Sperm ultrastructure of the spider crab Maja brachydactyla (Decapoda: Brachyura)

This study describes the morphology of the sperm cell of Maja brachydactyla, with emphasis on localizing actin and tubulin. The spermatozoon of M. brachydactyla is similar in appearance and organization to other brachyuran spermatozoa. The spermatozoon is a globular cell composed of a central acrosome, which is surrounded by a thin layer of cytoplasm and a cup‐shaped nucleus with four radiating lateral arms. The acrosome is a subspheroidal vesicle composed of three concentric zones surrounded by a capsule. The acrosome is apically covered by an operculum. The perforatorium penetrates the center of the acrosome and has granular material partially composed of actin. The cytoplasm contains one centriole in the subacrosomal region. A cytoplasmic ring encircles the acrosome in the subapical region of the cell and contains the structures‐organelles complex (SO‐complex), which is composed of a membrane system, mitochondria with few cristae, and microtubules. In the nucleus, slightly condensed chromatin extends along the lateral arms, in which no microtubules have been observed. Chromatin fibers aggregate in certain areas and are often associated with the SO‐complex. During the acrosomal reaction, the acrosome could provide support for the penetration of the sperm nucleus, the SO‐complex could serve as an anchor point for chromatin, and the lateral arms could play an important role triggering the acrosomal reaction, while slightly decondensed chromatin may be necessary for the deformation of the nucleus. J. Morphol., 2010. © 2009 Wiley‐Liss, Inc.     

Spermatogenesis in the black porgy,Acanthopagrus schlegeli (teleostei: Perciformes: Sparidae)

The ultrastructure of spermatogenesis and of the spermatozoon of Acanthopagrus schlegeli (Sparidae) are described. The testis is of the unrestricted type. Germ cells are surrounded by cyst cells. Spermiogenesis involves conspicuous modifications such as intracellular movements (diplosome and mitochondria migration, nuclear rotation, and depression) and structural changes (chromatin condensation, shape of mitochondria, and loss of cytoplasm). The mature spermatozoon has a spherical nucleus with a deep, axial nuclear fossa, and an unusual notch, shaped like a bow tie. The short midpiece contains four spherical mitochondria and encircles the basal body of the flagellum. It is concluded that the A. schlegeli spermatozoon is of a primitive type, but that it is characterized by a unique feature which may provide a useful systematic character. © 1993 Wiley-Liss, Inc.     

The penetration of the spermatozoon through the sea urchin egg surface at fertilization : Observations from the outside on whole eggs and from the inside on isolated surfaces

The external and cytoplasmic surfaces of the sea urchin egg at fertilization have been examined with the scanning electron microscope (SEM). The outside events were documented by glueing eggs to polylysine coated glass plates, adding sperm and fixing rapidly. To reveal the inner aspects of the surface as the sperm travels through it to reach the egg cytoplasm, the fertilized egg surface was isolated in 0.3 M KC1, 0.35 M glycine, 2 mM MgCl₂, 2 mM EGTA, pH 7.5, glued onto a polylysine-coated plate and processed for the SEM. The events of spermatozoon attachment, membrane fusion, sperm entry, rotation and detachment into the egg cytoplasm as well as the associated cortical changes are described. The egg cortex is revealed to be a uniform network of fibrous bundles.The spermatozoon initially attaches to the egg surface by the acrosomal filament. As membrane fusion occurs between the gametes, the plasma membrane of the egg engulfs the sperm, the cortical granules start to discharge and a spreading surface deformation, possibly caused by a cortical contraction, is initiated. The perpendicularly entering spermatozoon is surrounded by a cluster of elongate microvilli which appear to have 235 nm vesicles associated with their bases. The sperm is prevented by the cortex from directly entering the egg cytoplasm and lies upon the egg surface between the plasma membrane and the matrix of cortical fibers. It is subsequently rotated additionally to enter the egg cytoplasm with the posterior end first. A scar is left in the cortex where the spermatozoon penetrated. The egg cortex is shown to consist of 50–200 nm uniformly arranged fibers, and its thickness ranges from 0.2 to 0.5 μm. It is speculated that this structure may be contractile.     

