镇海棘螈的濒危原因探索及其保护建议

镇海棘螈极度濒危．柴桥中学研究小组在教师的指导下对镇海棘螈的濒危原因进行考察、分析和归类,并提出保护建议。     

镇海棘螈产卵场微生境选择

镇海棘螈(Echinotriton chinhaiensis)为国家一级重点保护野生动物,其种群面临着生境退化及丧失的重要威胁。产卵是决定镇海棘螈种群数量增长的关键环节之一,了解其产卵选择的微生境偏好可以更有针对性地保护该物种。本研究旨在确定影响镇海棘螈产卵场微生境选择的关键环境变量,同时为该物种的产卵生境保护、改造和重建提供科学基础。本文于2021年3-5月(繁殖期)在浙江省宁波市北仑区林场对镇海棘螈产卵位点(n=105)与非产卵位点(n=70)处的18个微生境变量进行调查。采用拟合优度卡方检验判断3种无序分类变量的差异性,并利用生境喜好系数对生境选择性进行分析。采用二元逻辑斯蒂回归模型对15个数值型变量进行分析,确定影响镇海棘螈产卵微生境选择的关键变量。结果显示镇海棘螈繁殖期间对产卵场微生境有明显偏好,通常产卵于朝向水坑、落叶层较厚(5.19±0.18 cm)、坡度较陡(18.64°±1.18°)和土壤含水量较低(33.51%±1.87%)的土壤基质上。此外,在大型遮蔽物中,镇海棘螈偏好选择体积较小的石块和乔木(2,994.63±316.17 cm³)作为遮蔽...     

镇海棘螈早期个体发育研究

根据野外观察和室内实验 ,对镇海棘螈的早期个体发育过程进行了研究。结果显示 ,镇海棘螈早期发育过程与琉球棘螈的个体发育过程非常相似 ,它们的平衡枝均不发达且在前肢芽晚期迅速消失。温度和湿度对胚胎发育具有显著影响。室温下 ,从受精卵到孵化需 2 9d左右。孵化幼体体长呈配合对数曲线增长 ,其体全长 (S)与日龄 (T)的关系可表示为S =1 2 974 9+6 2 398lnT(Sig.F <0 0 1 )。水生生活阶段为 58～ 88d。室内幼体的水生发育时间和变态个体大小与野外的情况有较大的差异 ,可能与营养条件和自然条件下的持续干旱有关。变态前的外鳃萎缩期是镇海棘螈早期个体发育过程中的死亡高峰期。     

我国棘螈属(Echinotriton)和疣螈属(Tylototriton)的研究概况

棘螈属（Echinotriton）和疣螈属（Tylototriton）均属于两栖纲有尾目蝾螈科.两属主要分布于我国西南、亚洲西南部、东南亚及日本.棘螈属在我国已知3种：Echinotriton andersoni（琉球棘螈）、E. Chinhaiensis（镇海棘螈）、E. Asperrimus（细痣棘螈）.疣螈属在我国已知4种：Tylototriton kweichowensis（贵州疣螈）、T. Shanjing（红瘰疣螈）、T.taliangensis（大凉疣螈）、T. Verrucosus（棕黑疣螈）.本文回顾了棘螈属和疣螈属物种的命名史,阐明了两属之间的关系,并从形态学、生态学、细胞遗传学和分子生物学方面对这两属的研究工作进行了综述.     

镇海棘螈及濒危现状

本文简述了镇海棘螈的研究概况、外部形态、生活习性及其研究价值。调查资料表明,棘螈分布区极窄,数量甚少,已处于极度濒危。同时对濒危原因作了分析,提出了保护措施和建议。     

探索标记镇海棘螈的方法

研究小组在教师指导下,积极探索标记镇海棘螈的方法,形成了若干方案,并对每个方案的操作方法和优缺点进行论述。     

镇海棘螈瑞岩寺种群年度繁殖量比较及分析

将镇海棘螈Echinotriton chinhaiensis瑞岩寺种群2008年的繁殖情况与1998、1999、2000年的繁殖量进行比较,发现虽然雌性平均窝卵数比较稳定,但2008年的繁殖总量小于其余3年任何一年总产卵量的50%.对其繁殖量大幅下降的原因进行了分析,发现2007年9、10月两次影响严重的强台风、瑞岩寺景区开发等因素可能是造成种群繁殖量大幅下降的原因,而2008年初50年不遇的低温是否影响镇海棘螈的繁殖值得进一步追踪研究.     

