Morphology of the spermatozoa of the Microhylidae (Anura, Amphibia)

Microhylid spermatozoa show the autapomorphic condition of possessing a thin post-mitochondrial cytoplasmic collar. Their spermatozoa are apomorphic in several respects. They have lost the distinct nuclear shoulder, endonuclear canal and axial perforatorium observed in urodeles, caecilians and primitive frogs, possess a conical perforatorium and apomorphically lack any fibres associated with the axoneme. The spermatozoa of Cophixalus , however, differ in several respects from those of the other microhylids examined. Cophixalus spermatozoa are longer in almost all measurements, the acrosome vesicle is cylindrical and does not completely cover the putative perforatorium, the perforatorium is asymmetrical and composed of fine fibres, the nucleus is strongly attenuated and narrower, and the mitochondria are elongate. The absence of fibres associated with the axoneme is an apomorphic condition shared with the Ranidae, Rhacophoridae and Pipidae.     

Spermiogenesis and introsperm ultrastructure of Scoloplax distolothrix (Ostariophysi: Siluriformes: Scoloplacidae)

Spermiogenesis in scoloplacids is characterized by initial lateral development of the flagellum, nuclear rotation, medial nuclear fossa formation, complex centriolar migration, and cytoplasmic channel formation. The scoloplacid spermiogenesis is similar to those found in Diplomystidae, the most primitive siluriform family. The scoloplacid spermatozoa have all the main characteristics of introsperm. They exhibit a conic head, a symmetric midpiece, a medial flagellum, and no acrosome. The conic forward‐elongated nuclei contain homogeneous chromatin. The thin extremity of the nuclei is strongly curved and along its internal face there is a well‐developed membranous compartment. The centrioles are completely inside the medial nuclear fossa, perpendicular to each other and with an electron‐dense material between them. In a cross view of the midpiece, the mitochondria form a ring surrounding internally the cytoplasmic channel, and in a longitudinal view they are organized in a row along it. Several elongated vesicles are distributed peripherally, mainly concentrated in the mid‐piece basal region. The flagellum contains the classical axoneme (9 + 2) and has two lateral projections or fins. The spermatozoa of scoloplacids share several characteristics with those of Auchenipteridae. Since these two families are not phylogenetically related this similarity seems to be due to convergence once both families are, until now, the only known siluriform families with introsperm.     

Optimization of fertilization success in Sander vitreus is influenced by the sperm : egg ratio and ova storage

Walleye Sander vitreum eggs could be stored at 7° C for up to 30 h with 60% fertilization rates. Fertilization rates at 1 and 12° C fell to 17 and 5%, respectively. At spermatozoa : egg ratios <25 000 : 1, survival to the eyed stage declined precipitously. Beyond a ratio of 25 000 : 1, survival to the eyed stage did not increase > c . ∼ 70%. Therefore, a standard spermatozoa : egg ratio of 25 000 : 1 was established and is recommended for studies concerning fertilizing success.     

Male reproductive strategy in Trichogramma evanescens: sperm production and allocation to females

Abstract.  The production and allocation of sperm among successive mates during a male's life largely determine its fitness. The sperm production pattern and sperm allocation to females is studied in Trichogramma evanescens , a short-lived egg parasitoid of several lepidopteran species. At emergence, virgin males have an average of 1607 ± 249 sperm stored in the seminal vesicles and no further production occurs during adult life. These males are able to mate with at least 20 females in rapid succession. During the first 10 matings, the males transfer approximately 100 sperm each time and then transfer fewer and fewer sperm per mating. The number of sperm stored in spermathecae of successively mated females remains relatively constant for the first 10 females, and decreases slowly for the subsequent females. The relationship between male reproductive strategy and some life-history traits are discussed.     

