峨眉髭蟾精子形态结构及分类学意义

应用透射电镜、扫描电镜和光学显微镜对峨眉髭蟾(Vibrissaphora boringii)精子的形态和超微结构研究的结果表明：峨眉髭蟾的精子具角蟾科物种精子基本的形态和结构特征，即精子头部呈螺旋状，尾部呈弯曲状；精子具锥形的顶体、纤维束构成的穿孔器、平行排列的中心粒和双轴丝；线粒体位于尾部；精子核窝不明显、无轴纤维和波动膜等特征。此外，对已有报道的角蟾科和无尾类物种精子的特征进行分析比较表明：(1)角蟾科精子细胞核呈螺旋状，中心粒平行排列，尾部具双轴丝等结构不同于无尾类其他科精子的结构，具有明显的科间差别；(2)角蟾科精子各部的量度，尾部线粒体的分布和数量，以及轴丝的排列等特征在属间和种间表现出明显的差异；(3)峨眉髭蟾和东南亚拟髭蟾指名亚种精子的形态和超微结构存在明显的差异。     

中国角蟾亚科精子形态比较及分类学讨论(两栖纲：锄足蟾科)

应用光镜和电镜对角蟾亚科短腿蟾属,拟角蟾属和角蟾属3属18种精子的形态结构进行观察和比较。并通过计算机对其各部位长度进行了测量和计算,结果表明角蟾亚科精子的共同特征是：精子主要由头和尾两部分组成,头部呈螺旋状,尾部为波状或弯曲状,但精子的量度和头部形状在属间和种间存在明显差别,角蟾亚科的精子可分为3类,该亚科精子形态的差异与成体形态学研究结果基本相吻合,精子形态的差异可为该亚科属,种的划分,亲缘关系研究提供新依据。     

挂榜山小鲵精子形态结构研究

应用扫描电镜和透射电镜对挂榜山小鲵(Hynobius guabangshanensis)精子的超微结构进行观察和研究.结果表明:1)挂榜山小鲵的精子形态具有小鲵科精子的共同特征:头部细直、有锥形顶体;颈部短而不明显,尾部细长、有波动膜;2)该种小鲵的精子超微结构具小鲵科物种精子的共同特征,即精子无顶体钩,顶体呈三叶草状,尾部无线粒体,轴纤维粗大呈圆柱状等;3)除精子全长(194.1±7.15μm)在小鲵科物种中属于中等外,挂榜山小鲵和本科其他物种的精子在形态量度学方面差异明显:其头部占全长的比例(22.98%)比其他已知小鲵科的物种都低,尾部占全长的比例(65.79%)比其他小鲵科物种都高.     

龙洞山溪鲵精子的超微结构

应用透射电镜和扫描电镜对龙洞山溪鲵(Batrachuperuslongdongensis)精子的超微结构进行观察和研究,探讨山溪鲵属以及小鲵科物种精子的结构特征,并探讨有尾两栖类精子结构特征的演化及其与生殖进化的关系。结果表明:1)龙洞山溪鲵的精子具小鲵科物种精子的共同特征,即精子无顶体钩,顶体呈三叶草状,尾部无线粒体,轴纤维粗大呈圆柱状等特征;2)龙洞山溪鲵精子核脊的形态结构与小鲵科其它属物种以及有尾类其它科物种精子的核脊存在明显的差别,由此可见核脊形态结构具有属或种的特异性。此外,与已有报道的有尾两栖类物种精子的结构特征进行分析比较表明:1)有尾类精子特征的分化与其受精方式的变化是一致的;2)精子结构特征支持隐鳃鲵超科为单系的推测     

中国角蟾科动物系统发育研究中的几个问题

根据外部形态、核型以技精子形态等特征,角蟾科分布于中国的角蟾属（Megophrys）较之拟髭蟾属（Leptobrachium）是更为原始的类群,角蟾科动物中拟角蟾属（Ophryophryne）与角蟾属亲缘关系较近,短腿蟾属（Brachytarsophrys）是较为原始的类群,且应为一无效属,无耳蟾属（Atympanophrys）应为一有效属。角蟾属可以分为两个种组,其是峨嵋角蟾种组比较进化,白颌大角蟾种组比较原始。     