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     

雌性瑶山肥螈泄殖腔显微结构研究

本文对雌性瑶山肥螈的泄殖腔进行了比较解剖学研究,发现雌性瑶山肥螈泄殖腔具有和其它有尾两栖类相似的皮肤形态。虽然不同部位皮肤组织的厚度存在一定差异,但都具有表皮和真皮等相同的基本结构。雌性瑶山肥螈的泄殖腔还具有和精子储藏相关的特殊腺体结构,即储精管腺和腹腺等腺体,以及与这些腺体结构相关的泄殖腔管。这表明该物种雌性个体的泄殖腔具备了精子储藏功能的形态结构,为其进行多次交配,分批产卵提供了基础。     

圆斑星鲽精子的超微结构及核前区特殊结构

分别以扫描电镜和透射电镜方法研究了圆斑星鲽 (Veraspervariegates)精子的超微结构。头部呈圆形、中段不发达 ,具有多个未分化的圆形小线粒体、鞭毛结构极简单 ,具有 9 2型的轴丝等特征 ,均表示圆斑星鲽的精子属于简单的原始类型 ,与多数其它高等鱼类精子结构相似。但是 ,圆斑星鲽精子核前部的凹陷 ,即不含染色质的电子透明区 ,以及核前区的囊泡结构却未见在其它新鳍鱼类精子中报道。圆斑星鲽精子核前凹陷开口于核前端稍偏处 ,凹陷呈不规则状 ,可深入到核的中央。凹陷处与染色质区无界膜分隔。凹陷区的前部及其开口处常有多个小的具单层界膜的囊泡 ,有时可见在凹陷区的开口处汇集成一个大泡。在某些处理过程中 ,例如在冷冻 -解冻过程中 ,囊泡可能丢失 ,而在精子顶端留下一个凹坑。尽管顶体在现存最早的新鳍鱼类 ,如雀鳝鱼及弓鳍鱼中已经完全消失 ,但考虑甚至在某些纯真骨鱼 ,如鲑形目中的螈鱼精子上 ,顶体仍然存在 ,而在其它一些纯真骨鱼精子或精子细胞中也存在顶体或顶体遗迹等事实。我们推测在圆斑星鲽精子中的上述特殊结构也是顶体的一种遗迹 ,但也不排除其是向外释放核内物质的一种途径的可能性     

Sperm morphology of salamandrids (Amphibia, Urodela): implications for phylogeny and fertilization biology

Mature spermatozoa belonging to four salamander species, Salamandrina terdigitata, Triturus alpestris, Triturus carnifex and Triturus vulgaris, have been investigated by electron microscopy. The sperm ultrastructure of these species was compared with that of previously examined urodeles (36 species and 20 genera) and with that of anurans and caecilians. Many phylogenetic considerations may be inferred as a consequence of comparative spermatology. Urodela appears to be a monophyletic order characterized by three sperm synapomorphies: the acrosomal barb, nuclear ridge and marginal filament. Cryptobranchoidea are confirmed to form a monophyletic suborder having two synapomorphic characters: absence of mitochondria in the tail, and cylindrical shape of the tail axial rod. Within the family Salamandridae, sperm morphology confirms the phylogenetic distance between Salamandrina and Triturus, as already pointed out on the basis of molecular and morphological characters. The very complex ultrastructure of spermatozoa confirms a previous opinion that internal fertilization is the ancestral condition of the Amphibia.     

