New ways and new hopes for IGR development

Insect Growth Regulators (IGRs) represent advanced, bio-rational insecticides. This Special Issue reflects progress in IGR development that has been enabled by insight into the molecular principles of biosynthetic or hormone signaling pathways. The unifying principle is aiming at processes and molecular targets that are unique to arthropods and ideally to narrower insect taxa representing pests or disease vectors. While some strategies of obtaining the desired compounds for chemical intervention rely on rational, structure-based design or computational power, others exploit technologies allowing automated, high-throughput screening of large chemical libraries. All avenues leading to selective and environmentally safe pest control are valid as we face the imminent threat of the declining world insect population.     

罗氏沼虾细胞色素P450家族CYP302a1基因克隆及其在蜕皮周期中的表达

蜕皮是甲壳动物重要的生理活动,与其蜕皮激素的合成密切相关,细胞色素P450(CYP)302a1是甲壳动物蜕皮激素合成通路中的关键酶之一。本研究克隆了罗氏沼虾CYP302a1基因(Mr-CYP302a1),cDNA全长1859 bp,开放阅读框(ORF)为1629 bp,编码543个氨基酸(aa),分子量大小为61.09 ku,等电点为8.42。氨基酸序列分析显示CYP302a1基因的保守结构域含有5个P450基因家族特征保守区域:heme-binding、helix-K、helixC、helix-I及PERF。系统进化分析结果显示Mr-CYP302a1首先与绿虾CYP302a1聚为一支,然后与凡纳滨对虾及三疣梭子蟹等十足目甲壳动物的CYP302a1聚为一支,与甲壳动物的亲缘关系最近。实时荧光定量PCR(qRT-PCR)检测表明Mr-CYP302a1在罗氏沼虾的多个组织中均有表达,其中在Y器官中的表达量最高,性腺中次之。同时研究发现,MrCYP302a1基因在罗氏沼虾的蜕皮后期(A期和B期)表达量很低,蜕皮间期(C期)表达量开始上升,在蜕皮前期D1亚期达到峰值。对Mr-CYP302a1进行蛋白表达及多克隆抗体制备,蛋白印迹法(Western blot,WB)检测表明Mr-CYP302a1蛋白在罗氏沼虾Y器官中的表达量最高,在蜕皮过程中的蜕皮前期D1亚期达到峰值。综上所述,该基因在罗氏沼虾的蜕皮过程中扮演着十分重要的角色。     

斑节对虾Chitinase-2基因的克隆及其在蜕皮和幼体发育过程中的表达分析

研究以斑节对虾(Penaeus monodon)转录组获得的几丁质酶基因(Pm Chi)片段,运用RACE技术克隆了PmChi基因c DNA全长,命名为PmChi-2。PmChi-2基因c DNA全长2 050 bp,其中5'-非编码区(5'-UTR)144 bp、3'-UTR 319 bp和开放阅读框(ORF)1 587 bp,编码528个氨基酸。生物信息学分析显示,PmChi-2与其他甲壳动物Chi-2的相似性为78%~97%。实时荧光定量PCR结果显示,PmChi-2在蜕皮前期(D期)表皮中表达水平最高,在胃、鳃、腹神经节和眼柄中表达水平依次降低,在其他组织(肝胰腺、肠、肌肉、心脏)中几乎不表达。不同蜕皮阶段,PmChi-2在鳃、胃和表皮3种组织中的表达变化模式基本一致,均在蜕皮期(E期)表达量最低,而最高表达量在鳃中出现在蜕皮后期(A期),在胃和表皮中为D期。幼体不同发育阶段分析揭示PmChi-2 mRNA在幼体发育阶段的无节幼体到糠虾幼体第二期表达量维持在一个低水平,在糠虾幼体第三期PmChi-2 mRNA表达水平显著升高,在仔虾期又显著下降,推测PmChi-2 mRNA可能与斑节对虾幼体发育密切相关。研究结果表明PmChi-2基因可能在斑节对虾蜕皮以及幼体的变态发育中发挥重要作用,为深入研究斑节对虾几丁质酶发育调控提供了重要信息。     

从江香猪生精上皮周期及睾丸发育的形态学分析

试验旨在探究初情期前后生精上皮周期差异及睾丸发育过程中的形态学变化。通过测定15、30、60和90 d睾丸相关指数，结合睾丸组织形态学特征，判断香猪初情期，划分从江香猪生精上皮周期。结果显示，30 d的睾丸指数较15 d极显著升高（P<0.01），睾丸重、长轴及短轴的增长率分别为298.05%、66.42%和65.45%，60和90 d两个阶段睾丸重增长率相对稳定。形态学观察表明，从江香猪30 d时生精小管出现游离精子，完成第一次生精并进入初情期；与15 d相比，30 d生精小管面积和生精上皮厚度极显著增加（P<0.01），增长率分别为136.12%和40.19%，在60和90 d均处于稳定增长状态。睾丸细胞数统计显示，日龄增加不影响支持细胞（setoli cells，SC）数量（P>0.05），而30 d生殖细胞数（germ cells，GC）较15 d极显著增加（P<0.01）。相关分析结果发现，生殖细胞数量增加与生精小管面积增大、生精上皮厚度变化之间呈明显正相关（r=0.994；0.96）。根据生殖细胞组合形式差异，将初情期前后生精上皮分为3和8个阶段。初情期前生殖细胞以第一次减数分裂前期为主，A、B型精原细胞、SC、初级精母细胞（primary spermatocyte，Ps）、前细线期（preleptotene，PI）、细线期（leptotene，L）等生殖细胞在初情期前后生精上皮中均存在，而圆形精子（round spermatids，R）、延伸精子（elongating spermatid，E）、精子细胞（spermatozoa，S）仅存在于初情期后的生精上皮。本研究结果表明，从江香猪30 d初情，睾丸发育以生殖细胞和生精小管面积的迅速增加为主，该结果对从江香猪早熟性状挖掘、种猪选育及开发利用等有重要的指导意义。     

