晶状体上皮细胞凋亡过程。本实验拟研究紫外线造成的氧化损伤中Caveolin-1表达变化与晶体上皮细胞凋亡之间的关系,同时探讨阿司匹林(ASA)作为抗氧化剂对Caveolin-1表达及晶状体上皮细胞凋亡的影响。方法 UVB(4.43 mw/cm2)照射人晶状体上皮细胞系SRA01/04(0 s,5 s,10 s,20 s),按照是否加入阿司匹林分为实验组和对照组。分别检测实验组和对照组细胞活性;利用AnnexinV-EGFP法检测细胞凋亡率;real time RT-PCR方法定量分析Caveolin-1 mRNA的量随照射剂量不同,及加入抗氧化剂前后所产生的变化。结果随UV照射时间延长,单纯紫外线照射组细胞活性,Caveolin-1 mRNA表达量下降,细胞凋亡率增加。加入阿司匹林后,细胞活性增加。5 s,10 s组Caveolin-1 mRNA的表达量较对照组升高,细胞凋亡率较对照组下降(P<0.05)。但是照射20 s的条件下,加药后的细胞凋亡率及Caveolin-1 mRNA的表达量较未加药组无变化(P>0.05)。结论 UV照射可造成人晶状体上皮细胞凋亡,Caveolin-1 mRNA表达量减少,推测Caveolin-1表达量的变化可能与凋亡有关。低剂量的阿司匹林(3μmol/L)可以在一定程度上减少UV照射造成的晶状体上皮细胞凋亡,增加Caveolin-1的表达;但大剂量阿司匹林(>30μmol/L)会起到相反的作用,因此找到一个合适安全的剂量范围具有一定临床意义。
赵芳坤赵江月李嶔于佳明刘秋芳赵恂张劲松
关键词:紫外线小窝蛋白1阿司匹林晶状体上皮细胞
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