银杏雌雄配子体发育及胚胎形成的研究进展

银杏(Ginkgo biloba)是现存最古老的裸子植物之一, 其生殖过程表现出许多原始特征和独特性状, 长期以来备受国内外专家的关注。经过近100年的研究取得了显著成果: (1) 银杏雄配子体发育周期长, 经历了从平周分裂到斜背式分裂,并最终垂周分裂形成带有鞭毛的游动精子; (2) 银杏雌配子体发育经历较长的游离核期和细胞化期, 分化形成颈卵器母细胞并经平周分裂、垂周分裂和斜向分裂形成成熟的颈卵器(包括有4个颈细胞和1个卵细胞); (3) 推测其精细胞中的液泡状结构为受精过程中的遗传物质载体; (4) 原胚的形成经历了游离核期和细胞化期。该文针对国内外最新银杏生殖生物学方面的研究进展, 对银杏雌雄配子体发育、受精过程以及胚胎形成等方面进行较为系统全面的分析和总结, 为进一步的银杏生殖生物学研究提供有价值的参考资料。     

金鱼精子质膜和核膜的区域特异性

本文结合采用扫描和透射电子显微镜(包括冷冻断裂-蚀刻、超薄切片以及细胞化学染色法),研究了金鱼精子的超微结构特征,结果表明金鱼精子的质膜和核膜都具有区域特异性:1)精子质膜内大部分区域含有许多蛋白颗粒,但在特定区域内,蛋白颗粒呈有序排列,构成晶格状结构。2)精子头颈部和尾部均有液泡密集之处,凡是覆盖着液泡区的质膜内,几乎都不含有蛋白颗粒。3)液泡区能被细胞化学方法染成致密色,表明内含糖蛋白。4)核膜孔只集中存在于靠近颈部的核膜上,而其他部分则没有。本文对上述诸点进行了讨论。     

Sperm cell architecture, insemination, and fertilization in the model fern, Ceratopteris richardii

Motile sperm cells of land plants are released directly into the environment and encounter numerous constraints on their way to the egg. Sperm cell organization, shape, size, and plasticity are crucial to the processes associated with fertilization. We conducted an ultrastructural investigation to detail insemination (sperm release, swimming and movement within the archegonium) and fertilization in the model fern Ceratopteris richardii. Gametophytes were grown from spores using sterile culture techniques and flooded in water when sexually mature. Materials were examined at different stages post-flooding. During insemination in C. richardii, the sperm cytoskeleton and flagella rearrange, and the coils of the cell extend while entering the neck canal. In this nearly linear configuration, the dense ridge, a densely compacted band of filaments presumed to be actin, expands to surround the leading edge of the sperm cell. This ridge fuses with the receptive site on the female gamete and is the sperm component that initiates contact with the egg nuclear envelope. All cellular components, except plastids, enter the egg cytoplasm. Sperm mitochondria are distinguishable from those of the egg because they are encased by two or three additional membranes and are sequestered from the zygote cytoplasm. During karyogamy, the sperm components, including the microtubule cytoskeleton (spline) and flagella, maintain their spatial integrity. Microtubules play key roles not only in sperm cell structure but also in facilitating karyogamy in this fern. After karyogamy is completed, microtubule arrays of the sperm cell and the components of the locomotory apparatus are disassembled. We provide the first demonstration of the likely involvement of sperm actin in egg penetration in land plants and new insights into the fate of paternal organelles. This study points to the roles sperm cell structure and dynamics play in the intricate processes of insemination and fertilization in land plants.     

金鱼精子质膜和核膜的区域特异性

Combined SEM and TEM technique including thin sectioning, freeze-fracture and etching as well as cytochemical staining have been used for ultrastructural study on goldfish (Carassius auratus) sperm. It has been shown that primitive sperm plasma membrane and nuclear membrane are differentiated with regional specificity. The results from this study can be summarized as follows: 1. Intercalated protein particles are highly organized in the plasma membrane in the certain region of the head to form crystalline-like structure, in contrast rest of the area is rich in randomly or clustered particles. 2. Many vesicles in different size are often tightly packed in the head, neck and tail regions. Only these plasma membrane covering the vesicles contain almost no protein particles. 3. The vesicles can be densely stained cytochemically, suggesting the existence of glycoprotein. 4. Most of the nuclear membrane have no nuclear pores on it except the area near the neck part where many nuclear pores concentrate.     

