Spermiogenesis and spermatozoa ultrastructure in Salminus and Brycon, two primitive genera in Characidae (Teleostei: Ostariophysi: Characiformes)

In Salminus, spermiogenesis is cystic and gives origin to a type I aquasperm. Spermatid differentiation is characterized by chromatin condensed into thick fibres, nuclear rotation, nuclear fossa formation, cytoplasmic channel formation, mitochondrial fusion producing long and ramified mitochondria, and the presence of several membranous concentric rings around the plasma membrane that encircles the cytoplasmic channel. In Salminus and Brycon, spermatozoa are very similar. They exhibit a spherical nucleus and chromatin condensed into fibre clusters, and a deep nuclear fossa. They show a long midpiece with few elongate mitochondria at the initial region and a cytoplasmic channel completely encircled by one or two membranous concentric rings. The flagellar axis is perpendicular to the nucleus and exhibits the classic axoneme (9 + 2). The very strong similarity observed between Salminus and Brycon spermatozoa supports the hypothesis that these subfamilies are likely to have a monophyletic origin.     

Ultrastructure of Thunnus thynnus and Euthynnus allettevatus spermatozoa

The spermatozoa of Thunnus thynnus and Euthynnus alletteratus consist of an acrosome-less head (comprising the ovoid nucleus and the short midpiece) and a long flagellar tail that contains the conventional 9 + 2 axoneme and lacks lateral fins. The centrioles are arranged at approximately right angles and lie outside of a shallow nuclear groove. The flagellum inserts laterally on the nucleus, therefore the spermatozoon is asymmetrical. The midpiece contains a few mitochondria which are separated from the axoneme by the cytoplasmic canal; they are spherical in T. thynnus and elongate, somewhat irregular in E. alletteratus . Although the main ultrastructural characteristics of the spermatozoa appear to indicate a great homogeneity in the sperm morphology within the family Scombridae, small species-specific divergences may be of use in systematics.     

Spermatozoon Ultrastructure of Two Holothurian Species of the Genus Cucumaria (Dendrochirotida,Holothuroidea) from the Sea of Japan

The ultrastructure of spermatozoa of Cucumaria japonica and a congeneric morphologically similar deep-sea species was studied. The spermatozoa of both C. japonica and C. conicospermium are similar to those of other holothurians: the acrosome is composed of an acrosomal granule and periacrosomal material; the centrioles lie at an acute angle to one another; and the proximal centriole is connected to the nuclear envelope by a flagellar rootlet. The spermatozoa of C. japonica differ from those of C. conicospermium in the shape of the head and the dimensions and position of the acrosome. In C. japonica, the acrosome is completely embedded in the nuclear fossa and measures 0.7 m. In C. conicospermium, only one-third of the acrosome is embedded in the nuclear fossa; this acrosome measures 1.3 m. A correlation between the structure of the sperm acrosome and that of the egg envelope is discussed.     

Spermatozoa ultrastructure in Sciaenidae and Polynemidae (Teleostei:Perciformes) with some consideration on Percoidei spermatozoa ultrastructure

Spermatozoa ultrastructure was studied in five marines (Paralonchurus brasiliensis, Larimus breviceps, Cynoscion striatus, Micropogonias furnieri, Menticirrhus americanus, Umbrina coroides, Stellifer rastrifer), and one freshwater (Plagioscion squamosissimus) species of Sciaenidae and one species of Polynemidae (Polydactylus virginicus). The investigation revealed that, in all species, spermatozoa display a round head, a nucleus containing highly condensed, filamentous chromatin clusters, no acrosome, a short midpiece with a short cytoplasmic channel, and a flagellum showing the classic axoneme structure (9+2) and short irregular lateral fins. In Sciaenidae, the spermatozoa are type II, the flagellar axis is parallel to the nucleus, the lateral nuclear fossa is double arched, the centriolar complex is outside the nuclear fossa, the proximal centriole is anterior and perpendicular to the distal centriole, and no more than ten spherical (marine species) or elongate (freshwater species) mitochondria are observed. Polynemidae spermatozoa are of the intermediate type with the flagellar axis eccentric to the hemi-arc-shaped nucleus, and exhibit no nuclear fossa, the centriolar complex close to the upper nuclear end, the proximal centriole lateral and oblique to the distal centriole, and one large ring-shaped mitocondrion. The data available show that no characteristic is exclusively found in the spermatozoa of members of the Sciaenidae family when compared to other Percoidei with type II spermatozoa. However, three characteristics were exclusively found in Polynemidae: (1) the hemi-arched nucleus; the positioning of the centrioles; and (2) the ring-shaped mitocondrion. The interrelationships between Sciaenidae and Polynemidae as well as between these two families and other Percoidei are herein discussed.     

