New findings on sperm ultrastructure in thrips (Thysanoptera, Insecta)

Sperm ultrastructure of several species in each of the two suborders of Thysanoptera Tubulifera and Terebrantia shows a distinctive and unusual architecture. Members of the whole order share a bizarre axoneme consisting of 27 microtubular elements derived from the amalgamation of 3 (9+0) axonemes present in each spermatid at the beginning of spermiogenesis. The reciprocal shifting of these axonemes along the length of the sperm, together with their possible shortening and overlapping for short distances, could explain why in some species it is never possible to observe the complete set of 27 microtubular elements in any one cross section. Tubuliferan sperm have a small elliptical (in cross section) acrosome extending the length of the sperm. In Bolothrips insularis and Compsothrips albosignatus this structure is larger and is associated with an external, flattened vesicle throughout its length. Terebrantian sperm lack an acrosome, but display for half their length a dense body running parallel to the nucleus. The sperm, in members of this suborder, are also characterized by possession of a small mitochondrion and by the unusual bilobed outline of cross sections through the anterior sperm region, with the nucleus located in one of the two lobes. Structures serving to anchor sperm to the inner surface of the cyst cell have been observed at their anterior tips in the testes of tubuliferans. In B. insularis, an anterior appendage is formed in immature sperm and is maintained in the mature spermatozoon parallel to its long axis in the most anterior region. Such an anchoring structure has not been observed in sperm of the terebrantian species examined, probably because the testis of terebrantians contains only a single cyst of developing gametes.     

Sperm acrosome antigen-1. Specific tissue distribution of immunoreactivity in a teleost fish,the swordtail ( Xiphophorus helleri)

The acrosome reaction is a fundamental requirement for mammalian fertilization. Its exact molecular mechanisms and cellular elements are still poorly understood. We have detected an acrosomal sperm antigen, SAA-1, by monoclonal antibodies directed against SAA-1, that appears to be critically involved in the regulation of the acrosome reaction. SAA-1 is conserved within a broad range of mammalian species, emphasizing its important role in mammalian reproduction. Here we demonstrate that SAA-1 is immunohistochemically detectable in a nonmammalian vertebrate whose sperm do not possess an acrosome. In the swordtail, a live-bearing teleost with special reproductive tactics, we were able to demonstrate immunoreactivity of sperm heads of spermatids and mature sperm in the testis using monoclonal antibodies against SAA-1. Due to the cystic spermatogenesis with synchronous sperm maturation, immunoreactive maturational stages could be clearly identified. Stored immunoreactive sperm were also identified in spermathecal tissue of the female genital tract. Interestingly, immunoreactivity was furthermore detected in defined cells of the compounded endocrine organs pituitary and endocrine pancreas. All these different cell systems are involved in paracrine regulation and exhibit exocytotic properties. The possible nature of SAA-1 is discussed. Additionally, some new aspects of the morphologic composition of the swordtail pituitary are described.     

Post-testicular change in the reptile sperm surface with particular reference to the snake, Natrix fasciata

Sperm surface changes occurring in the reptile Wolffian duct have been explored with particular references to the snake, Natrix fasciata. In the snake Wolffian duct there are several proteins not present in serum, the pattern of which changes in concert with the seasonal testicular cycle. Whereas testicular spermatozoa did not bind antibody to duct secretions, all Wolffian duct spermatozoa did so over both head and tail, according to immunofluorescence patterns. Thus, on entering the Wolffian duct, the entire surface of N. fasciata spermatozoa acquires one of more of the duct's secretory components. As indicated by immunofluorescence, immunoelectrophoresis, and immunodiffusion, epitopes on at least some molecules that bind to spermatozoa or that remain free in the duct fluid are shared with those in other Natrix species, but not in more distant reptiles (turtle, anole lizard), nor chicken, rat, or rabbit. In regard to glycoproteins, one prominent con A-reactive band was present in polyacrylamide gel electrophoresis (PAGE) of snake fluid and more were evident in fluid collected from the turtle duct. However, such lectin-reactive elements did not bind to spermatozoa as judged by an absence of any change in snake, turtle and lizard sperm lectin-binding patterns in passing from the testis into and through the Wolffian duct. In all, evidence from these and other species studied begins to suggest that the nature of the post-testicular sperm surface modification displayed in most vertebrates that fertilize internally may differ in sub-therian and therian groups, respectively. There appears to be a relative emphasis on glycosyl-rich surface elements in the latter. The possible significance of these changes for sperm function in the different groups is discussed briefly in terms of sperm survival/storage, as well as capacitation and sperm binding to the zona.     

