不同品级拟黑多刺蚁(Polyrhachis vicina)脑部雌激素相关受体基因的表达

采用原位杂交方法对不同品级拟黑多刺蚁(Polyrhachis vicina)脑部雌激素相关受体pvERRmRNA的表达进行了检测.结果表明,pvERRmRNA在不同品级的蕈形体、附叶、视叶、中脑、食道下神经节和球状细胞中都有表达.蚁后蕈形体和球状细胞中pvERRmRNA的阳性表达最为明显;工蚁中脑pvERR mRNA的阳性反应强度明显高于蚁后和雄蚁;pvERR mRNA在雄蚁视叶中有较强的表达.pvERR mRNA在不同品级的拟黑多刺蚁脑部的表达差异可能与它们各自在蚁群中的功能和行为有关.     

乙酰胆碱酯酶在拟黑多刺蚁脑部的分布

用组织化学方法研究乙酰胆碱脂酶(Acetylcholine esterase,AChE)在拟黑多刺蚁(Polyrhachisvicina Roger)脑部分布.结果显示,AChE在拟黑多刺蚁脑部分布广泛,主要分布于后脑、咽下神经节、中脑,其次是前脑的中央复合体、视叶、蕈形体冠,前脑的蕈形体柄和叶分布最少;品级间比较显示,工蚁视叶和嗅叶分布较雌蚁和雄蚁的少,雌蚁中央复合体的分布多于其他品级.由该结果推测AChE可能参与拟黑多刺蚁视觉和嗅觉信息传递;根据AChE组织化学在临床上着色判别标准确定蕈形体冠是蕈形体的感觉信息输入区,而草形体的柄和叶是蕈形体的运动信息输出区.     

一氧化氮合酶在拟黑多刺蚁脑中的分布

一氧化氮作为一种重要的信使分子,参与调节昆虫嗅觉、视觉、机械感受、发育和机体防御.本文采用NADPH-黄递酶组织化学技术,观察了一氧化氮舍酶(NOS)在拟黑多刺蚁脑内的分布,结果表明,NOS在雄蚁和雌蚁的蕈形体、视叶、中央复合体及中脑后脑区均有分布,在工蚁中主要分布在视叶、中脑及后脑区,推测可能对蚂蚁的视觉、嗅觉、记忆及行为产生影响.     

拟黑多刺蚁(Polyrhachis vicina)性腺雌激素相关受体基因的表达

采用原位杂交方法对拟黑多刺蚁(Polyrhachis vicina)蚁后卵巢中和雄蚁精巢中pvERR mRNA的表达进行了检测.结果表明,pvERR mRNA在卵子发生过程中滤泡细胞的胞质和生长期、卵黄发生期的卵母细胞有表达.在变形期的精细胞胞质和成熟精子头部有很强的表达.     

拟黑多刺蚁脑部结构的观察

运用Mallory染色方法,对拟黑多刺蚁(Polyrhachis vicina)脑部形态结构进行了显微观察.结果表明,拟黑多刺蚁脑由前、中、后脑三部分组成:前脑中具有较大的蕈形体,说明它可能具有比较发达的学习和记忆的能力;中脑中具有较大的嗅觉神经纤维球,表明拟黑多刺蚁具有发育良好的嗅觉系统.和其它膜翅目昆虫相比,拟黑多刺蚁前脑中的视叶较小.因此推测,视叶在其觅食、学习、记忆及其他行为中的作用不及中脑嗅叶.后脑在整个脑部占据的面积很小,它在蚂蚁脑部的作用目前还不清楚.     

拟黑多刺蚁体外抑瘤作用研究

目的　通过拟黑多刺蚁对体外S180 ,H2 2 ,YAC 1肿瘤细胞DNA合成影响及细胞毒性作用 ,研究该蚁抑瘤效应及机制 .方法　采用3H TdR掺入法和12 5I UdR释放法 .结果　拟黑多刺蚁对S180 ,H2 2 ,YAC 1肿瘤细胞DNA合成均有明显抑制作用 (P <0 .0 5 ) ,其抑制作用随剂量增加而增强 .以对S180 肿瘤细胞抑制作用最显著 .同时证实 ,该蚁对YAC 1,S180 肿瘤细胞均有毒性作用 .结论　拟黑多刺蚁对肿瘤细胞具有直接杀伤和抑制生长的作用 .     

