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.     

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.     

Ultrastructure of the biflagellate spermatozoon of tomopteris helgolandica greef, 1879 (annelida,polychaeta)

The spermatozoon of the polychaete Tomopteris helgolandica is of an aberrant type with two flagella, each measuring about 40μm. The nucleus is roughly conical and weakly bent. At the anterior end it is rounded and covered only by the nuclear and plasma membranes. Membraneous, electron-dense structures are applied laterally to the nucleus. These structures may have a helical arrangement. The middle piece contains about ten mitochondria, two centrioles, and two centriolar satellite complexes. The centriolar regions are connected with the posterior part of the nucleus. The axonemes of the two tail flagella lack the usual central complex with central tubules, radial spokes, or related structures. No arms seem to be present on the A tubules of the doublets. In the middle piece the tail flagella are surrounded by invaginations of the plasma membrane forming flagellar canals. The sperm has a bilateral symmetry whereas the primitive sperm has a radial symmetry. The occurrence of two tail flagella in this spermatozoon has no phylogenetical connection with biflagellate spermatozoa in other animal groups. A series of mutations has resulted in the development of two flagella emerging from the two centrioles, the lack of a central complex in the axoneme, and the lack of a typical acrosome. In the Polychaeta, sperm structure is generally more related to function that to phylogenetics. During swimming the spermatozoon of Tomopteris rotates around its longitudinal axis.     

FINE STRUCTURE OF SCIARA COPROPHILA SPERM

Though the fagellum of Sciara sperm arises from a blepharoplast and is characterized by doublet tubules with arms, it differs markedly from the familiar type of flagella in the number and arrangement of its tubules. The axial filament complex in sperm from the testis of Sciara consists of approximately 70 doublet tubules, each with an associated singlet tubule. Near the nucleus these tubules are displaced in an oval array. Posteriorly the oval breaks and coils from one free end so that the axial filament complex at posterior levels has the form of a spiral. The singlet tubules do not extend the full length of the sperm but terminate in order from inside the spiral. Farther posteriorly the axial filament complex reverses the direction of coiling, and the doublets terminate from outside the spiral. Four arms are specifically positioned on the singlet and doublet tubules. A single mitochondrial derivative extends most of the length of the sperm; it consists of a large mass of proteinacious material, a crystalloid located adjacent to the axial filament complex, and peripheral cristae. In the female genital tract, sperm undergo gross morphological changes which include sloughing of practically all the mitochondrial material except the crystalloid, repositioning of the crystalloid, and uncoiling and subsequent recoiling of the axial filament complex into a different configuration. From analysis of serial sections it was determined that the orientation of arms, when the axial filament is viewed from base to tip, is the same as in conventional flagella.     

Sperm structure of Trichoptera. I. Integripalpia : Limnephiloidea

Spermal ultrastructure in 16 caddisflies (Trichoptera) belonging to the suborder Integripalpia, superfamily Limnephiloidea, was examined in a search for apomorphic and plesiomorphic features. In all species examined, the sperm tail axoneme was of the 9 + 9 + 2 + type, the axonemal doublets lacked outer dynein arms but had inner ones, and the cell membrane was scalloped with a prominent glycocalyx. The number of protofilaments in the accessory tubules depended on the phylogenetic position: 18 in the family Leptoceridae, 19 in Limnephilidae, Goeridae and Odontoceridae (with a reduction in the number distally), and 20 in Sericostomatidae. Spermatozoa in Leptoccridae are further characterized by the 2 central microtubules being flattened and eccentric and not being surrounded by a central sheath. Spermatozoa of Sericostomatidae have an accessory body and a helicoidal array of the sperm tail.     

Ultrastructural spermatid and sperm morphology in Poecilocerus pictus (Fab.) with a reference to spermeiophagic cells in the testis and sperm duct

Electron microscopical studies were carried out on spermatid and sperm structure in P. pictus. The spermatid nuclear envelope possesses pores and is surrounded by microtubules which disappear on metamorphosis to sperm though centriolar adjunct, and its corresponding centriole comprising the basal body for flagellum. remains persistent in both. The mitochondria are arranged as two fused bodies with prominent cristae flanking the central axoneme and also contain curved end feet. In axoneme the microtubular complex is comprised of 9 + 9 (doublet) + 2 tubules + nine coarse fibres and also reveals nine radial links with electron-dense link heads. In P. pictus an alteration in temperature range, ambient for its rearing and generation of fertile spermatozoa, induces the production of sterile sperms which are characterized by multiple axonemes and mitochondrial bodies engirdled by a common plasma membrane. Presence of phagocytic cells is also an essential feature of its testis and vas deferens. These spermeiophagic cells engulf the neighbouring spermatozoa as evidenced by the fragments of axoneme, nuclei, and acrosomes in their cytoplasm.     

