Structure and energy pathways of spermatozoa of the rove beetle Aleochara bilineata (Coleoptera, Staphylinidae)

The morphology of mature spermatozoa of the rove beetle Aleochara bilineata was examined by using scanning and transmission electron microscopy. They are about 1000 mum long and filiform. The acrosome and the nucleus are elongate and each about 20 mum long. A well-developed centriole adjunct region connects the nucleus with the sperm tail. The axoneme reveals the 9 + 9 + 2 pattern of the pterygote sperm flagellum. Two accessory bodies and two mitochondrial derivatives with paracrystalline inclusions are present. Cristae are reduced to the cortical zone of the derivatives. Cytochrome-c oxidase activity was detected within the cristae by DAB-reaction. The energy metabolism of the spermatozoa was investigated by using different inhibitors affecting the mitochondrial and cytoplasmic metabolic pathways. Sperm movement was used as an indicator for the utilization of ATP by the axoneme. In control experiments, the duration of motility was longer than 45 min. In the presence of atractyloside or potassium cyanide the motility duration was not affected. On the other hand, iodoacetic acid in the medium stopped sperm motility within 15 min. This indicates that sperm energy metabolism mainly depends on the glycolytic pathway.     

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.     

Ultrastructure of apyrene and eupyrene spermatozoa from the seminal vesicle of Euptoieta hegesia (Lepidoptera: Nymphalidae)

The ultrastructure of the seminal vesicle's spermatozoa of the butterfly Euptoieta hegesia was analyzed. The apyrene spermatozoa measure about 300 microm in length and swim freely in a secretion. The anterior end consists in a cap with a cylindrical extension and a globular structure. The flagellum has a 9+9+2 axoneme, two mitochondrial derivatives with paracrystalline matrices and an external coat formed by concentric layers. The eupyrene spermatozoa measure about 550 microm in length and are grouped into bundles. The anterior end consists in an amorphous globule. Posterior to this globule, a coat with a dense material covers the spermatozoon where an acrosome and a nucleus appear. The flagellum has a 9+9+2 axoneme and two mitochondrial derivatives. External to the coat and attached to the dense material, there is a reticular appendage, which has a paracrystalline core and extends to the distal tip of the spermatozoon.     

Fine structure of scorpion spermatozoa (Buthus occitanus; buthidae,scorpiones)

The mature spermatozoa of Buthus occitanus are threadlike in shape and divided into sperm head, middle piece, and end piece. The sperm head is corkscrew shaped anteriorly and in this region bears an unusual acrosomal complex consisting of a ring-shaped acrosomal vacuole associated with a subacrosomal filament and a perinuclear amorphous component. The subacrosomal filament extends posteriorly into a tube-like invagination of the elongated nucleus. The middle piece is characterized by elongated mitochondria which spiral around the anterior part of the flagellum in an extended collar separated from the flagellum by an extracellular cleft, termed the central flagellar tunnel. In addition to the usual 9 × 2 + 2 axonemal pattern in flagella, 9 × 2 + 1 and 9 × 2 + 3 patterns also were observed. The end piece is represented by the free flagellum. Similarities and diversities of scorpionid spermatozoa are discussed with respect to systematic relationships.     

三角帆蚌精子的形态及超微结构

运用电子显微镜技术对三角帆蚌精子的形态和超微结构进行研究。结果发现,三角帆蚌精子为原生型,分为头部、中段和尾部,头部呈子弹头形,电子致密且均匀,主要是核所在的区域。核前端由3－4个小的电子致密颗粒组成一个浅弧形的囊泡,为顶体结构,中段具有5个球形线粒体,环绕着两个相互垂直的中心粒。中段末端具有的鞭毛质领结构（flagellar collar)为一电子致密环,与远端中心粒之间由9个分叉的电子致密小片连接。尾部为典型的9＋2结构。     

Ultrastructure and motility of the caudal epididymis spermatozoa from the volcano mouse (Neotomodon alstoni alstoni Merriam, 1898)

