Ultrastructural and functional characterization of circulating hemocytes from Plutella xylostella larva: Cell types and their role in phagocytosis

The hemocytes of different types encountered in the diamondback moth Plutella xylostella larvae of each instar and the development of the differential hemocytes counts were herein presented. Hemocytes classes/populations characterized based on their affinity with fluorescent dye (acridine orange) and ultrastructural differences comprised the prohemcoytes (<10–16%), plasmatocytes (22–65%), granulocytes (25–72%), oenocytoids (<1–9%), and spherulocytes (<1%). Prohemcoytes were the smallest cells with a comparatively tremendous nucleus. Plasmatocytes and granulocytes occupied the main proportion of total cell numbers. Oenocytoids were in a most stable presence, i.e. rotund in a diameter of 10 μm and with a nucleus deviated from the central location; however, sometimes with two nuclei which were adjoining with each other. Spherulocytes were rare and only could be observed occasionally. Ultrastructural investigation revealed that hemocytes in the diamondback moth larvae were of the typical model as in the Lepidoptera insect larvae. It is interesting to find that the cell which could phagocytize bacteria in vitro was granulocyte, not the other types of hemocytes, although plasmatocyte was usually declared to participate in this reaction in various previous studies.     

The identification of apolipoprotein genes in rare minnow (Gobiocypris rarus) and their expression following perfluorooctanoic acid exposure

Apolipoproteins play important roles in lipid transport and uptake in vertebrates, and they are associated with pathogenesis of many cardiovascular diseases. However, the diverse apolipoproteins in individual fish species have not been extensively characterized. Partial cDNA sequences encoding ApoA-IV, ApoE, ApoM, ApoL, and ApoO, and full-length cDNA sequences encoding ApoA-I were cloned from rare minnow (Gobiocypris rarus). Sequence analysis showed that these genes, as well as fragments of other known apolipoprotein genes (ApoC-I, ApoC-II, ApoB) of rare minnow had a high similarity (91–96%) to their orthologues in the spotted barbel Hemibarbus mylodon (Teleostei:Cypriniformes). The expression of these nine genes and their possible upstream genes, PPARα, PPARγ, and HNF4α, were investigated in rare minnow after subacute exposure to perfluorooctanoic acid (PFOA) for 14 days. Results showed that the expression of mRNA for ApoA-I, ApoC-II, and ApoM was significantly downregulated in all PFOA-treated animals. Only fish receiving the highest dose of PFOA showed downregulation of the expression of ApoA-IV and ApoC-I, while fish treated with 10 mg PFOA/L showed upregulation of expression of ApoE. Expression of ApoB, ApoO, and ApoL was unchanged between control and treated groups. In addition, the expression of PPARα was increased in all dosed fish, while the mRNAs for PPARγ and HNF4α were significantly altered with 30 and 3 mg PFOA/L doses, respectively. Therefore, subacute exposure to PFOA resulted in alteration of expression of apolipoproteins and related genes. These changes in gene expression may further influence lipid metabolism or other physiological functions in fish.     

Proteome comparison of hypopharyngeal gland development between Italian and royal jelly producing worker honeybees (Apis mellifera L.)

The hypopharyngeal gland (HG) of the honeybee (Apis mellifera L.) produces royal jelly (RJ) that is essential to feed and raise broods and queens. A strain of bees (high royal jelly producing bee, RJb) has been selected for its high RJ production, but the mechanisms of its higher yield are not understood. In this study, we compared HG acini size, RJ production, and protein differential expressions between the RJb and nonselected honeybee (Italian bee, ITb) using proteomics in combination with an electron microscopy, Western blot, and quantitative real-time PCR (qRT-PCR). Generally, the HG of both bees showed age-dependent changes in acini sizes and protein expression as worker behaviors changed from brood nursing to nectar ripening, foraging, and storage activities. The electron microscopic analysis revealed that the HG acini diameter of the RJb strain was large and produced 5 times more RJ than the ITb, demonstrating a positive correlation between the yield and HG acini size. In addition, the proteomic analysis showed that RJb significantly upregulated a large group of proteins involved in carbohydrate metabolism and energy production, those involved in protein biosynthesis, development, amino acid metabolism, nucleotide and fatty acid, transporter, protein folding, cytoskeleton, and antioxidation, which coincides with the fact that the HGs of the RJb strain produce more RJ than the ITb strain that is owing to selection pressure. We also observed age-dependent major royal jelly proteins (MRJPs) changing both in form and expressional intensity concurrent with task-switching. In addition to MRJPs, the RJb overexpressed proteins such as enolase and transitional endoplasmic reticulum ATPase, protein biosynthesis, and development proteins compared to the ITb strain to support its large HG growth and RJ secretion. Because of selection pressure, RJb pursued a different strategy of increased RJ production by involving additional proteins compared to its original counterpart ITb. To our knowledge, this morphological and proteomic comparison study on the HG of the two strains of worker honeybees associated with their age-dependent division of labor is the first of its kind. The study provided not only the quantity and quality differences in the HG from the RJb and the ITb, but also addressed the cellular and behavioral biology development question of how the RJb strain can produce RJ more efficiently than its wild type strain (ITb).     

