Ultrastructural characterization of spermatozoa in euglossine bees (Hymenoptera, Apidae, Apinae)

Summary. Euglossine spermatozoa are the longest described to date for the Hymenoptera. This cell includes a head and a flagellar region. In transverse sections, the acrosome is circular at the tip but has an oval contour along most of its length. The perforatorium penetrates into a deep cavity in the nuclear tip. The flagellum consists in an axoneme, a pair of mitochondrial derivatives, a centriolar adjunct and a pair of accessory bodies. The axoneme has a 9+9+2 microtubule pattern which becomes gradually disorganized in the final portion, with the central microtubules and the nine doublets terminating simultaneously, followed by the accessory microtubules. The mitochondrial derivatives are asymmetric both in length and diameter. Sectioned transversally, the derivatives are ellipsoidal or have a pear shape. The larger one has a more obvious paracrystalline region. The centriolar adjunct begins at the nuclear base and extends parallel to the axoneme until it encounters the smaller mitochondrial derivative, on which it fits, making a concave groove. In addition to these consistent euglossine features, species-specific differences that might be useful in phylogenetic work on the group are also noted.Received 18 October 2003; revised 4 September 2004; accepted 4 October 2004.     

The short spermatodesm of Arge pagana (Hymenoptera: symphyta)

In the seminal vesicle of the 'symphyta'Arge pagana the spermatozoa are stored in motile spermatodesm bundles, maintained by an anterior cap of extracellular material. This cap consists of a denser cortex and of an internal matrix, where part of the sperm heads are embedded. The number of spermatozoa per bundle is variable. The spermatozoa are short, only 30microm long, with a head region of about 23microm, and a very short flagellum of about 7microm. The head includes the acrosome, with a perforatorium, and the nucleus. The flagellum consists of an axoneme, with a 9+9+2 microtubule pattern, a centriolar adjunct, two mitochondrial derivatives and two accessory bodies. The mitochondrial derivatives are very slender and of different lengths. The longer begins at the base of the nucleus, while the shorter one starts just below the base of the centriolar adjunct. This latter is asymmetric and appears at the nuclear base, extending parallel to the axoneme up to the anterior end of the smaller mitochondrial derivative. The short spermatodesmata and the small mitochondrial derivatives characterize the A. pagana sperm. In addition, the centriolar adjunct asymmetry and the occurrence of spermatodesm bundles might be considered plesiomorphic states present in the basal Tenthredinoidea.     

Comparative ultrastructure of ant spermatozoa (Formicidae: Hymenoptera)

Mature spermatozoa from spermathecae of founding queens were obtained from 5 species of ants, representing the major subfamilies Myrmicinae (Acromyrmex versicolor, Crematogaster sp.) and Dolichoderinae (Tapinoma sessile, Conomyrma insana, Conomyrma wheeleri). The ultrastructure of ant spermatozoa has many features in common with that of higher insects and is similar to that of other Hymenoptera. Structural similarities to spermatozoa of other Hymenoptera include an acrosome containing an internal rod that extends into the nucleus, two elongate mitochondrial derivatives, a centriolar adjunct, and an axonemal arrangement of 9 + 9 + 2 that includes well-developed coarse, or accessory, tubules. Spermatozoa obtained from A. versicolor, a species that is known to store and utilize viable sperm from this supply for over 10 years, show greater development of the mitochondrial derivatives than do the other species. The most distinctive feature of ant spermatozoa in comparison to other Hymenoptera is the large size of the centriolar adjunct relative to the other organelles. The centriolar adjunct is located posterior to the nucleus, anterior to the mitochondrial derivatives, and opposite the axoneme.     

