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

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Análise da espermatogênese e do cariótipo da espécie Eptesicus furinalis (d’Orbigny, 1847) Vespertilionidae, Chiroptera

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Transplante interespecífico de espermatogônias-tronco de Brycon orbignyanus em Astyanax altiparanae (Teleostei: Characidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Reproductive apparatus of the male giant hermit crab Petrochirus diogenes (Anomura, Diogenidae): morphology and phylogenetic implications

We describe the male reproductive apparatus of the giant hermit crab Petrochirus diogenes, with morphological and biometric analyses of the spermatophore, the gonopore and the ultrastructure of the spermatozoa. Specimens were collected from the southern coast of Sao Paulo, Brazil. Morphological analyses were done using stereoscopic, light, transmission and scanning electron microscopy. The reproductive system of this hermit crab is composed of elongate and lobular testes followed by vasa deferentia that connect to the exterior via gonopores. The gonopores are ovoid and surrounded by setae, and each gonopore is composed of a membranous operculum that forms a depression constituting the gonopore opening. The gonopore constitutes a unique structure among the Diogenidae due to its number of setae. The spermatophores are tripartite, composed of a sperm-containing ampulla, a peduncle and a proximal foot. The spermatozoon has 3 main regions (acrosomal vesicle, nucleus and cytoplasm). The structure of the spermatophore indicates that this species can be considered an exception within Diogenidae with regard to spermatophore morphology and can therefore be used for phylogenetic inferences.

A novel spermatozoan ultrastructure in the shrimp Hippolyte obliquimanus Dana, 1852 (Decapoda: Caridea: Hippolytidae)

The aim of this study was to describe and illustrate the morphology of the spermatozoon of the Western Atlantic shrimp, Hippolyte obliquimanus. Individuals were sampled from Itagua Beach (Ubatuba, southern Brazil). The male reproductive system was dissected and morphological analysis was undertaken using a stereomicroscope, a light microscope, and transmission electron and scanning electron microscopes. When viewed from the nuclear or acrosomal poles, each spermatozoon has many translucent radiating arms (about 20) from a denser cell body, while laterally the cell body and arms resemble a "cnidarian medusa", with all the arms projecting away from the bell-like cell body. This sperm morphology is distinct from the "thumbtack"-shaped spermatozoa observed in the majority of carideans but has similarities to the spermatozoa of Rhynchocinetes spp. The morphology of sperm of several species of the genus Hippolyte resembles the spermatozoon of H. obliquimanus with the presence of posterior nuclear arms, but it is necessary to study other Hippolyte species to place these arms in the context of the genus.

Mitochondrial inheritance and germ line determination in a bivalve species with Doubly Uniparental Inheritance (DUI)

Mitochondria are inherited maternally in most metazoans. However, in some bivalves, two mitochondrial lineages are present: one transmitted through eggs (F), the other through sperm (M). This is called Doubly Uniparental Inheritance (DUI). During male embryo development, spermatozoon mitochondria aggregate and end up in the primordial germ cells, while they are dispersed in female embryos. The molecular mechanisms of segregation patterns are still unknown. In the DUI species Ruditapes philippinarum, I examined sperm mitochondria distribution by MitoTracker, microtubule staining and TEM, and I localized germ line determinants with immunocytochemical analysis. I also analyzed the gonad transcriptome, searching for genes involved in reproduction and sex determination. Moreover, I analyzed an M-type specific open reading frame that could be responsible for maintenance/degradation of M mitochondria during embryo development. These transcripts were also localized in tissues using in situ hybridization. As in Mytilus, two distribution patterns of M mitochondria were detected in R. philippinarum, supporting that they are related to DUI. Moreover, the first division midbody concurs in positioning aggregated M mitochondria on the animal-vegetal axis of the male embryo: in organisms with spiral segmentation this zone is not involved in further cleavages, so aggregation is maintained. Moreover, sperm mitochondria reach the same embryonic area where germ plasm is transferred, suggesting their contribution in male germ line formation. The finding of reproduction and ubiquitination transcripts led to formulate a model in which ubiquitination genes stored in female oocytes during gametogenesis would activate sex-gene expression in the early embryonic developmental stages (preformation). Only gametogenetic cells were labeled by in situ hybridization, proving their specific transcription in developing gametes. Other than having a role in sex determination, some ubiquination factors could also be involved in mitochondrial inheritance, and their differential expression could be responsible for the different fate of sperm mitochondria in the two sexes.