Scanning electron microscopic investigation of the spermatophore and spermatozoa of the shrimp Farfantepenaeus subtilis (Decapoda: Penaeidae)

Scanning electron microscopy was used to describe the structure of the spermatozoon and spermatophore of Farfantepenaeus subtilis. The spermatophore showed characteristics similar to those of members of the subgenus Farfantepenaeus. This included an extensive glandular epithelium and a lack of a wing. The sperm mass, which was distributed at the periphery of the spermatophore, was surrounded by a large amount of acellular material. The spermatozoon has a spherical main body and a well-defined acrosomal region with a single spike, which was bent in some cells. The immotile sperm cells have an average length of 7.1?±?0.6?μm. Information on sperm location within the spermatophore will assist in the efficient extraction of the sperm mass during dissection.     

The aflagellate spermatozoon of Diplozoon (Platyhelminthes: Monogenea: Polyopisthocotylea): a demonstrative case of relationship between sperm ultrastructure and biology of reproduction

Diplozoon is known to display an exceptional biology of reproduction: the hermaphroditic adults are permanently fused together and their genital ducts communicate. In contrast to all other polyopisthocotylean monogeneans in which the spermatozoa show an homogeneous biflagellate structure, the spermatozoon of Diplozoon is aflagellate. It is filiform, and composed of a cytoplasmic region and a nuclear region. The cytoplasmic region exhibits mitochondria, a well-developed smooth endoplasmic reticulum, and up to 450 longitudinal singlet microtubules. The microtubules show links between them; seen in cross section, they are arranged as rows or polygons. The spermatozoon nuclear region contains the nucleus surrounded by cortical longitudinal microtubules. The spermiogenesis shows no zone of differentiation, a typical structure found in all other parasitic Platyhelminthes. Diplozoon is the first case of aflagellate spermatozoon found in the parasitic Platyhelminthes. The atypical sperm structure is not linked with phylogeny, but is well correlated with the atypical biology of reproduction.     

Fine structure of the unistellate sperm of the shrimp, Sicyonia ingentis (Natantia)

Sperm of the prawn Sicyonia ingentis were studied cytochemically and ultrastructurally. Striking cytological differences were noted between these natantian sperm and previously studied reptantian sperm. In general, the S. ingentis sperm are composed of a spherical main body that is partially encompassed by a morphologically diverse cap region, from which extends a single appendage or spike. The main body houses an uncondensed, Feulgen-positive nuclear region that is partially surrounded by a cytoplasmic band. A single layer of small, 600 Å, vesicles lines the periphery of the cytoplasmic band. Large membranous vesicles extend from the inner surface of the cytoplasmic band into the nuclear region. The nucleus is separated from the cap or acrosomal complex by a dense plate and a highly organized crystalline lattice, which is composed of geometric squares that are approximately 350 Å in dimension. The cap region also contains convoluted membrane pouches; a central granular core; spherical bodies; an electron-dense, saucer-shaped plate; and a large anterior granule. The convoluted membrane pouches and anterior granule are periodic acid-Schiff (PAS) positive. The anterior granule also demonstrates RNAase-stable red fluorescence with acridine orange staining. A spiralled spike, approximately 6 μm long, extends from the anterior end of the cap. The cap and spike are bound by a double membrane, which results from the fusion of the plasma membrane and the convoluted pouch membrane. The sperm's acrosome is thought to be composed of the two PAS-positive cap components and the spike.     