Sperm structure and ultrastructure of the Melittobia hawaiiensis, Perkins and M. australica, Girault (chalcidoidea: eulophidae)

Spermatozoa morphology has, for some years, been used to help answer some phylogenetic questions for Hymenoptera. This is the second study describing spermatozoa morphology of an Eulophidae species in which important characteristics were observed. Melittobia spermatozoa are spiralled and measure approximately 270mum in length. The head contains a small acrosome, apparently formed only by an acrosomal vesicle, which, together with the initial nuclear region, is surrounded by an extracellular sheath, from which innumerable filaments irradiate. The nucleus is helicoidal and completely filled with compact chromatin. A centriolar adjunct is observed at the nucleus-flagellum transition; it associates laterally with the nucleus and exhibits two small expansions, which reach around the centriole. In the flagellum there are two mitochondrial derivatives, which in cross-sections are asymmetric. In the derivative with the larger diameter, two distinct regions are observed, a small one, near the axoneme, with a clear "fissure" inside, and a larger region where the cristae occur. Both derivatives initiate at the nuclear base, but the larger diameter derivative finishes first, before the flagellum extremity. At the end of the axoneme, the accessory microtubules are the first to finish.     

Structure and ultrastructure of the spermatozoa of Bephratelloides pomorum (Fabricius) (Hymenoptera: Eurytomidae)

The spermatozoa of Bephratelloides pomorum are very long and fine. Each spermatozoon measures about 620 μm in length by 0.38 μm in diameter and, when seen under the light microscope, appears to be wavy along its entire length. The head, which is approximately 105 μm, comprises a small acrosome and a nucleus. The acrosome is made up of a cone-shaped acrosomal vesicle surrounding the perforatorium and the anterior end of the nucleus. Innumerable filaments radiate from it. The perforatorium has a diameter equal to that of the nucleus at their junction, where it fits with a concave base onto the rounded nuclear tip. The nucleus is helicoidal and completely filled with homogeneous compact chromatin. It is attached to the tail by a very long and quite electron-dense centriolar adjunct that extends anteriorly from the centriole in a spiral around the nucleus for approximately 8.5 μm. The tail consists of an axoneme with the 9+9+2 microtubule arrangement pitched in a long helix, as well as a pair of spiraling mitochondrial derivatives (with regularly arranged cristae) that coil around the axoneme, and two small accessory bodies. As well as the spiraling of the nucleus, mitochondrial derivatives and axonemal microtubules, the sperm of B. pomorum present other very different morphological features. These features include the acrosome and centriolar adjunct, both of which differentiate the spermatozoa from the majority of sperm found in other Hymenoptera. In addition these structural variations demonstrate that the sperm of chalcidoids provide characteristics that can certainly prove useful for future phylogenetic analysis at the subfamily level and, possibly, the genus too.     

Sperm structure of Mecoptera and Siphonaptera (Insecta) and the phylogenetic position of<Emphasis Type="Italic"> Boreus hyemalis</Emphasis>

Sperm ultrastructure has been studied in three species of the taxa Mecoptera and Siphonaptera. The spermatozoon of the scorpion fly Panorpa germanica shows an apical bilayered acrosome, a helicoidal nucleus, a centriolar region and a 9+2 flagellar axoneme helicoidally arranged around a long mitochondrial derivative. A second mitochondrial derivative is very short and present only in the centriolar region. A single accessory body is present and it is clearly formed as a prolongation of the centriole adjunct material. Two lateral lamellae run parallel to the nucleus. The snow fly Boreus hyemalis has a conventional sperm structure and shows a bilayered acrosome, a long nucleus, a centriolar region, two mitochondrial derivatives and two accessory bodies. The axoneme is of the 9+2 type and is flattened at the tail tip. Both P. germanica and B. hyemalis have two longitudinal extra-axonemal rods and have a glycocalyx consisting of longitudinal parallel ridges or filaments. The spermatozoon of the flea Ctenocephalides canis has a long apical bilayered acrosome, a nucleus, a centriolar region, a 9+2 axoneme wound around two unequally sized mitochondrial derivatives, and two triangular accessory bodies. In the posterior tail end the flagellar axoneme disorganises and a few microtubular doublets run helicoidally around the remnant mitochondrial derivative. The glycocalyx consists of fine transverse striations. In all three species, the posterior tail tip is characterised by a dense matrix embedding the disorganised axoneme. From this comparative analysis of the sperm structure it is concluded that Mecoptera, as traditionally defined, is monophyletic and that B. hyemalis is a member of Mecoptera rather than of Siphonaptera.     

