摘要
为研究鱼类单倍体血液循环障碍产生机制,人工诱导获得金鱼(Carassius auratus)雌核发育单倍体胚胎并进行活体观察及邻联茴香胺染色,结果显示金鱼雌核发育单倍体胚胎存在不同程度的血液循环不良和红细胞生成缺陷。为进一步探讨其发生的分子机制,利用反义RNA整胚原位杂交技术比较分析了原始造血和血管发生关键基因scl(Stem cell leukemia)、gata-1(Gata binding protein-1)和flk-1(Fetal liver kinase-1)在雌核发育单倍体和自交二倍体胚胎中的表达模式。原始造血关键基因scl和gata-1的检测结果显示,在金鱼单倍体胚胎中,后侧板中胚层(Posterior lateral-plate mesoderm,PLM)不能正常迁移至胚体中线融合形成原始造血关键部位中间细胞群(Intermediate cell mass,ICM);同时,scl和gata-1的表达沿单倍体胚胎前后轴缩短,并在躯干区域逐渐减弱。血管内皮标记基因flk-1的检测结果显示,单倍体胚胎原始体轴血管结构紊乱,节间血管发育不良或者缺失,没有形成完整的血管系统。此外,活体观察及gsc(Goosecoid)和N-cadherin(Neural cadherin)原位杂交检测结果显示,金鱼雌核发育单倍体胚胎原肠胚期存在细胞运动缺陷。以上研究结果提示,细胞运动迁移异常导致ICM和原始体轴血管发育缺陷,同时红细胞分化减少及外周血管发育不全,可能是造成金鱼雌核发育单倍体胚胎血液循环障碍的重要原因。
Haploidy refers to the presence of only one set of chromosomes in a cell.In vertebrates,haploid cells can be cultivated in vitro and proliferate continuously,but artificially induced fish and amphibian haploid embryos show lethal“haploid syndrome”,and its mechanisms remain incompletely understood.The developmental defect of the circulatory system is the common morphological features in haploid fish embryos.Gynogenetic haploid embryos were produced when eggs were fertilized by UV-irradiated sperm of common carp.Live analysis of embryonic development showed that the blood circulation of goldfish gynogenetic haploid embryos had different phenotypes at the 65%otic vesicle closure(OVC)stage.We sort embryos into three categories:A embryos have normal blood circulation;B embryos have poor blood circulation;and C embryos have no blood circulation.The proportion of B embryos was significantly higher than that of the other two categories.The results of o-dianisidine staining showed that all goldfish gynogenetic haploid embryos contained hemoglobin,but the staining area and intensity of hemoglobin were lower than those of diploid embryos in the same period.These results suggested that goldfish gynogenetic haploid embryos had different degrees of blood circulation distemperedness and erythropoiesis defects.In order to further study the molecular mechanism of developmental defects in circulatory system of goldfish gynogenetic haploid embryos,the expression patterns of scl(stem cell leukemia),gata-1(gata binding protein-1)and flk-1(fetal liver kinase-1)during goldfish embryogenesis were compared in gynogenetic haploid and inbred diploid embryos by antisense RNA whole-mount in situ hybridization.Like inbred diploid embryos,the expression of scl and gata-1,the key regulatory genes required for primitive hematopoiesis,could be detected in the posterior lateral-plate mesoderm(PLM)of gynogenetic haploid embryos during segmentation stages.However,at the 14-somite stage,scl and gata-1 expression in PLM shortened along the anterio
作者
张琼宇
唐甲卉
彭娟
唐鹏程
孙远东
ZHANG Qiong-Yu;TANG Jia-Hui;PENG Juan;TANG Peng-Cheng;SUN Yuan-Dong(Department of Basic Medical Science,Yongzhou Vocational Technical College,Yongzhou 425100,China;Affiliated Hospital,Hunan Academy of Traditional Chinese Medicine,Changsha 410006,China;College of Life Science,Hunan University of Science and Technology,Xiangtan 411201,China)
出处
《水生生物学报》
CAS
CSCD
北大核心
2022年第1期126-136,共11页
Acta Hydrobiologica Sinica
基金
湖南省自然科学基金科教联合项目(2019JJ70038)
湖南省教育厅科学研究项目(17C1612)资助。