Die Spermatogenese der Zecke Ornithodorus moubata (Murr)

Zusammenfassung Die Genese der schwanzlosen Spermatozoen der Zecke Ornithodorus moubata wurde licht- und elektronenmikroskopisch untersucht.In der Keimzone des Hodens finden sich die kleinsten Zellen der Spermatogenese, die Spermatogonien A, die einzeln durch somatische Zellen voneinander isoliert sind. In einer Vermehrungsphase von 4 Teilungen entstehen Ballen von je 16 Spermatogonien B. Durch Wachstum gehen die Spermatogonien B in Spermatozyten I. Ordnung über. Dabei entsteht in ihnen eine Randstruktur, die sich nach den rasch ablaufenden Reifeteilungen in den Spermatiden zu einer Vakuole entwickelt, in die bandartige Zellfortsätze hineinhängen. Gleichzeitig mit einer Längsstreckung der Spermatide stülpt sich die spätere Spitze in die Vakuole ein und führt zu einer handschuhfingerartigen Umkrempung der Spermatide. Dieser letzte Schritt erfolgt erst nach der Kopulation im Weibchen. Dann liegen die ursprünglich in die Vakuole hängenden Fortsätze dem reifen Spermatozoon außen als Leisten an.Die Kernkondensation und die Akrosombildung werden im einzelnen beschrieben. Durch die komplizierte Umgestaltung der Spermatide erfahren auch Kern und Akrosom sowie die Zentriolen unfangreiche Ortsveränderungen, die erst im Weibchen abgeschlossen werden.

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The spermatogenesis of the tick Ornithodorus moubata

Summary The genesis of the tailless spermatozoa of the tick Ornithodorus moubata has been studied by light and electron microscopy.In the germ zone of the testis the smallest cells of the spermatogenesis are located, the spermatogonia A which are separated from each other by somatic cells. The result of a multiplication phase with 4 mitoses are clusters of 16 spermatogonia B. While growing up the spermatogonia B differentiate into spermatocytes I. They show a special structure of their border which, after the quickly passed maturation divisions, gets more developed in the spermatides and turns into a vacuole with ribbon-like cytoplasmic processes hanging into it. Simultaneously with an elongation of the spermatide the subsequent apex invaginates into the vacuole and induces an inversion of the spermatide like the finger of a glove. This last step takes place only after the copulation in the female. Then the processes previously hanging into the vacuole are lying as groins close to the outside of the mature spermatozoon. The condensation of the nucleus and the formation of the acrosome are described in particular. Because of the complicated transformation of the spermatide also the nucleus, the acrosome and the centrioles undergo extensive migrations being accomplished in the female.

     

Diacylglycerol species as messengers and substrates for phosphatidylcholine re-synthesis during Ca2+-dependent exocytosis in boar spermatozoa

We have investigated pathways of lipid metabolism in spermatozoa and generation of various metabolites with potential messenger functions during exocytosis stimulated with A23187/Ca²⁺. Stimulation of boar spermatozoa resulted in a considerable rapid increase in saturated/unsaturated 1,2-diacylglycerol (1,2-SU-DAG) and, concomitantly, a substantial reduction in disaturated 1,2-diacylglycerol (1,2-DS-DAG), and in phosphatidylcholine (PC). These changes preceded the onset of exocytosis. Phosphatidic acid was sometimes generated in parallel, but usually rose later, suggesting that 1,2-SU-DAG may be formed directly by phospholipase C action. Lipid changes observed in stimulated spermatozoa that have been prelabelled with several lipid precursors ([¹⁴C]palmitic acid, [¹⁴C]glycerol, [¹⁴C]choline, or [¹⁴C]arachidonic acid) suggested the existence of a unique process involving the utilization of the high basal levels of 1,2-DS-DAG to form 1,2-SU-DAG, with the latter being subsequently employed to replenish the PC pool. An ensuing generation of lysoPC and arachidonic acid, which paralleled the occurrence of exocytosis, revealed that the newly synthesized PC was hydrolyzed by phospholipase A₂. The highest levels of 1,2-SU-DAG, minimum levels of 1,2-DS-DAG, and the regeneration of the PC pool were tightly coupled to the beginning of visible exocytosis. These results suggest that changes in these lipid metabolites may be fundamental processes during acrosomal exocytosis occurring in response to physiological agonists. Mol. Reprod. Dev. 48:95–105, 1997. © 1997 Wiley-Liss, Inc.     

