克氏原螯虾消化道内分泌细胞的鉴别与定位

目的 对克氏原螯虾（Procambarus clarkii）消化道的嗜银细胞及5种内分泌细胞进行鉴别与定位。方法应用整块组织Grimelius银染法和过氧化物酶标记的链霉亲和素（SP法）免疫组织化学技术结合生物统计学分析。结果嗜银细胞在克氏原螫虾消化道除幽门胃外的各段均有分布,位于消化道上皮细胞间及结缔组织中。五羟色胺（5-HT）细胞在除幽门胃外的消化道各段均有分布。生长抑素（SS）细胞仅在食道和后肠中有分布。胃泌素（Gas）细胞分布于除幽门胃和中肠外的消化道各段。胰高血糖素（Glu）细胞在除幽门胃外的整个消化道均有分布,在食道和贲门胃中最多。胰多肽（PP）细胞仅在肠道中有较多分布。结论克氏原螫虾消化道中存在多种内分泌细胞,它们的分布情况与其它甲壳动物存在一定的共性,然而也有其一定的特异性。     

花鲈消化道内分泌细胞的鉴别和定位

应用免疫组织化学Elivision二步法 ,用 5 羟色胺 (5 HT)、生长抑素 (SST)、胃泌素 (Gas)、胰高血糖素 (Glu)、胰多肽 (PP)等 5种抗哺乳动物血清对花鲈 (Lateolabraxjaponicus)消化道内分泌细胞 ,进行免疫组织化学定位的研究。结果表明 :5 HT和SST细胞分布于食管、胃贲门部、胃体部和胃幽门部 ,而肠道各段均未检出。Gas细胞分布于胃幽门部和前肠、中肠 ;食管、胃贲门部、胃体部和后肠未检出。Glu细胞分布于前肠和中肠 ,其余各段均未检出。PP细胞在消化道各段均未检出。本文将对花鲈消化道内分泌细胞的分布密度、分布型及形态进行描述和讨论。     

中华稻蝗全消化道内壁显微结构观察

本文采用扫描仪和扫描电镜对中华稻蝗消化道内壁的细微结构进行了系统观察和研究。结果表明,中华稻蝗食道内壁由纵行脊组成,前端有齿。嗉囊包括两段,前段的一个小的膨大部分,由V-形区和两侧的V-形脊组成,只在前端内壁有齿;后段为一个大的膨大部分,由柳叶脊、扇形脊和不规则脊组成,脊的上缘有齿。前肠内壁的齿主要为单生齿,除贲门瓣上齿的齿尖指向前方外,全部齿的齿尖指向后方。后肠的前端为12个幽门瓣,内壁有齿。回肠和结肠由6条纵行脊组成,结肠内壁有齿。直肠的齿在除直肠垫外的直肠内壁上。后肠的齿主要为丛生齿,后肠除直肠内壁齿的齿尖指向附着环外,全部齿的齿尖指向后方。根据我们的观察,对前肠提出了新的分区。     

竹叶青蛇消化道内分泌细胞的免疫组织化学定位

应用8种特异性胃肠激素抗血清对竹叶青蛇（Trimersurus stejnegeri）消化道内分泌细胞进行了免疫组织化学定位。5－羟色胺细胞和生长抑素细胞在整个消化道中均有分布。5－羟色胺细胞以十二指肠和回肠密度为最高,空肠、直肠和幽门其次,胃体较低,食道和贲门中偶见;生长抑素细胞在十二指肠处数量最多,胃体、幽门和空肠其次,其余消化道各段偶见;胰高血糖素细胞只见于十二指肠和空肠;血管活性肠肽在贲门和胃体处有少量阳性细胞分布,食道中偶见。以上各处内分泌细胞的分布密度差异显著。未检出胃泌素、P－物质、胰多肽和胰岛素免疫反应阳性细胞。上述内分泌细胞的分布特点可能与竹叶青的摄食习性和生活环境有关。     

