不育症患者精子头部及尾部超微结构的研究

目的研究不育男性精子超微结构的形态特征。方法利用透射电镜对8例不育男性新鲜精液标本中的精子头及尾部超微结构进行观察。结果在电镜下不育男性精子存在多形态超微结构异常,有以下几种类型:(1)顶体异常精子,包括顶体膜受损,顶体发育不良、缺失,顶体内形成包涵体精子。(2)头部异常精子,包括其尖头精子、圆头精子、头部含空泡精子。(3)尾部异常精子:①尾部形态异常精子,包括无尾精子、短尾精子、卷尾精子、体尾胞质残余;②尾部结构异常精子,包括线粒体缺失精子、尾部线粒体多种形态和结构异常。结论不育男性精子存在顶体、头部、尾部线粒体、微管多种形态和结构异常。

黄精赞育胶囊优选方处理前后精子的超微结构研究

目的观察黄精赞育胶囊优选方作用前后精子的超微结构变化。方法运用原子力显微镜(AFM)技术对比观察正常精子和病理性精子在黄精赞育胶囊优选方作用前后超微结构的动态变化。结果获得精子头体、颈部和鞭毛等部位的实时超微结构图像,发现药物干预后活动力低精子超微结构的缺陷有逐步被修复的现象。结论黄精赞育胶囊通过修复活动力低精子超微结构的病理形态学缺陷可能是优选方改善弱精子质量的机制之一。

不同形态精子的超微结构研究

据报道,世界范围内约有15％～20％的育龄夫妇不能生育,且近半个世纪以来,随着全球污染问题的日益严重,社会竞争压力的日趋增加,生活步伐节奏的不断加快,男性生殖健康正受到严重威胁,男性精子数量几乎减少了一半,并且,还以每年2.1％的速度在减少.同时,畸形、劣质精子比例逐渐增多,精子活力、穿透力、致孕率在不断下降,致使男性不育的比例正在逐年上升.精液分析在诊断男性不育及辅助生殖方面起到越来越重要的作用,精液分析主要包括了精子形态、精子活力和精子密度等.随着染色技术的发展和畸形精子分类体系的形成,精子形态学检查对不孕不育症诊断、生育能力预测以及公共健康研究有重要意义,比精子密度、精液质量和运动性更能在受孕中起到预测作用[1].

UItrastructural Observation on Rat Spermatozoa Treatedwith Tripchlorolide(T_4) from Tripterygium wilfordii

Adult male rats were treated orally with monomer T_4(Tripchlorolide) isolated from Tripterygium wilfordii, at the dose of 50μg/kg/day,6 days/week for6 weeks.Ultrathin section and freeze etching replica of seminiferous tubules and epididymal spermatozoa were examined with elec-tron microscope. The results showed that the spemiogenesis was inhibited T_4 in seminiferous tu-bules. However,the damage and disruption of the spermatozoa were more serious in the epididymis.Damage of the structure and function ot microtubule and microfilament may be the chief reason for sperm damage. Sperm membrane was also very sensitive to the treatiment of monomer T_4.

Effects of Cryopreservation on the Quality and Ultrastructure of Tibetan Mastiff Sperm

[Objective] This study aimed to investigate the effects of cryopreservation on the quality and ultrastructure of Tibetan Mastiff sperm. [Method] The effects of cryopreservation on the quality of Tibetan Mastiff sperm were evaluated via motility, membrane integrity rate and acrosome intact rate. On that basis, the effects of cryopreservation on ultrastructure of sperm were observed under SEM and TEM. [Result] In Experiment 1, EG gave the best results not only in post-thaw motility rate （36.3%）, but also in low membrane integrity rate （38.0%） and acrosome intact rate （42.0% ）, but there was no significant difference between EG group and Glycerol group （P0.05）. In Experiment 2, the 5 cm freezing height obtained the best freezing-thawing results, but there was no significant difference between 2 and 5 cm height （P 0.05）, besides in acrosome intact rate. In Experiment 3, SDS and Vc added separately or together into extenders could improve freezing-thawing results, but there was not obvious difference between SDS group and Vc group （P0.05）, besides the lower motility of Vc group （P0.05）. Addition of SDS and Vc obtained the best results in post-thaw motility rate （44.1%）, and also in membrane integrity rate （48.0%） and acrosome intact rate （48.2%）. The ultrastructure of frozen-thawed sperm was also evaluated under SEM and TEM, results showed that cryopreservation caused various degrees of damage to Tibetan Mastiff sperm, more serious damages were observed in the acrosome such as swelling, vesiculation and even disappearance. [Conclusion] This study confirms that EG, horizontal height of 0.25 ml straw above LN 2 surface and additives SDS and Vc together can improve freezing effect. However, cryopreservation has certain damage to ultrastructure of sperm.