不同pH调节剂对产琥珀酸放线杆菌NJ113发酵产丁二酸的影响

在3L发酵罐中分别采用不同的碱性物质作为pH调节剂,考察其对产琥珀酸放线杆菌Actinobacillus succinogenes NJ113厌氧发酵制备丁二酸的影响。结果表明:Ca2+、NH4+调节剂对菌体生长代谢有较大阻碍作用,丁二酸产量较低;采用含Na+调节剂,在发酵中后期菌体出现絮凝现象严重,且产丁二酸能力骤降;采用含Mg2+调节剂,整个发酵过程菌体代谢旺盛,发酵效果较佳。根据各碱性物质的调节能力以及对菌体生长代谢的影响,选择NaOH、Mg(OH)2和Na2CO3、Mg(OH)2分别作为混合碱组分调节pH,并对两组混合碱中各物质的质量比例进行优化。结果表明,以NaOH、Mg(OH)2混合,两者质量比为1:1时,发酵效果最好,丁二酸质量浓度高达到69.8g/L,质量收率74.5%。该种混合碱配比可有效替代碱式MgCO3调节pH,既达到高产丁二酸的目的,又可降低生物制备丁二酸的成本。     

植物化感抑藻的作用机理

对植物化感物质抑藻机理,即化感物质影响藻细胞的光合作用、破坏细胞膜、改变细胞酶活性、影响细胞超微结构及基因表达等几个方面进行了综合评述。研究表明,化感物质主要通过降低叶绿素a含量,破坏光合系统Ⅱ,使细胞膜结构裂解,酶活性改变,抗氧化系统破坏,细胞膜内超微结构受损,基因表达异常等几个方面来抑制藻细胞的分裂或直接杀死藻细胞,从而减少藻细胞数量,抑制藻类的生长。未来应该从更微观角度,即基因表达、分子和遗传机制等方面对化感抑藻机理进行深入研究,以期为开发安全、高效的抑藻化感物质提供理论基础。     

原癌基因c-erbB_2在原始卵泡启动生长中的作用

目的:探讨原癌基因c-erbB2在原始卵泡启动生长中表达变化及可能的作用。方法:选用2日龄SD大鼠卵巢在Waymouth培养体系中进行体外培养,用原位杂交、RT-PCR和免疫组化方法检测c-erbB2mRNA和蛋白在原始卵泡启动生长中及在表皮生长因子(EGF)作用下的表达情况,用Westernblot方法同步测定卵泡活化生长的重要标志物——增殖细胞核抗原(PCNA)和磷酸化细胞外信号调节激酶1/2(p-ERK1/2)的表达情况,并分析p-ERK1/2与c-erbB2mRNA表达变化的相关关系。结果:伴随原始卵泡启动生长过程,PCNA表达逐渐增加,EGF能促进原始卵泡的增殖和分化;原始卵泡中有c-erbB2mRNA及蛋白的表达,且随原始卵泡的启动生长及在EGF作用下表达增强;RT-PCR结果显示,c-erbB2mRNA表达在2日龄大鼠卵巢培养8d后与培养0d相比显著增加(0.297±0.018vs0.178±0.011,P0.05),并在EGF作用下进一步增强;p-ERK1/2含量的变化与c-erbB2mRNA表达的变化呈显著的正相关关系(rs=0.900,P0.05)。结论:c-erbB2在原始卵泡启动生长中起重要促进作用,并为介导EGF促进原始卵泡启动生长的关键信号分子;ERK-MAPK信号通路可能在介导c-erbB2调控原始卵泡生长中起作用。     

温度对白条锦蛇代谢率和不同组织ATP酶活性的影响

测定了不同温度条件下白条锦蛇Elaphe dione的代谢率以及骨骼肌、心肌和脑3种组织ATP酶的活性.结果 表明.温度从5℃上升到35℃,白条锦蛇代谢率随着温度的升高逐渐升高;但不同组织ATP酶活性在温度从28℃上升到35℃均逐渐下降.ATP酶活性的最适温度为28℃.ATP酶最适温度与白条锦蛇所喜好的温度有很大的相关性.