Centriolar Complex Research Articles

Steps of spermiogenesis in the ostrich (Struthio camelus).

Few studies describe the sequence of morphological events that characterize spermiogenesis in birds. In this paper, the clearly observable steps of spermiogenesis are described and illustrated for the first time in a commercially important ratite, the ostrich, based on light microscopy of toluidine blue-stained plastic sections. Findings were supplemented and supported by ultrastructural observations, PNA labeling of acrosome development, and immunocytochemical labeling of isolated spermatogenic cells. Spermiogenesis in the ostrich followed the general pattern described in non-passerine birds. Eight steps were identified based on changes in nuclear shape and contents, positioning of the centriolar complex, and acrosome development. Only two steps could be recognized with certainty during development of the round spermatid which contributed to the fewer steps recorded for the ostrich compared to that described in some other bird species. The only lectin that displayed acrosome reactivity was PNA and only for the first three steps of spermiogenesis. This suggests that organizational and/or compositional changes may occur in the acrosome during development and merits further investigation. Immunological labeling provided additional evidence to support the finding of previous studies that the tip of the nucleus in the ostrich is shaped by the forming acrosome and not by the microtubular manchette. To our knowledge, this is the first complete description of spermiogenesis in ostrich and one of few in any avian species. In addition to comparative reproduction and animal science, this work has implications for evolutionary biology as the reported germ cell features provide a bridge between reptile and ratite-avian spermatogenesis.

Cryostorage of white cachama (Piaractus orinoquensis) sperm: Effects on cellular, biochemical and ultrastructural parameters

To date, little attention has been paid to identifying the effects of long-term cryopreservation on sperm quality for Piaractus orinoquensis. The object of this study was therefore to evaluate the effect of long-term cryopreservation (24 h, 1, 6 and 12 months) on sperm motility, viability, DNA integrity, ATP content, total antioxidant capacity (TAC), morphology and sperm ultrastructure in this species. Milt samples from six males were cryopreserved in a medium containing final concentrations of 7.5% Me2SO, 4.1% glucose and 9.0% egg yolk. The samples were frozen in liquid nitrogen (LN) vapor and stored in LN for periods of 24 h and 1, 6 and 12 months. After thawing, both the motility rate and the viability decreased significantly compared with fresh sperm; however, these parameters did not differ among the four cryopreservation times. The DNA integrity and ATP content decreased significantly after 6 months of cryopreservation. There were no significant differences in TAC values between fresh and cryopreserved sperm. The total sperm abnormalities in cryopreserved samples were about 5-fold higher than in fresh sperm; short tail was the most common defect occurring after cryostorage. The ultrastructural analysis reveals that P. orinoquensis spermatozoa consist of an ovoid head without acrosome, a cylindrical mid-piece, and a single flagellum. The nuclear fossa is located at the base of the nucleus and contains the centriolar complex. There are 1–2 ring-shaped mitochondria located in the mid-piece. The flagellum shows a 9 + 2 organization of microtubules in the axoneme. Post-thaw spermatozoa presented damage such as swelling and rupture of the plasma membrane, mitochondrial damage, loss of the electron-dense chromatin of the nucleus, and degeneration in the middle region.

Spermatozoon of the wild scalloped perchlet, Ambassis nalua (Hamilton, 1822): Ultrastructure and morphometric analysis

The description of sperm morphology is fundamental in the reproductive biology of fishes, but this information is limited in the family Ambassidae. Our report hence focused on the ultrastructure and morphometric analysis of spermatozoa in a pelagic fish Ambassis nalua. All fish (n = 75) were obtained during January and March 2017 from the Estuarine Pranburi River, Thailand. The standard length of fish used in this study was 3.4 ± 0.12 cm (mean ± standard deviation). All specimens were considered mature based on the abundance of spermatozoa in the testis. The testicular organs were collected and observed using standard histology and transmission electron microscopy (TEM). Ultrastructural observation associated with morphometric analysis showed that spermatozoa are structurally long cells of approximately 51.17 ± 4.54 µm total length, composed of a head, a midpiece and a tail. The head had no acrosome, and the granular structure of condensed chromatin was observed within the ovoid nucleus. The midpiece consisted of a short cylindrical region with the length of 1.29 ± 0.87 µm in diameter, having the centriolar complex organization and eight mitochondria (approx. 0.32 ± 0.02 µm each). The uniflagellar tail was clearly identified with a classical 9+2 arrangements of microtubules. Based on these characteristics, the spermatozoon of wild scalloped perchlet are considered as uniflagellate anacrosomal aquqsperm. The morphological features, including the number of mitochondria, may be used for further cryopreservation and in the evolutionary biology of this species.

