日本沼虾雄性生殖系统的研究：Ⅲ.输精管内精荚的结构与形成

于光镜和电镜下研究日本沼虾输精管内精荚的结构与形成。结果显示：粗荚呈索状,由精子群、精荚基质、粘液团和荚壁组成;精荚基质与粘液团内均含有交织的纤丝,其中散布着絮状泡和同心圆形泡,精荚壁呈“Ｃ”型,单层,由致密纤丝、絮状泡和沟形的网状结构组成,包裹精子群和粘液团;前、中、后输精管的上皮细胞均具合成、分泌精荚形成物质的功能。     

A structural analysis of the freshly extruded spermatophore from the lobster,Homarus americanus

Freshly extruded spermatophores from the lobster, Homarus americanus, were examined using light microscopy and scanning and transmission electron microscopy. The tubular spermatophore is trifoil in shape with two lobes tapered laterally from a third lobe situated ventrally. It is comprised of sperm surrounded by three acellular investments: (1) a primary spermatophore layer, (2) an intermediate layer, and (3) an outer bounding layer. The sperm are packed into a continuous tube contained largely within the ventral lobe and are embedded in a matrix of moderate electron density. The primary spermatophore layer is uniformly thick around the sperm mass and contains at its peripheral margins both ring structures and crystals in an amorphous matrix. The intermediate layer is thicker dorsally than ventrally. Dense granules dominate the ventral half of the intermediate layer while inclusions populate the dorsal half; both react positively to the periodic acid-Schiff (PAS) technique. The innermost portion of the outer bounding layer is composed of parallel fibrils; a flocculent material is present peripherally. This flocculent substance is presumed to impart stickiness to freshly extruded spermatophores. These observations provide a basis for the future understanding of the mechanisms involved in long-term storage of sperm in spermatophores.     

Oriented spermatozoa in the spermatophore of the palaemonid shrimp, Macrobrachium rosenbergii

Estimates of the degree of orientation of spermatozoa within spermatophores of the freshwater shrimp, Macrobrachium rosenbergii, were made using electron micrographs of ultrathin sections cut transverse and parallel to the longitudinal axis of the spermatophores. Counts of the total number of longitudinal and non-longitudinal profiles of sperm spikes in transverse sections of spermatophores indicated that 67% of the spermatozoa were oriented with their spikes approximately perpendicular to the long axis of the spermatophore. In longitudinal sections, ~30% of the sperm spike profiles were oriented parallel to the longitudinal axis of the spermatophore, leaving ~70% of the profiles oriented in planes not parallel to the longitudinal spermatophore axis. Stereologic analyses based on the number of intersections of longitudinally sectioned sperm spike profiles per unit length of test lines placed over photographic images of ultrathin sections cut in both transverse and longitudinal planes of spermatophores gave a Saltykov approximation of 0.607, favoring the view that the spermatozoa were oriented with their spikes perpendicular to the long axis of the spermatophore. This orientation brings many of the sperm bases approximately parallel to the surface of the spermatophore, an orientation which may facilitate normal sperm-egg interaction during fertilization in this species.     

Spermatozoa ultrastructure of the pink shrimp Farfantepenaeus paulensis (Decapoda: Dendrobranchiata)

Braga, A.L., Nakayama, C.L., Suita de Castro, L.A. and Wasielesky, W. 2011. Spermatozoa ultrastructure of the pink shrimp Farfantepenaeus paulensis (Decapoda: Dendrobranchiata). —Acta Zoologica (Stockholm) 00 : 1–6. The spermatozoa ultrastructure of the pink shrimp Farfantepenaeus paulensis was investigated in this morphological study. Spermatophores and spermatozoa were analyzed by electron microscopy. The pink shrimp spermatophore is divided into two regions: the appendage and the spermatophore main body, where spermatozoa are grouped in a spermatic mass. Pink shrimp spermatozoa are unistellate and are composed of main body and single spike. The spermatozoa body comprises a perinuclear cytoplasmic band, nucleus, acrosomal cap, and subacrosomal region. The spermatozoa cell mean total length was 10.71 μm, the mean body diameter was 5.56 μm, and the mean spike length and diameter were 5.15 μm and 0.85 μm, respectively.     