Effects of scrotal heating on sperm surface protein PH-20 expression in sheep

Sperm surface protein PH-20 expression was studied during spermatogenesis in pubertal and adult sheep, using molecular and histological methods. The effects of 24 hr of insulation raising scrotal temperatures to 39 degrees C on PH-20 expression in ejaculated sheep sperm were also determined. A 282 nt cDNA fragment of ovine PH-20 was identified in total RNA extracts of sheep testes, which exhibited 76% identity at the nucleotide level with the equivalent region of the human sequence. Ovine PH-20 mRNA and immunoreactivity were identified only in adult ram testis and not in peri-pubertal ram testis tubules lacking round spermatids, nor in adult sheep brain, pituitary, heart, spleen, lung, liver, kidney, epididymis, or ovary. Ovine PH-20 protein was distributed predominantly on the postacrosomal membrane and was also present on the anterior membrane of the sperm head in fresh, unheated sheep semen. Scrotal heating caused a significant, transient decrease in the percentage of PH-20 immunoreactive sperm, but did not change the pattern of PH-20 staining on the sperm head. The results strongly suggest that ovine PH-20 is postmeiotically expressed in haploid germ cells in sheep testis and is arrayed on the membrane of the mature ovine spermatozoon. Scrotal heating appears to have few effects on PH-20 expression and distribution on ejaculated sperm.     

Dirofilaria immitis: ultrastructural aspects of oocyte development and zygote formation.

Ultrastructural observations of the ovary and uterus of Dirofilaria immitis reveal some characteristics of oogonia, oocytes, and uterine sperm. Oogonia are confined to the distal portion of the ovary including a blind tip, where a morphologically distinct terminal cap cell was not observed. These cells contain a nucleus with a nucleolus, numerous dense bodies, scanty ribosomes, lipid droplets, and an occasional mitochondrion. Endoplasmic reticulum is lacking and Golgi complexes were observed only in fully grown oogonia. Primary oocytes located in the middle portion of the ovary are large, elongate, and have a complete set of organelles including many small mitochondria, fragmentary endoplasmic reticulum, ribosomes, Golgi complexes, and very few dense bodies. These cells are arranged into many rosettes about central cytoplasmic masses, the rachises, to which they maintain cytoplasmic continuity by pseudopodlike processes. The rachises contain no organelle except a few dense granules and are bound by winding membranes. Oocytes from the proximal portion differ from those of the middle portion of the ovary in their larger size, round shape, absence of many organelles, presence of small dense granules, and lacking a rachis. Dense bodies are specific to the oogonia and exhibit DNase susceptibility and a positive reaction for a mitochondrial enzyme. These findings together with their decreased number and a concomitant increase of mitochondria in the oocytes suggest a relationship between these bodies and mitochondria.Uterine sperm of D. immitis are of the amoeboid type and contain several chromatin masses without a nuclear envelope, many mitochondria, and specialized membranous organelles referred to as mesosomelike vesicles. The vesicles are probably originated from the sperm plasma membrane. Upon fertilization, the entire spermatozoon penetrates the oocyte and its contents are gradually dissolved in the ooplasm with a simultaneous appearance of large numbers of ribosomes at the site of dissolution. Ribosomes were later found in the nucleus. A pronucleus was not observed. These findings are basically in agreement with those described for Ascaris but differ in the morphologic features and number of rachises, presence of dense bodies, absence of refringent granules in the oocytes and the absence of a refringent body and presence of several chromatin masses in the sperm.     

Ultrastructure of spermiogenesis and synthesis of enigmatic cytoplasmic inclusions in the spirally shaped spermatozoon of the polychaete Spio setosa (Annelida: Spionidae)