Structural and ultrastructural studies of male reproductive tract and spermatozoa in Xylocopa frontalis (Hymenoptera,Apidae)

Fiorillo, B. S., Zama, U., Lino‐Neto, J. and Báo, S. N. 2010. Structural and ultrastructural studies of male reproductive tract and spermatozoa in Xylocopa frontalis (Hymenoptera, Apidae). —Acta Zoologica (Stockholm) 91 : 176–183. In Xylocopa frontalis the reproductive tract is composed of testes, deferent ducts, seminal vesicles, accessory glands and an ejaculatory duct. Each testis comprises four testicular tubules in which multiple cysts are present containing approximately 64 spermatozoa per cyst. The seminal vesicle consists of an epithelium, a thick basement lamina and a muscular external sheet. In the luminal region some vesicles can be observed; however, the epithelial cells of the seminal vesicle do not display morphological features associated with secretory functions. The spermatozoa, measuring approximately 260 µm long, are similar to the hymenopteran pattern. The head region consists of an acrosome with an inner perforatorium that penetrates an asymmetrical nuclear tip. The nucleus is linear, electron‐dense and its posterior tip projects into the beginning of the axoneme. The centriolar adjunct is asymmetric with many electron‐lucent lacunae interspersed throughout. The axoneme has the 9 + 9 + 2 pattern of microtubules and in the posterior region the central microtubules finish first, followed by the doublets and finally the accessory microtubules. The mitochondrial derivatives are asymmetric in both length and diameter with paracrystalline material present only in the larger one. These features may be useful characters for taxonomy and phylogenetic studies.     

The neck region of the spermatozoon of Discus rotundatus (Müller) (Pulmonata,Stylommatophora)

The neck region of the mature spermatozoon of Discus rotundatus is described. No evidence for a centriole or centriolar derivative is obtained. Nine striated coarse fibres and the two central fibres of the axoneme extend into the base of the implantation fossa. The axonemal doublet system is disrupted in the neck region. There are two fibrous accessory structures located between the central doublet and the striated coarse fibres.     

Sperm morphology of the Prorops nasuta (Waterston, 1923) (Hymenoptera: Bethylidae)

In the present study, spermatozoa of the Prorops nasuta (Hymenoptera: Bethylidae) parasitoid were described morphologically. This is the first publication to describe a species belonging to the superfamily Chrysidoidea. Light and transmission electron microscopy were used. The spermatozoa of P. nasuta are linear, with a mean length of 665 μm. The acrosome is composed of an acrosomal vesicle and a perforatorium. The nucleus measures approximately 17 μm in length and is circular at its cross-section; however, its anterior extremity is oval. The chromatin is electron-dense and compact, although there are clear areas in the posterior peripheral regions. In the nucleus-flagellum transition region, the cross-section of the centriole adjunct is oval, with a pleated border and an E-PTA-positive peripheral region. The axoneme shows a 9 + 9 + 2 microtubule arrangement. The microtubules are E-PTA positive and, at the posterior extremity, the accessories are the last to terminate. The diameters and shapes of the two mitochondrial derivatives are almost identical. One begins beside the nuclear base and the other after the centriole adjunct. Posteriorly, they terminate together, immediately before the axoneme. Both have mitochondrial cristae and a region of paracrystalline material; however, the format and arrangement of this material differs from those of all other species previously studied. The paracrystalline material is more strongly E-PTA positive than the cristae region. Accessories bodies are electron-dense and located between the mitochondrial derivatives and the axoneme. In general, P. nasuta spermatozoa are similar to those of the majority of Hymenoptera; however, they have various exclusive characteristics that may be useful for studying the phylogeny and taxonomy of the superfamily Chrysidoidea and of Hymenoptera in general.     

