Functional significance of seminal receptacle length in Drosophila melanogaster

Despite its central role in post-copulatory sexual selection, the female reproductive tract is poorly understood. Here we provide the first experimental study of the adaptive significance of variation in female sperm-storage organ morphology. Using populations of Drosophila melanogaster artificially selected for longer or shorter seminal receptacles, we identify relationships between the length of this primary sperm-storage organ and the number of sperm stored, pattern of progeny production, rate of egg fertilization, remating interval, and pattern of sperm precedence. Costs and benefits of relatively short or long organs were identified. Benefits of longer receptacles include increased sperm-storage capacity and thus progeny production from a single insemination. Results suggest that longer receptacles have not naturally evolved because of developmental time costs and a correlated reduction in longevity of mated females. This latter cost may be a consequence of sexual conflict mediated by ejaculate toxicity. Receptacle length did not alter the pattern of sperm precedence, which is consistent with data on the co-evolution of sperm and female receptacle length, and a pattern of differential male fertilization success being principally determined by the interaction between these male and female traits.     

Functional morphology of the female reproductive system of a crab with highly extensible seminal receptacles and extreme sperm storage capacity

We studied the functional morphology of the female reproductive system of the purple stone crab Danielethus crenulatus . The most remarkable feature is the relative storage capacity and extensibility of the seminal receptacles. These receptacles are a pair of simple sacs that lack internal structures dividing the internal lumen. Differences in seminal receptacle size and contents are accompanied by conspicuous changes in receptacle lining at a tissue level. Full seminal receptacles contain discrete sperm masses formed by hardened fluid and densely packed spermatophores. Different sperm masses are likely from different mates and their stratified disposition within the seminal receptacles is compatible with rival sperm displacement and last sperm precedence. Additionally, the anatomical structure of the vulva and vagina suggest active female control over copula. We discuss our results in the general context of sperm storage in brachyurans and the implications for the mating system of this species.     

Ejaculate expenditure and timing of gamete release in rainbow trout Oncorhynchus mykiss

Using a novel methodology, natural ejaculate volumes of 14 male rainbow trout Oncorhynchus mykiss were compared when males were housed with one female (absence of a rival male) or with another male and one female (rival male present). Contrary to theoretical predictions, male ejaculate expenditure was not influenced by the presence of a rival male. Male gape duration was positively correlated with the volume of sperm ejaculated. Release of sperm by the male always preceded release of eggs by the female. Analysis of the timing and duration of ejaculation suggests that males may rely on the timing and proximity of gamete release to enhance fertilization success. These results are discussed in the context of sperm competition theory.     

Studies of the seminal receptacle of the crab Portunus sanguinolentus (Portunidae: Brachyura)

The seminal receptacle or spermatheca of Portunus sanguinolentus consists of two parts--an anterior glandular and a posterior chitinous part. The chitinous part continues as the oviduct, which opens on the sternite of the sixth thoracic segment. Significant morphological and histological differences were observed between the spermatheca, as well as the oviduct, of mated and unmated crabs. In mated crabs the spermatheca is much more bulging, owing to receipt of a copious supply of seminal products, and its cells are hyperactive. Further stages of ovarian development were observed as indicators of sequential changes in the spermatheca. The secretory cells gradually disintegrate by way of holocrine secretion; this results in cellular stratification and the formation of distinct furrows in the chitinous posterior part.     

A new type of brachyuran seminal receptacle in the masked crab Ethusa mascarone (Brachyura,Ethusidae)

The reproductive system of the female Ethusa mascarone was studied with a combination of histological and MRI‐techniques. The study reveals a completely new type of eubrachyuran seminal receptacle. This receptacle consists of two largely separate chambers that engage with each other in a manner similar to shaking hands. One chamber facing the medial axis is lined by cuticle while the second chamber consists of a thick holocrine epithelium. Both chambers are connected by two openings of a unique structure. First, the glandular chamber opens ventro‐laterally to the cuticle chamber via a laterally flattened connective duct that is lined by a highly folded cuticle. A second opening connects both chambers dorsally with the oviduct orifice. A distinct character is the cuticular hook‐like projection that is situated in between the connection of oviduct opening, the glandular chamber and the cuticle chamber of the seminal receptacle. The complete seminal receptacle exhibits a combination of plesiomorphic and apomorphic characters. The arrangement of the receptacles featured in two separate chambers, including the ventro‐lateral connection of the glandular chamber to the cuticle chamber, presumably reflects an early evolutionary stage of an eubrachyuran receptacle. In contrast, the dorso‐lateral opening between both chambers, including the hook‐like projection, appears to be an apomorphic character of at least E. mascarone. J. Morphol. 277:1497–1508, 2016. © 2016 Wiley Periodicals, Inc.     

