Molecular cloning of the Bombyx mori prothoracicotropic hormone

Prothoracicotropic hormone (PTTH), a brain secretory polypeptide of insects, stimulates the prothoracic glands to produce and release ecdysone, the steroid essential to insect development. The complementary DNAs encoding PTTH of the silkmoth Bombyx mori were cloned and characterized, and the complete amino acid sequence was deduced. The data indicated that PTTH is first synthesized as a 224-amino acid polypeptide precursor containing three proteolytic cleavage signals. The carboxyl-terminal component (109 amino acids) that follows the last cleavage signal represents one PTTH subunit. Two PTTH subunits are linked together by disulfide bonds, before or after cleavage from prepro-PTTH, to form a homodimeric PTTH. When introduced into Escherichia coli cells, the complementary DNA directed the expression of an active substance that was functionally indistinguishable from natural PTTH. In situ hybridization showed the localization of the prepro-PTTH mRNA to two dorsolateral neurosecretory cells of the Bombyx brain.     

家蚕脑核蛋白抽提及其与PTTH基因启动子的凝胶阻滞分析

 【目的】转录调控对于明确昆虫许多生理过程如抗药性、抗病性、行为、进化及发育等方面的机理具有重要意义。家蚕PTTH是调控生长发育的重要神经肽,明确其转录调控机理对阐明其表达的发育和组织特异性具有重要价值。【方法】本文克隆并测定了家蚕PTTH启动子序列,利用Matinspector分析软件预测该片段可能包含的顺式作用元件,采用高糖抽提缓冲液抽提家蚕脑核蛋白;标记PTTH启动子片段进行凝胶阻滞分析。【结果】家蚕PTTH启动子分析含有MEF2、TATA box、pbx-1等元件;抽提获得理想的脑核蛋白,脑核蛋白可以与PTTH转录起始位点附件119bp的序列特异地结合,把TATA Box的TATATAA突变后,脑核蛋白与突变后的探针MBS2不能结合。【结论】表明本研究中抽提的脑核蛋白是有效的,首次检测家蚕PTTH启动子可以与转录因子特异结合,初步鉴定了TATA Box。     

印楝素对昆虫的毒理作用机制

印楝素对许多种昆虫具高效的拒食，胃毒，触杀及抑制生长发育的作用，其作用机制主要是扰乱昆虫的内分泌，作用靶标为脑神经分泌细胞，心侧体，前胸腺等，影响促前胸腺激素（ＰＴＴＨ）的合成与释放，减低前胸腺对ＰＴＴＨ的感应而造成２０－羟基蜕皮酮合成，分泌的不足，致使昆虫变态，发育受阻，印楝对其他一些与变态激素的合成，释放相关的组织的作用，在该文也有讨论。     

不同发育时期柞蚕雌蛾粘液腺的组织形态学观察

【目的】对柞蚕雌蛾粘液腺的生长发育过程和组织结构特征进行研究,并分析其内容物成分,为阐明粘液腺分泌胶着物质的机理提供参考。【方法】通过解剖获得不同发育时期的粘液腺并制备组织石蜡切片,观察其形态特征及组织显微结构。【结果】随着发育阶段的推进,粘液腺形态呈现出逐渐增大趋势,至羽化前达最大。而粘液腺内容物的颜色亦发生了显著变化,由透明色逐渐变为深黑色。光学显微镜观察结果显示,粘液腺的分泌部和贮藏部横截面结构存在差异,前者具备分泌细胞的特征,而后者不具备。【结论】粘液腺的形态结构特征在不同发育时期及组织部位存在差异。     

家蚕神经肽促前胸腺激素基因mRNA的发育表达

[目的]研究家蚕神经肽促前胸腺激素基因mRNA的发育表达。[方法]采用荧光定量PCR方法,研究家蚕5龄幼虫、预蛹期、蛹期和成虫4个阶段脑—咽下神经节复合体中PTTH基因的mRNA表达量变化。[结果]5龄幼虫PTTH基因mRNA表达量高于预蛹期、蛹期和成虫3个阶段;5龄第6天PTTH基因mRNA表达量最高,蛹期4～8d时持续维持较高表达水平,化蛾前表达量又有所降低,成虫表达量最低。[结论]家蚕PTTH基因mRNA的发育表达变化可能与其调控蜕皮激素合成的功能有关。     

Cytochemical localization of H_2O_2 in pigment glands of cotton(Gossypium hirsutum L.)

