In vitro acute cytotoxicity of neonicotinoid insecticide imidacloprid to gill cell line of flounder Paralichthy olivaceus

In vitro acute cytotoxicity of neonicotinoid insecticide imidacloprid (IMI) to the gill cell line of flounder (FG) that collected in the gill of Paralichthys olivaceus, was examined by 3 widely used endpoint bioassays: NR (neutral red), MTT (3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) and TCP (total cell protein). The result shows that the IMI increased at concentrations ≥0.5 μg/ml. The IC50 value of NR, MTT, and TCP was 41.86, 38.46, and 39.08 μg/ml, respectively. The ultrastructural observation revealed that the mitochondria of the cells exposed to 60 μg/ml IMI for 48 h were severely damaged, swollen or disrupted, while their nuclei and rough endoplasmic reticulum (RER) remained normal. This would suggest that the mitochondria are probably the primary target of IMI.     

Distribution and ultrastructure of pigment cells in the skins of normal and albino adult turbot, Scophthalmus Maximus

The distribution and ultrastructure of pigment cells in skins of normal and albino adult turbots were examined with transmission electron microscopy (TEM). Three types of pigment cells of melanophore, iridophore and xanthophore have been recognized in adult turbot skins. The skin color depends mainly on the amount and distribution of melanophore and iridophore, as xanthophore is quite rare. No pigment cells can be found in the epidermis of the skins. In the pigmented ocular skin of the turbot, melanophore and iridophore are usually co-localized in the dermis. This is quite different from the distribution in larvae skin. In albino and white blind skins of adult turbots, however, only iridophore monolayer still exists, while the melanophore monolayer disappears. This cytological evidence explains why the albino adult turbot, unlike its larvae, could never resume its body color no matter what environmental and nutritional conditions were provided. Endocytosis is quite active in the cellular membrane of the iridophore. This might be related to the formation of reflective platelet and stability of the iridophore.     

海洋无脊椎动物细胞培养概况

动物细胞培养技术萌芽于1885年Roux培养鸡神经板细胞的尝试,创建于1907年Harrison进行的蝌蚪髓管神经细胞的培养实验.医学上对生产抗病毒疫苗和研究肿瘤成因的需要,促进了细胞培养工作在人类和啮齿类动物中的广泛开展,并于1952年建立了人类的第一个肿瘤细胞系(Hela).     

3种对虾病毒囊膜蛋白对假型昆虫杆状病毒的包装与对虾组织亲嗜性的影响

为探究对虾白斑综合征病毒(WSSV)3种囊膜蛋白VP19、VP24和VP28对其假型昆虫杆状病毒的包装与对虾组织亲嗜性的影响,本研究首先分析了过表达的上述3种囊膜蛋白在包装细胞(Sf9)中的亚细胞定位,然后通过共转染的方法分别构建了其假型杆状病毒：Bacmid-GUS/VP19、Bacmid-GUS/VP24和Bacmid-GUS/VP28,比较了上述3种假型杆状病毒的包装效率及其在感染对虾组织上的特异性。结果表明,过表达的VP19、VP24和VP28均可定位于包装细胞的细胞膜上,均能促进假型病毒的包装效率,并分别提高了14.1%、11.5%和11.7%,且携带不同囊膜蛋白的假型病毒在感染对虾成体时具有不同的最低感染剂量和不同的组织特异性。其中,假型病毒Bacmid-GUS/VP19和Bacmid-GUS/VP24仅能感染对虾成体的心脏和肌肉组织,且在单个对虾肌肉组织中两者的最低感染剂量分别为1×10¹¹、1×10⁸个具有生物活性的病毒颗粒数。而假型病毒Bacmid-GUS/VP28不仅能感染对虾的心脏和肌肉组织,还能感染对虾的鳃组织,且其在...     

