Effects of inhibitors on 1-methyladenine induced maturation of starfish oocytes

1-methladenine (1-MA) induces starfish oocytes maturation via surface reaction followed by the appearance of a cytoplasmic maturation factor which in turn induces germinal vesicle breakdown (GVBD) to resume meiosis. Cellular mechanisms involved in GVBD were investigated by microinjection of metabolic inhibitors. Colchicine (Co) inhibited maturation, cytochalasin-B (CB) delayed GVBD and actinomycin-D-(Act-D) and puromycin (Pu) had no effect. It appears that the microtubule and the microfilament systems are associated with the nuclear membrane dissolution during the process of oocyte maturation of starfish.     

Comparison of lipids in organs of the starfish Asterias amurensis associated with different treatments

Lipids were extracted from organs of the starfish Asterias amurensis associated with different treatments(raw-control,boiling and heating),and then analyzed for lipid content,lipid oxidation index,lipid classes and fatty acid composition.Results showed that boiling softened the hard starfish shells,thus facilitating the collection of starfish organs.As compared with raw organs,the boiled organs had lower water content and higher lipid content,possibly due to the loss of water-holding capacity caused by protein denaturation.Both boiling and heating increased the peroxide value(PV),thiobarbituric acid(TBA) value and carbon value(CV) of lipids.Despite slight increases in the content of complex lipids,associated lipid composition had no substantial variations upon boiling and heating.For simple lipids,the content of 1,2-diglyceride decreased in boiled and heated organs,with free fatty acids observed on thin layer chromatography(TLC).However,neither boiling nor heating significantly changed the fatty acid compositions of simple or complex lipids in starfish organs,suggesting that these two treatments had no significant effects on complex lipids in starfish organs.Together,our results indicated that boiling of starfish soon after capture facilitated the handling and extraction of useful complex lipids consisting of abundant glucosylceramide and eicosapentaenoic acid(EPA)-bounded phospholipids.     

2种江蓠藻体切段的再生

初步研究了细基江蓠繁枝变种（Ｇ． ｔｅｎｕｉｓｔｉｐｉｔａｔａ ｖａｒ． ｌｉｕｉ）和细基江蓠（Ｇ． ｔｅｎｕｉｓｔｉｐｉｔａｔａ）切段培养的适宜培养基及其切段的再生特点。在此基础上,还研究了几种植物生长调节剂对江蓠藻体切段再生的影响。研究表明,江蓠藻体在改良ＰＥＳ培养基中生长良好。两种藻体的外皮层细胞均可产生长成新枝的生长点,并且表现出相同的再生极性。无论细基江蓠繁枝变种还是细基江蓠的切段都是在形态学上端切口处产生新枝,下端不再生;带分枝的切段,其分枝断口处亦可再生新枝。适宜的植物生长调节剂可明显促进藻体芽的产生和生长。１ ｍ ｇ／ ＬＢＡ 明显促进细基江蓠繁枝变种切段的再生,２ ｍ ｇ／ Ｌ ＢＡ＋ ０．５ ｍ ｇ／Ｌ ＮＡＡ 显著促进细基江蓠切段的再生     

Cytoplasmic regionalization in starfish oocyte occurrence and localization of cytoplasmic determinants responsible for the formation of archenteron and primary mesenchyme in starfish (asterias amurensis) oocytes

Starfish oocytes with intact germinal vesicles (GVs) were cut along desired planes with glass needles or ligated using silk thread loops into two parts and allowed to mature in vitro, and inseminated. The experimental results showed that (1) only the parts with GVs or partial GV contents (PGVCs) cleaved, those without any GV materials did not; but nucleated and non-nucleated fragments cut from mature eggs were able to divide; (2) the development of animal parts of oocytes containing GVs or PGVCs was like that of animal fragments of matured oocytes with female pronuclei; most of them gave rise to permanent blastulae, and just a few formed ectodermal vesicles with a little primary mesenchyme; (3) a large part of vegetal fragments with GVs or PGVCs, and the vegetal parts of mature eggs without female pronuclei developed into small but normal embryos; (4) the fragments containing GVs or PGVCs obtained from the oocytes along a plane parallel to the animal-vegetal (A-V) axis developed as normally as the halves (with or without female pronuclei) severed from mature eggs along the same axis. Based on the data above, it was concluded that (1) the non-chromatin materials in the oocyte GVs are indispensable for successful fertilization and cleavage of starfish eggs; (2) some factor (s) located asymmetrically in the vegetal hemispheres of starfish oocytes is (are) responsible for formation of the archenteron and primary mesenchyme. It is evident from the above findings that the oocyte cytoplasm of the starfish had already regionalized before the GV break-down. Contribution No. 1722 from the Institute of Oceanology, Academia Sinica     