黄姑鱼（ Nibea albiflora） 与大黄鱼（ Pseudosciaena crocea） 精子超微结构的观察与比较

应用扫描电镜（SEM）与透射电镜（TEM）观察了黄姑鱼和大黄鱼精子的超微结构。结果显示,黄姑鱼和大黄鱼精子无论在形态、大小还是超微结构上都十分相似。黄姑鱼和大黄鱼精子均由头部、中段和尾部（鞭毛）3部分组成。精子头部形状近似椭圆形,无顶体,细胞核呈肾形。中心粒复合体位于细胞核背侧,近、远端中心粒相互垂直,远端中心粒分化成基体并形成轴丝。中段的袖套呈筒状,4～5个圆形的线粒体围绕轴丝呈环形排列。精子尾部为单鞭毛,轴丝为典型“9＋2”结构,鞭毛表面质膜形成不规则侧鳍。     

重庆黔江发现尾突角蟾

2022年7月在重庆市黔江区武陵山市级自然保护区采集到1号两栖动物标本,经形态特征比较,与尾突角蟾（Boulenophrys caudoprocta）相似;基于线粒体16Sr RNA和COI基因联合构建布角蟾属（Boulenophrys）部分物种系统发育树,此次采集的角蟾标本与尾突角蟾聚为一支,具有较高的支持率（1/100,贝叶斯法/最大似然超快速引导支持分析值）;基于p-distance估算本次采集的角蟾标本与尾突角蟾湖南模式产地标本间的遗传距离为0%,远小于角蟾属物种间的遗传距离（8.7%～15.9%）。综合形态特征和分子系统发育比较,确定此次采集到的角蟾标本为无尾目（Anura）角蟾科（Megophryidae）布角蟾属的尾突角蟾,系重庆市两栖动物分布新记录种。重庆市尾突角蟾的发现,进一步扩展了该地区的物种名录,拓宽了对尾突角蟾分布范围的认知,为该属区系分类学和系统学提供了新信息,同时为当地生态系统和物种多样性的研究提供了基础信息,对制定更加全面和针对性的保护措施有着积极的促进作用。     

四种蝉科昆虫的精子形态(半翅目:蝉科)

【目的】为了明晰蝉科昆虫的精子形态及其在分类和系统发育分析方面的意义,本研究对蝉亚科的蒙古寒蝉Meimuna mongolica、黑蚱蝉Cryptotympana atrata、蛉蛄Pycna repanda及姬蝉亚科的蟋蝉Tettigetta sp.的精子进行了比较研究。【方法】分别通过光学显微镜和透射电子显微镜,观察这4种蝉科昆虫的精子形态特征。【结果】蒙古寒蝉、黑蚱蝉、蛉蛄和蟋蝉这4种蝉科昆虫精子形态基本相似,但精子长度在种内和种间都有明显不同,均表现出多态性。根据精子长度,蛉蛄精子可被分为长精子、中长精子和短精子3种类型;蒙古寒蝉、黑蚱蝉和蟋蝉的精子被分为长精子和短精子2种类型。4种蝉的精子结构也基本相似,头部包含顶体和细胞核,颈区由中心粒和中心粒侧体组成,尾部一般由一根轴丝和一对线粒体衍生物组成,轴丝微管为9+9+2模式。但蝉亚科3个物种的部分精子具有多个线粒体衍生物;首次在蛉蛄精子尾部发现一个电子致密的三角形区域,该结构在蝉科其他昆虫精子中未曾发现。蝉科不同类群的精子中心粒侧体存在显著差异,姬蝉亚科的蟋蝉精子中心粒侧体为片层状结构,蝉亚科昆虫则为鞘状结构。【结论】与蝉次目的角蝉总科和沫蝉总科昆虫精子相比,仅蝉科昆虫的精子表现出多态性,是该科的特有衍征。精子尾部可具多个线粒体衍生物的现象在蝉亚科物种中是否普遍存在有待进一步研究。蝉科不同类群在精子形态方面的差异,为蝉科昆虫分类及蝉次目系统发育分析提供了重要信息。     

毛蚶与青蚶精子超微结构及其所反映的蚶科进化关系

应用透射电镜技术,比较研究了毛蚶与青蚶精子的超微结构。毛蚶精子顶体为圆锥形,约为核长的1／2;精核无核前窝,具核后窝;中段横切面常见5个(偶见4个)线粒体环绕于中心粒周围;精子末段由轴丝及包绕轴丝的质膜组成,轴丝为典型的“9 2”结构。青蚶精子顶体轴向纵切面呈伞状,覆盖于细胞核前端,约为核长的1／3;精核具核前窝和核后窝;中段横切面常见有5个(偶见6个)线粒体环绕于中心粒周围;末段结构同毛蚶。顶体的形态、核前窝和核后窝的有无、中段线粒体的数量等是探索蚶科动物种间进化关系的线索。     