Morphology of the male system and spermatophores of the arrowworm Ferosagitta hispida (Chaetognatha)

Transmission electron microscopy reveals that the somatic testicular tissues and sperm ducts are elaborations of the epithelial lining of the tail coelom. The testes consist of closely packed spermatogonia embedded between specialized lateral field cells. These cells contain few organelles and appear to function mainly as a compartment boundary. Masses of spermatogenic cells are released into the tail coelom from the anterior end of the testes. The sperm ducts, lined by simple cuboidal ciliated epithelium, collect mature spermatozoa from the tail coelom and convey them to the blindly ending seminal vesicles. The sperm ducts also modify coelomic fluid entering them along with the spermatozoa. The seminal vesicles consist of a simple glandular lining epithelium embedded in the stratified epidermis. Secretions of the lining epithelium surround the enclosed sperm mass and correspond in position to a noncellular spermatophore coat visible by light microscopy around released sperm masses. Spermatophores leave the seminal vesicles through a temporary split that forms between microfilament-containing suture cells. Maturation of spermatozoa and filling of the seminal vesicles is cyclical, occurring late each day. © 1994 Wiley-Liss, Inc.     

Fine structure of the spermatophores and their ejaculated forms,sperm reservoirs,of the Japanese common squid,Todarodes pacificus

Spermatophores in a squid, Todarodes pacificus, were observed by light and electron microscopy and were further analyzed by X-ray microanalysis (XMA) of frozen thin sections. Each spermatophore consists of a sperm mass, a cement body, an ejaculatory apparatus, and some fluid materials, all of which are covered by an outer tunic. The outer tunic consists of about 20 membranous layers, each containing straight, parallel microgrooves. Each layer's microgroove pattern is roughly in an orthogonal arrangement with respect to the next layer's pattern. The sperm mass, which is the only cellular component, consists of a sperm rope which is coiled more than 500 times. Most of the spermatozoa in the rope are arranged regularly and are enveloped in materials which are well-stained by Alcian blue. The cement body is located between the sperm mass and ejaculatory apparatus and has a hard outer shell with an arrowhead-like structure, presumably for penetration into the tissue of the female. Calcium and phosphorus are present in the shell of the cement body, which also has an affinity for alizarin red. The ejaculatory apparatus consists of two tubes, designated as the inner tunic and the inner membrane. After the spermatophoric reaction, a sperm reservoir is formed at the anterior end of the extruded and inverted ejaculatory apparatus. The sperm reservoir, which encases the sperm mass, is composed of the cement body at the anterior end and the inner tunic of the ejaculatory apparatus at the posterior end.     

Spermatophores of the salamander Ambystoma texanum

Living spermatozoa were observed in freshly deposited spermatophores and in fluid from vasa deferentia. In the distal, but not proximal, vas deferens spermatozoa moved together in whorls with heads and tails in alignment. Around the entire periphery of the spermatophore cap, similar slowly undulating groups of spermatozoa had their heads aligned and directed outward. Over time, some individual spermatozoa left the cap of the spermatophore and moved into the surrounding water (cap deterioration). Microscopical observations were made on spermatophore squashes and paraffin sections of spermatophores and vasa deferentia. Spermatozoa around the periphery of the cap were underlain by a PAS-positive membrane-like material. Cytoplasmic droplets, which were attached to spermatozoan necks in the vas deferens, were accumulated in the center of the spermatophore cap deep to the PAS-positive membrane. Spermatophore stalks were strongly PAS and Alcian blue positive and showed positive reaction for RNA. Tests for lipids and proteins were negative in the whole spermatophore. Electron microscopic observations showed the stalk of the spermatophore to be composed of rounded ‘balls’ of fibrous material. At the juncture of the stalk and cap a less dense fibrous material impacted the stalk enclosing many sperm tails and some heads and, although no attachment devices were visualized, the sperm were closely apposed to this material as was the spermatophore stalk. This finely filamentous material encircled the cap and was more prominent in some regions than others. The PAS-positive material detected with the light microscope was also observed with the electron microscope. It was circumferentially oriented and was composed of 200 Å packed filamentous densities. Sperm heads and tails were found lying external to the membrane, whereas only tails and cytoplasmic droplets occupied the core of the spermatophore. Cytoplasmic droplets were usually free of the sperm tail and contained membranous sacs and two types of nuage material.     