Spermatogenesis in the inseminating African butterflyfish Pantodon buchholzi (Teleostei: Osteoglossiformes: Pantodontidae) with the revision of residual bodies formation.

The aim of this study was to analyse spermatogenesis in the African butterflyfish, Pantodon buchholzi, using transmission electron microscopy and scanning electron microscopy. P. buchholzi is the most basal teleost that exhibits insemination and produces a highly complex introsperm with the most elongate midpiece known in teleost fishes. Their early stages (spermatogonia and spermatocytes) do not differ greatly from those of other fishes, with the exception of Golgi apparatus degradation appearing as spindle-shaped bodies (SSBs). In round, early spermatids, the development of the flagellum begins after the migration of the centriolar complex towards the nucleus. Later, the elongation of the midpiece coincides with the displacement of the mitochondria and their fusion to produce nine mitochondrial derivatives (MDs). In these spermatids, the nucleus is situated laterally to the midpiece, with condensing chromatin in the centre of the nucleus. Within the midpiece, the flagellum is located within a cytoplasmic canal and is surrounded by a cytoplasmic sleeve containing fibres, MDs and a great amount of cytoplasm located on one side. During the next phase, nuclear rotation, the highly condensed chromatin is displaced to a position above the centriolar apparatus, whereas chromatin-free nucleoplasm is transferred to the cytoplasm. Later, this nucleoplasm, still surrounded by the nuclear membrane, is eliminated into the cyst lumen as the nucleoplasmic packet. Within the highly elongate spermatids, other excess organelles (SSBs, endoplasmic reticulum and mitochondria) are eliminated as residual bodies (RBs). Fully developed spermatozoa, which contain conical-shaped nuclei, eventually coalesce to form unencapsulated sperm packets (spermatozeugmata) that are surrounded by RBs at the level of the extremely elongate midpieces. Later, RBs are removed at the periphery of the cyst by means of phagocytosis by Sertoli cells.

Some Data on Reproductive Biology of Spotted Ghoul Inimicus sinensis (Synanceiidae)

Gonadal structure, cytological condition of oocytes, ultrastructure of egg envelope, and spermatozoa morphology are studied in the individuals of Inimicus sinensis 120–230 mm TL with the gonads mainly at maturity stage IV. The males possess hypertrophied urinary bladders. Based on oocytes’ size distribution in the ovaries, the oogenesis is continuous and the spawning is multiple. Oocytes of the maturation period are 600–700 µm in diameter. The batch fecundity is 15 380 oocytes on average. The zona radiata of oocytes that have completed growth is 10.2 µm in width on average and it consists of two layers. The narrow outermost layer of the egg envelope approximately 0.34 µm width is most likely chorion. Spermatozoon ultrastructure of I. sinensis does not notably differ from that in other representatives of the suborder Scorpaenoidei with external insemination. The centriolar complex is characterized by orthogonal position of centrioles. Based on ovarian structure, the representatives of the genus Inimicus do not differ from the majority of teleost fishes and viviparous species of the genera Sebastes and Sebastiscus. However, many representatives of the suborder Scorpaenoidei have unique ovarian structure with the central ovigerous stroma attached to the anterior part of the ovary. Based on the current phylogenetic schemes, this ovarian type evolved independently in several evolutionary lineages.

Spermatid differentiation, with particular reference to acrosomogenesis, in the passeridan bird, Carib grackle (Quiscalus lugubris)

