点带石斑鱼的亲鱼培育、产卵受精和胚胎发育

在水泥池人工养殖条件下,对人工诱导性逆转的“功能性雄性”、天然雄性及天然雌性点带石斑鱼(Epinephelus malabaricus)亲鱼进行强化培育,自然产卵受精。2002年繁殖季节,共获3cm以上的点带石斑鱼苗145万尾。亲鱼培育、产卵受精和胚胎发育的程序：(1)经过强化培育的亲鱼,自然产卵受精,受精率平均81％,最高97％;(2)成熟的“人工变性雄鱼”的精子头部呈球型,精子头径约3．7—6．1gm,尾长12．4～44．5μm;在水温26℃、盐度3％、pH7．9条件下,精子游动时间最长超过58min,精液中90％精子在31min停止游动,与天然雄亲鱼的精子无异,可作为生产雄亲鱼的来源;(3)在东山岛,产卵期由4月初持续到11月中旬,产卵高潮集中在4月底至5月底、9月底至10月初。在24h内,自然产卵通常从16：30持续到次日1：30;(4)产卵适宜水温为23．5—28．6℃,最适产卵水温为25．5—26．5℃;(5)水温26℃时,受精卵在盐度2．82％～2．96％,的海水中呈悬浮状态,盐度2．82％以下时下沉,盐度2．96％以上时上浮;(6)在水温24．9—27．6℃、盐度3％—3．3％、pH7．6—8．2的情况下,胚胎发育时间为21h。     

Improving heterologous protein expression in transfected Drosophila S2 cells as assessed by EGFP expression

Drosophila melanogaster S2 cells were co-transfected with plasmid vectors containing the enhanced green fluorescent protein gene (EGFP), under the control of metallothionein promoter (pMt), and the hygromycin selection gene, in view of establishing parameters for optimized gene expression. A protocol of transfection was worked out, leading after hygromycin selection, to ∼90% of S2MtEGFP fluorescent cells at day 5 after copper sulfate (CuSO₄) induction. As analyzed by confocal microscopy, S2MtEGFP cell cultures were shown to be quite heterogeneous regarding the intensity and cell localization of fluorescence among the EGFP expressing cells. Spectrofluorimetry kinetic studies of CuSO₄ induced S2MtEGFP cells showed the EGFP expression at 510 nm as soon as 5 h after induction, the fluorescence increasing progressively from this time to attain values of 4.6 × 10⁵ counts/s after 72 h of induction. Induction with 700 μM of CuSO₄ performed at the exponential phase of the S2MtEGFP culture (10⁶ cells/mL) led to a better performance in terms of cell growth, percent of fluorescent cells and culture intensity of fluorescence. Sodium butyrate (NaBu) treatment of CuSO₄ induced S2MtEGFP cell cultures, although leading to a loss of cell culture viability, increased the percent of EGFP expressing cells and sharply enhanced the cell culture fluorescence intensity. The present study established parameters for improving heterologous protein expression in stably transfected Drosophila S2 cells, as assessed by the EGFP expression.     

硝酸镧对霍霍巴多芽苗生长的促进作用及植株再生

在增殖培养基(改良MS 2mg／L6-BA 0．5mg／LG3)中添加1～3．5mg／L硝酸镧,对霍霍巴试管苗生长有显著促进作用,2．5mg／L硝酸镧对多芽分化有极显著的促进;在生根培养基(改良1/2MS 3mg／LIBA 1．5mg／LNAA)中,1～2mg／L的La(NO3)3对根的分化有显著的促进作用,生根率提高,最佳浓度为2．0mg／L。过高的浓度对试管苗生长有一定抑制。试验表明,不同器官对硝酸镧的敏感程度不同。取2cm高以上的霍霍巴试管苗,用25mg／L IBA或NAA处理30min,扦插于沙基质中。保持室温20～30℃。50d后揭去覆膜,保持光强3000 lx,相对湿度85％以上。正常管理条件下成活率达75％。     

几种化学物质对蟾蜍蝌蚪生存及生长发育的影响

在室内条件下,采用单因子急性和慢性毒性实验法,分别研究了水环境中的pH、洗涤剂、除草剂、重金属离子(Pb2 、Cu2 、Hg )对黑眶蟾蜍蝌蚪的毒性效应.结果表明,这些水体污染物不但对蝌蚪的生存造成危害,还对蝌蚪的红细胞有致畸作用;而其慢性毒害表现为蝌蚪身体畸形,肤色变浅,生长发育迟缓等.据此,可以利用黑眶蟾蜍蝌蚪对水体污染进行监测.     