Decompactization of chromosome 1 in the artificially decondensed human sperm nuclei: overall topology and non-random location of chromosome bends

We have used FISH with microdissected chromosome arm-specific DNA probes that allow direct visualization of chromosome 1 folding in human sperm nuclei. We described the overall topology and the mode of packaging of sperm chromosome 1 and thus established new elements of ordered genome architecture in these specialized cells.     

Sperm DNA fragmentation and oxidation are independent of malondialdheyde

Background  

There is clinical evidence to show that sperm DNA damage could be a marker of sperm quality and extensive data exist on the relationship between DNA damage and male fertility status. Detecting such damage in sperm could provide new elements besides semen parameters in diagnosing male infertility. We aimed to assess sperm DNA fragmentation and oxidation and to study the association between these two markers, routine semen parameters and malondialdehyde formation.     

Regulation of acrosomal exocytosis. II. The zona pellucida-induced acrosome reaction of bovine spermatozoa is controlled by extrinsic positive regulatory elements

The effects of accessory sex gland secretions on the zona pellucida-induced acrosome reaction of bovine spermatozoa were investigated. Soluble extracts of zonae pellucidae initiated exocytosis in ejaculated spermatozoa. This process had an ED50 of 20 ng/microliter zona pellucida protein and saturated at 50 ng/microliter (Florman and First, 1988. Dev. Biol. 128, 453-463). In epididymal sperm this dose-response relationship was shifted toward greater agonist concentrations by at least a factor of 10(3). Reconstitution of high potency agonist response was achieved in vitro by incubation of epididymal sperm with bovine seminal plasma. Reconstitution was dependent on the seminal plasma protein concentration. The ED50 of this process was 62 micrograms protein/10(8) sperm and saturation was observed with 124 micrograms protein/10(8) sperm. Agonist responses in reconstituted epididymal sperm and in ejaculated sperm were indistinguishable with regard to dependence on the zona pellucida protein concentration and the kinetics of induced acrosome reactions. Kinetic studies suggest that reconstitution is due to adsorption of regulatory factors from seminal plasma. In addition to the positive regulatory elements responsible for reconstituting activity, seminal plasma also contains negative regulatory elements which inhibit agonist response. These negative factors are inactivated during sperm capacitation, permitting the expression of positive regulators. Acting together, these regulatory elements could coordinate high affinity agonist response with the availability of eggs in vivo.     

Evaluation of the Relationship Between Height and Zinc,Copper, Iron,Calcium, and Magnesium Levels in Healthy Young Children in Beijing,China

Studies suggest a relationship between semen quality and the concentration of trace elements in serum or seminal plasma. However, trace elements may be linked to DNA and capable of altering the gene expression patterns. Thus, trace element interactions with DNA may contribute to the mechanisms for a trans-generational reproductive effect. We developed an analytical method to determine the amount of trace elements bound to the sperm DNA, and to estimate their affinity for the sperm DNA by the ratio: R = Log [metal concentration in the sperm DNA/metal concentration in seminal plasma]. We then analyzed the concentrations of 15 trace elements (Al, Cd, Cr, Cu, Hg, Mn, Mo, Ni, Pb, Ti, V, Zn, As, Sb, and Se) in the seminal plasma and the sperm DNA in 64 normal and 30 abnormal semen specimens with Inductively Coupled Plasma/Mass Spectrometry (ICP-MS). This study showed all trace elements were detected in the seminal plasma and only metals were detected in the sperm DNA. There was no correlation between the metals’ concentrations in the seminal plasma and the sperm DNA. Al had the highest affinity for DNA followed by Pb and Cd. This strong affinity is consistent with the known mutagenic effects of these metals. The lowest affinity was observed for Zn and Ti. We observed a significant increase of Al linked to the sperm DNA of patients with oligozoospermia and teratozoospermia. Al’s reproductive toxicity might be due to Al linked to DNA, by altering spermatogenesis and expression patterns of genes involved in the function of reproduction.     