Ultrastructure of the seminal receptacle and the dimorphic sperm in the commensal bivalve Mysella bidentata (Veneroida; Galeommatoidea; Montacutidae)

Jespersen, Å. and Lützen, J. 2001 . Ultrastructure of the seminal receptacle and the dimorphic sperm in the commensal bivalve Mysella bidentata (Veneroida: Galeommatoidea: Montacutidae). — Acta Zoologica (Stockholm) 82 : 107–115
The seminal receptacle and the euspermatozoa and paraspermatozoa of Mysella bidentata were examined at an ultrastructural level and the results were compared with earlier findings of the same and other species of the Montacutidae. The euspermatozoon has a slender 13 µm long nucleus and a 1.1 µm long bullet-shaped acrosome. The acrosome of the paraspermatozoon is almost identical in ultrastructure to that of the euspermatozoa but is longer (1.9 µm) and more slender and is bent at an angle to the diminutive nucleus (1.1 µm long). The unpaired seminal receptacle is lined by a heavily ciliated epithelium and a non-ciliated epithelium with short and broad microvilli. Euspermatozoa only are stored in the receptacle. They are densely packed and orientated with their heads towards the non-ciliated epithelium. In this position they develop numerous extremely fine microvilli from the acrosome which apparently serve to attach them to the epithelial microvillar surface. Stored sperm may presumably remain functional for at least six months. A possible function of paraspermatozoa could be to clump sperm into sperm bags to keep them in suspension.     

Changes in sperm stores, ejaculate size, fertilization success, and sexual motivation over repeated matings in the common toad, Bufo bufo (Anura: Bufonidae)

Fertilization is of central importance in the determination of reproductive success for both males and females. In species where males have the chance to mate repeatedly within a short period of time, sperm stocks may become depleted and males may have to carefully economize on their sperm reserves. Also, intensive intrasexual competition for females and repeated matings may lead to exhaustion on the behavioural level. To determine whether the reproductive potential of males is limited and if such a limitation is due to behavioural exhaustion or sperm depletion, we experimentally investigated changes in sperm stores, sperm expenditure, fertilization success, and sexual motivation over three repeated matings in the common toad, Bufo bufo , where the breeding season is short and sequential polygyny occurs. At the end of the breeding season, the number of sperm stored in the testes of males mated repeatedly was close to 50% lower than in testes of unmated males. Ejaculate size, which was estimated by applying a novel method allowing direct quantification, decreased by 88% from first to third matings. We also observed a drop in fertilization success from the first two to third matings by 65%, which was largest in males that had started the reproductive season in bad body condition. Some of these males also showed a decreased interest in females in the third mating round. Our results suggest that sperm depletion and loss of sexual motivation may together set a limit to the reproductive potential of common toad males. The present study draws attention to a limitation in reproductive potential, which may occur more often than currently anticipated and has the potential to strongly influence several aspects of reproductive behaviour.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 361–371.     

Functional evidence for differences in sperm competition in humans and chimpanzees

Sperm competition occurs when the gametes of or more males compete for opportunities to fertilize a given set of ova. Previous studies have demonstrated that certain morphological characteristics are affected by sperm competition intensity (e.g. relative testes size and sperm midpiece volume). This study examined whether aspects of sperm energetics may also be affected by sexual selection. We compared the membrane potential of mitochondria in live sperm between H. sapiens (single partner mating system) and P. troglodytes (multiple partner mating system). Flow cytometry of sperm stained with the carbocyanine fluorescent dye JC-1 (an assay for mitochondrial membrane potential) revealed marked differences in red fluorescence intensity. P. troglodytes sperm showed significantly higher mitochondrial membrane potential. Mitochondria provide a substantial part of the energy required for sperm motility. A higher mitochondrial loading may therefore be associated with enhanced sperm motility and/or longevity. Additionally, examination of JC-1 red fluorescence levels before and after in vitro capacitation revealed further differences. Whereas chimpanzee sperm showed maintenance of membrane potential after capacitation (in some cases even an increase), sperm from humans consistently showed reduction in membrane potential. These results indicate that the sperm of human beings and chimpanzees exhibit marked differences in mitochondrial function, which are affected by selection pressures relating to sperm competition and that these pressures differ significantly between humans and chimpanzees.     