斑鳢皮肤的组织学观察

本文对斑鳢皮肤的细微结构进行光学和电子显微镜的观察。研究结果表明:斑鳢的皮肤由表皮、基膜、真皮和皮下组织构成。表皮主要由复层的被覆上皮细胞和粘液细胞组成,电镜显示,上皮细胞彼此用指状突起和桥粒相联接。粘液细胞的内含物有三种不同形态,它们是处在不同功能时期的不同形态表现。真皮由稀疏层和致密层构成,其中细胞种类较多,结构复杂。真皮借基膜与表皮相联。电镜显示,基膜含有微丝状结构。皮下组织主要含胶原纤維,在与肌肉交界处,纤維向肌束间延伸。     

INDUCTION OF ECTODERMAL CELLS FROM VEGETAL-ENDODERMAL BLASTOMERES OF AMPHIOXUS(BRANCHIOSTOMA BELCHERI TSINGTAUNESE) EMBRYOS BY THE CALCIUM IONOPHORE A23187

INTRODUCnONAInPhioxusl6-celleInbryosconsistoftwotiersofblastomeres,eighboInalblas-tomeres,A8,andeightvegetalblastomeres,V8(Fig.l).Tungetal.(l958)shOWedthaV8isolatedatthel6-cellstage,whenculndexplan,gaVeriseInainlytointestineandoccasionalltonotochoal,muscleandneUraltube,butneverformedepidends-bendng-ly,thedeveloPmentalfateofV8blastomeresap~tobedetendnedatthel6-cellStag.HOWeVer,thesamauthors(l961)shoWedIaterthatifvegetal-mostblastomersatthe32-cellstage,thedescendansofV8,wereisolateda…     

Induction of ectodermal cells from vegetal-endodermal blastomeres of amphioxus (Branchiostoma belcheri tsingtaunese) embryos by the calcium ionophore A23187

Timing of vegetal-endodermal cell determination in amphioxus embryos remains uncertain. We tentatively tested effects of A23187, the calcium ionophore, on the development of vegetal blastomeres isolated at the 16-cell stage. It was found that when vegetal blastomeres committed to endoderm were treated with A23187 prior to gastrulation, they were transformed into ectodermal cells as evidenced by the cell morphology and function characteristic of epidermis. However, the developmental fate of the same blastomeres untreated or treated with DMSO at the same stage or of those treated with A23187 after gastrulation remained unchanged. Thus, vegetal-endodermal cells in amphioxus embryos are not irreversibly determined before the gastrula stage, and artificial increase in intracelluar Ca²⁺ concentration can induce transdetermination of the predetermined endodermal cells into ectodermal cells. Project 39470091 supported by NSFC and partly supported by Shandong Natural Science Foundation, Grant No.93D0140.     

马氏珠母贝珍珠囊发育的组织和组织化学研究

按照常规技术进行马氏珠母贝插核育珠,在手术后第1、3、7、15、20、30和60天解剖剪取珍珠囊,利用H E染色等组织化学方法研究马氏珠母贝珍珠囊形成过程中的组织学和组织化学变化。结果表明:珍珠囊表皮细胞来自于移植细胞小片的表皮细胞;在水温27℃条件下,插核后1~3 d来自育珠贝的游走细胞逐渐将细胞小片包围;插核后7~15 d,珠核表面密集来自育珠贝的游走细胞;插核后15～20 d,细胞小片外表皮细胞增殖形成珍珠囊,珍珠囊表皮细胞和细胞核均呈扁平状;插核后20～30 d,扁平状珍珠囊表皮细胞逐渐转变为高柱型,并分泌形成透明的壳皮层物质;在插核后第60天,已形成具有分泌珍珠质功能的扁平状珍珠囊表皮细胞,细胞核呈椭圆形或近圆形。     