棘胸蛙消化道内分泌细胞的免疫组织化学定位

We studied the distribution and density of the endocrine cells in the digestive tract of Paa spinosa using immunohistochemical method (streptavidin-peroxidase method) using eight gut hormone antisera.The 5-hydroxytryptamine immunoreactive cells were distributed throughout the digestive tract with the highest density in the stomachus pyloricus,the second highest in the duodenum,fewer in the oesophagus,stomachus cardiacus and rectum.The gastrin immunoreactive cells were located mainly in the stomachus pyloricus and occasionally in different parts of the intestine.The somatostatin immunoreactive cells occured mainly in the stomach,frequently in the stomachus pyloricus,and occasionally in different parts of the intestine.The pancreatic polypeptide immunoreaetive cells were found with the highest density in the duodenum,the second highest in the stomachus cardiacus,and rarely in the rectum.No immunoreactive cells were observed with the antisera to glucagon,substance P,growth hormone and calcitonin,but there were glucagon and substance P mucosal nerve plexus throughout the digestive tract,and both with the highest density in the duodenum[Acta Zoologica Sinica 49(6):858-864,2003].     

北京鸭消化道内分泌细胞的免疫组织化学研究

应用七种消化道激素抗血清，对北京鸭消化道内分泌细胞进行了免疫组织化学定位，促胃素释放肽细胞大量分布于腺胃和肌胃。生长抑素细胞在腺胃和肌胃数量很多，在幽门部密集，且偶见于十地二指肠，胃素细胞在幽门部非常密集，并较多分布于整个小肠，肌胃内亦有少量。５－羟色胺细胞大量见于肠管各段，并偶见于幽门，少量胰多肽细胞见于腺胃、十二指肠和空肠，未检出胃动素和抑胃肽细胞。     

中华稻蝗蝗蝻龄期识别

<正> 中华稻蝗Oxya Chinensis(Thunberg)是我市水稻的重要害虫,主要掌握在四龄蝗蝻前进行药剂防治。关于该虫的生活史,柳支英等曾作过报道。为了便于在田间识别其龄期,1989～1991年作者饲养其生活史时,对各龄蝗蝻的主要鉴别特征再作了较详细的观察比较,结果如下。     

重口裂腹鱼消化道内分泌细胞的免疫组织化学鉴别和定位

重口裂腹鱼（Schizothorax davidi）俗称“雅鱼”,属鲤科,裂腹鱼亚科,主要分布于大渡河、青衣江水系上游,以动物性食性为主。因其生长缓慢,肉质鲜嫩,深受人们喜爱。随着捕捞强度的增大和自然生态环境的破坏,其资源量急剧下降。目前大多数学者使用哺乳动物抗血清对鱼类消化道内分泌细胞进行了鉴别和定位,在硬骨鱼和软骨鱼的胃肠胰中分别发现了17种和20种内分泌细胞。[第一段]     

蓝狐消化道内分泌细胞的免疫组化研究

蓝狐又名北极狐(Alopex lagopus),属于食肉目(Carnivora),犬科(Canidae),北极狐属(Alopex).     

中华稻蝗几丁质脱乙酰基酶1基因的分子特性及功能

【目的】通过研究中华稻蝗Oxya chinensis几丁质脱乙酰基酶1 (chitin deacetylase 1, CDA1)基因的分子特性和生物学功能, 为新型农药靶标筛选提供理论基础。【方法】在中华稻蝗转录组数据库搜索获得几丁质脱乙酰基酶基因片段, 用生物信息学方法对其进行同源比对分析并作聚类关系图; 利用cDNA末端快速扩增（rapid-amplification of cDNA ends, RACE）获得该基因cDNA全长序列; 通过Real-time quantitative PCR(qPCR)方法检测OcCDA1在5龄若虫不同组织部位和不同发育时期的表达情况, 并采用RNAi技术研究OcCDA1在飞蝗生长发育中的功能。【结果】获得中华稻蝗几丁质脱乙酰基酶1基因全长cDNA,命名为OcCDA1（GenBank登录号：KP271171）。OcCDA1在前肠、体壁、后肠和脂肪体组织中高表达, 在5龄第1天表达量显著高于以后几天。RNAi结果显示, 注射该基因的dsRNA后, OcCDA1基因的表达量降低了93.3%。OcCDA1基因沉默后虫体出现进食减少、发育迟缓现象, 总死亡率达到95.2%,其中38.1%的试虫在蜕皮前死亡, 57.1%出现因蜕皮困难而死亡的表型。【结论】OcCDA1在中华稻蝗的发育中起着重要作用, 该基因沉默引起虫体进食减少和蜕皮异常从而导致死亡。     