Positive correlations between pre‐ and post‐copulatory sexual traits in warblers

Theoretical models predict that investment in pre‐copulatory and post‐copulatory sexually selected traits should trade‐off. At the macroevolutionary scale, the majority of studies to date have focused on male weaponry as the target of pre‐copulatory sexual selection, but the trade‐off should equally apply to traits used to attract females, such as bird song and plumage. We studied the Old World leaf warblers (Phylloscopidae), a group of socially monogamous songbirds that experience relatively high levels of sperm competition. We examined the relationships between song duration and number of elements in the song with sperm length across 21 species, and between the same song variables and combined testes mass in a subset of these species (n = 10). Across species, these song variables and testes mass/sperm length were generally positively correlated, albeit not statistically significantly so or with borderline significance. In contrast to theory, we found no evidence for negative associations between pre‐ and post‐copulatory traits. We argue that this is a consequence of males of some species investing more into overall fertilization success (i.e. the sum of pre‐ and post‐copulatory sexual selection) than males of other species, and high fertilization success is achieved through investment into both mate attraction and sperm competition.     

TO WHAT EXTENT DO DIFFERENT TYPES OF SEX RATIO DISTORTERS INTERFERE?

Within the Diptera, two different selfish genetic elements are known to cause the production of female-biased sex ratios: maternally inherited bacteria that kill male zygotes (male-killers), and X chromosomes causing the degeneration of Y-bearing sperm in males (meiotic drive). We here develop a mathematical model for the dynamics of these two sex-ratio distorters where they co-occur. We show that X chromosome meiotic drive elements can be expected to substantially lower the equilibrium frequency of male-killers and can even lead to their extinction. Conversely, male-killers can also decrease the equilibrium frequency of X drivers and cause their extinction. Thus, we predict that there will be some complementarity in the incidence of X chromosome meiotic drive and male-killing in natural populations, with a lower than expected number of species bearing both elements.     

SPERM COMPETITION AND REPRODUCTIVE MODE INFLUENCE SPERM DIMENSIONS AND STRUCTURE AMONG SNAKES

The role of sperm competition in increasing sperm length is a controversial issue, because findings from different taxa seem contradictory. We present a comparative study of 25 species of snakes with different levels of sperm competition to test whether it influences the size and structure of different sperm components. We show that, as levels of sperm competition increase, so does sperm length, and that this elongation is largely explained by increases in midpiece length. In snakes, the midpiece is comparatively large and it contains structures, which in other taxa are present in the rest of the flagellum, suggesting that it may integrate some of its functions. Thus, increases in sperm midpiece size would result in more energy as well as greater propulsion force. Sperm competition also increases the area occupied by the fibrous sheath and outer dense fibers within the sperm midpiece, revealing for the first time an effect upon structural elements within the sperm. Finally, differences in male–male encounter rates between oviparous and viviparous species seem to lead to differences in levels of sperm competition. We conclude that the influence of sperm competition upon different sperm components varies between taxa, because their structure and function is different.     

COMPARATIVE ANALYSIS OF MAMMALIAN SPERM MOTILITY

Spermatozoa of several mammalian species were studied by means of high-speed cinematography and electron microscopy. Three types of motile patterns were observed in mouse spermatozoa. The first type involved an asymmetrical beat which seemed to propel the sperm in circular paths. The second type involved rotation of the sperm and appeared to allow them to maintain straight paths. In the third type of pattern, the sperm appeared to move by crawling on surfaces in a snakelike manner. Spermatozoa of rabbit and Chinese hamster also had an asymmetrical beat which sometimes caused them to swim in circles. In spite of the asymmetry of the beat, these spermatozoa were also able to swim in straight paths by rotating around a central axis as they swam. Spermatozoa of some species appeared very flexible; their flagella formed arcs with a very small radius of curvature as they beat. Spermatozoa of other species appeared very stiff, and their flagella formed arcs with a very large radius of curvature. The stiffness of the spermatozoan appeared to correlate positively with the cross-sectional area of the dense fibers. This suggests that the dense fibers may be stiff elastic elements. Opossum sperm become paired as they pass through the epididymis. Pairs of opossum spermatozoa beat in a coordinated, alternating manner.     

Sperm methylation profiles reveal features of epigenetic inheritance and evolution in primates

During germ cell and preimplantation development, mammalian cells undergo nearly complete reprogramming of DNA methylation patterns. We profiled the methylomes of human and chimp sperm as a basis for comparison to methylation patterns of ESCs. Although the majority of promoters escape methylation in both ESCs and sperm, the corresponding hypomethylated regions show substantial structural differences. Repeat elements are heavily methylated in both germ and somatic cells; however, retrotransposons from several subfamilies evade methylation more effectively during male germ cell development, whereas other subfamilies show the opposite trend. Comparing methylomes of human and chimp sperm revealed a subset of differentially methylated promoters and strikingly divergent methylation in retrotransposon subfamilies, with an evolutionary impact that is apparent in the underlying genomic sequence. Thus, the features that determine DNA methylation patterns differ between male germ cells and somatic cells, and elements of these features have diverged between humans and chimpanzees.     