The sperm of Spio setosa (Polychaeta, Spionidae) are known to be very unusual in form; here, spermiogenesis and the structure of the spermatozoon in this species are described by transmission electron microscopy. While spermiogenesis is similar to that described for many other polychaetes, two notable exceptions to this process include the synthesis of abundant ring‐shaped and tubular, membrane‐bounded cytoplasmic inclusions in the midpiece, and the differentiation of a spirally shaped sperm head. Spermatids develop as free‐floating tetrads in the male's coelom. A microtubular manchette does not develop during chromatin condensation and nuclear elongation, and the spiral acrosome forms as a single Golgi‐derived vesicle that migrates anteriorly to become housed in a deep anterior nuclear fossa. Early in spermiogenesis, numerous Golgi‐derived, membrane‐bounded cytoplasmic inclusions appear in the cytoplasm; these ultimately occupy the sperm midpiece only. The mature spermatozoon in the male has a 15‐μm‐long head consisting of a nucleus coiled like a spring and a spiral acrosome with differentiated substructure, the posterior two thirds of which sits in an anterior nuclear fossa. The midpiece is wider than the rest of the spermatozoon and contains 9–10 spherical mitochondria surrounding the two centrioles, as well as numerous membrane‐bounded conoid and tubular cytoplasmic inclusions. The axoneme has a 9 + 2 arrangement of microtubules. By contrast, stored sperm in the female's seminal receptacles have lost the midpiece inclusions but contain an abundance of glycogen. The function of the midpiece inclusions remains unresolved, and the significance of their absence in stored sperm within the seminal receptacle and the appearance of midpiece glycogen stores remains unclear and requires additional investigation.     

THE SURFACE EVENTS AT FERTILIZATION OF THE SEA URCHIN EGG I. EVENTS ON THE SURFACE OF THE VITELLINE COAT1

Sperm-egg interaction during normal fertilization in the sea urchins, Strongylocentrotus intermedius and Hemicentrotus pulcherrimus, was studied by scanning and transmission electron microscopy. Several seconds after insemination, acrosome-reacted spermatozoa were found attached to the surface of the vitelline coat on each egg. Soon, several bulges of the vitelline coat appeared surrounding the fertilizing spermatozoon. These bulges then spread over the surface increasing in number, while they became fewer and disappeared around the sperm head. Thin sections of the bulging areas revealed discharging cortical granules. As the bulging vitelline coat was elevated, the sperm head was incorporated into the perivitelline space, passing through a small hole in the coat that resulted from penetration of the sperm acrosomal process immediately before fusion of the gametes. When the spermatozoon disappeared beneath the fertilization membrane, a hole was left in the membrane and the cortical reaction had finished on the other hemispheric surface. Mechanical removal of the membrane at that time exposed a spermatozoon protruding perpendicularly from the egg plasma membrane surface. The anterior tip of the sperm head was smoothly connected with the egg surface, and neither microvillous projections nor cytoplasmic covering of the egg cytoplasm could be found around the spermatozoon.     

CHANGES IN THE SPERMATOZOON DURING FERTILIZATION IN HYDROIDES HEXAGONUS (ANNELIDA) : II. Incorporation with the Egg

This, the last of a series of three papers, deals with the final events which lead to the incorporation of the spermatozoon with the egg. The material used consisted of moderately polyspermic eggs of Hydroides hexagonus, osmium-fixed at various times up to five minutes after insemination. The first direct contact of sperm head with egg proper is by means of the acrosomal tubules. These deeply indent the egg plasma membrane, and consequently at the apex of the sperm head the surfaces of the two gametes become interdigitated. But at first the sperm and egg plasma membranes maintain their identity and a cross-section through the region of interdigitation shows these two membranes as a number of sets of two closely concentric rings. The egg plasma membrane rises to form a cone which starts to project into the hole which the spermatozoon earlier had produced in the vitelline membrane by means of lysis. But the cone does not literally engulf the sperm head. Instead, where they come into contact, sperm plasma membrane and egg plasma membrane fuse to form one continuous membranous sheet. At this juncture the two gametes have in effect become mutually incorporated and have formed a single fertilized cell with one continuous bounding membrane. At this time, at least, the membrane is a mosaic of mostly egg plasma membrane and a patch of sperm plasma membrane. The evidence indicates that the fusion of the two membranes results from vesiculation of the sperm and egg plasma membranes in the region at which they come to adjoin. Once this fusion of membranes is accomplished, the egg cytoplasm intrudes between the now common membrane and the internal sperm structures, such as the nucleus, and even extends into the flagellum; finally these sperm structures come to lie in the main body of the egg. The vesiculation suggested above appears possibly to resemble pinocytosis, with the difference that the vesicles are formed from the plasma membranes of two cells. At no time, however, is the sperm as a whole engulfed and brought to the interior of the egg within a large vesicle.     