The volcano mouse Neotomodon alstoni alstoni is a genus endemic to the higher elevations of the Mexican transvolcanic belt. In the present study we examined for the first time the morphological features of the spermatozoa taken from the caudal epididymis of this species by transmission and scanning electron microscopy. Spermatozoan motility was studied in sucrose and bicarbonate solutions; vitality and morphology were observed by light microscopy. Transmission electron microscopy shows that the head of spermatozoon is asymmetric and possesses a large and curved hook. The axoneme of the spermatozoan tail is highly developed at fibers 1, 5, and 6. Absolute and relative measurements of the length of the head, the midpiece, and the rest of the tail were also obtained. N. alstoni alstoni spermatozoa were hyperactive in the presence of 290 mM sucrose and 10 and 20 nM bicarbonate solutions exhibited high motility (180-190 microm/sec), and high flagellum beating frequency (10-12 Hz). In contrast, the spermatozoa in 310 mM sucrose solution showed scarce motility (13.5 +/- 3.8 microm/sec) and low beating frequency (1.5 +/- 0.4 Hz). It is proposed that the volcano mouse spermatozoa possess some features very similar to other mammalian spermatozoa and that bicarbonate triggers caudal epididymal sperm motility of this species. J. Exp. Zool. 287:316-326, 2000.     

Morphology and ultrastructure of Siberian sturgeon (Acipenser baerii) spermatozoa using scanning and transmission electron microscopy

BACKGROUND INFORMATION: Available data concerning the sperm morphology of teleost fishes demonstrate wide variation. In the present study, the spermatozoa of Siberian sturgeon (Acipenser baerii Brandt, 1869), a chondrostean fish, was investigated. In contrast with teleost fish, chondrostean spermatozoa have a head with a distinct acrosome, whereas other structures, such as a midpiece and a single flagellum, are present in spermatozoa of most species. RESULTS: The average length of the head including the acrosome and the midpiece was 7.01+/-0.83 microm. Ten posterolateral projections derived from the acrosome were present on a subacrosomal region, with mean lengths of 0.94+/-0.15 microm and widths of 0.93+/-0.11 microm. The nucleus consisted of electrodense homogeneous nuclear chromatin. Three intertwining endonuclear canals, bound by membranes, traversed the nucleus longitudinally from the acrosomal end to the basal nuclear fossa region. There were between three and six mitochondria, two types of centrioles (proximal and distal) in the midpiece and two vacuoles composed of lipid droplets. The flagellum (44.75+/-4.93 microm in length), originating from the centriolar apparatus, had a typical 9+2 eukaryotic flagellar organization. In addition, there was an extracellular cytoplasm canal between the cytoplasmic sheath and the flagellum. CONCLUSIONS: A principal components analysis explained the individual morphological variation fairly well. Of the total accumulated variance, 41.45% was accounted for by parameters related to the head and midpiece of the sperm and the length of the flagellum. Comparing the present study with previous studies of morphology of sturgeon spermatozoa, there were large inter- or intra-specific differences that could be valuable taxonomically.     

The sperm structure of Cryptocercus punctulatus Scudder (Blattodea) and sperm evolution in Dictyoptera

Sperm of the dictyopteran key taxon Cryptocercus punctulatus was examined. It has largely maintained a blattodean groundplan condition, with a three‐layered acrosome, an elongate nucleus, a single centriole, a conspicuous centriole adjunct material, two connecting bands (=accessory bodies), and a long functional flagellum with a 9+9+2 axoneme provided with accessory tubules with 16 protofilaments and intertubular material. These sperm characters are shared with several other polyneopterans. The sperm of C. punctulatus is very similar to what is found in Periplaneta americana and species of other groups of roaches, including the sperm of Loboptera decipiens described here for the first time. The general sperm organization here described can be assumed for the groundplan of Insecta and Pterygota. The following evolutionary path can be suggested: after the split between Cryptocercidae and the common ancestor of Isoptera, the typical pattern of sperm formation was altered very distinctly, resulting in a duplication or multiplication (Mastotermitidae) of the centrioles. Mastotermes has maintained a certain sperm motility, but with a very unusual apparatus of multiple flagella with a 9+0 axoneme pattern. After the split into Mastotermitidae and the remaining Isoptera, sperm motility was completely abandoned, and different modifications of sperm components occurred, and even the loss of the sperm flagellum. J. Morphol. 276:361–369, 2015. © 2014 Wiley Periodicals, Inc.     