Spermatophore Transfer and Subsequent Sperm Development in a Homalorhagid Kinorhynch

The cuticular morphology and precise location of male and female gonopores and penile spines of the homalorhagid kinorhynch Kinorhynchus phyllotropis Brown & Higgins, 1983 are described and illustrated. In this species spermatozoa are transferred from male to female by a spermatophore. This is the first record of the mechanism of sperm transfer in a kinorhynch. The spermatophore is presumably extruded through the male gonopore and directed towards the female by the ductless penile spines. Spermatozoa in the spermatophore are rod-shaped and catenulate. The spermatophore is pressed directly against the cuticular plates of the female, and usually covers the female gonopores. The spermatophore contains a mass of intertwined spermatids and spermatozoa surrounded by clear material covered with a layer of debris. Spermatozoa are found in the female lodged in the seminal receptacle tissue applied to the dorsal aspect of posterior oocytes. There the spermatozoa complete their development. Nuclei change from filiform to geniculate, and oval corpuscles surrounding the nuclei disappear, so that the spermatozoa are seen as densely-packed, polyhedral cells. These observations conform with literature reports of aberrant spermatozoa of unknown origin seen in female Pycnophyes . The fertilization process remains unknown.     

Motility and dynamics of eupyrene and apyrene sperm in the spermatheca of the monandrous swallowtail butterfly Byasa alcinous

Lepidopteran males produce two sperm types: nucleated eupyrene sperm and non‐nucleated apyrene sperm. Although apyrene sperm are infertile, both sperm types migrate from the spermatophore to the spermathecal after copulation. As a dominant adaptive explanation for migration of apyrene sperm in polyandrous species, the cheap filler hypothesis suggests that the presence of a large number of motile apyrene sperm in the spermatheca reduces female receptivity to re‐mating. However, apyrene sperm are also produced in males of the monandrous swallowtail butterfly Byasa alcinous Klug. To identify the role of apyrene sperm in these males, the present study examines the number of spermatozoa produced and transferred and the dynamics and motility of spermatozoa in the spermatheca for each type of sperm. Apyrene sperm represents approximatey 89% of the sperm produced and transferred, which is comparable to polyandrous species. Two‐day‐old males transfer approximately 17 000 eupyrene and 230 000 apyrene spermatozoa to a spermatophore; approximately 5000 eupyrene and 47 000 apyrene spermatozoa arrive at the spermatheca. Eight days after copulation, most eupyrene spermatozoa remain in the spermatheca and a quarter of them are still active. However, the number of apyrene spermatozoa decreases and those remaining lose their motility after the arriving at the spermatheca. Consequently, 8 days after copulation, no motile apyrene sperm are found. The high proportion of apyrene sperm in the spermatophore, as well as in sperm migration, suggests that the production and migration of apyrene sperm is not simply an evolutionary vestigial trait. The possible functions of apyrene sperm in monandrous species are discussed.     