拟黑多刺蚁对H22荷瘤鼠抑制肿瘤生长的初探

通过建立动物肿瘤模型,采用对比实验法,探讨拟黑多刺蚁对H22荷瘤鼠的抑瘤作用.结果表明:拟黑多刺蚁具有抑制H22肿瘤细胞的作用,并增强荷瘤鼠的免疫功能.     

近暗散白蚁触角感受器扫描电镜观察

利用扫描电镜观察的方法,对近暗散白蚁触角形状和触角感受器的种类、形态、分布进行分析.结果显示,触角节数与长度:补充蚁后翅芽型补充生殖蚁兵蚁工蚁幼蚁;不同品级中发现12种感受器:短毛型感器、长刺形感器、短刺形感器、B?hm氏鬃毛、刺突状钟形感器、球状钟形感器、球状钟形感器与刺突状钟形感器间的中间型、半球形感器、单峰水滴形感器、双峰水滴形感器、单峰与双峰水滴形感器的中间型,其中,半球形感器、单峰水滴形感器、双峰水滴形感器、单峰与双峰水滴形感器的中间型为新发现感器;半球形感器仅存在于工蚁、兵蚁和翅芽型补充生殖蚁,水滴形感器仅存在于幼蚁.不同品级的触角节数和长度以及触角感器有明显差异,可以作为近暗散白蚁各品级的区分依据.     

白蚁与蚂蚁

白蚁和蚂蚁都是社会性昆虫，它们过着类似于人的生活，离开了集体，个体是无法生存的。它们的社会组织结构非常相似，分工非常明确。蚁王和雄蚁经过短暂的婚飞，落地脱翅，建巢。它们的巢穴可不是一个简单的洞穴，里面有很多住房，一层一层排列得井然有序，有点像我们人类居住的楼房。房间的分工也比较明确，有蚁王房间、工蚁房间、雄蚁房间、幼蚁房间、储藏室、垃圾室等。工蚁的工作非常辛苦，却从无怨言，在它们的一生中，默默无闻的负担着照料蚁王、幼蚁，寻找食物，打扫卫生等繁重的工作。兵蚁的主要任务是抵御外来侵略。白蚁和蚂蚁都会…     

类固醇激素受体在扬子鳄睾丸表达的免疫细胞化学研究

用免疫细胞化学方法对雄激素受体(AR)、雌激素受体(ER)和孕激素受体(PR) 在扬子鳄睾丸中进行定位研究.结果表明,三种类固醇激素受体在扬子鳄睾丸中都有分布:雄激素受体在睾丸的肌样细胞和支持细胞中呈阳性反应,雌激素受体在睾丸的间质细胞中呈 阳性反应,孕激素受体在睾丸的生精细胞中呈阳性反应.推测结果表明,雄激素受体在扬子 鳄精子发生过程有重要作用;雌激素受体可能在扬子鳄睾丸发育过程中影响睾丸间质细胞雄性激素的分泌,进而调节生精过程和精子发育成熟,孕激素受体可能通过转换成AR和ER来对精子发生进行调节.这些结果为证明性类固醇激素参与调节扬子鳄性腺生殖内分泌调控提供了重要的形态学新证据.     

Androgen receptor gene mutations in X-linked spinal and bulbar muscular atrophy

X-linked spinal and bulbar muscular atrophy (Kennedy's disease) is an adult-onset form of motorneuron disease which may be associated with signs of androgen insensitivity. We have now investigated whether the androgen receptor gene on the proximal long arm of the X chromosome is a candidate gene for this disease. In patient samples we found androgen receptor gene mutations with increased size of a polymorphic tandem CAG repeat in the coding region. These amplified repeats were absolutely associated with the disease, being present in 35 unrelated patients and none of 75 controls. They segregated with the disease in 15 families, with no recombination in 61 meioses (the maximum log likelihood ratio (lod score) is 13.2 at a recombination rate of 0). The association is unlikely to be due to linkage disequilibrium, because 11 different disease alleles were observed. We conclude that enlargement of the CAG repeat in the androgen receptor gene is probably the cause of this disorder.     