Sperm morphology, swimming velocity, and longevity in the house sparrow Passer domesticus

Sperm competition exerts strong selection on males to produce spermatozoa with an optimal morphology that maximizes their fertilization success. Long sperm were first suggested to be favored because they should swim faster. However, studies that investigated the relationship between sperm length and sperm competitive ability or sperm swimming velocity yielded contradictory results. More recently, ratios of the different sections of a spermatozoon (the head, midpiece, and flagellum) were suggested to be more crucial in determining swimming velocity. Additionally, sperm ability to remain and survive in the female storage organs may also influence fertilization success, so that optimal sperm morphology may rather maximize sperm longevity than velocity. In this study, we investigated how sperm morphology is related to sperm velocity and sperm longevity in the house sparrow Passer domesticus. Sperm velocity was found to be correlated with head/flagellum ratio. Sperm with small heads relative to their flagellum showed higher swimming velocity. Additionally, shorter sperm were found to live longer. Finally, we found sperm morphological traits to vary substantially within males and the head/flagellum ratio to be unrelated to total sperm length. We discuss the hypothesis that the substantial within-male variation in sperm morphology reflects a male strategy to produce a diversity of sperm from long, fast-swimming to short, long-living sperm to maximize their fertilization success in a context of sperm competition.

Morphological evaluation of spermatogenesis in Lake Magadi tilapia (Alcolapia grahami): a fish living on the edge

Spermatogenesis in Lake Magadi tilapia (Alcolapia grahami), a cichlid fish endemic to the highly alkaline and saline Lake Magadi in Kenya, was evaluated using light and transmission electron microscopy. Spermatogenesis, typified by its three major phases (spermatocytogenesis, meiosis and spermiogenesis), was demonstrated by the presence of maturational spermatogenic cells namely spermatogonia, spermatocytes, spermatids and spermatozoa. Primary spermatogonia, the largest of all the germ cells, underwent a series of mitotic divisions producing primary spermatocytes, which then entered two consecutive meiotic divisions to produce secondary spermatocytes and spermatids. Spermatids, in turn, passed through three structurally distinct developmental stages typical of type-I spermiogenesis to yield typical primitive anacrosomal spermatozoa of the externally fertilizing type (aquasperm). The spermatozoon of this fish exhibited a spheroidal head with the nucleus containing highly electron-dense chromatin globules, a midpiece containing ten ovoid mitochondria arranged in two rows and a flagellum formed by the typical 9 + 2 microtubule axoneme. In addition, the midpiece, with no cytoplasmic sheath, appeared to end blindly distally in a lobe-like pattern around the flagellum; a feature that was unique and considered adaptive for the spermatozoon of this species to the harsh external environment. These observations show that the testis of A. grahami often undergoes active spermatogenesis despite the harsh environmental conditions to which it is exposed on a daily basis within the lake. Further, the spermiogenic features and spermatozoal ultrastructure appear to be characteristic of Cichlidae and, therefore, may be of phylogenetic significance.

Sperm ultrastructure of the Actinocyclidae (Mollusca, Nudibranchia) and homology of the terminal region of nudibranch sperm

Sperm ultrastructure is examined and described for the actinocyclidid nudibranchs Actinocyclus verrucosus, Hallaxa iju and Hallaxa indecora. Although general characteristics were consistent with previously described heterobranch observations, present investigations revealed ultrastructural synapomorphies for the family based on the morphology of the terminal region of the spermatozoon. In actinocyclidids, the axonemal microtubules penetrate for some distance beyond the annulus, and the annular accessory body elongates to completely seal the terminal region. Chromodoris also has an annular accessory body that completely seals the axoneme and terminal region, but it does not extend far beyond the annulus, and it is possible that these states were derived independently. Cytochemical staining confirmed that there was no glycogen present in the posterior region of the sperm for H. indecora or Chromodoris kuniei. However, representatives of other chromodoridid genera (Noumea, Risbecia) have an axoneme that penetrates through the entire annular complex, after which it is sheathed by a glycogen deposit. Similarities in the acrosomal complex support the proposed sister group relationship between the Actinocyclidae and Chromodorididae.

Ultrastructure of the spermatid of Caprimuigus europaeus Linnaeus 1758, the European Nightjar (Aves; Caprimulgidae), with phylogenetic implications

The sperm of Caprimulgus europaeus is typical of other nonpasserines in many respects. Features shared with Paleognathae and Galloanserae are the conical acrosome, shorter than the nucleus; the presence of a perforatorium and endonuclear canal; the presence of a proximal as well as distal centriole; the elongate midpiece with mitochondria grouped around a central axis (here maximally six mitochondria in similar to 10 tiers); and the presence of a fibrous or amorphous sheath around the principal piece of the axoneme. A major (apomorphic) difference from paleognaths and galloanserans is the short distal centriole, the midpiece being penetrated for most of its length by the axoneme and for only a very short proximal portion by the centriole. Nonpasserines differ from paleognaths in that the latter have a transversely ribbed fibrous sheath, whereas in nonpasserines it is amorphous, as in Caprimulgus, or absent. The absence of an annulus is an apomorphic feature of Caprimulgus, apodiform, psittaciform, gruiform, and passerine sperm, homoplastic in at least some of these. In contrast to passerines, in Caprimulgus the cytoplasmic microtubules in the spermatid are restricted to a transient longitudinal manchette. The structure of the spermatid and spermatozoon is consistent with placement of the Caprimulgidae near the Psittacidae, but is less supportive of close proximity to the Apodidae, from DNA-DNA hybridization and some other analyses.