Hamster sperm antigen P26h is a phosphatidylinositol‐anchored protein

We have previously identified a hamster sperm protein, P26h, proposed to be involved in the interaction between spermatozoa and the egg's zona pellucida. In this study we investigated the mechanism of P26h accumulation on hamster spermatozoa during epididymal maturation. Immunocytochemical studies showed an accumulation of P26h on the acrosomal cap of hamster spermatozoa during epididymal transit. To document the anchoring mechanism of P26h, cauda epididymal spermatozoa were exposed to different treatments. High‐salt buffered solutions were unable to remove P26h from the surface of intact spermatozoa. P26h was released in a dose‐dependent manner when live spermatozoa were treated with a solution of phospholipase C specific to phosphatidylinositol. In contrast, the P26h remained associated to the sperm surface following treatment with trypsin. To document the transfer mechanisms of P26h on the maturing spermatozoa, prostasomes were isolated from the epididymal fluid and subjected to immunodetection. Western blots and immunogold studies showed that P26h was associated to epididymal prostasomes. Phospholipase C treatment performed on epididymal prostasomes, indicated that P26h also is anchored to these vesicles via a phosphatidylinositol. These results suggest that epididymal sperm maturation involves a cell to cell transfer of a phosphaditylinositol‐anchored protein and that prostasomes may be implicated in this process. Mol. Reprod. Dev. 52:225–233, 1999. © 1999 Wiley‐Liss, Inc.     

Adenovirus‐mediated introduction of DNA into pig sperm and offspring

The ability of adenoviral vectors to transfer DNA into boar spermatozoa and to offspring was tested. Exposure of spermatozoa to adenovirus bearing the E. coli lacZ gene resulted in the transfer of the gene to the head of the spermatozoa. Treatment did not affect either viability or acrosomal integrity of boar sperm. Of the 2‐ to 8‐cell embryos obtained after in vitro fertilization with adenovirus‐exposed sperm, 21.7% expressed the LacZ product. Four out of 56 piglets (about 7%) obtained after artificial insemination with adenovirus‐exposed spermatozoa were positive in PCR analyses, even though none of the piglets showed the LacZ gene after southern blot analysis. RT‐PCR analysis performed in tissues from two positive stillborn piglets showed the presence of the LacZ mRNA in all of the tissues tested. The offspring obtained after mating two positive animals did not show LacZ gene presence. Our results indicate that adenovirus could be a feasible mechanism for the delivery of DNA into spermatozoa, even though the transfer of the transgene may be limited to the first generation. Mol. Reprod. Dev. 53:149–158, 1999. © 1999 Wiley‐Liss, Inc.     

Novel,testis‐specific mRNA transcripts encoding N‐terminally truncated choline acetyltransferase

Previous studies reported the presence of choline acetyltransferase (ChAT) mRNA and protein in the mammalian testis. We have now found that none of the ChAT mRNAs produced in the testis is capable of encoding a full‐length ChAT protein. Two ChAT cDNAs were isolated from an adult rat testis cDNA library encoding N‐terminally truncated ChAT proteins of 450 and 414 amino acids (aa), respectively, the former containing a novel N‐terminal extension of 69 residues. Rapid Amplification of cDNA Ends (RACE) analysis revealed a complex pattern of 5′ untranslated mRNA termini generated from the ChAT gene locus in the testis, all representing truncated versions of the ChAT enzyme. Two of these proteins were produced in transfected fibroblasts and found to lack ChAT activity. Neither did they show binding to the ChAT substrates, acetyl CoA and choline, in a competition assay. These results indicate that mammalian testis lacks a bona fide ChAT enzyme but expresses truncated ChAT proteins with a possible unique function to the testis. Mol. Reprod. Dev. 53:274–281, 1999. © 1999 Wiley‐Liss, Inc.