Sperm ultrastructure and a new type of spermiogenesis in two species of Pimelodidae, with a comparative review of sperm ultrastructure in Siluriformes (Teleostei: Ostariophysi)

During spermiogenesis, the spermatids of the pimelodid species Pimelodus maculatus and Pseudoplatystoma fasciatum show a central flagellum development, no rotation of the nucleus, and no nuclear fossa formation, in contrast to all previously described spermatids of Teleostei. These characteristics are interpreted as belonging to a new type of spermiogenesis, named here type III, which is peculiar to the family Pimelodidae. In P. maculatus and P. fasciatum, spermatozoa possess a spherical head and no acrosome; their nucleus contains highly condensed, homogeneous chromatin with small electron-lucent areas; and a nuclear fossa is not present. The centriolar complex lies close to the nucleus. The midpiece is small, has no true cytoplasmic channel, and contains many elongate and interconnected vesicles. Several spherical to oblong mitochondria are located around the centriolar complex. The flagellum displays the classical axoneme (9+2) and no lateral fins. Only minor differences were observed among the pimelodid species and genera. Otherwise, spermiogenesis and spermatozoa in the two species of Pimelodidae studied exhibit many characteristics that are not found in other siluriform families, mainly the type III spermiogenesis.     

Structure and function of the spermathecal complex in the phlebotomine sandflyPhlebotomus papatasi Scopoli (Diptera: Psychodidae): II. Post-copulatory histophysiological changes during the gonotrophic cycle

The spermathecal complex ofPhlebotomus papatasi Scopoli (Diptera: Psychodidae) undergoes histological and physiological changes during its gonotropic cycle. The present histochemical study revealed a mucopolysaccharide secretory mass in the spermathecae of the newly emerged sandfly. Sperm competition occurs when two or more males compete to fertilize an ovum in the female reproductive tract. In this study, spermatophores of two or more competing males were deposited at the base of the spermathecal ducts, which originate from the female bursa copulatrix. This suggests that females play a role in sperm displacement, which is defined as any situation in which the last male to mate with a female fertilizes maximum number her eggs. A blood meal ingested by the female for ovary development and egg laying stimulates the release of sperm from the spermatophore. The spermatozoa then migrate to the lumen of the spermatheca. The ultrastructure of spermatozoa comprises a head with double-layered acrosomal perforatorium, an elongate nucleus, and the axoneme with a 9 + 9 + 0 flagellar pattern. This axomene differs from the aflagellate axoneme of other Psychodinae. Morphological changes, such as the casting off of the acrosomal membrane, and histological changes in the spermatophore are also described. Mating plugs that have been described previously in sandflies appear to be artefacts. Females ofP. papatasi may be inseminated more than once during each gonotrophic cycle, and additional inseminations may be necessary for each cycle. The relationships between the volumes of the sperm and the spermatheca were calculated to determine sperm utilization and fecundity ofP. papatasi. As the females ofP. papatasi mate polyandrously, the anatomical and physiological complexity of the spermathecal complex may be related to post-copulatory sexual selection.     