Only a few studies on the development of the passerine spermatozoon are available, yet species variations in the conformation as well as structure of the generally helical acrosome have been reported. This study of spermiogenesis in the Carib grackle (Quiscalus lugubris) intended to provide a deeper understanding of the development of the sperm, and in particular to investigate the bi-partite nature and development of the acrosome as well as its relationship with the nucleus, in the absence of a perforatorium that is found in most non-passerine birds. The acrosomal vesicle already displays a bi-partite nature in the acrosomal granule within the Golgi complex, and the attachment of the dense granule (future acrosomal core) within the crest part (future acrosomal crest) establishes polarity as it approaches and attaches to the nucleus. Thereafter, they develop variably. The acrosomal crest leads the elongation and spiraling of the acrosome, and the core portion contributes significantly to the formation of the keel of the crest part. The rounded, core-bearing part of the base of the acrosome progressively indents and fits into the concavity, thus formed, at the anterior part of the nucleus. The possible homology of the acrosomal complex (including the perforatorium) and the nucleus between non-passerine and passerine birds was discussed. The centriolar complex comprises both the proximal and distal centrioles in all spermatids and spermatozoa. The mitochondria undergo a number of morphological changes, including size and electron-density, from the round spermatid through to the mature spermatid; changes that are probably influenced by their functional states in the different evolving phases of the spermatids.

Ultrastructure of spermatophoral sperm in the freshwater gastropod Thiara amarula (Linnaeus, 1758) (Cerithioidea: Thiaridae): potential taxonomic features including eusperm nuclear content differentiation

The ultrastructure of spermatophoral sperm is investigated for the first time in a cerithioidean gastropod. Thiara amarula (Linnaeus, 1758), a widespread Indo-West Pacific species of the Thiaridae, produces uniaxonemal euspermatozoa and multiaxonemal paraspermatozoa. Euspermatozoa possess a laterally-flattened acrosomal vesicle (with eccentrically positioned invagination and subacrosomal material), a tubular, laterally-compressed nucleus, an elongate midpiece (with four, straight, equal-sized mitochondria each with parallel cristae), an elongate glycogen piece and an end piece. The eusperm nucleus exhibits or is associated with a number of unusual features including: (1) nuclear contents differentiated into two components (fibrous, highly electron-dense material, enclosing a pair of less-dense, finely granular tracts); (2) nuclear invagination with two elongate grooves aligned at 90 degrees to the axonemal central microtubules and parallel with the 1 and 5 axonemal doublets; (3) a periodically-banded rootiet associated with the centriolar complex near the nuclear apex; (4) an electron-dense structure attaching the nucleus to the axoneme at the nucleus-midpiece junction. Feature (1) has been reported elsewhere in Melanoides tuberculata (Thiaridae), but in no other examined gastropods (or other molluscs) and its purpose is unknown. Paraspermatozoa of T. amarula possess an elongate head region (up to 14 axonemes and large, irregular mitochondria enclosed by sheath of spherical, dense vesicles), a short 'glycogen' region and a posterior tuft of glycogen tails. The two sperm types do not form spermatozeugmata and observations suggest that neither type undergoes any noticeable structural change while inside the spermatophore. Sperm features suggest a close relationship between T. amurula and M. tuberculata and more broadly with other 'Group T cerithioidean families exhibiting four equal-sized eusperm mitochondria, including Potamididae, Modulidae, Scaliolidae, Melanopsidae, Semisulcospiridae and Pleuroceridae.

Ultrastructure of spermatozoa in cobia, Rachycentron canadum (Linnaeus, 1766)

Ultrastructure and development of spermatozoa in cobia, Rachycentron canadum are described. Sections through the testis show different developmental stages viz, Spermatocytes, spermatids and sperm. Spermatozoa of R. canadum exhibit the configuration of uniflagellated, anacrosomal Type I aquasperm, typical for externally fertilizing fish. Mature spermatozoon is seen with a prominent head and long cylindrical flagellum. Ultrastructure of sperm shows invaginated 'U' shaped nucleus and other organelles. The mitochondrial matrix is electron-dense with irregular arrangement of the cristae. The nucleus reveals a deep invagination (nuclear fossa) in which the centriolar complex is located. The centriolar complex lies inside the nuclear fossa and is composed of a proximal and a distal centriole. The two centrioles are placed perpendicular to each other. The flagellum has a typical eukaryotic organization (microtubule doublets 9 + 2 pattern) and measures around 36.21 ± 0.42 μm in length. This study for the first time provides a comprehensive detail on the ultrastructure and developmental process of sperm in cobia, R. canadum.

Morphology and ultrastructure of Brachymystax lenok tsinlingensis spermatozoa by scanning and transmission electron microscopy.