Ultrastructure of the Blepharoplast and the Multilayered Structure in Spermatogenesis in Osmunda cinnamomea var. asiatica

The ultrastructure of the blepharoplast and the multilayered structure (MLS) in the fern Osmunda cinnamomea var. asiatica Fernald have been studied by electron microscopy with respect to spermatogenesis. The blepharoplast appears in the young spermatid. The differentiating blepharoplast is approximately a spherical body, which is composed of densely stained granular material in the center and some cylinders outside of it. The differentiated blepharoplast is also a sphere, but without the densely stained material in the center, consisting of scattered or radially arranged cylinders. The MLS seen in the spermatid lies between the basal bodies and the giant mitochondrion. In the early developmental stage, the MLS only consists of lamellar layers, each of which runs parallel to one another and forms a strip. In the mid stage, the MLS is composed of the microtubular ribbon (MTr), the lamellar layers and a layer of plaque. In the late stage, the MLS forms accessory band, osmiophilic crest and a layer of osmiophilic material. The MTr grows out from the MLS and extends along the surface of the nucleus to unite with the nuclear envelope in a complex. The basal body coming from the cylinder produces the axoneme of the flagella in the distal end and the wedge-shaped structure in the proximal end, respectively. In the present study, the ultrastructural features of blepharoplast and the MLS of the protoleptosporangiopsida fern, O. cinnamomea var. asiatica, have been described and compared with those of other kinds of pteridophytes in detail. The lamellar layers appearing before the formation of the MTr was found and reported for the first time.

A Novel Type of Spermiogenesis in the Nile Catfish Chrysichthys auratus (Siluriformes: Bagridae) in Egypt, with Description of Spermatozoon Ultrastructure

Spermiogenesis and spermatozoon ultrastructure in the Nile catfish Chrysichthys auratus are described using transmission electron microscopy. Spermiogenesis involves some unique peculiarities such as ： the development of the centriolar complex and the initial segment of the flagellum in a position directly perpendicular to the basal pole of the nucleus, as a result of absence of nuclear rotation ; lack of a cytoplasmic canal during differentiation of the spermatids into spermatozoa; the base of the basal body is not traversed by the basal plate; a basal foot anchors the basal body to the nucleus; and the presence of numerous vesicles around the midpiece and base of the flagellum. In addition, spermiogenesis includes some common features such as： chromatin compaction; formation of a medial shallow nuclear fossa; and elimination of excess cytoplasm. The mature spermatozoon has an elongate conical-shaped head with no acrosome or acrosomal vesicle, a long midpiece with numerous vesicles that continue backwards around the base of the flagellum and a long tail or flagellum, which has no lateral fins or a membranous compartment. The mitochondria lie close to the nucleus basal pole and surround the initial segment of the axoneme and are separated from the flagellum by the inner mitochondrial envelope due to disappearance of the cytoplasmic canal. The flagellum has the classical axoneme structure of a 9 ＋ 2 microtubular pattern. On the basis of the peculiar features mentioned above, it is concluded that spermiogenesis in this Nile catfish is a synapomorphic type derived from types Ⅰ and Ⅱ spermiogenesis, which are common among teleosts. Accordingly, this type could be considered as a novel type of spermiogenesis and could be termed as ＂type Ⅲ＂.

