Gene cloning and characterization of novel antinociceptive peptide from the brain of the frog, Odorrana grahami

Amphibian opiate peptides including dermorphins and deltorpins have been recently found only in the skin of South American frogs belonging to the subfamily Phyllomedusinae (Phyllomedusa, Agalychnis and Pachymedusa species). No opiate peptides have ever been identified from other amphibians or organs except skin. Here we report the purification and characterization of a novel antinociceptive peptide named odorranaopin from the homogenates of the frog brains, Odorrana grahami, which is also the first antinociceptive peptide found in Ranidae amphibian. Odorranaopin comprises 17 amino acid residues with the sequence of DYTIRTRLHQESSRKVL (Mr 2102 Da). The cDNA encoding odorranaopin was cloned from the frog brain cDNA library, and it was confirmed to be a specific gene. The odorranaopin precursor deduced is composed of 61 amino acid residues including the predicted signal peptide, acidic spacer peptide and mature odorranaopin positioned at the C-terminus. Odorranaopin could inhibit nociceptive responses induced by formalin and acetic acid. It also inhibited the contractile responses of ileum smooth muscle induced by bradykinin, implying that the antinociceptive activity of odorranaopin possibly results from its blockade on bradykinin or bradykinin receptor functions. Odorranaopin is the first antinociceptive peptide found in Ranidae amphibian.     

竹叶臭蛙的分类学研究Ⅱ.两新种记述(两栖纲:蛙科)

据记载竹叶臭蛙Odorrana versabilis分布于广西,贵州,湖南,福建,浙江,安徽,广东,海南,作者对比研究了7个省的13个居群标本的形态特征,可分为3个形态型,这3个形态型标本的特征彼此差异明显,被确定为3个不同的种,其中包括2个新种,即小竹叶臭蛙O.exiliversabilis sp.nov。和鸭嘴臭蛙O.nasuta sp.nov.,为印证形态分类的结果,又采用形态数值性状标准差别分析和DNA指纹实验分析,其结果与形态特征分类研究结果一致。本文对2个新种的形态和生物学资料进行了记述。     

基于12S rRNA基因序列探讨凹耳蛙(无尾目,蛙科)的分类地位

为探讨凹耳蛙Rana tormotus的分类地位,测定了凹耳蛙线粒体12S rRNA基因序列,并从GenBank中下载了蛙科33种蛙的同源序列进行分析,用BI法、NJ法和MP法构建分子系统树.结果表明:3种树拓扑结构基本一致,都显示凹耳蛙与长吻胡湍蛙Huia nasica亲缘关系最近,凹耳蛙与长吻胡湍蛙相聚后再与臭蛙Odorrana组成单系群,而与湍蛙Amolops亲缘关系较远.据此,支持将该蛙从湍蛙属划出,并建议归入臭蛙属.     

Characterization of an insulinotropic peptide from skin secretions of Odorrana andersonii

Insulinotropic peptide agents are regarded as potential candidates for anti‐diabetic treatment. In the present study, a novel insulinotropic peptide, termed OA‐A1, was purified from frog skin secretions of Odorrana andersonii. Mature OA‐A1 was determined to be a 1965.049 Da peptide with an amino acid sequence of LVGKLLKGAVGDVCGLLPIC, in which an intramolecular disulfide bridge was formed by two cysteine residues. At the cellular level, OA‐A1 exhibited potent proliferation promoting effects on mouse‐derived pancreatic β‐TC‐6 cells and significantly stimulated insulin release in β‐TC‐6 cells at a minimum concentration of 1 nM. In the animal model, OA‐A1 also showed a dose‐dependent insulin‐releasing role in mice. At concentrations ranging from 1 nmol/kg to 1 μmol/kg, OA‐A1 had a significant acute hypoglycemic effect on streptozotocin (STZ)‐induced diabetic mice. The pancreatic islet areas of diabetic mice increased dose‐dependently after 21 days of OA‐A1 treatment (1–100 nmol/kg) compared with those of the saline control group. Moreover, OA‐A1 significantly improved the oral glucose tolerance of STZ‐induced diabetic mice. Taken together, these results suggest that OA‐A1 provides an excellent template for the development of novel anti‐diabetic therapeutic agents. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.     