DETECTION OF WHITE SPOT SYNDROME VIRUS(WSSV) OF PENAEUS CHINENSIS BY IN SITU HYBRIDIZATION

ＩＮＴＲＯＤＵＣＴＩＯＮＷｈｉｔｅＳｐｏｔＳｙｎｄｒｏｍｅＶｉｒｕｓ (ＷＳＳＶ)ｉｎＰｅｎａｅｕｓｃｈｉｎｅｎｓｉｓｉｓｔｈｅｃａｕｓａｔｉｖｅａｇｅｎｔｏｆａｖｅｒｙｓｅｖｅｒｅｅｐｉｚｏｏｔｉｃｄｉｓｅａｓｅｗｈｉｃｈ,ｓｔａｒｔｉｎｇｆｒｏｍ 1 993 ,ｈａｄｒｅｓｕｌｔｅｄｉｎｍｏｒｅｔｈａｎ 80 %ｍｏｒｔａｌｉｔｙｔｈｒｏｕｇｈｏｕｔｓｈｒｉｍｐｃｕｌｔｕｒｅｆａｒｍｓｉｎＣｈｉｎａ (Ｚｈａｎｅｔａｌ.,1 995;1 998) .Ｎｏｗ ,ｆｉｖｅｙｅａｒｓｌａｔｅｒ,ｔｈｅｖｉｒｕｓｄｉｓｅａｓｅｉｓｓｔｉ…     

Gene cloning and prokaryotic expression of recombinant outer membrane protein from Vibrio parahaemolyticus

Gram-negative Vibrio parahaemolyticus is a common pathogen in humans and marine animals. The outer membrane protein of bacteria plays an important role in the infection and pathogenicity to the host. Thus, the outer membrane proteins are an ideal target for vaccines. We amplified a complete outer membrane protein gene (ompW) from V. parahaemolyticus ATCC 17802. We then cloned and expressed the gene into Escherichia coli BL21 (DE3) cells. The gene coded for a protein that was 42.78 kDa. We purified the prote...     

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.     

Interaction of Haematopoietic Tissue Cultures of the Dublin Bay Prawn,Nephrops Norvegicus(L.),with the Causal Agent of Luminous Vibriosis Vibrio Harveyi

Vibrio harveyi cells (dose = ＞ 103 cells mL 1) and extracellular products (ECP; ＞25μg m L-1 of total protein concentration) destroyed haematopoietic cultures of Nephrops norvegicus within 24 h of exposure. Cytopathic effects (CPE)started after 4 h of exposure to the bacterial cells, with some granularity in the cytoplasm, mostly in cells in the outer periphery of the explant growth. At the end of the infection, a considerable number of nuclei remained attached to the substrate,apparently unaffected. Following exposure to ECP, initial deterioration was observed at 2 h with the presence of granularity in the cytoplasm of＜ 20% cells, and few cells displayed small vacuoles around the nuclei. Parallel results were obtained using whole animal experiments, with V. harveyi cells being lethal to nephrops within 24 h.     

Detection of white spot syndrome virus (WSSV) ofPenaeus chinensis by in situ hybridization

White Spot Syndrome Virus (WSSV) was purified from hemolymph of infected shrimp. After nucleic acid extraction from the purified virus particles, EcoR I-digested fragments of the WSSV genome were cloned; three of these fragments were used as non-radioactive probes labeled with DIG-11-dUTP. The probes hybridized in situ, with sections located in the nuclei of all WSSV-infected tissues. The virus was detected in the gill, stomach, epidermis, and connective tissue and so on, but not detected in healthy shrimp tissues and epithelial cells of hepatopancreatic tubules of diseased shrimp. This work was supported by SRF for ROCS, SEM and the National 863 Project (Grant 819-Q-08) and Project under major State Basic Research Development Program (Grant G1999012002).     

