Male gonads morphology,spermatogenesis and sperm ultrastructure of the seahorse Hippocampus guttulatus (Syngnathidae)

Testes morphology, spermatogenetic process and mature sperm ultrastructure were analysed in Hippocampus guttulatus, using both light and transmission electron microscopy. Both testes were organized in a single large germinal compartment, with a central lumen. Spermatocysts only contained spermatogonia and primary spermatocytes. Inside the testis lumen, together with mature sperm, two types of large mono‐nucleate cells, flagellate and aflagellate, were present. Both types of cells were interpreted as developing germ cells precociously released inside the testis lumen, where their maturation was completed. According to the different morphological features of the nuclei, such as chromatin condensation degree, aspect of the nuclear fossa and others, the flagellate cells were unquestionably developing spermatids. On the contrary, the developmental stage of the aflagellate was more difficult to interpreted. They could be secondary spermatocytes or young spermatids. No dimorphic sperm were recognizable, the only sperm type observed have features typical of the intro‐sperm reports in other syngnathids species. They had a cylindrical head, a short midpiece, characterized by two mitochondrial rings housed inside a cytoplasmic collar, and a long flagellum. These and previous data about the same topic reported on other syngnathids species were compared and discussed.     

Ultrastructure and histochemistry of the testicular efferent duct system and spermiogenesis in Opistognathus whitehurstii (Teleostei, Trachinoidei)

 Testis organization and spermatogenesis, with the emphasis on spermiogenesis, in Opistognathus whitehurstii are described by ultrastructural and histochemical methods. The germinal epithelium is extremely reduced and restricted to the periphery of the testis, while most of the organ is occupied by a highly developed system of testicular efferent ducts. A semicystic type of spermatogenesis is observed and in the germinal epithelium spermatogenesis occurs only until the spermatidal stage. Young spermatids are released into the lumen of the testicular lobules and mature to sperm within the efferent duct system. The epithelial cells of these ducts are involved in protein and glycogen secretion and in phagocytosis of degenerating germ cells and residual bodies cast off by developing spermatids. On the basis of these functions, the testicular efferent duct system cells are considered to be homologous to the Sertoli cells. A correlation between a highly developed testicular efferent duct system and semicystic spermatogenesis is examined and a possible functional meaning of this apparently unusual mode of sperm production is proposed. Accepted: 18 March 1997     

Ultrastructural aspects of spermatogenesis in Calyptogena pacifica Dall 1891 (Vesicomyidae; Bivalvia)

The process of spermatogenesis and spermatozoon morphology was characterized from a deep‐sea bivalve, Calyptogena pacifica (Vesicomyidae, Pliocardiinae), a member of the superfamily Glossoidea, using light and electron microscopy. Spermatogenesis in C. pacifica is generally similar to that in shallow‐water bivalves but, the development of spermatogenic cells in this species has also some distinguishing features. First proacrosomal vesicles are observed in early spermatocytes I. Although, early appearance of proacrosomal vesicles is well known for bivalves, in C. pacifica, these vesicles are associated with electron‐dense material, which is located outside the limiting membrane of the proacrosomal vesicles and disappears in late spermatids. Another feature of spermatogenesis in C. pacifica is the localization of the axoneme and flagellum development. Early spermatogenic cells lack typical flagellum, while in spermatogonia, spermatocytes, and early spermatids, the axoneme is observed in the cytoplasm. In late spermatids, the axoneme is located along the nucleus, and the flagellum is oriented anteriorly. During sperm maturation, the bent flagellum is transformed into the typical posteriorly oriented tail. Spermatozoa of C. pacifica are of ect‐aqua sperm type with a bullet‐like head of about 5.8 μm in length and 1.8 μm in width, consisting of a well‐developed dome‐shaped acrosomal complex, an elongated barrel‐shaped nucleus filled with granular chromatin, and a midpiece with mainly four rounded mitochondria. A comparative analysis has shown a number of common traits in C. pacifica and Neotrapezium sublaevigatum.     