Sperm ultrastructure and spermiogenesis of Coniopterygidae (Neuroptera, Insecta)

The spermiogenesis and the sperm ultrastructure of several species of Coniopterygidae have been examined. The spermatozoa consist of a three-layered acrosome, an elongated elliptical nucleus, a long flagellum provided with a 9+9+3 axoneme and two mitochondrial derivatives. No accessory bodies were observed. The axoneme exhibits accessory microtubules provided with 13, rather than 16, protofilaments in their tubular wall; the intertubular material is reduced and distributed differently from that observed in other Neuropterida. Sperm axoneme organization supports the isolated position of the family previously proposed on the basis of morphological data.     

Identification of the 58-kDa phosphoprotein associated with motility initiation of hamster spermatozoa

In our previous paper [M. Fujinoki et al. (2001) BIOMED: Res. 22, 45-58], we reported that the 58-kDa protein obtained from hamster sperm flagella was phosphorylated at serine residues in association with the start of motility. In the present experiments, we identified and localized the 58-kDa protein. The 58-kDa protein was assumed to exist in the acrosomal region domain of the sperm head and the whole sperm flagellum. In particular, a large amount of 58-kDa protein was localized in the equatorial segment of the acrosomal region domain of the sperm head and the middle piece of the sperm flagellum. In the next step, the 58-kDa protein was identified by peptide mass finger printing and LC-MS/MS analysis. The results suggested that the 58-kDa protein was ATP synthase H(+) transporting F1 beta, which is one of the mitochondrial components. Therefore, it is likely that the 58-kDa protein is associated with ATP production in the mitochondrial sheath in the middle piece of the sperm flagellum, and H(+) transport in the sperm head and the sperm flagellum except for the middle piece, since ATP synthase also acts as an H(+) pump.     

Effects of osmolality on sperm morphology, motility and flagellar wave parameters in Northern pike (Esox lucius L.)

Northern pike (Esox lucius L.) spermatozoa are uniflagellated cells differentiated into a head without acrosome, a midpiece and a flagellar tail region flanked by a fin structure. Total, flagellar, head and midpiece lengths of spermatozoa were measured and show mean values of 34.5, 32.0, 1.32, 1.17 μm, respectively, with anterior and posterior widths of the midpiece measuring 0.8 and 0.6 μm, respectively. The osmolality of seminal plasma ranged from 228 to 350 mOsmol kg⁻¹ (average: 283.88 ± 33.05). After triggering of sperm motility in very low osmolality medium (distilled water), blebs appeared along the flagellum. At later periods in the motility phase, the tip of the flagellum became curled into a loop shape which resulted in a shortening of the flagellum and a restriction of wave development to the proximal part (close to head). Spermatozoa velocity and percentage of motile spermatozoa decreased rapidly as a function of time postactivation and depended on the osmolality of activation media (P < 0.05). In general, the greatest percentage of motile spermatozoa and highest spermatozoa velocity were observed between 125 and 235 mOsmol kg⁻¹. Osmolality above 375 mOsmol kg⁻¹ inhibited the motility of spermatozoa. After triggering of sperm motility in activation media, beating waves propagated along the full length of flagella, while waves appeared dampened during later periods in the motility phase, and were absent at the end of the motility phase. By increasing osmolality, the velocity of spermatozoa reached the highest value while wave length, amplitude, number of waves and curvatures also were at their highest values. This study showed that sperm morphology can be used for fish classification. Sperm morphology, in particular, the flagellar part showed several changes during activation in distilled water. Sperm motility of pike is inhibited due to high osmolality in the seminal plasma. Osmolality of activation medium affects the percentage of motile sperm and spermatozoa velocity due to changes in flagellar wave parameters.     