Distribution of intramembranous particles and filipin-sterol complexes in mouse sperm membranes: polyene antibiotic filipin treatment

The distribution of intramembranous particles (IMPs) and membrane filipin-sterol complexes (FSC) was examined ultrastructurally in mouse spermatozoa from the male reproductive tract and ejaculates. IMPs were qualitatively analyzed on freeze-fracture replicas of glutaraldehyde-fixed tissue, while membrane FSC were quantitatively analyzed on replicas of filipin-treated cells. The distribution pattern of IMPs of mouse spermatozoa was fundamentally similar to that of other mammalian spermatozoa. 1) In the head, the plasma membrane had a heterogeneous population density, e.g., few IMPs on the acrosomal region, particularly few on the marginal segment, and somewhat regularly arranged IMPs on the postacrosomal region. The acrosomal membrane had many IMPs in hexagonal arrays. The nuclear membrane had many IMPs on the P-face, few IMPs on the variegated E-face, and an intense population density on the P-face of the basal plate. 2) In the neck, the plasma membrane had many IMPs with square arrangements of small IMPs in some areas on the P-face; the redundant nuclear membrane had a few IMPs on both P- and E-faces. 3) In the tail, the plasma membrane had diagonal rows of IMPs in some areas amongst larger IMPs on the middle piece, while it had "zippers" composed of IMPs running parallel to the axis on the principal piece. The distribution of sperm membrane FSC may be summarized as follows: 1) In the head, the acrosomal plasma membrane, which was heavily labeled with filipin, had much more FSC in the equatorial segment than in the marginal segment throughout the study. The postacrosomal plasma membrane generally had no FSC, but some sperm in ejaculates were slightly positive to filipin. The acrosomal membranes (both outer and inner) had no FSC. The nuclear membrane in the main part of the head had less FSC in vas deferens and ejaculated sperm than in the epididymal sperm. The nuclear membrane on the basal plate had no FSC. 2) In the neck, the plasma membrane had little FSC. The redundant nuclear envelope had scattered FSC with a higher incidence in the epididymal sperm than in those from the vas deferens and ejaculates. The membrane scroll, which was elongated from the extreme caudal end of the redundant nuclear envelope, had abundant FSC in the vas deferens and ejaculated sperm. 3) The tail plasma membrane (both middle and principal piece), which was weakly labeled with filipin, had less FSC in sperm from the vas deferens and ejaculates than in those from the epididymis. The limiting membrane covering the mitochondria had no FSC.     

Female sperm storage in amphibians.

The three orders of extant amphibians are Gymnophiona, Anura, and Urodela. Although all gymnophionans apparently have internal fertilization and many are viviparous, female sperm storage is unknown. Internal fertilization has convergently evolved in a few anurans, but females of just one species, Ascaphus truei, are known to possess oviductal sperm storage tubules (SSTs). The SSTs of A. truei are similar anatomically to such glands in squamate reptiles. This similarity is convergence due to similar functional adaptations and/or internal design constraints. In salamanders and newts (Urodela), absence of sperm storage in females is the ancestral condition (three families). In the derived condition, sperm storage occurs in cloacal glands called spermathecae, and their possession is a synapomorphy for females in the suborder Salamandroidea (seven families). Salamandroids are the only vertebrates with cloacal sperm storage glands. In this paper, a phenetic analysis of variation in spermathecal characters reveals patterns of convergence in certain spermathecal characters in unrelated taxa that breed in similar habitats. In the family Salamandridae, a role in sperm nutrition for the spermathecal epithelium is questioned, and the widespread occurrence of spermiophagy is related to other reproductive strategies. I propose how the packaging of sperm in structurally different types of spermathecae may influence male paternity.