Spermatogenesis,sperm cell morphology and accompanying secretions from two interstitial marine mites

The sperm cell morphology and spermatogenesis of Halacaroides antoniazziae Pepato Tiago and da Rocha 2011 and Acaromantis vespucioi Pepato and Tiago 2004 was investigated. Halacaroides sperm cells have a complete acrosomal complex, dense tubules crossing the cytoplasm and modified mitochondria. Mature sperm cells are surrounded by two kinds of secretions. Inside the ejaculatory duct, they lie upon a centre composed of a secretion structured as heaps of elongated bodies. Acaromantis spermatozoa are spindle shaped and lack an acrosomal complex. The plasmalemma is deeply folded; the cytoplasm is very reduced and devoid of organelles. A single kind of globular secretion was found. The sperm mass is surrounded by two layers of amorphous secretions. These species share a peripheral pattern of nuclear condensation during spermatogenesis, a possible apomorphy for most halacarids, and no special adaptation to the interstitial environment could be related to their sperm cell morphology.     

Structure of the Testes,Spermatozoa and Spermatozeugmata of Mimagoniates barberiRegan, 1907 (Teleostei: Characidae), an Internally Fertilizing,Oviparous Fish

Abstract The testis of Mimagoniates barberiis bipartite. Spermatogenic tissue is restricted to the anterior part. The posterior part of the testis is devoid of spermatogenic tissue and contains numerous efferent ducts filled with mature sperm. Cells in germinal cysts develop synchronously, sperm nuclei and flagella become oriented parallel in the late stages of spermiogenesis. In the caudal portion of the aspermatogenic part all sperms are arranged into unencapsulated sperm bundles — spermatozeugmata. Two types of spermatozeugmata are found both in the caudal portion of the testis and in milt. In the larger, spindle–shaped type, sperm flagella form the spindle tips. In the smaller ones, which have approximately a length of spermatozoon, the sperm are parallel and approximately in register. In both types sperm heads are arranged parallel. A mature spermatozoon is flail–shaped. The sperm head is highly elongated and situated alongside the flagellum, the tip of the head is directed backwards. Large mitochondria are situated on one side of the elongated nucleus only and form the tip of the head. Live spermatozoa move with the centriolar part ahead. Both testis and spermatozoon structure as well as formation of spermatozeugmata in M. barberiare highly derived features which perhaps evolved as adaptations to internal fertilization.     

Differences in sperm morphology in foam‐nesting leptodactyline frogs (Anura,Leptodactylidae)

Sperm morphology is diverse among vertebrates and is influenced by the reproductive strategies adopted by species. In anurans, sperm morphology is associated with reproductive modes and mating systems. Here, we describe the sperm morphology of 11 frog species in the genus Leptodactylus and that of Lithodytes lineatus and discuss the relationship between sperm morphology and species' mating systems. We observed two distinct sperm morphotypes among the leptodactyline species, which differed mostly in head morphology. Type I sperm had triangular head, discrete acrosome vesicle with posterior margin not clearly visible; type II sperm had elongated head, clear acrosomal vesicle with posterior margin clearly visible. These sperm types do not seem to be associated with phylogeny; instead, type II sperm was observed in all polyandrous species analysed and in species with evidences of polyandry. Moreover, sperm of all species presented tail with undulating membrane connected to the axial fibre. We suggest that differences in sperm morphology might be associated with sperm competition to what polyandrous species are subjected. However, natural history observations on polyandrous mating in some species presenting type II sperm and phylogenetic comparative studies are need to elucidate the role of mating systems in the evolution of sperm morphology in leptodactylines.     

Ultrastuctural and cytochemical study of spermatogenesis in Scrobicularia plana (Mollusca,Bivalvia)

The ultrastructure of the mature spermatozoa and spermatogenesis of the bivalve Scrobicularia plana are described. Support cells extend from the basal lamina to the lumen of the testis and are laterally connected to the germinal epithelium. Germ cells present intercellular bridges and flagella since the spermatogonial stage. While spermatogonia and spermatocytes appear connected to support cells by desmosome-like junctions, elongated spermatids are held at the acrosomal region by support cell finger-like processes. During spermiogenesis, the acrosomal vesicle differentiates from a golgian saccule and then migrates to the nuclear apex. A microtubular manchette arising from centrioles surrounds the acrosomal vesicle, the nucleus, and the mitochondria at the time these three organelles start their elongation, disappearing after that. The mature spermatozoon of S. plana lacks a distinct midpiece because the mitochondria extend from the region of the pericentriolar complex along the nucleus anteriorly for approximately 1.4 μm. The features of this bivalve type of modified spermatozoon are compared with those of other animal groups having similar modifications.     