氟虫脲对东亚飞蝗和中华稻蝗几丁质合成酶基因表达的影响

为了探讨氟虫脲可能的作用靶标及毒性机制, 本研究以重要农业害虫东亚飞蝗Locusta migratoria manilensis (Meyen)和中华稻蝗Oxya chinensis (Thunberg)为材料, 采用简并引物扩增中华稻蝗几丁质合成酶1基因（OcCHS1）的部分cDNA序列; 以氟虫脲浸渍法处理2龄中期中华稻蝗及1, 2和3龄东亚飞蝗若虫为处理组, 丙酮处理为对照组, 使用RT-PCR和实时荧光定量PCR方法分析氟虫脲对蝗虫几丁质合成酶基因mRNA表达的影响。结果获得的OcCHS1部分cDNA序列, 其长度为312 bp, 编码104个氨基酸, GenBank登录号为HM214491, 与东亚飞蝗几丁质合成酶1基因（LmCHS1）在氨基酸水平上相似度达95％。RT-PCR结果显示, 处理组几丁质合成酶1扩增带均强于对照组。实时荧光定量PCR结果表明: 与对照组相比, 处理组中华稻蝗2龄中期若虫OcCHS1 mRNA表达提高了1.02倍, 东亚飞蝗1, 2, 3龄若虫LmCHS1 mRNA表达分别提高了34%, 82%和89%, 差异显著（P<0.05）。分析基因表达提高的原因是几丁质合成受阻后基因表达水平的一种代偿性增加, 由此推测几丁质合成酶可能是氟虫脲作用的靶标之一。     

Complete mitochondrial genome of Oxya chinensis （Orthoptera, Acridoidea）

The complete sequence of Oxya chinensis （0. chinensis） mitochondrial genome is reported here. It is 15,443 bp in length and contains 75.9% A＋T. The protein-coding genes have a similar A＋T content （75.2%）. The initiation codon of the cytochrome oxidase subunit I gene in the mitochondrial genome of O. chinensis appears to be ATC, instead of the tetranucleotides that have been reported in Locusta migratoria （L migratoria） mitochondrial genome. The sizes of the large and small ribosomal RNA genes are 1319 and 850 bp, respectively. The transfer RNA genes have been modeled and showed strong resemblance to the dipteran transfer RNAs, and all anticodons are identical to those of dipteran. The A＋T-rich region is 562 bp, shorter than that of other known Orthoptera insects. The six conserved domains were identified within the A＋T-rich region by comparing its sequence with those of other grasshoppers. The result of phylogenetic analysis based on the dataset containing 12 concatenated protein sequences confirms the close relation-ship of O. chinensis with L migratoria.     

Toxicological and biochemical characterizations of malathion sensitivity in two field populations of Oxya chinensis （Orthoptera： Acridoidea）

We evaluate comparative toxicity of malathion in the two populations of the grasshopper Oxya chinensis, collected from Daixian and Fanshi of Shanxi province, China. General esterases and acetylcholinesterase （ACHE） from the two populations were characterized and compared. LD50 of the Daixian population （7.58 μg/g body weight） was 2.02-fold higher than that of the Fanshi population （3.75μg/g body weight）. General esterase-specific activities in the Daixian population were 1.91,130 and 1.85-fold higher than those in the Fanshi population, when α-NA, α-NB and β-NA were used as a substrate, respectively. Kinetic studies of general esterase showed that Vmax values of general esterases hydrolyzing α-NA,α-NB and β-NA in the Daixian population were 2.15-, 1.12-, and 1.47-fold, respectively, higher than those in the Fanshi population. The AChE activity of the Fanshi population was 1.54-fold higher than that of the Daixian population. Kinetic analysis of AChE showed that significant differences were presented between the two populations in the Km values; and the Vmax value in the Fanshi population was higher than that in the Daixian population. Inhibition studies of AChE indicated that AChE from the Daixian population was 2.56-, 2.80-, and 2.29-fold less sensitive to inhibition by paraoxon, chlorpyrifos-oxon, and demeton-S-methyl, respectively, than that from the Fanshi population. These biochemical characterizations of general esterases and AChE were consistent with malathion bioassay in the two populations. It is inferred that the reduced sensitivity of altered AChE and increased general esterase activities play an important role in the differences of insusceptibility of Oxya chinensis to malathion between the two populations.     