中华蟾蜍卵巢性类固醇激素受体的免疫细胞化学研究

为探讨两栖动物卵巢内不同性类固醇激素受体在卵子发育中的调控作用.采用免疫细胞化学方法,对中华蟾蜍不同发育时期卵泡中的雌激素受体、孕激素受体和雄激素受体进行了定位检测.结果发现,三种受体在不同发育时期的卵泡中均有阳性反应:雌激素受体在卵黄合成期和卵黄合成后期的滤泡细胞表达为强阳性,在卵母细胞胞质和核膜中呈弱阳性反应;孕激素受体在卵黄合成早期的滤泡细胞和卵母细胞质、核膜中表达均较弱,在卵黄合成后期表达均增强;雄激素受体在卵黄合成早期的滤泡细胞和卵母细胞胞质、核膜中表达也均较弱,在卵黄合成后期卵母细胞胞质和核膜中表达略有增强.     

Common non-hormone binding component in non-transformed chick oviduct receptors of four steroid hormones

Steroid hormones produce a response in target cells by binding to hormone-specific soluble receptors, which undergo a transformational change, leading to their interaction with chromatin and to modified gene expression. In a previous paper, we described a monoclonal antibody, BF4, that specifically recognizes and binds the non-transformed '8S' form of chicken oviduct progesterone receptor (8S-PR). We now show that BF4 does not form an immune complex with the 4S transformed form of 3H-progestin-labelled progesterone receptor, but does interact with the 8S non-transformed forms of the oestrogen, androgen and glucocorticosteroid receptors. Our results suggest that the antigenic determinant recognized by BF4 is present on a non-hormone binding unit, which we identify as a polypeptide of molecular weight (MW) 90,000 in the case of the progesterone receptor, and that this unit is common to other 8S non-transformed chicken steroid receptors.     

Immunorecognition of estrogen and androgen receptors in the brain and thoracic ganglion mass of mud crab, Scylla paramamosain

The brain and the thoracic ganglion of a crustacean can synthesize and secrete gonad-stimulating hormone (GSH) which stimulates the maturation of gonad. In the previous experiments, sex steroid hormones (estradiol, testosterone, progesterone, etc.) have been detected from the crustacean. However, the feedback regulation of sex steroid hormones on the brain and the thoracic ganglion of the crustacean has not been reported so far. In the present experiment, monoclonal antibodies were applied to investigate the immunorecognition of estrogen receptor (ER) and androgen receptor (AR) in the brain and the thoracic ganglion mass of Scylla paramamosain. The results showed that the distribution of the immunopositive substances of ER and AR was extremely similar. They distributed in the protocerebrum, deutocerebrum and tritocerebrum of the brain, and mainly in protocerebrum. In the thoracic ganglion mass, immunopositive substances distributed in the subesophageal ganglion, thoracic ganglion and abdominal ganglion, and mostly in subesophageal ganglion. Immunopositive substances of ER and AR mostly existed in the cytoplasm of neurons. The present study will provide morphological evidence for the origin and the evolution of ER and AR. In addition, the immunoreactivities of ER and AR suggested that the estrogen and androgen may be involved in the feedback regulation of crustacean neuroendocrine.     

Immunorecognition of estrogen and androgen receptors in the brain and thoracic ganglion mass of mud crab, Scylla paramamosain