The interstitial origin of germinal cells in the testis of the stickleback

The brook stickleback, Culaea inconstans (Kirtland), in common with other bony fishes, lacks a germinal epithelium in the tubules of the testis, and the tubule wall is composed of a thin, discontinuous layer of myoid cells and collagenous fibers. Labelling of germ cells with tritiated thymidine has shown that the germ cells are derived from clumps of spermatogonia in the interstitial area. Large companion cells within the lumina of the tubules extend their processes to engulf spermatogonia from the interstitium which then enter the lumen of the tubule. Subsequent development of the germ cells takes place within individual compartments formed by folds of the plasma membrane of a companion cell. The companion cell, together with its complement of germ cells, constitutes a cyst. A companion cell may surround spermatogonia in the interstitium and at the same time encompass residual sperm of the previous season within the lumen. The plasma membranes of the germ cells and the companion cells remain discrete. Mature sperm are released into the lumen of the tubule and the companion cell again extends its processes into the interstitium and engulfs more spermatogonia for the following year. Companion cells may be homologous to the Sertoli cells of higher vertebrates although their processes penetrate the interstitium during the initial stages of spermatogenesis and they do not contain a permanent stock of spermatogonia.     

Derived sperm morphology in the interstitial sea cucumber Rhabdomolgus ruber,with observations on oogenesis and spawning behavior

Some life history features of the interstitial sea cucumber Rhabdomolgus ruber are described from intertidal specimens collected from the northern coast of Maine. Histological studies suggest that the population consists of hermaphrodites with gametogenesis being initiated in April and reproduction beginning in May and continuing through the summer months. Sexually mature adults possess a single, blind‐ended gonadal tubule that functions as an ovotestis by producing both eggs and sperm. The ovotestis wall consists of an outer peritoneum composed of flagellated epithelial cells and muscles; an inner germinal epithelium of germ and somatic cells; and a middle connective tissue (hemal) compartment bounded by the basal laminas of the peritoneum and germinal epithelium. During the reproductive season, the gonadal tubule contains all stages of oocyte development. Vitellogenesis appears to involve the biosynthetic activities of the Golgi complex and rough endoplasmic reticulum. A few specimens had transitional ovotestes with mature sperm in the gonad lumen and asynchronously developing oocytes and a small number of spermatocytes within the germinal epithelium. The mature spermatozoon is an ent‐aquasperm with ultrastructural features significantly different from those described from other echinoderm classes including a highly elongated acrosome, a large periacrosomal region between the acrosome and nucleus, numerous unfused mitochondria in the midpiece, and a cytoplasmic sleeve or collar extending posteriorly along the proximal portion of the flagellum. The sperm head reaches 11.5 μm in length (combined midpiece, nucleus, periacrosomal region, acrosome), making it the longest yet reported from the Holothuroidea and among the longest in the Echinodermata. Some elements of this derived morphology could be attributed to fertilization biology, but others may have phylogenetic significance. Spawning behavior was observed in which two individuals appeared to pseudocopulate by intertwining their oral tentacles for several minutes before one of them abruptly secreted an egg mass containing three eggs.     

Annual changes in germinal epithelium determine male reproductive classes of the cobia

Five reproductive classes of cobia Rachycentron canadum , caught along the Gulf of Mexico and the south-east Atlantic coast of the U.S.A., are described during the annual reproductive cycle. These are based upon changes in the testicular germinal epithelium and the stages of germ cells that are present: early maturation, mid maturation, late maturation, regression and regressed. During early maturation, the germinal epithelium is continuous from the testicular ducts to the periphery of the testis and active spermatogenesis occurs throughout the testis. In mid maturation, the germinal epithelium near the ducts becomes discontinuous, but it remains continuous distally. In late maturation, a discontinuous germinal epithelium extends all along the lobules to the testicular periphery; lobules are swollen with sperm and there is minimal spermatogenesis. The regression class is characterized by a discontinuous epithelium throughout the testis, sperm storage and widely scattered spermatocysts. Spermatogonial proliferation also occurs along the lobule walls and at the periphery of the testis. In regressed testes, spermatogonia exist only in a continuous or discontinuous germinal epithelium, although residual sperm are nearly always present in the lobules and ducts. The presence or absence of sperm is not an accurate indicator of reproductive classes. At the periphery of the testis in the regression and regressed classes, the distal portions of lobules elongate as cords of cells containing spermatogonia and Sertoli cells. All reproductive classes can be identified in paraffin sections, although plastic sections provide better resolution. Using maturation classes defined by changes in the germinal epithelium to describe testicular development and spermatogenesis gives a more accurate picture than does using the traditional terminology.     