This study was conducted to investigate Brachymystax lenok tsinlingensis spermatozoa cell morphology and ultrastructure through scanning and transmission electron microscopy. Findings revealed that the spermatozoa can be differentiated into three major parts: a spherical head without an acrosome, a short mid-piece, and a long, cylindrical flagellum. The mean length of the spermatozoa was 36.11±2.84μm, with a spherical head length of 2.78±0.31μm. The mean anterior and posterior head widths were 2.20±0.42μm and 2.55±0.53μm, respectively. The nuclear fossa was positioned at the base of the nucleus that contained the anterior portion of flagellum and a centriolar complex (proximal and distal centrioles). The short mid-piece was located laterally to the nucleus and possessed just one spherical mitochondrion with a mean diameter of 0.65±0.14μm. The spermatozoa flagellum was long and cylindrical, and could be separated into two parts: a long main-piece and a short end-piece. The main piece of the flagellum had short irregular side-fins. The axoneme composed the typical '9+2' microtubular doublet structure and was enclosed by the cell membran e. This study confirmed that B. lenok tsinlingensis spermatozoa can be categorized as teleostean "Type I" spermatozoa; 'primitive' or 'ect-aquasperm type' spermatozoa. To the best of the authers knowledge, this was the first study conducted on the morphology and ultrastructure of B. lenok tsinlingensis spermatozoa.

A comparative overview of the sperm centriolar complex in mammals and birds: Variations on a theme

This paper presents an overview of the structure, function and anomalies of the sperm centriolar complex (CC) on a comparative basis between mammals and birds. The information is based on selected references from the literature supplemented by original observations on spermiogenesis and sperm structure in disparate mammalian (cheetah and cane rat) and avian (ostrich, rhea and emu) species. Whereas the basic structure of the CC (a diplosome surrounded by pericentriolar material) is similar in Aves and Mammalia, certain differences are apparent. Centriole reduction does not generally occur in birds, but when present as in oscines, involves the loss of the proximal centriole. In ratites, the distal centriole forms the core of the entire midpiece and incorporates the outer dense fibres in addition to initiating axoneme formation. The elements of the connecting piece are not segmented in birds and less complex in basic design than in mammals. The functions of the various components of the CC appear to be similar in birds and mammals. Despite obvious differences in sperm head shape, the centrosomal anomalies afflicting both vertebrate groups demonstrate structural uniformity across species and display a similar range of defects. Most abnormalities result from defective migration and alignment of the CC relative to the nucleus. The most severe manifestation is that of acephalic sperm, while angled tail attachment, abaxial and multiflagellate sperm reflect additional defective forms. The stump-tail defect is not observed in birds. A comparison of defective sperm formation and centrosomal dysfunction at the molecular level is currently difficult owing to the paucity of relevant information on avian sperm.

A comparative gametogenesis and biochemical differentiation of goatfish (Family: Mullidae) from the Red Sea

Comparative study between few species of goatfishes (Mullidae), locally called Sultan Ibrahim, has been investigated from Jeddah coast. The present study compares gonad structure, oogenesis and spermatogenesis in three species of goatfish. The various stages are similar in the different species, but differ in time of spawning. The reproductive biology of three goatfishes, Mulloidichthys flavolineatus (1 black blotch), Mulloidichthys flavolineatus (3 black blotches), Mulloidichthys vanicolensis, were examined. A macroscopic and histological examination of the gonads demonstrated that all the three species are typically gonochoristic. Species-specific reproductive parameters including male stages, changes in gonado-somatic index (GSI), classification of oocyte stages were analyzed for each of the three targeted species. All three species have asynchronous ovary. The spawning season for the three studied species differ from one species to another i.e., the spawning season for M. vanicolensis females and males from April to July; M. flavolineatus, (1 black blotches) for both sexes from May until December; for M. flavolineatus (3 black blotches) showed a peak in May to early October for both sexes. Ultrastructurally the mature spermatozoon’s head shape appears like a computermouse, the tiny mid-piece consist two mitochondria and a flagellum. The chromatin, within the nucleus is electron-dense and homogenous in M. flavolineatus (with one black blotch), while the chromatin is granular in (M. vanicolensis and M. flavolineatus (3 black blotches)). The posterior region of the nucleus is deeply penetrated by the nuclear fossa which contains, the basal part of flagellum and centriolar complex. The ultrastructure of the eggs of the three Mullidae species is circular in shape and contains a simple, circular micropyle. The ovarian follicular epithelium of the three Mullidae species consists of two layers, an outer theca layer and an inner granulosa layer with zona radiata under them. Zona radiata consists of 2 layer-interna and externa. Zona radiata interna is thicker than zona radiata externa. Differences in protein patterns of muscles and gonads were observed in the present study.