The spermatozoon of arthropoda. XXX. The multiflagellate spermatozoon in the termite Mastotermes darwiniensis

In this paper the spermatozoon of the termite Mastotermes darwiniensis is described. It is the first example of a multiflagellate sperm cell in animals. The sperm consists of a conical head and 100 flagella. Other remarkable features of this sperm cell are the absence of an acrosome, the presence of centrioles containing doublet microtubules instead of triplets, and the presence of axonemes devoid of central tubules and with doublets bearing only one arm. The flagella are feebly motile.     

Accessory tubules and axonemal microtubules of Apis mellifera sperm flagellum differ in their tubulin isoform content

In the insect sperm flagellum, an extra set of nine additional microtubules, named accessory tubules, is present surrounding the axoneme. Using a sarcosyl/urea extraction, we were able to fractionate the microtubular cytoskeleton of the sperm flagellum of the insect Apis mellifera resulting in the dissociation of the axonemal microtubule protein components and the accessory tubules. This has allowed us to compare the tubulin isoform content of axonemal microtubules and accessory tubules by immunoelectron microscopy and immunoblotting using a panel of monoclonal antibodies directed against different tubulin post-translational modifications (PTMs). All the PTMs occurring in axonemal tubulin are also present in accessory tubules, which indicates the close relativeness of accessory tubules to axonemal rather than to cytoplasmic microtubules. However, our results demonstrate the presence of significant differences in the tubulin isoform content of axonemal microtubules and accessory tubules. First, the tubulin tyrosination extent of accessory tubules is far lower than that of axonemal microtubules, thus confirming at the molecular level their morphogenetic origin as outgrowths from the B-subtubule of each microtubular doublet. Second, although polyglycylation seems to occurr at the same extent in both microtubular systems, alpha-tubulin exhibits a larger amount of monoglycylated sites in axonemal microtubules than in accessory tubules. Third, a greater amount of beta-tubulin molecules is glutamylated in axonemal microtubules than in accessory tubules. Moreover, highly acidic isoforms, likely molecules with longer polyglutamate side chains, are present only in axonemal microtubules. Taken together, our data are indicative of a higher level of tubulin heterogeneity in axonemal microtubules than in accessory tubules. They also show a segregation of post-translationally modified isoforms between accessory tubules and axonemal microtubules and suggest the implication of PTMs in the functional specialization of the two microtubular systems.     

蓝蝽对马铃薯甲虫低龄幼虫的捕食潜能初探

室内研究了蓝蝽Zicrona caerula(Linnaeus)对马铃薯甲虫Leptinotarsa decemlineata(Say)低龄幼虫的捕食功能反应和选择效应。HollingII型圆盘方程和HollingⅢ型新模型拟合结果共同显示,蓝蝽若虫对马铃薯甲虫1龄幼虫的捕食量随着龄期的增加而增大;4、5龄若虫及成虫的捕食量均在17头以上。由HollingII模型获得蓝蝽对马铃薯甲虫1龄幼虫的控制能力为2龄<3龄<4龄<5龄<成虫;随着食物密度的增加,蓝蝽各虫态的寻找效应逐渐下降。田间防治马铃薯甲虫1龄幼虫时,2～5龄蓝蝽若虫及成虫的最佳益害比应该分别为1∶2、1∶4、1∶13、1∶14、1∶13,然而,食物选择效应显示蓝蝽原生境中猎物-兰叶甲可能会对防治产生影响。     

Ultrastructural patterns of the flagellar axoneme in the non-motile part of the mole-cricket sperm

The mole-cricket spermatozoon (Gryllotalpa gryllotalpa) has a motile anterior tail region and an immotile posterior region. The posterior portion appears stiff and its microtubular doublets and central singlet microtubules are swollen, apparently due to an excess of material within them. In particular, doublet number 6 is of an unusually large size. The general organization of the axoneme is also modified by a loss of dynein arms and spokes in the posterior portion. When studied by a fixation technique that involves tannic acid to outline the protein molecules and PA-TCH-Ag method for staining polysaccharides it could be seen that the microtubular doublets and accessory microtubules contain rounded globules surrounded by polysaccharides. The arrangement of protofilaments within the microtubular walls is visible both in the anterior tail region with normal doublets and in the posterior region with degenerated doublets.     

Characteristics of the sperm flagellum in fungus gnats (Insecta,Diptera, Mycetophiloidea)

Spermatozoa from several members of the closely related Mycetophilidae and Keroplatidae were examined by electron microscopy using a fixative that contains glutaraldehyde and tannic acid, followed by a post-fixative that consists of uranyl acetate rather than osmium tetroxide. With this fixative, the detailed architecture of the flagellar axoneme and its various microtubules could be resolved. The so-called accessory tubules, which surround the central 9+2 unit of the sperm axoneme, were found to have 16 protofilaments in several examined Mycetophiloidea, but in no other Diptera. As 16 is the common number in holometabolic insects, it is presumably the plesiomorphic condition in Diptera. Other fungus gnats have accessory tubules with 15 or 14 protofilaments. The intertubular material situated between the accessory tubules is smaller in the examined members of the Mycetophilidae than in the Keroplatidae. The acrosome consists of an apical vesicle, which in one species, Macrorhyncha ancae, has three microtubular doublets in its anterior part and two large and three small extensions which extend posteriorly along the sperm axoneme.     