New insights in the male anatomy,spermatophore formation,and sperm structure in Atyidae: The red cherry shrimp Neocaridina davidi

This study aims to analyze the functional anatomy of the male reproductive system in Neocaridina davidi, a very popular ornamental species of caridean shrimp. Mature males were cold‐anaesthetized and their reproductive systems were dissected for histological and histochemical analysis, while the spermatozoa and spermatophore wall ultrastructure were analyzed under transmission electron microscopy. The male reproductive system consisted of two coiled testes, which were continuous with the vasa deferentia. Testes were positioned on the dorsal side of the cephalothorax above the hepatopancreas, and comprised seminiferous tubules where spermatogenesis occurred. Each vas deferens (VD) was a long tube dorsolaterally positioned with respect to the hepatopancreas, and increased in diameter at the distal end. Three regions could be recognized in the VD: proximal, middle, and distal. The proximal region had a cylindrical epithelium with secretory cells. The middle region (or typhlosole) had a dorsal fold (or typhlosole) with a thick columnar epithelium and high secretory activity. The spermatophore was a continuous cord with three acellular layers, which were mainly characterized by the presence of neutral glycoconjugates and proteins. The sperm morphology was distinct from the inverted cup‐shaped spermatozoa observed in the majority of caridean shrimps. The spermatozoa in specimens of N. davidi were spherical in shape, with a cross‐striated, single, short spike, and arranged in clusters of three or four sperm cells. The composition of the spermatophore, and the arrangement and form of the spermatozoa, seem to be unique in comparison to other species of Caridea.     

Ultrastructure of the male nephridium and its role in spermatophore formation in spionid polychaetes (Annelida)

Summary The mature male nephridia ofPolydora ligni andP. websteri (Polychaeta: Spionidae) are segmental organs composed of a ciliated nephrostome connected to a nephridial canal that crosses the intersegmental septum, expands into a large modified part extending dorsally through the coelom and subsequently narrows into a canal terminating in a dorsal nephridiopore. The nephridial canal is ciliated throughout and is composed of several cell types. Cells in the expanded region of the nephridia of both species contain large urn-shaped depressions filled with long microvilli. InP. ligni, one section of a nephridium contains cells packed with electron-dense granules that are not observed inP. websteri.The spermatophores ofPolydora ligni are composed of a central sperm mass surrounded by a layer of randomly oriented tubules that form a capsule around the sperm and taper into a long thin tail. These tubules are identical in dimensions to the microvilli present in parts of a nephridium and apparently are derived from these microvilli. The spermatophore capsule ofP. websteri is composed of similar tubules also presumed to originate from nephridial microvilli.The microvilli in nephridia of both species are surrounded with a glycocalyx that may function as an adhesive to hold the spermatophore capsule together. This glycocalyx may also function as a species specific message when encountered by a receptive female.Contribution Number 179 from Harbor Branch Foundation, Inc.     

罗氏沼虾精子棘状部的结构生化组成和功能

作者探讨了罗氏沼虾精子棘状部的组成,生化成分和受精时所起的作用。棘状部主要由具周期性横纹的纤丝组成,在近尖端横切面的中央有絮状颗粒存在。棘状部Ｆｅｕｌｇｅｎ反应呈阴性,不含核物质,而ＨｇＢｐＢ反应呈强阳性,蛋白质含量很高。     

Eupyrene sperm migrates to spermatheca after apyrene sperm in the swallowtail butterfly, Papilio xuthus L. (Lepidoptera: Papilionidae)