A histological study of testis development and ultrastructural features of spermatogenesis in cultured Acrossocheilus fasciatus.

Testis development and ultrastructural features of spermatogenesis in Acrossocheilus fasciatus (Cypriniformes, Barbinae), a commercial stream fish, were studied using light and electron microscopy. The reproduction cycle in A. fasciatus testes is classified into six successive stages from Stage I to Stage VI. Based on an analysis of previous results, May to July can be confirmed as the best breeding season for A. fasciatus males. During this time, the A. fasciatus testes are in Stage V and the sperm in males is most abundant. In the first reproductive cycle, sexually mature male testes return to Stage III in October, subsequently overwintering at this stage. In the lobular-type testes of A. fasciatus, cystic type spermatogenesis occurs with restricted spermatogonia. All spermatogenic cells at different stages are distributed along the seminiferous lobules, which contain spermatogonia, spermatocytes, spermatids and spermatozoa. At the end of spermatogenesis, spermatogenic cysts open to release spermatozoa into the lobule lumen. Ultrastructural observation of A. fasciatus spermiogenesis reveals that electron-dense substances appear at the different stages of germ cells, from primary spermatogonia to secondary spermatocytes. We have termed these dense substances as "nuage" when free in the cytoplasm or adjacent to the nuclear envelope, while those close to the mitochondria are called inter-mitochondrial cement. The spermatozoa in A. fasciatus can be classified as type I due to the presence of nuclear rotation. Although the nuclear chromatin in the head of sperm was highly condensed, no acrosome was formed. The cytoplasmic canal, a common ultrastructural feature of Teleostei spermatozoa, was also present in the midpiece. In addition, numerous fused mitochondria were observed. The distal centriole and proximal centriole constituting the centriolar complex were oriented incompletely perpendicular to each other. The flagellum showed a typical 9+2 arrangement pattern. Conversely, our study on A. fasciatus yielded no information concerning the lateral fins although an enlarged saclike area was present at the end of some flagella.

Ultrastructure of spermatogenesis and spermatozoa in agoutis during sexual development.

The aim was to study the ultrastructure of testicular parenchyma and define the morphological ultrastructure of spermatozoa of agoutis kept in captivity. Segments of testes from eight agouti males at prepubescence, prepuberty, pubescence and sexual maturity were fixed in glutaraldehyde. Laboratory procedures were performed for transmission electron microscopy. Spermatogonial cells of Type A - pale, Type A - dark, intermediate and Type B were found. Spermatocytes in the pachytene phase were abundant among primary spermatocytes. From the prepubertal phase, Sertoli cells exhibited invaginations in the nuclear membrane and lipid inclusions in the cytoplasm due to their phagocytic function. Leydig cells displayed higher metabolic activity during puberty as evidenced by the presence of lipid droplets. Spermatozoa were fully formed morphologically at prepuberty. The centriolar complex had partially degenerated and featured a centriolar space as in rodents. Sperm heads were tapered, without prominence of the acrosome or evidence of the perforatorium, differing from cavies, rats and mice. This is the first study to describe the ultrastructure of agouti spermatozoa. This research may assist as a basis for future work related to fertility and other biotechnologies applied to reproductive biology in agoutis.

Ultrastructure of sperm of the Spotted scat (Scatophagus argus, Linnaeus, 1766) observed by scanning and transmission electron microscopy

An investigation was conducted to understand the sperm cell morphology and ultrastructure of Spotted scat (Scatophagus argus) through scanning and transmission electron microscopy. The present study reveals that the sperm of S. argus can be differentiated into three major parts – an acrosome-less spherical head, a short mid-piece, and a cylindrical flagellum. The scat sperm cell had a mean total length of 21.32±1.80μm with the presence of ovoid electron-dense nucleus. The mean length and width of ovoid nucleus measured 1.44±0.34 and 1.54±0.33μm, respectively. The structural characteristics of the nucleus were found to be a shallow axial nuclear fossa and centriolar complex. The two centrioles were positioned nearly perpendicular to each other with a conventional “9+0” pattern in the proximal centriole. The short mid-piece was located laterally to the nucleus and contains 5 or 6 spherical and unequal-sized mitochondria. The mitochondria were separated from the axoneme by a cytoplasmic canal. The flagellum was inserted at the base of the nucleus with the presence of an axoneme structure of 9+2 paired micro tubules. The sperm flagellum had short irregular lateral fins. The present study reveals that Spotted scat sperm can be categorized as being of a “primitive or ect-aquasperm type” and belongs to the teleostean “type I” sperm. This is the first report on the morphology and ultrastructure of sperm in Scatophagidae family.