湖南省花臭蛙复合体分类及分布格局

运用形态特征系统聚类和分子系统学分析,研究先前报道的湖南省花臭蛙(Odorrana schmackeri)各地理种群的分类组成及其分布格局。结果显示,原认定的花臭蛙湖南省各地理种群已分化为花臭蛙和黄岗臭蛙(O.huanggangensis)两个物种。分布于湘西南及南部雪峰山和南岭的花臭蛙种群应修订为黄岗臭蛙,为湖南省臭蛙属物种新发现,湘西北及湘东南罗霄山脉的种群为花臭蛙,黄岗臭蛙与花臭蛙在张家界市武陵源景区同域分布。湘西北武陵山与雪峰山之间以及湘中丘陵地带花臭蛙种群的分类归属,黄岗臭蛙和花臭蛙在湖南省分布格局的形成以及同域分布的机制值得探讨。     

Sexual Dimorphism in Mass of the Hindlimb Muscles of the Piebald Odorous Frog （Odorrana schmackeri）

Male-biased sexual dimorphism in hind limb muscles is widespread in anuran species where scramble competition is common among males. Such sexual difference is thought to result from sexual selection. In this view, we tested the differences in muscle mass between the sexes and between amplectant and non-amplectant males by quantifying the mass of four hindlimb muscles （triceps femoris, sartorius, gracilis and plantaris longus） of females and males of Odorrana schmackeri. The results showed that females significantly exceeded males for muscle triceps femoris, gracilis, plantaris longus and total mass when controlled for body size. There are no significant differences between amplectant and non-amplectant males. It is probable that the maintenance of the amplectant position in O. schmackeri may depend on the strength of hindlimb muscles in females to support the pair.     

竹叶臭蛙的分类学研究Ⅰ.各地理居群的分类探讨(两栖纲:蛙科)

采用形态特征比较 ,对 7省 13个居群的竹叶臭蛙Odorranaversabilis的标本进行了研究 ,结果表明 :它们的形态特征差异明显 ,可以分为 3个形态型 ;同时经数值性状标准判别分析和DNA指纹分析印证其结果与形态学研究结果相吻合。据此将 3个形态型标本确定为 3个种 ,其中包括2新种。本文重新界定了 3个种的分布范围。     

不同物种雄性鸣声刺激下雌性凹耳蛙脑组织中ZENK的表达

通过免疫组化SABC法,研究在背景噪音刺激、雄性凹耳蛙(Odorrana tormota)声音刺激以及雄性大绿臭蛙(O. graminea)声音刺激下ZENK蛋白在雌性凹耳蛙脑部的表达。结果显示,上述三种声音刺激下雌性凹耳蛙的端脑、间脑和小脑区域均未发现ZENK蛋白的阳性细胞。在雄性凹耳蛙声音刺激下,雌性凹耳蛙中脑的视顶盖、脚间核、中脑深部核、被盖、半环隆枕以及延脑的孤束核、中缝核、网状核、网状结构、上橄榄核部位均有阳性细胞分布,在大绿臭蛙声音刺激下,这些部位的阳性细胞数量明显减少。本实验中ZENK蛋白阳性细胞表达部位的一部分是位于与两栖类处理听觉信息有关的中脑半环隆枕、被盖以及延脑的上橄榄核,由此可推断,在雌性凹耳蛙ZENK蛋白与听觉信息处理有着密不可分的联系。     

重庆市发现光雾臭蛙和南江臭蛙

2012年6月23日,在重庆市城口县大巴山国家级自然保护区综合科考过程中采集到臭蛙属(Odorrana)标本5号,经鉴定为光雾臭蛙(O.kuangwuensis)3号、南江臭蛙(O.nanjiangensis)2号,均为重庆市两栖类新纪录.     

中国四川省蛙科一新种——南江臭蛙(两栖纲:无尾目)

对分布于四川、重庆市、贵州、湖北、湖南、安徽和福建的花臭蛙[Odorrana(Odorrana)schmackeri(Boettger,1892)]标本进行了形态比较,结果发现四川省南江和万源县的花臭蛙标本与其他产地的花臭蛙标本在形态特征上存在明显区别,主要是:1)成体雄雌体长之比值较小,约为1∶1.34;2)犁骨齿列短,两内侧间距宽,距内鼻孔远;3)背面深色斑点周围无浅色边缘;4)股后部深色斑大而稀疏;5)趾间蹼较弱,第四趾蹼达远端关节下瘤;6)腹内成熟卵的动植物极均为乳白色。故将四川省南江和万源的标本订为新种--南江臭蛙[Odorrana(Odorrana)nanjiangensis,sp.nov.]。