Structural features of the abalone egg extracellular matrix and its role in gamete interaction during fertilization

Abalone eggs are surrounded by a complex extracellular coat that contains three distinct elements: the jelly layer, the vitelline envelope, and the egg surface coat. In this study we used light and electron microscopy to describe these three elements in the red abalone (Haliotis rufescens) and ascribe function to each based on their interactions with sperm. The jelly coat is a spongy matrix that lies at the outermost margin of the egg and consists of variably sized fibers. Sperm pass through this layer with their acrosomes intact and then go on to bind to the vitelline envelope. The vitelline envelope is a multilamellar fibrous layer that appears to trigger the acrosome reaction after sperm binding. Next, sperm release lysin from their acrosomal granules, a nonenzymatic protein that dissolves a hole in the vitelline envelope through which the sperm swims. Sperm then contact the egg surface coat, a network of uniformly sized filaments lying directly above the egg plasma membrane. This layer mediates attachment of sperm, via their acrosomal process, to the egg surface. © 1995 Wiley-Liss, Inc.     

Detection of Sperm Histone Diversity Among Vertebrates by Alkaline Fast Green Staining

Testis sections from fifteen species from six classes of vertebrates were stained with alkaline fast green (AFG) to correlate staining differences with the known biochemical diversity of histones in the spermatozoa. After trichloroacetic acid (TCA) hydrolysis the sperm of some species known to contain sperm-specific histones did not stain. This correlation held if fixation in neutral buffered formalin was limited to 3 to 6 hr and hydrolysis was done at 90 C. The species whose sperm did stain after TCA hydrolysis could be divided into three groups. In some species the sperm no longer stained if, after TCA, the sections were treated with thioglycollic acid. These sperm contained basic proteins that were rich in cysteine. In turn, the group of species whose sperm continued to stain after TCA and thioglycollic acid treatments could be subdivided. The sperm of some were stained specifically without DNA hydrolysis if the AFG was made up with sodium chloride. These sperm contained sperm-specific histones. In other species the sperm did not stain under these conditions, and these sperm had a basic protein complement similar to that found in somatic cell nuclei. These correlations suggest that AFG staining can be used to detect sperm histone diversity in a wide range of organisms.     

Detection of sperm histone diversity among vertebrates by alkaline fast green staining

Testis sections from fifteen species from six classes of vertebrates were stained with alkaline fast green (AFG) to correlate staining differences with the known biochemical diversity of histones in the spermatozoa. After trichloroacetic acid (TCA) hydrolysis the sperm of some species known to contain sperm-specific histones did not stain. This correlation held if fixation in neutral buffered formalin was limited to 3 to 6 hr and hydrolysis was done at 90 C. The species whose sperm did stain after TCA hydrolysis could be divided into three groups. In some species the sperm no longer stained if, after TCA, the sections were treated with thioglycollic acid. These sperm contained basic proteins that were rich in cysteine. In turn, the group of species whose sperm continued to stain after TCA and thioglycollic acid treatments could be subdivided. The sperm of some were stained specifically without DNA hydrolysis if the AFG was made up with sodium chloride. These sperm contained sperm-specific histones. In other species the sperm did not stain under these conditions, and these sperm had a basic protein complement similar to that found in somatic cell nuclei. These correlations suggest that AFG staining can be used to detect sperm histone diversity in a wide range of organisms.     

Sperm whales (Physeter macrocephalus), found stranded along the Adriatic coast (Southern Italy,Mediterranean Sea), as bioindicators of essential and non-essential trace elements in the environment

In September 2014, a pod of seven sperm whales (Physeter macrocephalus) was stranded along the Adriatic coast of Southern Italy. Concentrations of 20 essential and non-essential trace elements were quantified in the brain, muscle, liver and kidneys of three female sperm whales, which died in this event.The essential elements copper, iron, manganese and zinc showed low ranges of variability, suggesting a homeostatic physiological control, while selenium concentrations were associated with age. Molybdenum, nickel and chromium showed low ranges of concentrations and no evidence of preferential accumulation in selected organs. Very low concentrations of the non-essential elements cadmium, lead, tin and vanadium were detected in all tissues, suggesting a minor impact of these pollutants on the sperm whale populations of the Mediterranean Sea. Aluminum was revealed to have relatively high concentrations, together with a high variability between tissues and individuals, reaching the highest values in the kidneys and muscle of the oldest female, which was pregnant; the rare earth elements – lanthanum and cerium – were also detected in the kidneys of this female, indicating that pregnancy probably influenced metal concentrations in body tissues.     