The Mechanism of Dynein Light Chain LC8-mediated Oligomerization of the Ana2 Centriole Duplication Factor

Centrioles play a key role in nucleating polarized microtubule networks. In actively dividing cells, centrioles establish the bipolar mitotic spindle and are essential for genomic stability. Drosophila anastral spindle-2 (Ana2) is a conserved centriole duplication factor. Although recent work has demonstrated that an Ana2-dynein light chain (LC8) centriolar complex is critical for proper spindle positioning in neuroblasts, how Ana2 and LC8 interact is yet to be established. Here we examine the Ana2-LC8 interaction and map two LC8-binding sites within the central region of Ana2, Ana2M (residues 156-251). Ana2 LC8-binding site 1 contains a signature TQT motif and robustly binds LC8 (KD of 1.1 μm), whereas site 2 contains a TQC motif and binds LC8 with lower affinity (KD of 13 μm). Both LC8-binding sites flank a predicted ~34-residue α-helix. We present two independent atomic structures of LC8 dimers in complex with Ana2 LC8-binding site 1 and site 2 peptides. The Ana2 peptides form β-strands that extend a central composite LC8 β-sandwich. LC8 recognizes the signature TQT motif in the first LC8 binding site of Ana2, forming extensive van der Waals contacts and hydrogen bonding with the peptide, whereas the Ana2 site 2 TQC motif forms a uniquely extended β-strand, not observed in other dynein light chain-target complexes. Size exclusion chromatography coupled with multiangle static light scattering demonstrates that LC8 dimers bind Ana2M sites and induce Ana2 tetramerization, yielding an Ana2M4-LC88 complex. LC8-mediated Ana2 oligomerization probably enhances Ana2 avidity for centriole-binding factors and may bridge multiple factors as required during spindle positioning and centriole biogenesis.

Sperm evolution in the family Alestidae with comparative data for the genus Chalceus (Ostariophysi: Characiformes)

Spermiogenesis and spermatozoa in six genera of the African family Alestidae plus the Neotropical genus Chalceusare described. Spermiogenesis is quite similar in all Alestidae and is identified as Type I and its variants. In Type I spermiogenesis, the flagellum of earliest spermatids lies lateral to the nucleus, and rotation of the nucleus towards the centriolar complex is observed. Nuclear rotation is complete reaching 90 degrees in Bryconalestes longipinnis, Brachypetersius altus, Brycinus imberi, B. lateralis, and Alestopetersius compressus; and is incomplete reaching 20 degrees in Micralestes acutidensand Rhabdalestesrhodesiensis. Spermatozoa morphology varies from a medial nucleus with fibrillar chromatin in the most basal genus Brycinusto a strongly eccentric nucleus with highly condensed chromatin in the more derived Rhabdalestesand Micralestes. Chalceushas a very similar spermatozoon to that found in Brycinussharing the fibrillar aspect of the chromatin in the nucleus. This feature is so far only observed in these two genera among African and Neotropical characiform fishes.

Testicular structure in three viviparous species of teleosts in the genus Jenynsia (Anablepidae).