Spermatogenesis in a Free-Living Marine Nematode Neochromadora poecilosoma (Chromadorida,Chromadoridae) Studied Using TEM

Spermatogenesis and the structure of mature spermatozoa were studied using TEM in a free-living marine chromadorid nematode Neochromadora poecilosoma from the Sea of Japan. In spermatocytes, fibrous bodies (FB) develop; in spermatids, the synthetic apparatus lies in the residual body, while the nucleus, mitochondria, and FB are located in the main cell body (MCB). The nucleus consists of a diffuse chromatin of fibrous structure, which is not enclosed in a nuclear envelope. In the spermatid stage, the development of FB is completed, and immature spermatozoa from the proximal region of the testis do not show any structural differences from the MCB of spermatids. The mature spermatozoa are polarized cells. They attach to the uterus wall by a pseudopod filled with filaments of the cytoskeleton; in the MCB of spermatozoon, there is a nucleus surrounded by mitochondria and osmiophilic bodies. The spermatozoa of N. poecilosoma show typical ultrastructure features of sperm cells found in most studied nematodes (amoeboid nature and the absence of axoneme, acrosome, and nuclear envelope). However, no aberrant organelles characteristic of nematode spermatozoa were found throughout sperm development in N. poecilosoma and other chromadorids.     

Spermatogenesis and sperm structure of Acerella muscorum, (Ionescu, 1930) (Hexapoda, Protura)

The general organization of the male genital system, the spermatogenesis and the sperm structure of the proturan Acerella muscorum have been described. At the apex of testis apical huge cells are present; their cytoplasm contains a conventional centriole, a large amount of dense material and several less electron-dense masses surrounded by mitochondria. Spermatocytes have normal centrioles and are interconnected by cytoplasmic bridges. Such bridges seem to be absent between spermatid cells and justify the lack of synchronization of cell maturation. Spermatids are almost globular cells with a spheroidal nucleus and a large mass of dense material corresponding to the centriole adjunct. Within this mass a centriole is preserved. Mitochondria of normal structure are located between the nucleus and the plasma membrane. The spermatids are surrounded by a thick membrane. No flagellar structure is formed. Sperm have a compact spheroidal nucleus, a large cap of centriole adjunct material within which a centriole is still visible. A layer of mitochondria is located over the nucleus. The cytoplasm is reduced in comparison to spermatids; many dense bodies are interspersed with sperm in the testicular lumen. The sperm are small, immotile cells of about 2.5-3 μm in diameter.     

Penetration of spermatozoon into the ovum and transformation of the sperm nucleus into the male pronucleus in the domestic fowl,Gallus gallus

Summary The apex of the sperm head which has undergone the acrosome reaction comes in contact with the plasma membrane of the ovum. After the entire surface of the inner acrosomal membrane has come into close contact with the plasma membrane of the ovum, the two membranes fuse to form a continuous membrane. All parts of the spermatozoon that are devoid of plasma membrane penetrate into the ooplasm. As the head of the spermatozoon moves deeper into the ooplasm, the chromatin begins to disperse, and the head of spermatozoon is transformed into a large spherical nucleus with low electron density. At a later stage of the transformation, many small vesicles appear around the nucleus and subsequently fuse to form two continuous membranes. These membranes represent the male pronuclear envelope. The condensation of the chromatin occurs in places in the nucleus, so that the male pronucleus is formed. During the course of the formation of the male pronucleus, the subacrosomal rod and tail become detached from the head and disintegrate.The authors are greatly indebted to assoc. Prof. Dr. Osamu Koga for his valuable advices. The authors also wish to thank Mr. Takayuki Mori for his helpful suggestions and technical advices. This investigation was supported by a grant from the Ministry of Education of Japan (156185)     