Comparative morphological and photosynthetic studies on three Malaysian species of Pogonatum from habitats of varying light irradiances

Abstract

Comparative morphological and physiological studies were carried out on three species of Malaysian Pogonatum collected from habitats exposed to different light intensities ranging from 28±4 W/m² for P. cirratum subsp. macrophyllum (Dozy & Molk.) Hyvönen, to 230±39 W/m² for P. subtortile (Müll.Hal.) A. Jaeger to 751±45 W/m² for P. neesii (Müll.Hal.) Dozy. Total chlorophyll and beta-carotene content were higher in P. cirratum subsp. macrophyllum and P. subtortile than in P. neesii when calculated on a fresh weight basis. However, soluble protein content was higher in the sun species compared to its shade-adapted counterparts, and the soluble protein to total chlorophyll ratio was highest in P. neesii. The chloroplasts in the leaves of P. cirratum subsp. macrophyllum were significantly larger than those in the other two species, and had more grana and thylakoids per chloroplast profile. The numbers of starch grains in P. cirratum subsp. macrophyllum and P. subtortile were more than double that observed in P. neesii. Morphological studies of the leaves showed specific differences in the shapes and heights of the lamellae found on the adaxial surfaces of the leaves. The lamellae of P. cirratum subsp. macrophyllum were rudimentary whilst those of P. subtortile were 2–3 cells high and P. neesii possessed lamellae 5–7 cells high. These findings indicate a direct correlation between the height of the lamellae and the light irradiances received by the plants. Determination of the in vitro Photosystem II photochemical rates, with an oxygen electrode, of chloroplasts isolated from these plants, showed that P. neesii and P. subtortile exhibited higher rates than P. cirratum subsp. macrophyllum. Similarly in vivo light saturation studies with an infrared gas analyzer showed that CO₂ assimilatory rates were highest in P. neesii, even at low light intensities, suggesting a relationship between photosynthesis and light irradiance in these mosses that is different from vascular plants.     

LIGHT AND ELECTRON MICROSCOPICAL OBSERVATIONS OF THE DINOFLAGELLATE ACTIMSCUS PESTASTERIAS (DINOPHYCEAE)1

I examined the heterotrophic non-armored dinoflaget-late Actiniscus pentasterias (Ehrenberg) Ehrenberg by light and electron microscopy. Actiniscus pentasterias contains an internal skeleton consisting of two star-like siliceous elements. Special emphasis is given to the flagellar apparatus, the nucleus, and a new type of extrusome, named a docidosome. A three dimensional model of the flagerllar apparatus includes a fibrous nuclear connnective, a posterior striated root, and a dorsal striated component of the longitudinal microtabular root. The nucleus is surrounded by a conspicuous fibrous lamina, also visible in the light microscope. The nuclear pores are situated in annulated invaginations of the nuclear envelope, increasing the nuclear surface area by 15–25%. The docidosomes are rod-shaped membrane-bound structures that terminate in a distinct proximal head. They show very complex substructure, consisting of an inner medulla with highly ordered paired ribbons and an outer cortex.     

A comparison of the structure of the spermatozoa and spermatogenesis of 16 species of patellid limpet (Mollusca: Gastropoda: Archaeogastropoda)

Light and transmission electron microscopy of the spermatozoa and spermatogenesis of 16 species (in three genera, Patella, Helcion, Cellana) of patellid limpet have shown that head lengths of the sperm range from 3 to 13 μm, and each species has a sperm with a unique morphology, indicating that the spermatozoa can be used as a taxonomic character. Although spermatozoon structure is species specific, five types can be recognized, based on the size, shape, and structure of the nucleus and acrosome. The occurrence of five morphological types of sperm, one of which (Cellana capensis) is particularly different from other patellids, suggests that the taxonomy of the family Patellidae be re-examined. The morphological changes that occur during spermatogenesis are very similar in all species, although two patterns of chromatin condensation are found. Those species with sperm that have short squat nuclei (length:breadth < 3.5:1) have a granular pattern of condensation. Species with sperm that have more elongate nuclei (length:breadth > 5:1) have an initial granular phase followed by the formation of chromatin fibrils. These fibrils become organized along the long axis of the elongating nucleus. The absence of a manchette suggests that nuclear elongation is brought about from within the nucleus.     