Histopathological Observation of Lymphocystis Disease and Lymphocystis Disease Virus （LCDV） Detection in Cultured Diseased Sebastes schlegeli

Lymphocystis nodules occurring in the cultured sting fish Sebastes schlegeli were observed under light and electron microscope. Lymphocystis disease virus (LCDV) in the tissues of diseased fish was detected with indirect immunofluorescence test (IFAT). Results showed that lymphocystis cells had overly irregular nuclei, basophilic intracytoplasmic inclusion bodies with virions budding from the surface, and hyaline capsules outside the cell membrane. Numerous virus particles about 200 nm in diameter scat- tered in the cytoplasm, electron-dense particles 70-80 nm in diameter filled in perinuclear cisterna, and membrane-enveloped parti- cles with electron-dense core of 70-80 nm appeared around cellular nucleus. IFAT using monoclonal antibody against LCDV from Paralichthys olivaceus revealed that specific green fluorescence was present in the cytoplasm of lymphocystis cells, epithelium of stomach, gill lamellae, and muscular fibers under epidermis of S. schlegeli, just as that in the cytoplasm of lymphocystis cells of P. olivaceus, suggesting the presence of LCDV in these tissues.     

DYNAMIC STUDIES ON FLAGELLAR REGENERATION IN DUNALIELLA SALINA

INTRODUCTIONMicroalgalbiotechnologyhasattractedmoreandmoretheinterestofscientistsandpeopleex ploitingitscommercialpossibilitiesasitisthebiotechnologythatcanmatchbacterialbiotechnologyinthecomingcenturyintheaspectofeconomicreturnoninvestment.Dunaliellasalinaisoneofthemostdetailedlystudiedunicellulargreenalgae,hassomeuniquescientificvalues,andgreatestpotentialformasscultureasafoodsource ,andisrichestalgalsourceofglycerolandβ carotene(Borowitaka,1 988;Ginzburg,1 987) .Optimizingperformanceo…     

Culture of yeast cells immobilized by alginate-chitosan microcapsules in aqueous-organic solvent biphasic system

Immobilization biocatalysis is a potential technology to improve the activity and stability of biocatalysts in nonaqueous systems for efficient industrial production. Alginate-chitosan(AC) microcapsules were prepared as immobilization carriers by emulsification-internal gelation and complexation reaction,and their contribution on facilitating the growth and metabolism of yeast cells were testified successfully in culture medium-solvent biphasic systems. The cell growth in AC microcapsules is superior to that in alginate beads, and the cells in both immobilization carriers maintain much higher activity than free cells,which demonstrates AC microcapsules can confer yeast cells the ability to resist the adverse effect of solvent.Moreover, the performance of AC microcapsules in biphasic systems could be improved by adjusting the formation of outer polyelectrolyte complex(PEC) membrane to promote the cell growth and metabolic ability under the balance of resisting solvent toxicity and permitting substrate diffusion. Therefore, these findings are quite valuable for applying AC microcapsules as novel immobilization carriers to realize the biotransformation of value-added products in aqueous-solvent biphasic systems.     

九孔鲍外套膜表皮细胞的超微结构

利用透射电镜观察九孔鲍（Haliotidaediversicoloraquatilis）的外套膜表皮细胞,结果表明细胞可分为4大类,即普通柱状表皮细胞、嗜碱性粒细胞、嗜酸性粒细胞和感觉细胞。它们在不同区域的分布、形态和数量变化与外套膜的功能分化密切相关。     

Ontogeny of the Lymphoid Organs of Japanese Flounder, Paralichthys olivaceus

Histological study on the ontogeny of the lymphoid organs, kidney, thymus and spleen of Japanese flounder, Paralichthys olivaceus, from hatching to 40d was carried out. The pronephric kidney duct appeared early in hatching although the primordial haemopoietic stem cells wereobserved within a week after hatching. The spleen was first seen after 8d of hatching. The thymus appeared after 15d, situated near the pronephric kidney. Small lymphoid cells appeared during the later phase of the post-larval stage in the sequence of thymus, kidney and spleen. During the 40d of observations, there were no distinct inner or outer zones in thymus and no red or white pulp in spleen. These results suggest that the nonspecific defense immune system plays a very important role in the early larval stage of Japanese flounder.     

Dynamic studies on flagellar regeneration inDunaliella salina

Flagella, the basic locomotive organ in algae, as well as bacteria and some cells of animals or high plants, would be damaged in the well stirred mass culture due to the strong forces caused by the fast mixing impellers. The dynamic regeneration of the flagella in deflagellatedDunaliella salina was studied microscopically by using a bench-top flat-bottom photobioreactor. The results showed that 90 minutes was necessary for the repair of flagella, after which half of the cells became motile as their flagella generated within 120 minutes and nearly all of the cells could swim freely within 180 minutes. Contribution No. 3503 from the Institute of Oceanology, Chinese Academy of Sciences.