Reproductive morphology of the male Tuatara,Sphenodon punctatus

Over the past decade, studies on reproductive morphology in the Squamata (snakes and lizards) have expanded tremendously. With the accumulation of these studies and revisions of the terminology based on structural similarities and differences, it is imperative to review the work on tuataras to determine whether the structural organization fits the revised terminology of vertebrates. We investigated the morphology of the male reproductive system in the Tuatara, Sphenodon punctatus (Rhynchocephalia), the sister taxon to the Squamata. Previous studies on the Tuatara used a nomenclature for the testicular ducts different from the current terminology for amniotes. The reproductive system in the Tuatara is consistent with reports in the Squamata. Two rete testis tubules exit the testis within a connective tissue sheath similar to that shown in other squamate species and the protherian Echidna. Each rete testis divides into multiple ductuli efferentes that fuse with the epididymis. The epididymis transitions into the ductus deferens where the sperm become more concentrated into spherical bundles. The ductus deferens enters the cloacal urodeum separately from the ureter. An ampulla ureter or ampulla urogenital papilla was not observed, which differs from previous studies of lepidosaurians. Furthermore, a sexual segment of the kidney (SSK) was not observed, consistent with previous studies on the Tuatara.     

Functional morphology of the female reproductive apparatus of Stephanitis pyrioides (Heteroptera,Tingidae): A novel role for the pseudospermathecae

At mating, female insects generally receive and store sperm in specific organs of their reproductive tract called spermathecae. Some Heteroptera, such as Cimicomorpha, lack a true spermatheca; some have receptacles of novel formation where sperm cells can transit or be stored. In Tingidae, there are two sac‐like diverticula, the “pseudospermathecae,” each at the base of a lateral oviduct, which previously were considered to function as spermathecae. However, this role has never been documented, either by ultrastructural studies or by observations of sperm transit in the female reproductive tract. In this article, we investigate the morphology and the ultrastructure of the female reproductive apparatus in the economically important tingid species Stephanitis pyrioides, focusing our attention on the functional role of the pseudospermathecae in an evolutionary perspective. Each ovary consists of seven telotrophic meroistic ovarioles, the long pedicels of which enlarge into a bulb‐like structure near the terminal oocyte. The ovarioles flow into two long lateral oviducts, which join to form a very short common oviduct. Basally, each lateral oviduct is connected through a short duct to one of two pseudospermathecae. The ultrastructure of the ectodermal epithelium of the pseudospermathecae is dramatically different in sexually immature or mated females. In virgin females, cells delimit a very irregular lumen, filled with a moderately electron‐dense granular material. The large nucleus adapts to their irregular shape, which can have long projections in some regions and be flattened in others. After mating, epithelial cells generally elongate and display an apical layer of microvilli extending beneath the cuticle, often containing mitochondria. In the lumen of the pseudospermathecae there is a dense brownish secretion. No sperm cells were ever found inside this organ. After mating, sperm move upward along the lateral oviducts and the ovarioles, accumulating in the bulb‐like structure of the pedicels, and proceeding into the distal region between the follicle cells surrounding the oocyte and the ovariole wall. The egg, most likely fertilized in the bulb‐like region of the ovariole, moves through the lateral oviduct, entirely enters the pseudospermatheca and is smeared with its secretion just before oviposition. We exclude a function of sperm storage for the pseudospermathecae, and instead suggest a novel role for these organs as reproductive accessory glands. J. Morphol., 2010. © 2009 Wiley‐Liss, Inc.     