Peculiarities in the organization of testis of Ophidian sp. (Pisces Teleostei). Evidence for two types of spermatogenesis in teleost fish

The walls of lobules in the testis of Ophidion sp. are composed of Scrtoli cells and young germinal cells (spermatogonia and spermatocytes). Spermatocytes are linked by cytoplasmic bridges. The associations of Sertoli cells and spermatocytes constitute true cysts. Meiosis takes place in the cysts. When meiosis is complete, cysts open. Spermatids are released into the lumen of the lobules and the cyloplasmic bridges break down. Spermiogenesis occurs in the lumen. Spermatids at various levels of spermiogenesis are then mixed with ripe spermatozoa. In teleosts we thus recognize two types of spermatogenesis: a cystic type where spermatogenesis is completed within cysts, and leads to synchronous development of germ-cells; and a semi-cystic type, where spermatogenesis occurs partly outside cysts. This may produce asynchronous spermatogenesis.     

Electron microscopic study of spermatogenesis in the lung fluke (Paragonimus miyazakii)

Summary The characteristics of spermatogenesis in a type of pulmonary parasite, Paragonimus miyazakii have been observed using the electron microscope. Groups of several spermatocytes revealed mutual cytoplasmic connection. That degree of this fusion increased as spermatogenesis progressed, and finally developed into a so-called cytophore. Then, this cytophore remained joined with a spermatid by a short stalk until the spermatid changed into a sperm. The nucleus of the spermatid became elongated with a string-like arrangement of the chromatin, which, in turn, showed increased electron density. At the pole of the spermatid, linearly arranged microtubules developed just below the plasma membrane. Close to an elongated portion at the pole, two separate flagella start growing and later fuse with the sperm itself. In the sperm tail a couple of tail filament complexes, longitudinally oriented slender mitochondria, and a tubular structure were present.     

A light and electron microscopic study on the organization of the testis and the semicystic spermatogenesis of the genus Scorpaena (Teleostei,Scorpaenidae)

The testicular organization and semicystic spermatogenesis of Scorpaena porcus and Scorpaena scrofa are analyzed by means of optic and electron microscopy and immunohistochemical techniques. The testicular structure of S. porcus and S. scrofa belongs to the unrestricted spermatogonial type, but has typical features of the restricted type. Moreover, the structure presents an epithelioid arrangement of Sertoli and germ cells rather than the germinal epithelium that appears in the majority of teleosts. After the cysts open, Sertoli cells hypertrophy and remain on the basement membrane, linked by interdigitations and tight junctions and bordering the lumen of the lobule, which at this moment works as an efferent duct. Secretions of Sertoli cells usually function in the nutrition of germ cells, and they seem to contribute in it even in this kind of spermatogenesis in which the free lumen spermatids do not have any connection with Sertoli cells. In addition, Sertoli cells can divide after the cysts have broken apart and hypertrophied, suggesting that they are still important for the final maturation of spermatozoa and seminal fluid formation. J. Morphol. 2009. © 2009 Wiley‐Liss, Inc.     

Fine structure of the male reproductive system in two species ofHaplognathia sterrer (Gnathostomulida,Filospermoidea)

Summary The structure of the male reproductive systems of two species ofHaplognathia cf.lyra andH. cf.rosacea was described. The structure of the testes and the anterior portions of the sperm ducts in both species was found to be similar. However, considerable species differences were found between the structures of the glands and muscles associated with the reproductive systems. These were more elaborate inH. cf.lyra than inH. cf.rosacea. The former species possessed an H-shaped sperm duct gland, three distinct groups of penis muscles and a penis with two cell types and with a lumen. The latter species had paired sperm duct glands, no specialized penis muscles and a penis with only one cell type and without a detectable lumen. No open gonopore was observed in either species. The sperm presumably exit through a ventral tissue connection observed connecting the penis and the ventral epidermis. These findings were discussed in the light of Mainitz's (1977) theory concerning the primitive penis type within the Gnathostomulida.Abbreviations ap anterior-posterior penis muscles - bm basement membrane - csd common sperm duct - dl dorsal lumen of the penis - dp dorsal gland cells of the penis - dv dorsoventral muscles anterior to the penis - dw sperm duct wall cell - e epidermis - ex exit cell - g intestine - gl gut lumen - n nerve - p penis - sd sperm duct - sdg sperm duct gland - tw testes wall cell - vl ventral lumen of the penis - vp ventral gland cells of the penis This project was supported by NSF grant #GB 42211 (R.M. Rieger P.I.). The line drawings have been executed after our design by Ms. Linda McVay     

