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采用同工酶技术和流式细胞术研究了刀鲚肝脏、眼睛、肾脏、肌肉、鳃和血清6个组织的乳酸脱氢酶(LDH)及尾鳍、鳃、肌肉、性腺和肝脏5个组织的细胞核DNA含量,旨在为刀鲚遗传背景和种质标准的建立提供技术参数。结果显示:(1)刀鲚不同组织的LDH同工酶呈现一定的组织特异性,眼睛是LDH表达较为典型的组织;除肌肉和血清没有LDH2条带外,其余组织均出现了5条同工酶酶带,并且相对迁移率一致,各组织中均未见LDH-c基因的表达。(2)刀鲚鳃、肌肉和肝脏3个组织的细胞核DNA含量不存在显著差异(P>0.05),但都显著低于尾鳍、卵巢细胞DNA含量(P<0.05);测定的鳃、肌肉和肝脏组织细胞核的DNA含量与单倍体的精子细胞的比值依次为2.16、2.19和2.22,即刀鲚为二倍体鱼类。此外,肝脏细胞容易从组织上脱落,较易制成单细胞悬液,是用流式细胞仪进行倍性分析的适宜试材;如果要保持生物体健康存活状态,则剪取小部分尾鳍进行倍性分析为宜。研究亮点:首次报道了刀鲚的乳酸脱氢酶同工酶酶带相对迁移率、组织分布、活性扫描及细胞核DNA含量。研究发现,刀鲚眼睛是LDH表达较为典型的组织;刀鲚是二倍体鱼类,肝脏细胞是进行倍性分析的适宜试材,这为刀鲚遗传背景和种质标准的建立提供了科学依据。
The lactate dehydrogenase (LDH) and the five tissues of the caudal fin, gill, muscle, gonad and liver in 6 tissues of the liver, eye, kidney, muscle, gill and serum of Daphniphyllum were studied by isoenzyme technology and flow cytometry Nucleus DNA content, designed to provide technical parameters for the establishment of the genetic background and germplasm standards. The results showed that: (1) LDH isozymes in different tissues showed some tissue specificity, and the eyes were typical tissues with LDH expression. Enzyme band, and the relative mobility of the same, no expression of LDH-c gene in all tissues. (2) There was no significant difference in DNA content in the gill, muscle and liver (P> 0.05), but both were significantly lower than that in caudal fin and ovary (P <0.05) The ratio of DNA content in liver tissue nuclei to haploid sperm cells was 2.16, 2.19 and 2.22, respectively. In addition, the liver cells easily shed from the tissue, easy to make single cell suspension, is the use of flow cytometry ploidy analysis of suitable materials; if you want to maintain the healthy state of living organisms, then cut a small part of the caudal fin Ploidy analysis is appropriate. Research highlights: For the first time, the relative mobility, tissue distribution, activity of nuclear and lactate dehydrogenase isoenzyme bands in Daphniphyllum scoparium were reported. The study found that the knife 鲚 eye is a typical expression of LDH tissue; knife 鲚 is a diploid fish, liver cells is a suitable ploidy analysis of the material, which for the knife 鲚 genetic background and establishment of germplasm standards provide a scientific in accordance with.