Impact of ivermectin on the ultrastructure of the testis of <Emphasis Type="Italic">Argas (Persicargas) persicus</Emphasis> (Ixodoidea: Argasidae)

Mated male Argas persicus were dissected 1 and 2 weeks after feeding on untreated and ivermectin (IVM)-treated pigeons. One week after feeding, testes of untreated ticks were filled with rounded spermatids with subplasmalemmal vesicles and cytoplasmic organelles, but lacking in treated ticks. Two weeks after feeding, testes were crowded with elongated spermatozoa supported by double-walled cisternal tubes. The tubes consisted of two opposite walls, each with outer-fringed processes and inner elongated cisternae. Both were supported with electron dense striated plates in the middle of the spermatozoon. Internally, the cisternal tubes contained mitochondria and vacuoles. The nuclei were elongated dense masses between the tubes and the cell membranes. Subcutaneous inoculation of IVM at the dose 400 μg/kg pigeon resulted in extensive alterations in the testis of A. persicus. IVM prevented the development of new spermatids. There was a break down of cell membranes and cytoplasmic organelles of spermatozoa. Multivesicular bodies and numerous vacuoles were noticed in their cytoplasm. Double membranes of elongated cisternae and striation of electron dense plates became indistinct. IVM caused granulation and vacuolization of the nucleus as well as injury of mitochondrial cristae. The results suggest that IVM may bind to the neurotransmitter or the hormone involved in the process of sperm development or may be toxic to the germinal cells of A. persicus testis.     

拟异蚖和古蚖精子的超微形态和精子形成的研究(原尾目:古蚖科)

余山拟异蚖和3种古蚖的精子均为扁圆形,未见顶体,线粒体集中在一侧;核呈环形、边位、中部由膜状体分布其间.领结古蚖的早期精细胞为球形,染色质凝集成团,继而核中裂并沿细胞赤道逐渐围绕成环,染色质呈细沙状,胞间有“桥”相通.核膜一端开始内陷,出现黑点.待发育到中期精细胞,这些黑点逐渐形成奇特的管状核膜陷体;染色质变成短线形,随后排成4—5行.线粒体颗粒状,细胞间仍有“桥”连通.晚期精细胞的染色质凝集成粗带,最后形成光滑质密的核,而多余的核物质,一段一段从精子一端脱离,形成一串孢囊状体夹在精子之间,待精子成熟游离时,这些孢状体分散开来.从观察结果表明拟异蚖精子与古蚖的非常相近.     

Morphological changes in eupyrene and apyrene spermatozoa in the reproductive tract of the male butterfly Atrophaneura alcinous Klug

Summary

Eupyrene and apyrene spermatozoa are contained in separate cysts in the testis of the butterfly Atrophaneura alcinous. Spermatozoa of both types from various parts of the male reproductive tract were examined with particular reference to their morphological characteristics. All spermatozoa collected from the vas deferens and the vesicula seminalis were found to be immotile under a dissecting microscope. No spermatozoa of either type were recognized in any part of the ejaculatory duct. Within the testis, eupyrene spermatozoa are present in bundles and each spermatozoon has a slender nucleus with an acrosome and a long flagellum containing mitochondrial derivatives. Two kinds of appendages, lacinate and reticular, are present on the surface of the sperm membrane. They are replaced with an extracellular sheath during passage through the vas deferens. In contrast, apyrene spermatozoa have neither nucleus nor acrosome, whereas a cup-shaped structure was found at the sperm tip instead of the acrosome. Unlike eupyrene spermatozoa, they are surrounded by a concentric sheath outside the sperm membrane in the vas deferens. Individual apyrene spermatozoa and coiled bundles of eupyrene spermatozoa were both found to accumulate in the vesicula seminalis before mating. These morphological changes during passage through the male reproductive tract suggests the occurrence of a kind of maturation and capacitation process reminiscent of mammalian spermatozoa.     