Ultrastructure and Protein Composition Changes during Acrosome Reaction in the Sperm of Chinese Shrimp, Fenneropenaeus Chinensis

Egg water was used to induce acrosome reaction in mature sperm of Fenneropenaeus chinensis .Transmission electron microscope and SDS PAGE were used to study the ultrastructure and protein changes of the sperm in F.chinensis ,which has undergone acrosome reaction.The results demonstrated that the sperms from female thelycum could be induced acrosome reaction in vitro by egg water,which was a kind of egg jelly released from oocyte into seawater during oocyte activation.More than fifty percent of sperm finished acrosome reaction during the first 30 min, in vitro .The whole event consists of the retraction of the spike and acrosome exocytosis.Spike was retracted and fused to acrosome cap followed by the release of acrosome granules.When the original egg water was diluted 5 and 10 times,The diluted egg water also have the ability to induce sperm acrosome reaction,but the acrosome reaction rate is much lower than that of the original egg water in the same time.Capillary zone electrophoresis was used to detect the biochemical components of egg water.Egg water was composed of two different components,which were also demonstrated by SDS PAGE.The molecular weights of the two kinds of egg jellies are both about 200kDa.Gelatin substrate SDS PAGE results did not show any hydrolytic enzyme activity in egg water.After acrosome reaction,many sorts of proteins in sperm are degraded.When the reacted sperms were examined with gelatin substrate SDS PAGE,there were six major peptide bands with hydrolytic enzyme activity were detected.Their molecular weights are 200kDa,130 kDa,66 kDa,53 kDa,48 kDa and 41 kDa respectively.These hydrolases cannot be detected in the sperms before acrosome reaction.The acrosome reaction of Chinese shrimp, F.chinensis is much different to that of the sperm in the shrimp, Sicyonia ingentis ,which the acrosome reaction was clearly studied.There is not filament formation during the acrosome reaction in Chinese shrimp.The time of exocytosis in the sperm of Chinese shrimp is also much longer than that of the sperm of Sicyonia ingentis .

The ultrastructure of the spermatozoon of Octopus ocellatus Gray, 1849 (Cephalopoda: Octopoda)

Morphology of the spermatozoon of Octopus ocellatus was studied by light, scanning electron, and transmission electron microscopes. Sperm are 600-700 um long, with a large number of granules in diameter about 130 um. Each spermatozoon is composed of a head, neck, and tail. The head is made up of an acrosomal complex anterior to the nucleus. The spiral acrosomal complex consists of an electron-lucent vesicle, lacuna, and an electron-dense acrosomal vesicle. Additionally, the spiral acrosomal vesicle has numerous equidistant striations, and is surrounded by many small granules （20 nm diameter）. A long straight nucleus, which is electron-densed, has a deep posterior concavity, the nuclear vacuole. At the terminal end of the nucleus is a sleeve-like structure with a concave posterior nuclear fossa （PNF）. The neck is short connecting the PNF. The basal body is located in the PNF and gives rise to the axoneme. This structure connects the head, neck, and tail. The tail is divided into a middle piece and a principal piece. The middle piece, having a 9＋9＋2 arrangement, is surrounded by a mitochondrial sheath and terminates by an electron-dense fibrous sheath. The principal piece is the longest part of the sperm with coarse fibers tapering posteriorly. The results of this study shall provide some useful information for artificial breeding of this species.

Ultrastructure of spermatozoa of Bullacta exarata （philippi） and its significance on reproductive evolution and physio-ecological adaptation

The morphology and ultrastructure of Bullacta exarata spermatozoa observed by light and transmission electron microscopy are presented in this paper. The spermatozoon is composed of head with a simple acrosomal complex and an elongated nucleus, and tail with a midpiece, principal piece and an end piece. The midpiece consists of a mitochondrial ring, and the principal piece is composed of axoneme and lateral fin. The structure of 5. exarata spermatozoa differs significantly from that of other gastropods, especially in the lateral fin and the principal piece, which was described scarcely before. A comparison is made between B. exarata and other gastropods, and its significance on reproductive evolution and physio-ecological adaptation is preliminarily discussed.

Characteristics of the Sertoli Cells of Salmonidae from Ohrid Lake during Spermatogenesis-Ultrastructural Analysis

Ultrastructual characteristics of Sertoli cells of Salmonidae from Ohrid Lake during the spermatogenetic cycle have been analysed. Sertoli cells being an integral part of the seminiferous lobules underwent considerable changes, which influenced their cytomorphological features. The degenerative changes of Sertoli cells were manifested by an extreme vacuolisation, mitochondria in degeneration with widened crysts and thickened matrix, desorganised ER, autophagosomes, ＂myeline-like＂ structures and lysed cytoplasmic regions. The above mentioned changes were followed by karyopycnisis, comlete degeneration and delamination of cells from the wall of the seminiferous lobules, lysis and detritus formations （Sertoli necrotic material） in the lumen of the lobules. The aim of this paper is special research of the ultrastructural characteristics, i.e. the changes on a level with testes which happen in the postspawning period in the two species of Teleostei of Ohrid Lake, Ohrid trout （Salmo letnica Kar.） and Ohrid belvica （Acantholingua ochridana Steind.）. The postspawning period is emphasized in Teleostei in this relatively short period, when one reproductive cycle finishes and the following has to start, on a level of testicular parenhyma very important histological changes are going on which give special histological identification, i. e. in the postspawning period there is a complete reorganization of the testes.