Role of kisspeptin/GPR54 signaling pathways in prompting formation of precocious puberty

AIM: To explore the role of kisspeptin/GPR54 signaling pathway in the pathogenesis of precocious puberty based on female precocious puberty rat model induced by the single dose of danazol. METHODS: Female SD rats aged 3 d were randomly divided into normal group, vehicle group and model group. On the 5th day, the model rats were given a single subcutaneous injection of danazol with ethanol and ethylene glycol mixture. All rats were executed on 15 d, 25 d, 30 d, 35 d and 40 d, and the samples were collected to observe the sexual organ development. The levels of E₂, FSH and LH in peripheral blood were measured by ELISA. The Kiss-1 mRNA expression of Kiss-1, GPR54 and GnRH in hypothalamus was detected by real-time PCR. The kisspeptin expression in rat hypothalamus was observed by immunofluorescence. RESULTS: The time for puberty onset and sexual maturation in the model rats was significantly earlier than that in normal group and vehicle group. On days 25 and 30, the levels of peripheral sex hormones and uterine coefficients in model group were significantly higher than those in normal group and vehicle group. On days 25 and 30, the ovarian morphological development in the model rats was significantly earlier than that in normal group. On day 25, the mRNA expression of hypothalamic Kiss-1 and GnRH, and kisspeptin expression of hypothalamic arcuate nucleus in the model rats significantly increased compared with normal group and vehicle group. On day 30, kisspeptin expression of hypothalamic arcuate nucleus in the model rats decreased compared with normal group and vehicle group. On day 35, the mRNA expression of Kiss-1 and GnRH in the model rats decreased compared with normal group and vehicle group. The mRNA expression of GPR54 had no obvious difference among all groups. CONCLUSION: The Kiss-1 mRNA and kisspeptin expression in the model rats with precocious puberty is significantly increased in the hypothalamus during onset of puberty, suggesting that kisspeptin is an initiating factor for precocious puberty. Kisspeptin/GPR54 signaling pathway may play an important role in the occurrence of precocious puberty.     

西农萨能奶山羊母羔初情期的研究

对30只西农萨能奶山羊母羔的初情期体重、日龄、发情表现,发情周期及17β—雌二醇(17β-E_2)和孕酮(P_4)水平变化进行了研究。研究结果表明:1.初情期体重和日龄分别为10.2(8.0—12.9)kg和54.7(43—71)天,初情期体重和日龄的相关系数为0.798,初情期日龄和出生日期的相关系数则为0.963;2.第一个发情周期平均为8.3(2—37)天,其中短周期(2—8天)羊占62.4％;3.发情表现的特点是阴道和阴门下角有灰白色粘液;4.初情期17β—E_2(pg/ml)/P_4(ng/ml)比值为141.1,17β-E_2和P_4的分泌关系有同步、反步和部分同步部分反步现象。     

哺乳动物性晚熟相关基因的研究进展

人和哺乳动物性的发育和成熟源于下丘脑性腺激素释放激素（gonadotropin releasing hormone,GnRH）脉冲式释放。GnRH释放受到抑制或破坏,就会导致性腺机能减退,人在发育前和发育期就会出现发育迟缓和性发育不良,表现为无精症或闭经;动物则表现为初情期延迟、生殖能力下降等表型。编码促性腺激素及其受体基因的突变可能引起哺乳动物性晚熟。笔者简要介绍了GPR54、GnRH/GnRHR、FSH/FSHR、LH/LHR基因与哺乳动物性晚熟的关系。     

双抗虫基因三倍体毛白杨对舞毒蛾幼虫的抗性研究

用室内群体饲养舞毒蛾幼虫的方法,对15个双抗基因三倍体毛白杨株系进行饲虫试验。结果表明:校正死亡率划分了高抗、中抗、低抗虫植株,舞毒蛾校正死亡率在95以上为高抗株系;40～75为中抗株系,30以下为低抗株系。舞毒蛾的发育历期、蜕皮指数和毒效反映的规律相一致。     

脱皮激素对木薯蚕吐平板丝的影响

本在脱皮激素对木薯蚕吐平板丝的影响方面进行了探讨。脱皮激素在合适的浓度范围内对木薯蚕吐平板丝的生理活动有一定的影响，可减少蔟中爬动，缩短吐丝时间，并可提高吐丝量。这些影响是有利用价值的。因而脱皮激素在木薯蚕吐平板丝的应用上值得进一步探讨。