黑眶蟾蜍幼蛙适应生境的探讨

报道人为设置6种不同生境对黑框蟾蜍（Bufo melanostictus Schneider)幼蛙存活时间的影响研究。结果表明：水深超过体长或干燥环境对都幼蛙不利,存活时间短。水深低于幼蛙体长并设有陆地或保持环境泥沙湿润为适宜的生境。在适应生境条件下,幼蛙平均存活674．86h,最长达35d。     

The neuralized homology repeat 1 domain of Drosophila neuralized mediates nuclear envelope association and delta-dependent inhibition of nuclear import

Signaling by the Notch (N) pathway is critical for many developmental processes and requires complex trafficking of both the N receptor and its transmembrane ligands, Delta (Dl) and Serrate. neuralized encodes an E3 ubiquitin ligase required for N ligand internalization. Neuralized (Neur) is conserved from worms to humans and comprises two Neur homology repeat (NHR) domains, NHR1 and NHR2, and a carboxyl-terminal RING domain. We have previously shown that the RING domain is required for ubiquitin ligase activity and that NHR1 mediates binding to Dl, a ubiquitination target. In Drosophila, Neur associates with the plasma membrane and hepatocyte responsive serum phosphoprotein-positive endosomes. Here we demonstrate that Neur also exhibits nuclear envelope localization. We have determined that Neur subcellular localization is regulated by nuclear trafficking and that inhibition of chromosome region maintenance 1, a nuclear export receptor, interferes with Neur nuclear export, trapping Neur in the nucleus. Moreover, we demonstrate that nuclear envelope localization is mediated by the Neur NHR1 domain. Interestingly, Dl expression in Schneider cells is sufficient to inhibit Neur nuclear import and inhibition occurs in an NHR1-dependent manner, suggesting that Neur nuclear localization occurs in contexts where Dl expression is either low or absent. Consistent with this, we found that Neur exhibits nuclear trafficking and associates with the nuclear envelope in the secretory cells of the larval salivary gland and that overexpression of Dl can reduce Neur localization to the nucleus. Altogether, our data demonstrate that Neur localizes to the nuclear envelope and that this localization can be negatively regulated by Dl, suggesting a possible nuclear function for Neur in Drosophila.     

A Systematic Cell‐Based Analysis of Localization of Predicted Drosophila Peroxisomal Proteins

Peroxisomes are membrane‐bound organelles found in almost all eukaryotic cells. They perform specialized biochemical functions that vary with organism, tissue or cell type. Mutations in human genes required for the assembly of peroxisomes result in a spectrum of diseases called the peroxisome biogenesis disorders. A previous sequence‐based comparison of the predicted proteome of Drosophila melanogaster (the fruit fly) to human proteins identified 82 potential homologues of proteins involved in peroxisomal biogenesis, homeostasis or metabolism. However, the subcellular localization of these proteins relative to the peroxisome was not determined. Accordingly, we tested systematically the localization and selected functions of epitope‐tagged proteins in Drosophila Schneider 2 cells to determine the subcellular localization of 82 potential Drosophila peroxisomal protein homologues. Excluding the Pex proteins, 34 proteins localized primarily to the peroxisome, 8 showed dual localization to the peroxisome and other structures, and 26 localized exclusively to organelles other than the peroxisome. Drosophila is a well‐developed laboratory animal often used for discovery of gene pathways, including those linked to human disease. Our work establishes a basic understanding of peroxisome protein localization in Drosophila. This will facilitate use of Drosophila as a genetically tractable, multicellular model system for studying key aspects of human peroxisome disease.      

矮生栒子的分类学研究

在有关模式和产地标本研究基础上,结合叶表皮微形态和细胞学资料,对矮生Ｘｕｎ子（Ｃｏｔｏｎｅａｓｔｅｒ ｄａｍｍｅｒｉ Ｓｃｈｎｅｉｄ．）进行了分类学修订,结果将Ｃ．ｄａｍｍｅｒｉ Ｓｃｈｎｅｉｄ．ｖａｒ．ｒａｄｉｃａｎｓ Ｓｃｈｎｅｉｄ．（即Ｃ．ｒａｄｉｃａｎｓ（Ｓｃｈｎｅｉｄｅｒ）Ｋｌｏｔｚ）归并作该种的同物异名;并描述了矮生Ｘｕｎ子的１个新亚种,Ｃ．ｄａｍｍｅｒｉ ｓｓｐ．ｓｏｎｇｍｉｎｇｅｎ     

Myogenic differentiation of Drosophila Schneider cells by DNA double-strand break-inducing drugs

Drosophila melanogaster has been widely used as a model organism to study various aspects of development. Apart from the whole Drosophila embryo, there are a number of cultured cell lines derived from Drosophila embryo that have also been used for elucidating various aspects of development. Drosophila Schneider line 2 cells were derived from the late stages of the embryo (Schneider, 1972). We found that the Schneider cells undergo myogenic differentiation upon treatment with neocarzinostatin (NCS), DNA double-strand break (DSB)-inducing drug, as indicated by elongated morphology, myosin heavy chain protein expression, multinucleation and exit from the cell cycle. No induction of differentiation was observed when cell proliferation was inhibited with drugs that do not cause DNA DSBs. Pre-treatment of Schneider cells with inhibitors of PKC, PP 1/2A, p38 MAPK, JNK and proteasomes resulted in the inhibition of morphological differentiation induced by NCS. These results indicate that DNA DSBs can turn on the myogenic program in Drosophila Schneider cells and the process is dependent on PK C-, PP 1/2A-, p38 MAPK-, and JNK- mediated signaling and proteasomal activity. The molting hormone, 20-hydroxyecdysone (20-HE), also showed an anti-myogenic effect on the process. This is the first report of insect cells undergoing differentiation by DNA DSB-inducing drugs as far as we know, and it provides a very useful and convenient in vitro system to study various aspects of Drosophila myogenesis.