Spermatogenesis and the structure of the testes in Nemertodermatida

The ultrastructure of the testes in two representatives of the enigmatic taxon Nemertodermatida was studied using transmission electron microscopy. Nemertoderma westbladi has paired testes, which are delineated by lining cells. Within each testis, different follicles, each surrounded by a membrane-like structure, are found. Flagellophora apelti has genuinely follicular testes, consisting of several follicles, each containing a certain stage of spermatogenesis. As the gametes are not enclosed by a structure that can be called a true gonad, the structure of the testes differs from most bilaterian animals, but resembles the organization of gametogenic areas of ctenophores. Each stage of spermatogenesis in F. apelti is described, enabling the inference of the origin of the structures seen in mature spermatozoa. The overall structure of the mature spermatozoa is similar in all nemertodermatids and unique within the Metazoa: an elongated head containing the nucleus; a middle piece containing an axoneme, mitochondrial derivatives and in F. apelti granular derivatives; and a flagellar tail.     

Cauda epididymal spermatozoa of the rufous hare wallaby, Lagorchestes hirsutus (Metatheria, Mammalia) imaged by electron and confocal microscopy

The ultrastructure of mature Lagorchestes hirsutus spermatozoa is described for the first time, revealing unusual aspects of sperm structure in macropodid species. The sperm head is ovoid rather than cuneiform, lacks a ventral nuclear groove and has an acrosomal distribution over approximately 85–90% of its dorsal surface. Immediately adjacent to the nuclear membrane the peripheral nucleoplasm in most spermatozoa form an irregular series of distinctive evaginations previously not described in the spermatozoa of any other marsupial. The midpiece is extremely thickened and short, containing no helical network or peripheral plasma membrane specializations. Axonemal structure is unspecialized with no connecting lamellae; dense outer fibres are closely adherent to axonemal doublets. The sperm morphology of this species is highly aberrant in comparison to other macropod taxa and supports the retention of Lagorchestes as a distinctive genus. In light of this new information, skeletal and serological data should be re‐evaluated to determine the true taxonomic and phylogenetic position of this species.     

Juxtanuclear changes during the early spermatogenesis in Lebistes reticulatus (Guppy)

Summary The sperm cell of Lebistes reticulatus during Spermatogenesis is studied. The nuclear differentiation correlate to the flagellar outgrowth. Differential distribution in the chromatin condensation corresponds to a new membrane synthesis at the centriolar side of the nucleus. The fan-shaped microtubular bundle in Juxtanuclear position is described and its function as a stabilizing entity is discussed. At the centriolar complex satellites and an intercentriolar lamellated body are investigated. In submicroscopic details the sperm cell of Lebistes distinctively differs from other species previously described.The present investigation was supported by a grant from the Pehr Oscar Klingendahl Foundation.The authors wish to thank Mr. Mauri Nyholm, M. Sc., for his technical advices and Miss Maria Sjöstedt for preparing the specimens.     

The spermatozoa of ascidians: Acrosome and nuclear envelope

Summary The spermatozoon of Ascidia callosa has a head with a wedge-shaped tip. Between the nuclear envelope and the plasmalemma, at the tip of the head, there are one or two previously undescribed vesicles, 45 to 55 nm in diameter. These vesicles have the characteristics of an acrosome. Their role in the process of fertilization has not been determined. Ultrastructural studies of sperm activation are needed, but claims that the spermatozoa of ascidians do not have an acrosome should be reconsidered.Behind the tip of the sperm there are pores in the nuclear envelope. This part of the envelope also contains a dense band of amorphous material that may have a supportive function. A nearly identical structure, associated with pores has been found in the spermatozoon of Boltenia villosa. An analysis of the nuclear envelope of Ascidia callosa indicates that the same structure has previously been misinterpreted as an acrosome in the spermatozoon of Ascidia nigra.     