Ultrastructure of the sperm and spermatogenesis and spermiogenesis of Dina lineata (hirudinea,erpobdellidae)

The mature sperm of Dina lineata is of the modified type. The sperm are 48 μm long and 0.3 μm wide. The sperm are filiform and helicoidal cells with a distinct head, a midpiece, and a tail. There are two distinct regions in the head: the acrosome and the posterior acrosome, each with its own characteristic morphology. The midpiece is the mitochondrial region and has a single mitochondrion. Two distinct portions can be observed in the tail: the axonematic region and the terminal piece. In the process of spermatogenesis the early spermatogonia divide to form a poliplast of 512 spermatic cells. In the spermiogenesis the following sequential stages can be distinguished: elongation of the flagellum; reciprocal migration of mitochondria and Golgi complex; condensation of chromatin and formation of the posterior acrosome; spiralization of nuclear and mitochondrial regions; and, finally, formation of the anterior acrosome. The extreme morphological complexity of the Dina spermatozoon is related to the peculiar hypodermal fertilization which characterizes the erpobdellid family. Correlation between sperm morphology and fertilization biology in the Annelida is revised.     

The passage of spermatozoa through the vitelline membrane in the domestic fowl,Gallus gallus

Summary The developing outer layer of the vitelline membrane of the ovum in the posterior part of the infundibulum of the domestic fowl contains many spermatozoa in nearly parallel orientation with its inner layer. When the acrosomal region of a spermatozoon approaches or contacts the inner layer, promptly undergoes the acrosome reaction. The outer acrosomal membrane and overlying plasma membrane fuse together and the apical region of the acrosome opens, so that the acrosomal contents are released. Meanwhile the spermatozoon remains a time in contact with the surface of the inner layer, and the network of the inner layer just under the tip of the sperm head begins to be dissolved. This dissolution extends downward forming a tunnel, approximately 9 m in diameter. The spermatozoon then passes through the inner layer obliquely via the central region of the tunnel and arrives at the perivitelline space.The authors are greatly indebted to assoc. prof. Dr. Osamu Koga for his valuable advices. The authors also wish to thank Mr. Takayuki Mori for his helpful suggestions and technical advices. This investigation was supported by a grant from the Ministry of Education of Japan (156185)     

SPERMIOGENESIS IN CANCER CRABS

Spermiogenesis in Cancer crabs was studied by light and electron microscopy. The sperm are aflagellate, and when mature consist primarily of a spherical acrosome surrounded by the nucleus with its short radiating arms. The acrosome forms by a coalescence of periodic acid-Schiff-positive (PAS-positive) vesicles. During spermiogenesis one edge of the acrosomal vesicle invaginates to form a PAS-negative central core. The inner region of the acrosome bounding the core contains basic proteins which are not complexed to nucleic acid. The formation of an elaborate lattice-like complex of fused membranes, principally from membranes of the endoplasmic reticulum, is described. These membranes are later taken into the nucleus and subsequently degenerate. In late spermatids, when most of the cytoplasm is sloughed, the nuclear envelope and the cell membrane apparently fuse to become the limiting boundary over most of the sperm cell. In the mature sperm the chromatin of the nucleus and arms, which is Feulgen-positive, contains no detectable protein. The chromatin filaments appear clumped, branched, and anastomosed; morphologically, they resemble the DNA of bacterial nuclei. Mitochondria are absent or degenerate in mature sperm of Cancer crabs, but the centrioles persist in the nucleoplasm at the base of the acrosome.