Computational identification and characterization of microRNAs and their targets in Penaeus monodon

Study on shrimp miRNAs was limited and just 7 mature miRNA sequences of Marsupenaeus japonicus are deposited in mir Base database. In this study, miRNAs and their target gene candidates were computationally identified from shrimp Penaeu s monodon and then experimentally validated. Using 39 908 expressed sequence tags(ESTs) and 21 124 genome survey sequences(GSSs) of P. monodon(pmo) as reference dataset, a comprehensive approach based on inter-species homolog search was employed to investigate the candidate miRNAs(i.e. pmo-miRNA). A total of eight miRNAs belonging to 7 families were computationally identified and five out of them were subsequently validated by PCR and sequencing. Of these, pmo-miR-4961a, pmo-miR-4961b, pmo-miR-4979 and pmo-miR-3819 were first identified from shrimps. Both the mature pmo-miRNAs and the corresponding precursors were conserved among different species. Based on perfect or near-perfect match to the target region, the target gene candidates of pmomiRNAs were predicted from 10 331 mRNA sequences of P. monodon. A total of 20 genes were predicted as the targets of pmo-miR-4961a, pmo-miR-4961b, pmo-miR-4979 and pmo-miR-6492. Experimental validation by dual luciferase reporter assay confi rmed the targeting between 3 pmo-miRNAs and one or two of their target genes, especially the pmo-miR-4979 which could significantly down-regulate the expression of target gene(JR226772). This study updates the miRNAs and their targets in P. monodon and lays a solid foundation for future RNAi study.     

CYTOTOXICITY AND GENOTOXICITY OF POLYETHYLENIMINE AND NICKEL CHLORIDE IN RED SEA BREAM (Pagrosomus major) FIN CELL LINE RSBF

ＩＮＴＲＯＤＵＣＴＩＯＮＡｃｃｕｍｕｌａｔｅｄｅｖｉｄｅｎｃｅｓｓｕｇｇｅｓｔｅｄａｃｌｏｓｅｃｏｒｒｅｌａｔｉｏｎｂｅｔｗｅｅｎｎｅｏｐｌａｓｔｉｃｄｉｓｅａｓｅｏｆａｑｕａｔｉｃｏｒ ｇａｎｉｓｍｓａｎｄｔｈｅｉｎｃｒｅａｓｉｎｇｒｅｌｅａｓｅｏｆｇｅｎｏｔｏｘｉｎｓｂｙａｎｔｈｒｏｐｏｇｅｎｉｃａｃｔｉｖｉｔｉｅｓｉｎｔｏａｑｕａｔｉｃｅｎｖｉｒｏｎｍｅｎｔ(Ｍｉｘ ,1 986;Ｇａｒｄｎｅｒｅｔａｌ.,1 991 ;ＧＥＳＡＭＰ ,1 991 ) .Ｉｔｈａｓ,ｔｈｅｒｅｆｏｒｅ,ｂｅｃｏｍｅｎｅｃｅｓｓａｒｙｔｏｅ…     

CYTOTOXICITY AND GENOTOXICITY OF POLYETHYLENIMINE AND NICKEL CHLORIDE IN RED SEA BREAM（Pagrosomus major）FIN CELL LINE RSBF

A continuous marine fish cell line RSBF(i.e.Red Sea Bream Fin)was utilized to screen the cytotoxicity and genotoxicity of polyethylenimine(PEI)and nickel cholride(NiCl2)in this study on the deleterious effects of aquatic genotoxins on fish.At the 0.01 to 1μgml concentration tested,PEI had acute toxicity to the treated RSBF cells(IC50=1.12,0.92,0.88 and 0.64μg/ml PEI for time 0 h,24 h,48 h and 72 h after treatment,respectively)and markedly inhibited their proliferation in a dose-dependent manner.At the 0.001 to 5 μmol/L concentration tested,NiCl2 posed no acute toxicity but significantly stimulated their growth(107?214?of control).Random amplified polymorphic DNA(RAPD)technique was used to detect the genotoxic effects of PEI and NiCl2 by comparing the RAPD banding patterns of the control and treated cells.RAPD analysis indicated that at the concentrations tested,PEI was more genotoxic than NiCl2 to RSBF cells;that there was a slight dose-dependent response in the genotoxic effect of PEI bue not NiCl2;and that RAPD technique might provide a sensitive,non-specif-ic gentoxic endpoint.And the potent cytotoxicity and genotoxicity of PEI on fish cells showed that we should be cautious in utilizing it as gene vector in fish gene transfer and human gene therapy.