中华稻蝗两地理种群酯酶特性的比较研究

对采自江苏徐州和山西临猗两个种群中华稻蝗进行了马拉硫磷敏感性的生物测定,同时对两个种群的酯酶特性进行了比较研究。生物测定结果表明,徐州种群的LD₅₀值（13.00 μg/g虫重）是临猗种群（4.64 μg/g虫重）的2.8倍;用对氧磷、马拉氧磷、西维因及毒扁豆碱等四种抑制剂对该两个种群的酯酶的体外抑制研究表明,两个种群所含酯酶大都为B型酯酶;酯酶动力学研究结果表明,徐州种群动力学参数米氏常数（K_m值）和最大反应速度（V_max值）均较临猗种群为高;用α-乙酸萘酯（α-NA）、α-丁酸萘酯（α-NB）和β-乙酸萘酯（β-NA）三种底物测定酯酶活性,在雌性稻蝗中,徐州种群比临猗种群分别高2.02、1.58和1.28倍,雄性中则分别高2.71、1.67和1.33倍;对两个种群酯酶活性频率分布进行比较,徐州种群中酯酶活性高的个体数远大于临猗种群。我们推测徐州种群酯酶的生化特性可能不同于临猗种群,这可能与地理分布、生态环境和食物条件不同有关,杀虫剂选择压力不同可能也起一定的作用。     

青海沙蜥消化道组织结构及嗜银细胞研究

为揭示青海沙蜥消化系统的组织结构,探索其高海拔适应的组织学基础,应用解剖学与石蜡切片、H.E染色和Grimelius银染法对青海沙蜥消化道组织结构和嗜银细胞进行研究。结果显示:青海沙蜥的消化道管壁结构分为4层,从内到外依次是粘膜层、粘膜下层、肌层和浆膜。消化道各段的长度和壁厚均存在显著差异,其中小肠最长,胃幽门部的管壁最厚。粘膜皱襞和绒毛的分布也存在差异,空肠部位的小肠绒毛数量最多,其次是十二指肠和回肠。嗜银细胞形状多样,广泛分布在消化道各段的黏膜上皮基部和黏膜上皮之间。胃体是嗜银细胞分布密度最高的部位,其次是贲门,回肠最低。与栖息在低海拔的有鳞类相比,青海沙蜥为适应高海拔环境,小肠的相对长度变长,胃体部嗜银细胞增多。     

中华稻蝗五地理种群的卵滞育强度及生活史模式变异

为调查中华稻蝗Oxya chinensis的地理适应,自北纬42.3°至20.0°采集了5个地理种群的标本,比较了其子代的卵滞育发生、滞育强度及成虫虫体大小。结果表明: 不同的中华稻蝗地理种群显示了不同的光周期和温度反应模式。铁岭、济宁和泗洪种群的滞育不受光周期和温度的影响,滞育率均为100%。南方的长沙和海口种群仅部分卵进入滞育,海口种群在短日条件下的滞育率高于长日条件,卵期温度亦对其滞育有较大的影响,而长沙种群的卵滞育仅受卵期温度的调控。长沙和海口种群的卵滞育强度最弱,其次是铁岭种群,中间纬度的济宁和泗洪种群的滞育强度最高。在北方的一化性种群中,滞育强度随着纬度的降低而增强。成虫的虫体大小因栖息地纬度的变化而呈现复杂的变异,北方种群的虫体大小与栖息地纬度存在显著的负相关关系,而长沙和海口种群的虫体小于济宁和泗洪种群。结果提示中华稻蝗各种群已形成各自的地理适应机制,纬度梯度在其生活史的形成和发育中起着重要的作用。     

Cellular and humoral immune defenses of Oxya japonica (Orthoptera: Acrididae) to entomopathogenic fungi Metarhizium anisopliae

One Indonesian isolate of the fungus Metarhizium anisopliae, named Majalengka strain, was evaluated not only for its virulence but also for the immune response of rice grasshopper Oxya japonica (Orthoptera: Acrididae) as a target organism. Five aqueous suspensions with different conidia concentrations in logarithmic series were prepared. The fungus showed high virulence as it caused 100% mortality at low conidia concentration (1.5 × 10² conidia/mL). Remarkable changes in the cellular and humoral responses were also observed when adult grasshoppers were infected with the fungus. The number of hemocytes decreased significantly within 12 h after infection. In addition, the total number of granulocytes increased rapidly in the first 12 h then gradually decreased 24 and 48 h after infections, while the number of coagulocytes fluctuated over time. The infection influenced the humoral response by increasing the phenoloxidase activity.