Ultrastructural study of spermatozoa of the paddlefish, Polyodon spathula

Paddlefish, Polyodon spathula, spermatozoa were examined by transmission electron microscopy. Their structure has the same characteristic architectural features as sturgeon spermatozoa. Paddlefish spermatozoa are of the primitive type and consist of a rod-shaped head, a midpiece and a long flagellum. The head is about 5.15 mm in length and contains the nucleus and an apical acrosomal complex. Inside the nucleus there are three nuclear channels that begin in the subacrosomal area and have a triple helical arrangement. An nuclear fossa is present centrally, at the posterior end of the nucleus. The midpiece contains a pair of centrioles in a perpendicular arrangement, mitochondria and a narrow cytoplasmic sleeve. The flagellum has a central axoneme with a 9 + 2 pattern and two lateral projections or fins.     

Calcium regulation of flagellar curvature and swimming pattern in triton X-100--extracted rat sperm

Free Ca2+ changes the curvature of epididymal rat sperm flagella in demembranated sperm models. The radius of curvature of the flagellar midpiece region was measured and found to be a continuous function of the free Ca2+ concentration. Below 10(-7) M free Ca2+, the sperm flagella assumed a pronounced curvature in the same direction as the sperm head. The curvature reversed direction at 2.5 x 10(-6) M Ca2+ to assume a tight, hook-like bend at concentrations of 10(-5) to 10(-4) M free Ca2+. Sodium vanadate at 2 x 10(-6) M blocked flagellar motility, but did not inhibit the Ca2+-mediated change in curvature. Nickel ion at 0.2 mM and cadmium ion at 1 microM interfered with the transition and induced the low Ca2+ configuration of the flagellum. The forces that maintain the Ca2+-dependent curvature are locally produced, as dissection of the flagella into segments did not significantly alter the curvature of the excised portions. Irrespective of the induced pattern of curvature, the sperm exhibited coordinated, repetitive flagellar beating in the presence of ATP and cAMP. At 0.3 mM ATP the flagellar waves propagated along the principal piece while the level of free Ca2+ controlled the overall curvature. When Ca2+-treated sperm models with hooked midpieces were subjected to higher concentrations of ATP (1-5 mM), some cells exhibited a pattern of movement similar to hyperactivated motility in capacitated live sperm. This type of motility involved repetitive reversals of the Ca2+-induced bend in the midpiece, as well as waves propagated along the principal piece. The free Ca2+ available to the flagellum therefore appeared to modify both the pattern of motility and the flagellar curvature.     

Morphology of the male reproductive system and sperm ultrastructure of Leucoptera coffeella (Lepidoptera: Lyonetiidae)

The male reproductive tract of Leucoptera coffeella was processed for light and transmission electron microscopy. In the testis, the eupyrene cells are arranged in individual cysts, while the apyrene cysts form aggregates, never observed in other Lepidoptera. Both cysts contain 128 spermatozoa, which differ from the typical pattern. In the seminal vesicle, both types of spermatozoa are dispersed in the lumen, also different from other Lepidoptera. The apyrene spermatozoa are similar to those observed for other Lepidoptera. They present an anterior region covered by a dense cap and the flagellum is composed of a 9 + 9 + 2 axoneme and two mitochondrial derivatives. The eupyrene spermatozoa, however, differ from the typical pattern for Lepidoptera. Their anterior region contains a nucleus, an acrosome and a peculiar arc of eight accessory microtubules connected to the plasma membrane by dense bridges. In the nucleus–flagellum region, the ninth accessory microtubule is assembled between both mitochondrial derivatives, to participate in the axoneme. The flagellum comprises a 9 + 9 + 2 axoneme and two mitochondrial derivatives with paracrystalline cores. External to the plasma membrane and close to the accessory microtubules, there are tufts of an amorphous material, suggesting reduced lacinate appendages, while the reticular ones are absent. The reduction of lacinate appendages and the absence of sperm bundles in the seminal vesicle support the concept that the appendages of other Lepidoptera could be associated with the eupyrene aggregations. The characters ‘number of spermatozoa per cyst’ and ‘absence of bundles’ should be considered plesiomorphic, supporting the position of this taxon in the base of the Ditrysia.     