Programmed cell death(PCD) plays a critical role in the development of plant pigment glands, while H_2O_2, which is a kind of reactive oxygen species(ROS) produced by the aerobic metabolism of cells, acts as an important signal in this process. Here, we investigated the temporal and spatial dynamics of accumulated H_2O_2 in pigment glands of Gossypium hirsutum L. with 3,3-diaminobenzidine(DAB) staining, 2',7'-dichlorodihydrofluorescein diacetate(DCFH2)-DA fluorescent labeling and Ce Cl3 cytochemical localization techniques. The results showed that the pigment glands of G. hirsutum could generate H_2O_2, and the amount and localization of H_2O_2 varied at different developmental stages. At the early developmental stage, a small amount of H_2O_2 accumulated in the vacuole membrane of pigment gland cells. At the intermediate stage, a large number of H_2O_2 appeared in the vacuole membrane, while cell walls started to accumulate a small amount of H_2O_2. When pigment gland cell degraded, H_2O_2 mainly accumulated on the chloroplast envelope membrane of inner sheath cells. With the degradation of the sheath cells, H_2O_2 was detected in cell wall and the membrane of secretory vesicles which contains the preliminary contents of pigment gland. With the pigment glands completely maturation, H_2O_2 would disappeared. The accumulation sites of H_2O_2 are consistent with the process of PCD of individual gland cells, which started from the degradation of intracellular membrane and ended with the degradation of cell walls. Thus H_2O_2 probably plays an important role in the development of pigment glands. In addition, the development of pigment glands and the generation of H_2O_2 are not associated with the light, and no H_2O_2 was detected in the secretions of pigment glands.     

Human T-cell clones from autoimmune thyroid glands: specific recognition of autologous thyroid cells

The thyroid glands of patients with autoimmune diseases such as Graves' disease and certain forms of goiter contain infiltrating activated T lymphocytes and, unlike cells of normal glands, the epithelial follicular cells strongly express histocompatibility antigens of the HLA-DR type. In a study of such autoimmune disorders, the infiltrating T cells from the thyroid glands of two patients with Graves' disease were cloned in mitogen-free interleukin-2 (T-cell growth factor). The clones were expanded and their specificity was tested. Three types of clones were found. One group, of T4 phenotype, specifically recognized autologous thyroid cells. Another, also of T4 phenotype, recognized autologous thyroid or blood cells and thus responded positively in the autologous mixed lymphocyte reaction. Other clones derived from cells that were activated in vivo were of no known specificity. These clones provide a model of a human autoimmune disease and their analysis should clarify mechanisms of pathogenesis and provide clues to abrogating these undesirable immune responses.     

Insensitivity to Ethylene Conferred by a Dominant Mutation in Arabidopsis thaliana

Ethylene influences a number of developmental processes and responses to stress in higher plants. The molecular basis for the action of ethylene was investigated in mutants of Arabidopsis thaliana that have altered responses to ethylene. One mutant line, which has a dominant mutation at a locus designated etr, lacks a number of responses to ethylene that are present in the wild-type plant. These include inhibition of cell elongation, promotion of seed germination, enhancement of peroxidase activity, acceleration of leaf senescence, and feedback suppression of ethylene synthesis by ethylene. These diverse responses, which occur in different tissues of Arabidopsis, appear to share some common element in their transduction pathways-for example, a single receptor for ethylene. Results of ethylene binding experiments in vivo indicate that this receptor may be affected by the etr mutation.     

昆虫脑神经肽的研究进展:抑前胸腺肽PTSP

1999年发现和鉴定的新型昆虫脑神经肽--抑前胸腺肽,对昆虫的变态起着重要的作用.本文介绍它的研究背景、结构、功能、与促前胸腺素的相互作用关系及其分子生物学研究的结果,并对它的研究作一评价和展望.     