The brain and the thoracic ganglion of crustacean can synthesize and secrete gonad-stimulating hormone (GSH) which stimulates the maturation of gonad. In the previous experiments, sex steroid hormones (estradiol, testosterone, progesterone, etc.) have been detected from crustacean. However, the feedback regulation of sex steroid hormones on the brain and thoracic ganglion of crustacean has not been reported so far. In the present experiment, monoclonal antibodies were applied to investigate the immunorecognition of estrogen receptor (ER) and androgen receptor (AR) in the brain and thoracic ganglion mass of Scylla paramamosain. The results showed that the distribution of the immunopositive substances of ER and AR was extremely similar. They distributed in the protocerebrum, deutocerebrum and tritocerebrum of the brain, and mainly in protocerebrum. In the thoracic ganglion mass, immunopositive substances distributed in the subesophageal ganglion, thoracic ganglion and abdomen ganglion, and mostly in subesophageal ganglion. Immunopositive substances of ER and AR mostly existed in the cytoplasm of neurons. The present study will provide morphological evidence for the origin and evolution of ER and AR. In addition, the immunoreactivities of ER and AR suggested that the estrogen and androgen may be involved in the feedback regulation of crustacean neuroendocrine.     

Immunorecognition of estrogen and androgen receptors in the brain and thoracic ganglion mass of mud crab, Scylla paramamosain

The brain and the thoracic ganglion of crustacean can synthesize and secrete gonad-stimulating hormone (GSH) which stimulates the maturation of gonad. In the previous experiments, sex steroid hormones (estradiol, testosterone, progesterone, etc.) have been detected from crustacean. However, the feedback regulation of sex steroid hormones on the brain and thoracic ganglion of crustacean has not been reported so far. In the present experiment, monoclonal antibodies were applied to investigate the immunorecognition of estrogen receptor (ER) and androgen receptor (AR) in the brain and thoracic ganglion mass of Scylla paramamosain. The results showed that the distribution of the immunopositive substances of ER and AR was extremely similar. They distributed in the protocerebrum, deutocerebrum and tritocerebrum of the brain, and mainly in protocerebrum. In the thoracic ganglion mass, immunopositive substances distributed in the subesophageal ganglion, thoracic ganglion and abdomen ganglion, and mostly in subesophageal ganglion. Immunopositive substances of ER and AR mostly existed in the cytoplasm of neurons. The present study will provide morphological evidence for the origin and evolution of ER and AR. In addition, the immunoreactivities of ER and AR suggested that the estrogen and androgen may be involved in the feedback regulation of crustacean neuroendocrine.     

Human CMTM2/CKLFSF2 enhances the ligand-induced transactivation of the androgen receptor

CKLF (chemokine-like factor)-like MARVEL (MAL and related proteins for vesicle trafficking and membrane link domain) transmembrane domain containing (CMTM) is a novel gene family. One member of this family, CMTM2, also named chemokine-like factor superfamily 2 (CKLFSF2), is expressed highly in the testis and moderately in the prostate, marrow and peripheral blood cells. However, the function of human CMTM2 remains unknown. Here, we found that CMTM2 was upregulated in 5α-dihydrotestosterone (DHT)-treated LNCaP cells. We investigated the relationship between CMTM2 and the androgen receptor. Our results showed that CMTM2 enhanced DHT-mediated androgen receptor (AR) transactivation and the expression of prostate specific antigen (PSA). We also observed that CMTM2 enhanced the AR protein level, which was reversed by silencing endogenous CMTM2 expression, which suggested that CMTM2 might play an important role in maintaining the AR protein level. We also found that CMTM2 suppressed Akt activation. A previous study showed that Akt could phosphorylate AR at Ser210 and Ser790 and lead to AR ubiquitylation and degradation as well as suppression of AR activity. Taken together, suppressing Akt activation and increasing the AR protein level might be one of the mechanisms for the CMTM2-mediated enhancement of AR transactivation. Supported by National High Technology Research and Development Program of China (Grant Nos. 2006AA02A305 and 2002BA711A01) and National Natural Science Foundation of China (Grant Nos. 30271203 and 30671907)     

昆虫全基因组DNA的保存及提取

以蚂蚁、天牛、蝗虫等昆虫为实验材料,建立了75%酒精常温浸泡密封保存和福尔马林常温浸泡密封保存昆虫样品2~4周后进行全基因组DNA提取的方法。基因组DNA经紫外分光光度法(260 nm、280 nm)、琼脂糖凝胶电泳、PCR检测,结果表明:从两种方法保存的样品中均能提取到纯净完整的基因组DNA,适用于分子生物学实验。