Polymorphism of spermatozoa in Largus rufipennis Laporte 1832 (Heteroptera: Pyrrhocoroidea: Largidae)

The production of polymorphic spermatozoa has been registered in various insect orders such as Diptera, Lepidoptera, and Hemiptera. In this work, morphology of two types of spermatozoa produced by Largus rufipennis was reported for the first time in the Largidae family. For this, techniques including optical and transmission electron microscopy were used. Spermatozoa measured, on the average, 260 and 200 μm, and both types possessed a nucleus measuring on the average 65 μm. No ultrastructural differences were observed between the two spermatozoa types from L. rufipennis. The head region is composed of an acrosome, a nucleus, and part of the centriolar adjunct. The centriolar adjunct is in parallel with the nucleus and followed by mitochondrial derivates. The flagellum consists of an axoneme (9 + 9 + 2 microtubules) and two mitochondrial derivatives; no other accessory bodies were observed. The mitochondrial derivatives are symmetric in size and diameter. A similar quantity of the two spermatozoa types was observed in the seminal vesicle (57% of the large type and 43% of the small type), while in the spermatheca of the female, the larger spermatozoa were preferentially stored (87%). These results permit discussions concerning of the species biology reproduction, most specifically sperm competition strategies.     

Sperm structure of Limoniidae and their phylogenetic relationship with Tipulidae (Diptera, Nematocera)

The sperm ultrastructure of a few species of Limoniidae (Limonia nigropunctata; L. nubeculosa; Chionea n. sp.; C. alpina; C. lutescens) was studied. The two species of Limonia have a monolayered acrosome with crystallized material, a three-lobed nucleus in cross section, a ring of centriole adjunct material and a flagellum which consists of a 9+9+1 axoneme and a single mitochondrial derivative. The central axonemal tubule is provided with 15 protofilaments in its tubular wall, while the accessory tubules have 13 protofilaments and are flanked by the electron-dense intertubular material. The three species of Chionea share a monolayered acrosome, a nucleus with two longitudinal grooves, a centriole adjunct material which surrounds the centriole and the initial part of the axoneme. The axoneme is of conventional type, with 9+9+2 microtubular pattern, with accessory tubules provided with 13 protofilaments and intertubular material. However, in C. lutescens the accessory tubules start with 15 protofilaments and transform into a tubule with 13 protofilaments. These data are discussed in the light of the phylogenetic relationship between Limoniidae and Tipulidae. For this purpose, the sperm ultrastructure of Nephrotoma appendiculata was also considered comparatively.     

New findings on sperm ultrastructure of Xenos vesparum (Rossi) (Strepsiptera,Insecta)

The systematic position of insect order Strepsiptera is still under debate. It was, therefore, thought of interest to examine the ultrastructure of a strepsipteran in a search for synapomorphies shared with Coleoptera, Diptera, or any other insect order. The fine structure of spermatozoa and the spermatid from Xenos vesparum (Rossi) was re-examined using scanning and transmission electron microscopy and a fixation technique that permits the visualization of the macromolecular organization of the organelles. The spermatozoon was shown to possess several traits that are characteristics of insects in general, such as a 9 + 9 + 2 axoneme, two mitochondrial derivatives containing a crystalline material and two 'zipper lines' present along the sperm tail. Seventeen protofilaments occurred along most of the accessory tubules, which reduced to 16 posteriorly. An acrosome is absent. The neck region contains a prominent centriolar adjunct, which gives rise to two accessory bodies which adhere to the mitochondrial derivatives, and to slender strands of the so-called intertubular material found between the accessory tubules. Of interest is the finding that the glycocalyx consists of prominent filamentous strands, similar to those found in siphonapterans, mecopterans and basal dipterans.     

Ultrastructure des spermatozoides des males haploides et diploides de Diadromus pulchellus wesmeal (Hymenoptera : Ichneumonidae)

The mature spermatozoa were described in the haploïd and diploïd males of Diadromus pulchellus Wesmeal (Hymenoptera : Ichneumonidae). Diploïd males produce spermatozoa, which do not seem to be different from those produced by haploïd males. The spermatozoon is about 100 μm long, and consists of a head, 0.8 μm in diameter, and a tail 0.3 μm in diameter. Its anterior part shows an acrosomal complex, including a perforatorium and a compact and electron-dense fusiform nucleus. The postnuclear region includes a longitudinal axoneme with 2 mitochondrial derivatives. The axoneme shows 2 typical central units, 9 peripheral doublet microtubules, 9 accessory internal tubules, and 9 external microtubules with dense contents. In the testes of diploïd males, a great number of abnormal spermatozoa were observed. These spermatozoa with degenerative structures are probably not implicated in egg fertilization.     