赤眼蜂属(Trichogramma)精子的超微结构研究

利用透射电子显敢镜研究了赤眼蜂属的松毛虫赤眼蜂和玉米螟赤眼蜂精子的超微结构。精子呈线形,由头部、颈部和尾部组成。头部前端的顶体较小,为梭形,顶端稍有弯曲,它和核呈楔状拼接排列。颈部的中心粒以45°方向斜插入核内,在中心粒的外周不被中心粒侧体所包围。尾部的轴丝为9+9+2型。在副微管之间有九条粗纤维存在。一对线粒体衍生物内的晶体物质不明显。两种赤眼蜂的精子结构差异很小,松毛虫赤眼蜂精子尾部轴丝结构部分的九个链头之间没有横向联系,而在玉米螟赤眼蜂上能清楚见到。     

中国蟾蜍属精子形态比较

采用扫描电镜和光学显微镜观察了采自我国的蟾蜍属(Bufo)7种(亚种)的精子形态,对精子各部位量度进行了测量和计算。结果表明,该属7种(亚种)精子的形态基本相同,精子由头部、中片和尾部组成,头部细长微弯且前端渐尖,中片有球状突起,尾部长,由轴纤维、轴丝和波动膜构成。与已有报道的两栖动物的精子形态相比较,蟾蜍属精子与无尾类其他科精子形态差别较大,而与有尾类精子形态相似。本文认为两栖动物精子形态和量度在科间存在明显差异;两栖动物精子形态的差异可能与其繁殖模式有关。     

峨眉髭蟾在重庆的分布——蝌蚪在无尾类分类和多样性调查中的应用一例

2008年5月在重庆市酉阳县采集到7号体尾背部具有"Y"形斑纹的蝌蚪,对其形态特征和口部结构予以描述,经与峨眉髭蟾Vibrissaphora boringii和崇安髭蟾瑶山亚种V. liui yaoshanensis蝌蚪比较,确定其为峨眉髭蟾,为重庆市新纪录.现作为应用蝌蚪进行无尾两栖动物分类鉴定和多样性调查实例予以报道.     

The spermatogenesis and the sperm structure of Terebrantia (Thysanoptera, Insecta)

Spermatogenesis and the sperm structure of the terebrantian Aeolothrips intermedius Bagnall are described. Spermatogenesis consists of two mitotic divisions; the second is characterized by the loss of half of the spermatids, which have pyknotic nuclei. Early spermatids have two centrioles, but when spermiogenesis starts, a third centriole is produced. The three basal bodies give rise to three flagella; later these fuse into a single flagellum which contains three 9 + 0 axonemes. The basal bodies are surrounded by a large amount of centriole adjunct material. During spermiogenesis this material contributes to the shifting of the three axonemes towards the anterior sperm region parallel to the elongating nucleus, and it is transformed into a dense cylinder. In the mature spermatids the three axonemes amalgamate to create a bundle of 27 doublet microtubules. Near the end of spermiogenesis the dense cylinder of the centriole adjunct lies parallel to the nucleus and the axonemes. It ends where the mitochondrion appears at half-sperm length. We confirm that Terebrantia testes have a single sperm cyst; their sperm are characterized by a cylindrical nucleus, three axonemes fused into one, a small mitochondrion and a short cylindrical centriole adjunct which corresponds to the dense body described in a previous work. The acrosome is lacking. At the midpoint of the anterior half of the sperm the outline of the cross-section is bilobed, with the nucleus contained in a pocket evagination of the plasma membrane. These characters are discussed in light of a comparison between Tubulifera and Terebrantia.     

Spermatogenesis and spermiogenesis in Didymocystis wedli Ariola, 1902 (Didymozoidae, Digenea).

The ultrastructure of the male reproductive system of Didymocystis wedli was studied for the first time, demonstrating spermiogenesis and spermatogenesis at different cell stages. The spermatozoa morphology was compared with that of other Digenea species. It was observed that the different cells of the spermatogenesis process follow the classic pattern reported for the majority of the parasitic platyhelminthes. During spermiogenesis, rootlet fibers, electrondense bodies and median cytoplasmic process were not observed. The mature spermatozoa of D. wedli were filiform, presenting nucleus, mitochondrion and two 9+1 axonemes, with a biflagellate distal extremity.     