Arrangement of tubulin subunits and microtubule-associated proteins in the central-pair microtubule apparatus of squid (Loligo pealei) sperm flagella

This study provides a comprehensive, high-resolution structural analysis of the central-pair microtubule apparatus of sperm flagella. It describes the arrangement of several microtubule-associated "sheath" components and suggests, contrary to previous thinking, that microtubules are structurally asymmetric. The two microtubules of the central pair are different in several respects: the C2 tubule bears a single row of 18-nm-long sheath projections with an axial periodicity of 16 nm, whereas the C1 tubule possesses rows of 9-nm globular sheath components with an axial repeat of 32 nm. The lumen of the C2 tubule always appears completely filled with electron-dense material; that of the C1 tubule is frequently hollow. The C2 tubule also possesses a series of beaded chains arranged around the microtubule; the beaded chains are composed of globular subunits 7.5-10 nm in diameter and appear to function in the pairing of the C1 and C2 tubules. These findings indicate: that the beaded chains are not helical, but assume the form of lock washers arranged with a 16-nm axial periodicity on the microtubule; and that the lattice of tubulin dimers in the C2 tubule is not helically symmetric, but that there are seams between certain pairs of protofilaments. Proposed lattice models predict that, because of these seams, central pair and perhaps all singlet microtubules may contain a ribbon of 2-5 protofilaments that are resistant to solubilization; these models are supported by the results of the accompanying paper (R. W. Linck, and G. L. Langevin. 1981. J. Cell Biol. 89: 323-337.     

Characteristics of the flagellar axoneme in Neuroptera,Coleoptera, and Strepsiptera

Spermatozoa from representatives of the five insect orders in superorder Neuropteroidea were examined by electron microscopy following a new fixation method that includes tannic acid in the primary fixative but has uranyl acetate rather than osmium tetroxide as the secondary fixative. The sperm axoneme was found to be similar in the four orders Megaloptera, Raphidioptera, Neuroptera, and Coleoptera, and is characterized above all by its so-called intertubular material being divided into two portions, one located outside, but in contact with the doublet, and the other projecting from the accessory tubule and having a beak-like shape. These features have not been seen in insects from other orders and may be a synapomorphy for these neuropteroid orders. The accessory tubules in these four orders have 16 protofilaments. The shape of the accessory bodies adjacent to the mitochondrial derivatives is nearly the same in insects from the more primitive neuropteroid orders and in Coleoptera. The sperm tail of the examined strepsipteran deviates in several respects from that of other neuropteroids: the particle row in the wall of accessory tubules is incomplete, an intertubular material is missing, and the mitochondria contain no crystal. © 1994 Wiley-Liss, Inc.     

Ultrastructure of the male nephridium and its role in spermatophore formation in spionid polychaetes (Annelida)

Summary The mature male nephridia ofPolydora ligni andP. websteri (Polychaeta: Spionidae) are segmental organs composed of a ciliated nephrostome connected to a nephridial canal that crosses the intersegmental septum, expands into a large modified part extending dorsally through the coelom and subsequently narrows into a canal terminating in a dorsal nephridiopore. The nephridial canal is ciliated throughout and is composed of several cell types. Cells in the expanded region of the nephridia of both species contain large urn-shaped depressions filled with long microvilli. InP. ligni, one section of a nephridium contains cells packed with electron-dense granules that are not observed inP. websteri.The spermatophores ofPolydora ligni are composed of a central sperm mass surrounded by a layer of randomly oriented tubules that form a capsule around the sperm and taper into a long thin tail. These tubules are identical in dimensions to the microvilli present in parts of a nephridium and apparently are derived from these microvilli. The spermatophore capsule ofP. websteri is composed of similar tubules also presumed to originate from nephridial microvilli.The microvilli in nephridia of both species are surrounded with a glycocalyx that may function as an adhesive to hold the spermatophore capsule together. This glycocalyx may also function as a species specific message when encountered by a receptive female.Contribution Number 179 from Harbor Branch Foundation, Inc.     