An ultrastructural examination of developing and mature paraspermatozoa in Pyrazus ebeninus (Mollusca,Gastropoda, Potamididae)

Summary The mesogastropod Pyrazus ebeninus, produces true spermatozoa (here termed euspermatozoa) and multi-flagellate, mobile cells (here termed paraspermatozoa). The mature paraspermatozoon consists of an elongateconical head (6.5–8.5 m in length), constructed of an electron-dense mosaic sheath surrounding a similarly dense, rod-shaped nuclear core (which runs almost the full length of the head). An acrosome-like structure forms the apex of the head. Five to eight axonemes are fixed to the posterior extremity of the nuclear core, each by means of an attachment complex (dense attachment rod, centriolar cap and centriole). A short (3–4 m) midpiece zone follows the head and consists of the multiple axonemes interspersed with very elongate mitochondria. A tuft of short (20 m) tails (termed minor tails) emerges from the midpiece in addition to one very long tail (termed the major tail) ensheathed in dense granules which resemble glycogen granules. A single membrane surrounds head, midpiece and tails whilst the nuclear core retains the original double nuclear membrane.Developmentally, the multiple axonemes arise from one of a pair of wheel-shaped arrangements of centrioles and attach to posterior indentations in the nucleus prior to its transformation into the nuclear core. Dense vesicles, derived apparently from the endoplasmic reticulum, accumulate along and around the developing nuclear core and (in the presence of microtubules) condense into the mosaic head sheath. Cytoplasmic mitochondria elongate and collect at the posterior axis of the cell, where, together with the axonemes, they form the midpiece.Features not previously reported in other ultrastructural studies of paraspermatozoa include the acrosome-like structure of the head, the structure of the midpiece zone, the glycogen sheath of the major tail, the dense annular structure at the junction of the midpiece and major tail and the presence of microtubules in the final phase of head and midpiece maturation. Some features of the euspermatozoon are also described and the comparative ultrastructure of mature and developing paraspermatozoa and their possible functions in the Gastropoda, are reviewed.Abbreviations ac euspermatozoon acrosomal cone - ar euspermatozoon axial rod - ax axoneme - b dense block of mosaic sheath - c centriole - cc centriolar cap - co cone of acrosome-like structure - dr dense attachment rod - dv dense vesicle - g glycogen granules - G Golgi complex - GER granular endoplasmic reticulum - H head of paraspermatozoon - m mitochondrion - M midpiece (euspermatozoon, paraspermatozoon) - maj major tail - min minor tails - mt microtubules - n nucleus - nc nuclear core - p dense plug of acrosome-like structure - pm plasma membrane - sGv small Golgi vesicles - Z transition of centriole to centriolar cap of attachment complex     

The restructuring of the flagellar base and the flagellar necklace during spermatogenesis of Ephestia kuehniella Z. (Pyralidae,Lepidoptera)

Summary Transmission electron microscopy was used to study the development of the flagellar base and the flagellar necklace during spermatogenesis in a moth (Ephestia kuehniella Z.). Until mid-pachytene, two basal body pairs without flagella occur per cell. The basal bodies, which contain a cartwheel complex, give rise to four flagella in late prophase I. The cartwheel complex appears to be involved in the nucleation of the central pair of axonemal microtubules. In spermatids, there is one basal body; this is attached to a flagellum. At this stage, the nine microtubular triplets of the basal body do not terminate at the same proximal level. The juxtanuclear triplets are shifted distally relative to the triplets distant from the nuclear envelope. Transition fibrils and a flagellar necklace are formed at the onset of axoneme elongation. The flagellar necklace includes Y-shaped elements that connect the flagellar membrane and the axonemal doublets. In spindle-containing spermatocytes, the flagellar necklace is no longer detectable. During spermatid differentiation, the transition fibrils move distally along the axoneme and a prominent middle piece appears. Our observations and those in the literature indicate certain trends in sperm structure. In sperms with a short middle piece, we expect the presence of a flagellar necklace. The distal movement of the transition fibrils or equivalent structures is prevented by the presence of radial linkers between the flagellar membrane and the axonemal doublets. On the other hand, the absence of a flagellar necklace at the initiation of spermiogenesis enables the formation of a long middle piece. Thus, in spermatozoa possessing an extended middle piece, a flagellar necklace may be missing.     