When swallowtail butterflies, Papilio xuthus, are mated by the hand-pairing method, both types of sperm, eupyrene and apyrene sperm, are transferred from the male to the spermatheca via the spermatophore in the bursa copulatrix. This mechanism is demonstrated by two different kinds of experiments. The first set of experiments employed interrupted copulation, and the second set was examination of the sperm in the spermatophore and spermatheca after the termination of copulation. The sperm was transferred 30 min after the start of copulation. The eupyrene sperm was still in the bundle; the number of the bundles ranged from 9 to 108 (mean, 42.7; n = 27). The bundles were gradually released after the completion of copulation, and the free eupyrene spermatozoa then remained in the spermatophore at least 2 h before migrating to the spermatheca. On the other hand, about 160 000 apyrene spermatozoa were transferred to the spermatophore and remained there for more than 1 h. We observed 11 000 apyrene spermatozoa in the spermatheca 12 h after the completion of copulation, but most of this type of sperm disappeared shortly thereafter. In contrast, the eupyrene sperm arrived in the spermatheca more than 1 day after the completion of copulation and remained there at least 1 week. Therefore, these findings suggest that apyrene sperm migrate from the spermatophore to the spermatheca earlier than eupyrene sperm. Accordingly, if females mated multiply, the time difference might avoid the mixing of sperm. In addition, the predominance of sperm from the last mating session may occur not in the bursa copulatrix but in the spermatheca. Received: January 7, 2000 / Accepted: May 24, 2000     

On the spermatophore of ixodid ticks

Structure and formation of the spermatophore and the extrusion of its contents in ixodid ticks were studied. The structure of the ectospermatophore resembles that of argasid ticks, whereas that of the endospermatophore differs completely; it is composed of several organelles which are absent in argasids.The chemical nature of the components of the spermatophore was studied by histochemical methods. The male forms the spermatophore by ejaculating the components in a definite order. The components of the ectospermatophore are deposited one into another—while those of the endospermatophore one on top of the other and are linked together in a chain. The last component to be ejected is a plug that closes the endospermatophore.During extrusion of the endospermatophore all of the components burst out of the ectospermatophore. The endospermatophore sheath evaginates and forms the capsule into which the sperm is inserted. Each capsule has two tube-like appendices.     

Ultrastructure of the sperm of the deep-sea decapod Aristeus antennatus

The sperm of Aristeus antennatus presents notable differences in relation to the two basic models of decapod crustaceans considered to date. Basically, it does not present a single appendage, or spike, characteristic of the so-called unistellate sperm of the suborder Dendrobranchiata and the infraorder Caridea of the suborder Pleocyemata. Nor does it have arms or spikes characteristic of the multistellate sperm that all belong to the Pleocyemata group. The spermatozoa of A. antennatus are composed of a nucleus and an electron-dense acrosome, which have the polarity of multistellate sperm. A number of mitochondria and vesicles are present in the cytoplasm, located between the acrosome and the nucleus. In accordance with the fine structural details, the morphology of the sperm has been described at two different levels of the male gonad, the vas deferens and terminal ampulla, and in the spermatophore placed in the thelycum of the female. Three ultrastructural changes in the acrosome (unorganized structures, tubular organization, disintegration process) and the nucleus (uncondensed, condensed, and compact) are present along the male reproductive apparatus. They first appear in a non-organized manner at the level of the vas deferens, subsequently undergo a process of structural configuration in the ampulla, and finally show disorganized structures in the spermatophore. J. Morphol. 234:79–87, 1997. © 1997 Wiley-Liss, Inc.     

日本沼虾精子的形态和超微结构研究

应用电子显微镜技术结合细胞化学方法对日本沼虾精子进行子形态和超微结构研究。结果显示,日本沼虾精子琪似外翻的伞状,无鞭毛,不运动的精子主要由前端棘突,中间帽状体和后主体部分组成,精子的核属非浓缩型,内布许多絮状物质,外无核膜包被,呈Ｆｅｕｌｇｅｎ阳性反应,位于后主体部内;中间帽状体的细胞质内含有一对中心粒和１５－２０根放射状排列的纤丝,纤丝在帽状体凸面中央汇聚并延伸成精子的棘突,棘突和放射状纤丝皆具     

Ultrastructure of the male reproductive system and spermatophore formation in Labidocera aestiva (Crust.: Copepoda)