葱蝇非滞育蛹期的发育形态学研究

葱蝇是昆虫蛹滞育研究的理想模式种,其发育形态学特征对推断滞育发育过程至关重要,目前还没有花蝇科蛹期发育形态学的相关报道.本研究通过解剖和拍照观察,系统地研究了葱蝇老熟幼虫期到羽化的发育形态学特征,特别注重头外翻前后的形态变化.研究结果将老熟幼虫期划分为5个发育阶段;将预蛹期分为白色围蛹期和棕色围蛹期2个亚时期,13个发育阶段;将蛹期分为隐头蛹期和显头蛹期2个亚时期,30个发育阶段.其中,显头蛹期又可细分为显头蛹初期、发育形态停滞期、半透明眼期、浅黄色到琥珀色眼期、红褐色眼期、鬃毛蛹期和预成熟期.本研究还详细地描述了各时期及发育阶段的发育过程、历期和形态特征,并详细地观察和描述了头外翻的发育过程,把蛹壳外可见幼虫口钩确定为头外翻发生的外部标志特征.同时,还描述了蛹主要发育阶段的长宽比例,成熟雌、雄蛹间的形态差异,结果表明雌虫体型(体长和体宽)比雄虫略大,雄虫的复眼比雌虫大,雌虫两复眼间的距离超过雄虫的2倍.本文是花蝇科昆虫蛹期发育形态学的首次报道,对认知蝇类昆虫蛹期发育形态,了解葱蝇滞育与非滞育蛹发育过程、历期和形态变化有重要的意义,为葱蝇滞育的分子机理研究奠定了形态学基础.     

棉蝗的形态研究：Ⅴ.消化系统

棉蝗消化系统由消化道和唾腺组成，消化道分为前肠，中肠和后肠三段，前肠由口，咽喉，食道，嗉囊和前胃组成。中肠为一简单的直管。后肠由回肠，结肠和直肠组成。中肠为简单的直管。后肠由回肠，结肠和直肠组成。前肠与中肠交界处处面着生胃盲囊，内有贲门瓣，中肠与后肠交界处外面着生有马氏管，内面有幽门瓣。前肠内壁通常具齿。在内壁不同部倍，齿的排列方式和形状有所差异。直肠内具有直肠垫。唾腺形状似葡萄状，由许多分泌球体     

番荔枝花发育不同阶段的差异蛋白质组分析

【目的】从蛋白质组学的角度研究番荔枝花发育不同阶段差异表达的蛋白质,分析相关信号通路,为揭示番荔枝花发育的分子机制提供理论基础。【方法】以番荔枝不同发育阶段的花为材料,采用双向电泳技术（Two-dimensional gel electrophoresis,2-DE）分离获得花芽（1 mm＜花芽＜2 mm）、花蕾（5 mm＜花蕾＜6 mm）、开花前期（花瓣轻微张开）及开花期（花瓣张开）等不同阶段表达丰度不同的蛋白点,利用MALDI-TOF-MS质谱鉴定技术分析相关差异蛋白质,并对其进行功能注释。【结果】通过蛋白图谱比较分析发现,番荔枝花双向电泳图谱中的每张凝胶图谱中都可以检测到800多个重复蛋白点,其中在不同花发育阶段存在表达差异的有50个蛋白点。经质谱鉴定分析,并采用MASCOT软件在线检索,共检测到36个表达量相差2倍以上的蛋白点。36个蛋白质可分为7个主要的功能类别,这些功能类别包括：呼吸与能量代谢（11个蛋白质）,蛋白合成与代谢（8个蛋白质）,转录与翻译（3个蛋白质）,胁迫与防御（2个蛋白质）,细胞分化与增殖（3个蛋白质）,次生物质代谢（8个蛋白质）和未知功能蛋白（1个蛋白质）。通过GO分析,发现36个已鉴定的蛋白,分别归属于20个分类（term）。通过对差异蛋白进行定量分析,发现呼吸与能量代谢相关的蛋白中,3个蛋白（A07、C28、C42）随着花发育表达量不断下降,4个蛋白（A36、A37、B13、B29）表达则显著上升。在蛋白合成与代谢相关蛋白中,2个蛋白（A13和A22）在花发育的4个阶段都处于低水平表达,而另5个蛋白（B17、B20、A19、A21和C21）表达量显著上升。一个胁迫与防御相关蛋白（B32）在开花期,表达量达到最大值。2个细胞分化与增殖相关蛋白（B27和C57）表达量也呈逐渐升高的趋势。5个次生物质代谢相关蛋白（A06、A29、A34、C100和C110）随着花发育,表达量逐渐降低,而蛋白质C79和D38在花蕾阶段表达量达到最大值后,又逐渐下降。【结论】通过比较番荔枝花发育4个阶段的蛋白图谱,发现36个蛋白具有不同的差异表达谱。其中呼吸与能量代谢相关蛋白质占最大比例,诸如两个ATP合酶α亚基和丙酮酸脱羧化同工酶,都在番荔枝花发育过程呈现出显著的差异表达,这可能与花发育过程需要耗费大量的能量有关。此外,发掘了两个含有三角状五肽重复蛋白,其表达随着番荔枝花发育阶段的变化而改变,这些蛋白的功能还有待进一步研究。能量代谢、次生代谢、蛋白质合成等生物过程可能都参与了对番荔枝开花过程的调控。     