Differential basic nucleoprotein kinetics in the two kinds of lepidoptera spermatids: Nucleate (eupyrene) and anucleate (apyrene)

Normal lepidopteran males produce two kinds of spermatozoa: nucleate (eupyrene) and anucleate (apyrene). Eupyrene spermatozoa have the usual type of elongate nuclei. But in apyrene spermatids, the nuclei never elongate and the chromatin remains in a telophase-like condition until enucleation occurs. The study of the differential nucleoprotein kinetics of the two types of spermatids, using the fluorescent dye sulfoflavine, shows that: (1) In the elongate eupyrene nuclei, lysine-rich nucleoproteins are replaced by arginine-rich ones, while in the non-elongating apyrene nuclei only lysine-rich nucleoproteins are detected. However, nuclear elongation is not causally related to nucleoprotein transitions as transitions occur in the eupyrene spermatids after nuclear elongation. (2) The replacement of the nucleoproteins occurs in the eupyrene nuclei in a polarized manner. This may be correlated with the heterogeneous ultrastructural configuration of the chromatin fibers in elongating spermatid nuclei, as shown in other insect species. (3) Concomitantly with the eupyrene spermatid nucleoprotein transition, the cytoplasm of the head cyst cell shows an increasing amount of cytoplasmic lysine-rich proteins, while no such a phenomenon occurs in apyrene cysts. This differential pattern distribution may reflact functional differences among the two types of cysts and is probably related to the regulation of the dichotomy in lepidopteran spermatogenesis.     

A new species of saw‐scaled viper of the Echis coloratus complex (Ophidia: Viperidae) from Oman,Eastern Arabia

A study of 353 museum specimens of the Echis coloratus complex from its entire range of distribution revealed an undescribed species in the United Arab Emirates and northern Oman. The results of UPGMA clustering and principal coordinate analysis of 138 male and 142 female specimens yielded for both sexes two major clusters, one with specimens from the UAE and northern Oman and one from southern and western Arabia, the Levant and Egypt. The new species has a longer tail with higher subcaudal counts; the lower prenasal scale is often missing and the upper prenasal is frequently fused with the nasal; the subnasal is often missing or fused with the nasal. The gular scales between the chin‐shield and the preventrals are round or only slightly elongate, not elongate as in Echis coloratus, and their number is higher. Other differences in characters of the gular area indicate a different scale structure of the ventral surface of the head. The new species is allopatric or parapatric with E. coloratus, but sympatric with Echis carinatus sochureki.     

Ultrastructure of spermatids and spermatozoa in the cyclostomatous bryozoan Tubulipora (Bryozoa,Cyclostomata)

Summary Differentiation of spermatids to mature spermatozoa in the bryozoan Tubulipora liliacea was studied by transmission electron microscopy. The spermatozoon of Tubulipora is of a filiform, modified type, and has evolved from the primitive type as an adaptation to a specialized biology of fertilization. The head of the spermatozoon consists of a small, conical acrosome capping an elongated, cylindrical, anteriorly tapering nucleus. A basal invagination in the nucleus contains the proximal portion of the axoneme and a dense attachment matrix. The flagellar axoneme has the typical 9+2 structure. Four elongated rodshaped mitochondria with typical cristae surround the axoneme in the cylindrical middle piece. Granular electron-dense material is accumulated in the form of four columns alternating with four long cylindrical mitochondria. The mitochondrial middle piece is separated externally from the tail region by an involution of the plasma membrane. The tail region contains a cytoplasmic sheath with accessory fibers surrounding the axoneme. Nine outer, coarse fibers extend posteriorly paralleling the nine doublets of the axoneme. The coarse fibers develop from electron-dense plate-like structures associated with the doublets of the axoneme. A characteristic feature in spermiogenesis is that spermatozoa develop in tetrads. There seem to be significant differences in spermatozoan ultrastructure between the three bryozoan classes Stenolaemata, Gymnolaemata, and Phylactolaemata. The differences indicate different lines of evolution of fertilization biology in these groups.Abbreviations used in the figures a acrosome - av acrosomal vesicles - ax axoneme - c coarse fiber - d electron dense rod - m mitochondrion - mp middle piece - Scale bars=0.5 m - mt microtubule - n nucleus - ne nuclear envelope - p nuclear protrusion - pm plasma membrane - t tail