短翅华癞蝗精子发生中尾部的演化过程及与飞蝗有关内容的比较（直翅目：蝗总科）

本文以精子发生中线粒体衍生体的演化为主要线索系统描述了短翅华癞蝗（Ｓｉｎｏｔｍｅ－ｔｈｉｓｂｒａｃｈｙｐｔｅｒｕｓ）精子尾部演化过程,并参照Ｓｚｏｌｌｏｓｉ（１９７５）的文章将整个过程分为１０个阶段。与飞蝗（Ｌｏｃｕｓｔａｍｉｇｒａｔｏｒｉａ）精子发生中尾部演化过程（Ｓｚｏｌｌｏｓｉ,１９７５）相比较,两者间１至６期及９期精细胞尾部结构惊人地相似,而７、８两期精细胞尾部线粒体衍生体周围微管排列及分布有所差异,第１０期即精子的中心粒侧体及线粒体衍生体超微结构有明显不同。     

Ultrastructural studies on euspermatozoa and paraspermatozoa in Mantispidae (Insecta, Neuroptera)

Previous studies have demonstrated the presence of sperm dimorphism in the Mantispidae Perlamantispa perla. We extended the study on several other mantidflies. In all the examined species the occurrence of euspermatozoa (typical) and paraspermatozoa (atypical) was established. The euspermatozoa are characterized by the presence of a cylindrical nucleus surrounded by an envelope that fans out laterally into two thin wings of different length. The acrosome seems to be missing. The nucleus is surrounded by extracellular material. The flagellum is provided with a 9 + 9 + 2 axonemal pattern; the accessory tubules contain 16 protofilaments and the intertubular material has the distribution typical of the taxon. Two elongated accessory bodies flank partially the axoneme and connect this structure with the mitochondrial derivatives. The flagellar axoneme of paraspermatozoa consists of an axoneme and two giant mitochondrial derivatives filled with large globular units. The axoneme exhibits a 9 + 9 + 2 pattern, in which the central 9 + 2 units have a normal structure, in that the microtubular doublets are provided with both dynein arms and radial links. On the contrary, the nine accessory microtubules have a large diameter and their tubular wall consists of 40 protofilaments. This comparative study provided evidences about the uniformity of sperm ultrastructure in Mantispidae. The function of non-fertilizing giant sperm in mantidflies is discussed.     

Sperm ultrastructure of five species of the Neotropical ant genus Pseudomyrmex (Hymenoptera: Formicidae)

The seminal vesicles of adult males of five species of Pseudomyrmex were prepared for light and transmission electron microscopy. The Pseudomyrmex spermatozoa are long and slender with similar morphology. The head region has an acrosome and a nucleus. In all the studied species, two morphologically distinct types of acrosomal vesicles were observed, a long structure, as observed in all known ants, and a pear‐shaped one, never before observed in ants. The nucleus is elongated and both condensed and loose chromatin are present. The flagellum has an axoneme, a centriolar adjunct, two mitochondrial derivatives and two accessory bodies. The centriolar, the mitochondrial derivatives and the accessory bodies are similar to observations in most ant species that have been studied. The axoneme presents an uncommon 9 + 9 + 1 microtubule arrangement and the central microtubule has 13 protofilaments. The acrosomal dimorphism and the different levels of chromatin organization are exclusive characteristics of Pseudomyrmex. Furthermore, the 9 + 9 + 1 microtubule arrangement is different from all Hymenoptera, as well as from most insects, which present a 9 + 9 + 2 arrangement. These new morphological characters that are specific to Pseudomyrmex, are valuable synapomorphies of the genus and can be used in taxonomic characterization of the Pseudomyrmecinae subfamily and in phylogenetic analyses in Formicidae family.