Summary The male reproductive system of Labidocera aestiva produces a flask-shaped spermatophore connected to a chitin-like coupling apparatus. As immature spermatozoa leave the anterior region of the testis, they pass through the lumen of a long, sinuous duct composed of a ductus deferens and seminal vesicle. Ultrastructural examination of the ductus deferens reveals a highly glandular, columnar epithelium. The cells contain arrays of rough endoplasmic reticulum and abundant, well-developed Golgi complexes. This region produces and releases into the lumen, a flocculent substance and two granular secretions that constitute the seminal fluid. In its terminal part, the ductus deferens synthesizes another secretion that forms the spermatophore wall enclosing the spermatozoa and seminal fluid. Final synthesis of the spermatophore wall occurs within the thin-walled seminal vesicle, although this region functions primarily as a storage organ. Contiguous to the seminal vesicle is an elongate, highly glandular spermatophore sac. The chitin-like coupling apparatus, which functions to attach the spermatophore to the female, is formed in the anterior region of the sac by secretions from eight cell types. The posterior region of the sac stores the flask-shaped spermatophore and produces secretions that aid ejaculation of the entire spermatophore complex.Contribution No. 236, Harbor Branch Foundation, Inc.     

Sperm penetration into immature mouse oocytes and nuclear changes during maturation: an EM study

The ultrastructure of oocyte and sperm nuclei was studied in mouse ovarian oocytes inseminated in vitro and cultured for 1 1/2 and 3 h in a medium containing dbcAMP or lacking the maturation inhibitor. In oocytes blocked at the germinal vesicle (GV) stage, certain maturation-linked changes were noted. Sperm apposition and sperm-oocyte fusion were similar to that during fertilization of ovulated oocytes. The sperm nucleus and its nuclear envelope remained intact after penetrating into the ovarian oocyte. One and a half h after removal of the drug (time 0 of maturation) the germinal vesicle (GV) and sperm nucleus remained intact. In oocytes maturing for 3 h, the nuclear envelopes of the GV and sperm nucleus had fragmented. The NE of the oocyte formed quadruple membranes while the NE of the sperm remained as flat vesicles. Oocyte chromatin condensed to form chromosomes, whereas at the same time the sperm chromatin was in the process of decondensation and was surrounded by fragments of the sperm NE. The sperm chromatin, composed of DNA complexed with protamines, consisted of thin fibrils; the individual fibrils measured 3.8 nm in diameter. Near the penetrated spermatozoa only occasional Mts were detected which were not related to the proximal centriole which was recognizable in the neck-piece of the flagellum. Thus in mouse oocytes the introduced sperm centriole is not capable of behaving as a centrosome and organizing microtubules in the form of an aster.     

Correlation between the spermatozoal characteristics and sperm penetration distance in polyacrylamide gel and bovine cervical mucus

Correlation between the spermatozoal characteristics and the sperm penetration distance in polyacrylamide gel was assessed, utilizing frozen thawed semen samples obtained from 6 bulls, and it was compared with the correlation between sperm penetration in bovine cervical mucus and spermatozoal characteristics. In vitro sperm penetration tests were performed with mucus and gel. The sperm penetration in gel and mucus was significantly and positively correlated with post-thaw motility (r=0.81; r=0.89:P<0.01) and acrosome integrity (r=0.88; r=0.94:P<0.01). A significant negative correlation with abnormal spermatozoa (r=-0.84;r=0.83:P<0.01) was observed. Both sperm concentration and post-thaw live spermatozoa were not significantly correlated. A significant multiple regression between sperm penetration and the spermatozoal characteristics both in gel (R2=0.87; F=40.27; P<0.01) and mucus (R2=0.91; F=60.48; P<0.01) was observed. The major spermatozoal characteristics determining the capacity of spermatozoa to penetrate gel were post-thaw motility, percentage of abnormal spermatozoa and acrosome integrity. The acrosome integrity has a more significant contribution. The correlation established with sperm penetration in gel was very similar to that of sperm penetration in mucus. The utility of gel as a mucus substitute in in vitro sperm penetration tests was discussed.