Exceptions to the prevailing pattern of tubules (9 + 9 + 2) in the sperm flagella of certain insect species

Various deviations from classical 9 + 2 flagellar structure are found in sperm of insect species. In mature spermatozoa of a psocid, Psocus, the outer flagellar tubules are not straight, but are disposed in a long-pitched helix such that they form an angle of about 8° with a single dense rod located in the position usually occupied by the central pair. In young spermatids of Psocus the outer tubules are straight; thus, spiraling of the flagellar tubules occurs during the course of spermiogenesis. Spiraling of flagella also occurs in the cat flea Ctenocephalides felis. Variations in the number and morphology of the central element or elements occur in other insect species besides Psocus. Among the observed deviations from a central pair of tubules are a 9 + 0 tubule pattern in the sperm of three species of mayflies, a 9 + 1 tubule pattern in the sperm of two species of mosquitoes, and 9 + 7 tubules in sperm of two species of caddis flies. Spermatozoa of treehoppers vary in yet another respect from the typical 9 + 9 + 2 insect flagellum. These sperm tails branch into four long tails, three of which each contain two doublet and two singlet tubules while the fourth branch contains three doublet and three singlet tubules. The wide distribution of insects with aberrant flagella suggests that the variant forms have evolved independently.     

Three-dimensional motion of avian spermatozoa

Observations have been made on spermatozoa from the domestic fowl, quail and pigeon (non-passerine birds) and also from the starling and zebra finch (passerine birds). In free motion, all these spermatozoa roll (spin) continuously about the progression axis, whether or not they are close to a plane surface. Furthermore, the direction of roll is consistently clockwise (as seen from ahead). The flagellar wave has been shown to be helical and dextral (as predicted) for domestic fowl sperm when they swim rapidly in low viscosity salines. Calculations have shown that their forward velocity is consistent with their induced angular velocity but that the size of the sperm head is suboptimal for progression speed under these conditions. Dextrally helical waves also occur on the distal flagellum of fowl, quail and pigeon sperm in high viscosity solutions. But in other cases, the mechanism of torque-generation is more problematical. The problem is most profound for passerine sperm, in that typically these cells spin rapidly while seeming to remain virtually straight. Because there is no evidence for a helical wave on these flagella, we have considered other possible means whereby rotation about the local flagellar axis (self-spin) might be achieved. Sometimes, passerine sperm, while maintaining their spinning motion, adopt a fixed curvature; this must be an instance of bend-transfer circumferentially around the axonemal cylinder-though the mechanism is obscure. It is suggested that the self-spin phenomenon may be occurring in non-passerine sperm that in some circumstances spin persistently, yet without expressing regular helical waves. More complex waves are apparent in non-passerine sperm swimming in high viscosity solutions: added to the small scale bends is a large scale, sinistrally helical curvature of the flagellum. It is argued that the flagellum follows this sinistrally helical path (i.e. "screws" though the fluid) because of the shape of the sperm head and the angle at which the flagellum is inserted into it. These conclusions concerning avian sperm motility are thought to have relevance to other animal groups. Also reported are relevant aspects of flagellar ultrastructure for pigeon and starling sperm.     

Sperm Bundles in the Seminal Vesicle of the Crematogaster victima (Smith) Adult Males (Hymenoptera: Formicidae)

This study establishes the presence of spermatodesm in the seminal vesicles of sexually mature males of Crematogaster victima (Smith). In this species, the spermatozoa are maintained together by an extracellular matrix in which the acrosomal regions are embedded. This characteristic has not yet been observed in any other Aculeata. However, the sperm morphology in this species is similar to that described for other ants. The spermatozoa measure on average 100 μm in length, and the number of sperm per bundle is up to 256. They are composed of a head formed by the acrosome and nucleus; this is followed by the flagellum, which is formed by the centriolar adjunct, an axoneme with a 9?+?9?+?2 microtubule pattern, two mitochondrial derivatives, and two accessory bodies. The acrosome is formed by the acrosomal vesicle and perforatorium. The nucleus is filled with compact chromatin with many areas of thick and non-compacted filaments. Both mitochondrial derivatives have the same shape and diameters. The presence of sperm bundles in sexually mature males differentiates C. victima from other ants; however, the similarities in the sperm ultrastructure support the monophyly of this insect group.     

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.