Extracellular localization of cyclic GMP in the house cricket male accessory reproductive gland and its fate in mating

The male accessory reproductive gland (ARG) of the house cricket, Acheta domesticus (L.), contains an exceedingly high concentration of cyclic GMP, about 1,000 pmol/mg protein. Immunofluorescent localization and radioimmunoassay measurements show that cyclic GMP is concentrated in a small number of tubules. It accumulates in the tubule lumina where it is protected from degradation by phosphodiesterases. Cyclic GMP is secreted by the ARG and is incorporated into spermatophores. Over 80% of spermatophore cyclic GMP is found in the handle-capillary tube, a thin conduit through which sperm pass during transfer to the female. The concentration of cyclic GMP in the insemination fluid is about 20 microM but does not appear to be specifically associated with the sperm. Cyclic GMP enters the female spermatheca during insemination but disappears rapidly. Physiological effects of cyclic GMP on sperm were not observed nor was an effect of cyclic GMP observed on egg laying by mated females. Cyclic AMP was localized on sperm flagella in the spermatophore and in the spermatheca. These studies indicate that cyclic nucleotides have important roles in insect reproduction and that the house cricket is a good model for elucidating these functions.     

The dipteran sperm tail: ultrastructural characteristics and phylogenetic considerations

The sperm tail from representatives of several families of Diptera has been examined by high resolution electron microscopy and a computer analysis that improved the visualization of recorded patterns. A considerable variability in sperm tail structure is found within Diptera, and is actually greater than that of any other insect order. The 'generalized insect sperm axoneme'. which is characterized as a 9+9+2 axoneme and by the accessory microtubules having 16 protofilaments, was found only in some dipterans; these are members of Mycetophilidae. From this fact we conclude that Mycetophilidae is likely to be the most primitive extant dipteran group. Another mycetophilid, Boletina , was seen to have accessory tubules with 15 protofilaments as have members of families Dixidae, Chironomidae, Culicidae, and Bibionidae. The last two families have spermatozoa of a type designated as 9+9+'1' there is a central rod rather than two microtubules. We regard this 9+9+'1'pattern with 15 protofilaments to represent a synapomorphic feature. Representatives of the neatoceran families Tipulidae and Trichoceridae have accessory tubules with 13 protofilaments as do examined members of several brachyceran families. Brachycera is hence likely to be derived from the vicinity of the tipulid family. The intertubular material is small in Mycetophilidae and most nematoceran groups, whereas in Tipulidae and Brachycera it is enlarged; here it bridges the space between the accessory tubules and contains various inclusions.     

Deficiency of SPAG16L causes male infertility associated with impaired sperm motility

The axonemes of cilia and flagella contain a "9+2" structure of microtubules and associated proteins. Proteins associated with the central doublet pair have been identified in Chlamydomonas that result in motility defects when mutated. The murine orthologue of the Chlamydomonas PF20 gene, sperm-associated antigen 16 (Spag16), encodes two proteins of M(r) approximately 71 x 10(3) (SPAG16L) and M(r) approximately 35 x 10(3) (SPAG16S). In sperm, SPAG16L is found in the central apparatus of the axoneme. To determine the function of SPAG16L, gene targeting was used to generate mice lacking this protein but still expressing SPAG16S. Mutant animals were viable and showed no evidence of hydrocephalus, lateralization defects, sinusitis, bronchial infection, or cystic kidneys-symptoms typically associated with ciliary defects. However, males were infertile with a lower than normal sperm count. The sperm had marked motility defects, even though ultrastructural abnormalities of the axoneme were not evident. In addition, the testes of some nullizygous animals showed a spermatogenetic defect, which consisted of degenerated germ cells in the seminiferous tubules. We conclude that SPAG16L is essential for sperm flagellar function. The sperm defect is consistent with the motility phenotype of the Pf20 mutants of Chlamydomonas, but morphologically different in that the mutant algal axoneme lacks the central apparatus.     

Partial purification of a novel insect antidiuretic factor from the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), which acts on Malpighian tubules

In this study, we report for the first time the presence of an antidiuretic factor in the head of the Colorado potato beetle (Leptinotarsa decemlineata (Say)) which acts directly on Malpighian tubules. Biologically active fractions were isolated from the head and separated using molecular weight filtration and high-performance liquid chromatography (HPLC). The resulting fractions were tested for their antidiuretic activity on single isolated Malpighian tubules. Antidiuretic activity was found in the 25% acetonitrile Sep-Pak fraction and the Cn-2 (3000-10,000 MW) and Cn-3 (<3000 MW) fractions, suggesting that the antidiuretic factor was probably a peptide of 25 to 50 amino acids. The antidiuretic factor was very potent, since after five successive fractionations on two different HPLC columns, a high level of inhibition (63%) of fluid secretion by Malpighian tubules could be observed at low dose (0.14 head-equivalent/microl). The antidiuretic factor isolated from the head of the Colorado potato beetle was not affected by repeated freezing and thawing but was sensitive to heat. The differences observed between the Colorado potato beetle antidiuretic factor and other insect diuretic and antidiuretic factors may indicate the possibility of a novel family of water regulation hormones in insects.