Ultrastructure of the spermatozoa of the flatworms Phaenocelis peleca and Boninia divae (Platyhelminthes, Polycladida)

The ultrastructure of spermatozoa of the acotylean Phaenocelis peleca and the cotylean Boninia divae is described. All spermatozoa are filiform and biflagellate with a 9+"1" microtubular pattern in the axoneme. Sperm characters in P. peleca follow the morphologies described for other acotyleans, with axonemes exiting the sperm shaft at the distal end and remaining in close contact with the sperm membrane. The nucleus occupies the proximal region of the shaft, and two types of dense bodies and mitochondria are located at the distal end. Unlike other members of the Cotylea, the axonemes of B. divae spermatozoa are incorporated into the sperm shaft, leaving the shaft at some distance from the distal end and then remaining free. This type of morphology is characteristic for acotyleans. Additionally, the spermatozoa of B. divae contain only one type of dense bodies plus a unique structure, which we call a central core. The nucleus in this species is unique as well; it shows periodic constrictions and rings of electron-dense granules, characters that further contribute to the distinct status of Boniniidae.     

Ultrastructure of the sperm of blue sprat, Spratelloides gracilis; Teleostei, Clupeiformes, Clupeidae

The general sperm structure of the investigated clupeoids possess an oliviform head with a distinct deep nuclear fossa, a midpiece with one mitochondrion and a posterior flagellum. They are characterized by two apomorphies: an annular or C-shaped mitochondrion and ITDs (intratubular differentiation) in the A tubules of the axonemal doublets. The fine structure of the spermatozoa of blue sprat, Spratelloides gracilis, was investigated to see if resulting data conformed to the current hypotheses of the taxonomy of the Clupeidae. The mature sperm is characterized by the following features: (1) the nucleus is oliviform; its prominent deep nuclear fossa encloses the initial portion of flagellum, (2) a proximal centriole has not been identified, (3) a single spherical mitochondrion is located laterally in relation to the flagellum and (4) no cytoplasmic canal is present. Our blue sprat-spermatozoan morphology data suggest that the blue sprat has Clupeomorpha affinities, while indicating no close affinity with Elopomorpha. Clupeoid sperm exhibit morphological variations with the nucleus and mitochondrion being particularly variable. This study provides useful systematic characters to the existing knowledge of comparative spermatology and may provide additional clue to Euteleost phylogeny.     

基于16S rRNA序列的角蟾科部分属间系统关系

采用最大似然法(ML法)、最大简约法(MP法)、邻接法(NJ法)对来自角蟾科Megophryidae(Anura)8属17种及外群2种共25条400bp左右的线粒体16S rRNA序列进行系统发育关系分析。结果表明,用于分析的所有角蟾科物种形成两大支,第一支包括Megophrys,Brachytarsophrys和Atympanophrys,第二支包括Leptobrachium,Leptolalax,Scutiger,Oreolalax和Vibrissaphora。支持将角蟾科划分为两大类群,即第一支属于Megophryini族．第二支属于Leptobrachiini族。根据分子钟模型初步推测两类群问的分歧时间大致为14．2～18．7百万年前。在Megophryini族中,基因树的拓扑结构不支持Atympanophrys建立后的Megophrys为单系;而在Leptobrachiini族中,Leptolalax为一有效属,其系统发生较早。     

Ophryophryne属在角蟾科系统发育中所处的位置及拟髭蟾族四属间系统关系

采用最大似然法、贝叶斯法、最大简约法对来自角蟾科Megophryidae(Anura)9属20种及外群2种共30号标本的DNA序列进行了系统发育分析,所用片段为线粒体16S rRNA和cytb基因部分序列。结果表明,用于分析的所有角蟾科物种形成两大支。Ophryophryne、Brachycarsophrys、Atympanophrys同属第一支,前两者的系统关系较近。第二支5属间系统发育关系为(Leptolalax,((Leptobrachium,Vibrissaphora),(Oreolalax,Scutiger)))。     

Spermatozoon structure and motility in the anuran Lepidobatrachus laevis

Synthetic human gonadotropin releasing hormone (GnRH) injections were used for induction of spermatozoon release followed by cloacal lavage or mechanical stimulation of sperm release in Lepidobatrachus laevis . Light microscopic observations of Lepidobatrachus laevis spermatozoa indicated an acrosomal segment with a length of 4.1 μm delineated by an indentation, a nuclear region of 12.6 μm in length and a midpiece of 0.87 μm in length. The tail was 54.9 μm long by 1.35 μm wide with two lateral axial fibers and a central undulating membrane. At the electron microscopic level, the unusual tail had two complete axonemes that emanated from the distal centriole. The tail also contained two axial fibers 77 nm in diameter medial to the axonemes and was connected by an undulating membrane. An unusual accessory cell adherent to the head of the spermatozoon was noted in freshly obtained suspensions of spermatozoa. Spermatozoa with the accessory cell were motile and a subsequent loss of motility was correlated with the shedding of the accessory cell.