Clustering and methylation of repeated DNA: persistence in avian development and evolution.

In the chicken genome, clusters of repeated DNA sequences occur which have alternate arrangements of the component sequence elements. Many of these clustered, repeated sequences are extensively methylated. We have established that both their arrangement and their methylation are invariant regardless of the source of chicken DNA. Comparisons included DNA from sperm, from a series of embryonic stages, from tissues of single adult individuals, and from thirty individual chickens of two strains. These same sequences are found in the DNA of some avian species related to chickens, and there they show the same clustered, methylated form. In related species, some of the arrangements found in chicken DNA are different or missing.     

Enigmas of mammalian gamete form and function

The gametes of man and some other Eutheria have been manipulated successfully for practical reasons, but many gaps remain in our basic understanding of the way that they function. This situation stems not least from a failure to recognize the extent to which eutherian spermatozoa and eggs, and elements related to their operation, have come to differ from those of other groups. Novel features in the male that reflect this include a radical design of the sperm head with the acrosome seeming to function primarily in egg-coat binding rather than its lysis, a multifaceted post-testicular sperm maturation and an androgen/low-temperature-regulated system of sperm storage--both tied to the epididymis, a variable male accessory sex gland complex, and descent of the testis and epididymis to a scrotum. In the female, such novelties are represented in a need for sperm capacitation, in an unusual regulation of sperm transport within the oviduct, in the cumulus oophorus and character of the zona pellucida around the small egg, and in a unique configuration of gamete fusion. The collective evidence now suggests that many of these features reflect a new fertilisation strategy or its consequences, with most being causally linked. One initial 'domino' in this regard appears to be the small yolkless state of the egg and its intolerance for polyspermy, as determinants of the unusual mode of oviductal sperm transport and possibly the existence and form of the cumulus oophorus. However, a particularly influential first 'domino' appears to be the physical character of the eutherian zona pellucida. This differs from the egg coats of other animal groups by virtue of a resilient elasticity and thickness. These qualities allow this primary and often only coat to stretch and so persist during later expansion of the blastocyst, usually until close to implantation. At the same time, the dimensions, physical character, and particularly the relative protease-insensitivity of the zona appear to have had profound effects on sperm form and function and, more indirectly, on sperm-related events in the male and the female tract. Marsupials display some similarities and also some strikingly different features, against which the enigmas of the eutherian situation can be evaluated.     

Substructure of the axoneme of pterygote insect spermatozoa: Phylogenetic considerations

Spermatozoa from a great number of insect species were fixed in a tannic acid-containing fixative and the ultrastructure of the flagellar axoneme was examined in a search for apomorphies. Most of the examined species, representing a majority of insect orders. have accessory tubules outside the axoneme (hence a 9 + 9 + 2 pattern), and these consist of 16 protofilaments. Some important apomorphies concern the number of protofilaments in the accessory tubules: 13 (plus 7 inner elements) in Ephemeroptera, 13 in the (elliptic) tubules of Psocoptera + Anoplura + Mallophaga (thus a synapomorphy), 13 in Tipulidae + Brachycera, 15 in the dipteran families Dixidae + Chironomidae (with a 9 + 9 + 2 axoneme) and Culicidae + Bibionidae (with a 9 + 9 + “1” axoneme), 17 in Phasmatodea, and 17–20 in Trichoptera. Other apomorphies concern the appearance of the so-called intertubular material outside the microtubular doublets, the appearance of the interior of the various microtubules, and the loss, in some taxa, of outer or inner dynein arms of both dynein arms. In some cases, the flagellum is completely abnormal; the sperm tail of Thysanoptera, for example, consists of 27 elements of 3 different kinds. The different taxa within orders Diptera and Trichoptera have sperm tail axonemes of different appearances, where those from other orders have a rather uniform appearance. The conclusions that can be drawn from this spermatological study, generally agree with data from classical studies, except with some variations, in some cases.