FURTHER OBSERVATION OF THE SPERMATOZOA OF LEFT-EYE FLOUNDER PARALICHTHYS OLIVACEUS BY ELECTRONIC MICROSCOPY

The spermatozoon ultrastructure of left-eye flounder, Paralichthys olivaceus, was observed by electronic microscopy, and shown to consist of head, midpiece and tail. Some nuclear vacuoles were observed inside and along the outer part of the nucleus and double-membrane structures were observed between the nuclear membrane and plasma membrane. The sperm sleeve consisted of four independent parts and had more than four mitochondria. Along the sperm tail there were wing-like lateral fins.

Ultrastructure Developments During Spermiogenesis in Polydora ciliata(Annelida: Spionidae),a Parasite of Mollusca

Spionid worms of Polydora ciliata inhabit the shells of many commercially important bivalves and cause disease in molluscan aquaculture. Their sperm structure is closely related to their fertilization method. To give an insight into the sperm structure and spermatogenesis, ultrastructure details of the subcellular components of germ cells during spermiogenesis of Polydora ciliata are detected by transmission electron microscopy(TEM). In P. ciliata, during spermiogenesis, chromatin is regularly arranged as dense fibrils and becomes more condensed when the nucleus elongates. Microtubules do not surround the nucleus during its elongation. The Golgi phase is characterized by the formation of proacrosomal granules within the Golgi apparatus. The proacrosomal granules fuse to form a single, spherical acrosomal vesicle that migrates to the anterior pole of the cell. At the time of nuclear condensation, mitochondria become reduced in number but increased in size, causing deep indentation at the base of the nucleus. The mid-piece has a few mitochondria. The cap phase includes the spreading of the acrosomal granule over the surface of the nucleus of the differentiating spermatid. The acrosomal phase of spermiogenesis is typically associated with changes in the shape of the nucleus, acrosome and tail. The relationship of sperm ultrastructure to spermiogenesis in spionidae species was discussed.

精子纤维鞘发育不良的形态学和遗传学初步研究(附3例报道)

目的 分析纤维鞘发育不良患者精子形态学特点,探寻其致病基因.方法 对3例表现为严重弱精子症的患者进行光镜下精液分析,通过扫描电镜和透射电镜进一步明确其超微结构特点.候选基因精子鞭毛蛋白2（SPEF2）和减数分裂特异性蛋白1（MNS1）全外显子测序,分析可能的致病突变位点.结果 3例患者均表现为100％（或接近）不活动精子,精子存活率26.0％～80.0％.光镜和扫描电镜下可见无尾、粗短尾、卷尾和不规则尾等严重畸形,DFS缺陷精子分别占99.5％,97.5％和79.5％.透射电镜表现为精子鞭毛纤维鞘等多种结构组装异常,伴有中心微管缺失（59％～78％）和动力蛋白臂缺失.MNS1和SPEF1基因外显子未见病理性突变.结论 DFS是严重弱精子症原因之一,根据形态学特点可以诊断;遗传学病因有待研究.

Ultrastructure of sperm in wild yak

The ultrastructure in sperm of wild yak can be used to examined and assess truly and all round the function and fertilization ability of the sperms. The ultrastructure in the sperm of wild yak was investigated under different states, such as fresh semen, frozen semen and capacitation sperm, and its characteristics were described by electron microscope. The sperm consisted of the head, neck and tail, the length was 78.34±7.24 μm. After capaeitation, the acrosome in sperm swelled and vesiculated, there were obvious vesicu]ation in the acrosome. The evagination on the ectoblast of acrosome formed the large vesiculation and inner membrane swelled. The evagination on the ectoblast of acrosome folded and tbrmed catenular vesiculation. The plasma membrane on the tail swelled. The head, neck and tail of the abnormal sperm were abnormal.