Histological structure of the testes and development of spermatozoa in Jenynsia species is described using light, scanning and transmission electron microscopy. The testis type is restricted spermatogonial, wherein spermatogonia are restricted to the distal ends of lobules, typical of the Atherinomorpha, and spermatogenesis is continuous throughout the year in wild-caught fish. Within the testicular lobes there are lobular germinal compartments wherein the functional units are spermatocysts, whose borders are formed by Sertoli cells. Spermatocysts may contain meiotic primary spermatocytes, secondary spermatocytes, spermatids, undergoing spermiogenesis, or spermatozoa. Spermatocysts with later stages of developing sperm are located proximal to the testicular ducts. During spermiogenesis, spermatid nuclei become elongated. As this occurs, the nucleus develops a deep, central fossa that contains the centriolar complex. As the flagellum grows, enlarging spermatid mitochondria migrate posteriorly alongside the flagellum but remain separated from it by the cytoplasmatic canal, an indentation of the plasma membrane. Between the enlarged mitochondria and plasma membrane, a sub-mitochondrial net develops. In longitudinal sections, the enlarged mitochondria are stacked in a zig-zag fashion, and in transverse sections they appear as a ring surrounding the flagellum, but separated from it by the cytoplasmic canal. Spermatozoa of the 3 jenynsiid species have an introsperm complex composed of a long mid-piece whose flagellum has a single "wing." Within the efferent ducts and the tubular gonopodium, sperm are lightly packed in a side by side fashion which facilitates their transfer into the female reproductive tract. This study presents detailed descriptions of testicular organization and cytological characterization of the stages of spermatozoa differentiation in 3 species of Jenynsia from northwestern Argentina (J. alternimaculata, J. multidentata and J. maculata), in order to contribute to the understanding of testicular structure and development of spermatozoa in the context of evolution of viviparity in this fish lineage.

Gamete ultrastructure in two species of the genus Upeneus (Mullidae) from the South China Sea

Ultrastructure of envelopes of ovulated oocytes and spermatozoon ultrastructure are described in two closely related species of the family Mullidae, Upeneus tragula and U. cf. margarethae. Oocyte envelope is represented by the thin zona radiata and weakly expressed chorion. The spermatozoon is characterized by the presence of elongated head with a pointed bend in its apical part and nuclear fossa reaching approximately 90% of the head length. The centriolar complex and basal part of the flagellum are located inside of the nuclear fossa.

Ultrastructure features of spermiogenesis and spermatozoa of sparid fish Rhabdosargus sarba (Forsskål, 1775) from Red SEA, Jeddah coast

Transmission and scanning electron microscopy were used to investigate the ultrastructure of spermatogenesis of the Sparid fish Rhabdosargus sarba. The testis is lobular in shape and spermatogensis is of unrestricted type. Spermatogonia occur isolated or in clusters within the seminiferous lobules. The germ cells are found in cysts formed by sertoli cell processes. Cells within cysts are found in the same developmental stage. Spermiogenesis is characterized by chromatin condensation, movement of the centrioles, flagellum development, nuclear rotation, nuclear indentation and nuclear fossa formation, reduction of the cytoplasm and differentiation of the flagellar complex. Ultrastructurally the spermatozoa of R. sarb belonges, like that of other sparid fish, to the teleosean type 1 spermatozoan with the flagellum axis perpendicular to the nuclear fossa. It has an ovoidal head, a short, cylindrically shaped midpiece and a long tail region. The nucleus reveals a deep invagination (nuclear fossa) in which the centrriolar complex is located, and a satellite nuclear noch shaped like a bell. The centriolar complex lies inside the nuclear fossa and is composed of a proximal and a distal centriole. The two centerioles are perpendicular to each other. The distal centriole is traversed by a conspicuous basal plate in its base. At the neck of the flagellum the necklace is observed. A nuclear noch is located above the proximal centeriol. The short midpiece houses two spherical mitochondria. The flagellum is inserted medio-laterally into the head, contains the conventional 9+2 axoneme.

Sperm characteristics as additional evidence of close relationship between Lebiasina and Piabucina (Characiformes: Lebiasinidae: Lebiasininae)

Spermiogenesis and spermatozoa of representatives of the genera Lebiasina and Piabucinaare described. Spermiogenesis is quite similar among all analyzed species of these genera and is identified as Type II. In this type of spermiogenesis, the flagellum of earliest spermatids lies lateral to the nucleus. A slight movement of the nucleus towards the centriolar complex is observed. In late spermatids, the nucleus slightly elongates towards the flagellum. The formation of two striated rootlets apparently occurs in early/intermediate spermatids. The spermatozoa of species of Lebiasinaand Piabucina share (1) a drop-shaped slightly elongated nucleus that is laterally positioned relative to the flagellum; (2) a superolateral centriolar complex; (3) two striated rootlets; (4) a basolateral midpiece; (5) oblong mitochondria and (6) a few spherical vesicles. The spermatic characteristics of Lebiasinaand Piabucinaindicate that spermiogenesis is a homologous process among all species of both genera examined to date and can be considered, along with the spermatozoa morphology, additional evidence indicating a close relationship between Lebiasina and Piabucina.