桃果实生长素结合蛋白ABP1的克隆及表达分析

ABP1是一种能够快速响应生长素信号的结合蛋白,参与细胞伸长、质膜极化、网格蛋白内吞及果实发育等生理过程。以‘24号’桃果实为试验材料,应用荧光实时定量PCR分析PpABP1在桃果实发育过程中的表达量变化,以及外施NAA对桃果实PpABP1表达量的影响。结果表明:PpABP1在桃果实中果皮和种子不同发育时期均有表达,花后52dPpABP1表达量均达到最大值;外施NAA对桃果实发育过程中PpABP1表达量的影响与发育时期相关,在桃果实硬核期,外施NAA会降低PpABP1在中果皮的表达。本试验初步证实桃果实发育过程中存在由ABP1介导的信号转导途径,PpABP1在桃果实中的表达存在发育特异性。     

An ancient developmental induction: heat-shock proteins induced in sporulation and oogenesis

Every eukaryotic and prokaryotic organism tested to date synthesizes a small number of heat-shock proteins in response to heat and other forms of stress. A particular pattern of heat-shock gene expression was observed during ascospore development in Saccharomyces: heat-shock proteins hsp26 and hsp84 were strongly induced nor inducible by heat shock. Instead, two proteins related to hsp70 were induced. A strikingly similar pattern of expression occurs during oogenesis in Drosophila, suggesting that it may be one of the earliest developmental pathways to evolve in eukaryotic cells.     

Larval development and dispersal at deep-sea hydrothermal vents

Deep-sea hydrothermal vent communities exhibit an array of reproductive strategies. Although a few vent species undergo planktotrophic, high-dispersal modes of development, most exhibit relatively low dispersal, but probably free-swimming nonplanktotrophic development. This predominance of nonplanktotrophy may be largely a reflection of phylogenetic constraints on the vent colonizing taxa; intervent dispersal among these forms may be facilitated by reduced developmental rates in the cold abyssal waters away from the vents. It is proposed that for those vent species with nonplanktotrophic development, larval dispersal is a stepwise process with oceanic ridge axes serving as discrete dispersal corridors.     

大麦小孢子原位发育过程中二态性现象的研究

对大麦不同原位发育时期小孢子直径测量和小孢子活力镜检的结果表明 ,小孢子原位发育过程中小孢子群体存在二态性现象———活力小孢子和衰败小孢子同时存在。对小孢子二态性现象的研究 ,有助于透彻地理解小孢子不同原位发育时期的特点 ,同时为大麦单雄生殖的研究提供帮助。给出的大麦穗子的I L值和小孢子发育时期之间的关系 ,是选择包含某一原位发育时期小孢子的大麦穗子的直观可靠的参数     

蛋白质组学在水稻生长发育研究中的应用研究进展

水稻不同生育时期的蛋白质组学是近年来的研究热点,对揭示水稻发育的分子机制具有重要价值.通过分析蛋白质组学在水稻生长发育研究中的应用现状,提出了亚细胞蛋白质组学和翻译后修饰蛋白质组学将是未来几年水稻蛋白质组学研究的热点,而技术手段的多样化、分析层次的多元化及与其他学科的紧密融合是水稻发育蛋白质组学的主要发展趋势.     

Nuclear number in the rotifer asplanchna: intraclonal variation and environmental control

The gastric glands and vitellarium of Asplanchna are exceptions to the rule of constancy of the number of nuclei in rotifers. The two glands of a female may have different numbers of nuclei. Nuclear number in both organs varies widely among different individuals in a single culture. The mean nuclear number characteristic of a clone may be modified by dietary changes.