Ultrastructure of sperm development and mature sperm morphology in three species of commensal bivalves (Mollusca: Galeommatoidea)

Ultrastructural features of the ovotestes, spermatogenesis, and the mature sperm are described for three galeommatid bivalves, Divariscintilla yoyo, Divariscintilla troglodytes, and Scintilla sp., from stomatopod burrows in eastern Florida. All three species yielded similar results except with respect to mature sperm dimensions. The ovotestis contains three types of somatic cells within the testicular portion: flattened myoepithelial cells defining the outer acinal wall; underlying pleomorphic follicle cells containing abundant glycogen deposits; and scattered, amoeboid cells containing lysosomal-like inclusions which are closely associated with developing sperm. Early spermatogenesis is typical of that reported from other bivalves. In contrast, the late stages of spermiogenesis involve the migration and gradual rotation of the acrosomal vesicle, resulting in a mature acrosome tilted about 70° from the long axis of the cell. The mature sperm possesses an elongated, slightly curved nucleus; a subterminal, concave acrosome with a nipple-like central projection; five spherical mitochondria and two centnoles in the middlepiece; and a long flagellum. The rotational asymmetry and the presence of perimitochondrial glycogen deposits in these sperm are unusual in the Bivalvia and may be associated with fertilization specializations and larval brooding common among galeommatoideans.     

Luteinizing hormone receptor-mediated effects on initiation of spermatogenesis in gonadotropin-deficient (hpg) mice are replicated by testosterone

Testosterone (T) is an absolute requirement for spermatogenesis and is supplied by mature Leydig cells stimulated by LH. We previously showed in gonadotropin-deficient hpg mice that T alone initiates qualitatively complete spermatogenesis bypassing LH-dependent Leydig cell maturation and steroidogenesis. However, because maximal T effects do not restore testis weight or germ cell number to wild-type control levels, additional Leydig cell factors may be involved. We therefore examined 1). whether chronic hCG administration to restore Leydig cell maturation and steroidogenesis can restore quantitatively normal spermatogenesis and testis development and 2). whether nonandrogenic Leydig cell products are required to initiate spermatogenesis. Weanling hpg mice were administered hCG (0.1-100 IU i.p. injection three times weekly) or T (1-cm subdermal Silastic implant) for 6 weeks, after which stereological estimates of germinal cell populations, serum and testicular T content, and testis weight were evaluated. Human CG stimulated Leydig cell maturation and normalized testicular T content compared with T treatment where Leydig cells remained immature and inactive. The maximal hCG-induced increases in testis weight and serum T concentrations were similar to those for T treatment and produced complete spermatogenesis characterized by mature, basally located Sertoli cells (SCs) with tripartite nucleoli, condensed haploid sperm, and lumen development. Compared with T treatment, hCG increased spermatogonial numbers, but both hCG and T had similar effects on numbers of spermatocytes and round and elongated spermatids per testis as well as per SC. Nevertheless, testis weight and germ cell numbers per testis and per SC remained well below phenotypically normal controls, confirming the involvement of non-Leydig cell factors such as FSH for quantitative normalization of spermatogenesis. We conclude that hCG stimulation of Leydig cell maturation and steroidogenesis is not required, and that T alone mostly replicates the effects of hCG, to initiate spermatogenesis. Because T is both necessary and sufficient for initiation of spermatogenesis, it is likely that T is the main Leydig cell secretory product involved and that additional LH-dependent Leydig cell factors are not essential for induction of murine spermatogenesis.     

Ultrastructure of sperm morphology ofTrilobodrilus axi andT. heideri (Dinophilidae,Polychaeta)

The ultrastructure of the sperm of the dinophilidsTrilobodrilus axi andTrilobodrilus heideri was investigated. The sperms are extremely modified and show an elongated head, middle piece, and tail region, which are overlapping. This condition is a synapomorphous feature for the dinophilids. The sperm structure has to be related to the specialized mode of sperm transfer by hypodermal injection in these species.