Functional morphology of the sperm-containing chambers of the sea slug Okenia polycerelloides in the context of sexual selection

Sea slugs are interesting models to study post-copulatory sexual selection in simultaneous hermaphrodites due to the enormous variation of their reproductive systems. However, the knowledge of the functional morphology of their reproductive system is limited to few species, and it is rarely discussed in the context of sexual selection theory. In this study, we investigated the functional morphology of the sperm-containing chambers (i.e., ampulla, seminal receptacle, and bursa copulatrix) of the reproductive system of Okenia polycerelloides (Ortea & Bouchet, 1983), based on light, confocal, and electron microscopy. Although the morphology of the ampulla is similar to other species, indicating that it is a site for autosperm storage, we found some sperm facing the ampullar epithelium, a feature commonly regarded as characteristic of the seminal receptacle of sea slugs. The seminal receptacle of O. polycerelloides showed secretory activity and contained sperm with distribution and orientation suggestive of stratification of allosperm from distinct mating events, a feature that would affect sperm competition. The bursa copulatrix had epithelial cells with secretory and absorptive characteristics, and contained degraded sperm and yolk granules within its lumen. Comparative analyses of the contents of each organ demonstrated that sperm digestion occurs in the bursa copulatrix and affects sperm heads first, changing their morphology from slender and curved to shorter and ellipsoid before complete lysis. Although digestion and absorption of surplus sperm are currently the main hypothesized functions for the bursa copulatrix, its role in cryptic female choice should not be ruled out. The close structural connection between the seminal receptacle and bursa copulatrix, as well as their muscular walls, would enable control over the fate of the sperm received in each mating event, that is, storage or digestion.     

Sperm midpiece length predicts sperm swimming velocity in house mice

Evolutionary biologists have argued that there should be a positive relationship between sperm size and sperm velocity, and that these traits influence a male''s sperm competitiveness. However, comparative analyses investigating the evolutionary associations between sperm competition risk and sperm morphology have reported inconsistent patterns of association, and in vitro sperm competition experiments have further confused the issue; in some species, males with longer sperm achieve more competitive fertilization, while in other species males with shorter sperm have greater sperm competitiveness. Few investigations have attempted to address this problem. Here, we investigated the relationship between sperm morphology and sperm velocity in house mice (Mus domesticus). We conducted in vitro sperm velocity assays on males from established selection lines, and found that sperm midpiece size was the only phenotypic predictor of sperm swimming velocity.     

Spermatozoon head size – the main differentiating feature between spermatozoa of blue and white Arctic fox (Vulpes lagopus)

Morphology and sperm morphometry, this is an important determinant of male reproductive capacity. Morphometric data may provide relevant information in studies focused on evolutionary biology, sperm quality assessment, including prediction of the potential fertility, semen cryopreservation, or the effect of reprotoxicants. The paper presents the morphometric analysis of spermatozoa from two colour morphs of Arctic fox (Vulpes lagopus), and attempts to determine the relationship between selected quality indicators and dimensions and shape of spermatozoa. The research material consisted of ejaculates collected once by manual stimulation from 20 one-year-old Arctic foxes (10 individuals of the blue morph and 10 of the white morph). Ejaculates were analysed for standard parameters (volume, sperm concentration, total number of spermatozoa in the ejaculate) and used for the preparation of microscopic specimens. It was found that, the dimensions of spermatozoa from Arctic foxes depend on the male colour morphs. Spermatozoa from white Arctic foxes were significantly longer (by 1.82 µm) and had larger heads (0.32 µm longer and 0.15 µm wider) compared to spermatozoa from blue Arctic foxes (P<0.05). The interactions between particular sperm dimensions indicated the occurrence of gametes differing in shape. The all correlation coefficients between the morphometric traits of spermatozoa were statistically significant. Our research proved that in the blue Arctic foxes, sperm dimensions (tail length and total sperm length) can be related to the percentage of spermatozoa with primary changes (respectively: r = -0.68 and r = -0.75; at P <0.05). However, in the case of white Arctic foxes, these characteristics depend on the ejaculate volume (respectively: r = 0.65 and r = 0.68; at P <0.05).     

Effect of mating on sperm distribution in the reproductive tract of the male rat

Extragonadal sperm reserves in male rats were measured in different regions of the genital tract before and subsequent to normal ejaculation. In sexually rested rats, the sperm count (million spermatozoa for the paired organs) in different regions was: distal vas, 18; proximal vas, 9.8; cauda epididymidis, 229; caput + corpus epididymidis, 154. Following mating, the sperm count was reduced in the proximal and distal vas deferens and in the cauda epididymidis. The reproductive tract of mated females was found to contain 29% (no copulatory plug) or 59% (with copulatory plug) of the estimated mean ejaculate, which was estimated from the difference between the sperm counts in the sexually rested rat and following ejaculation. It is concluded that in the rat the immediate source of spermatozoa for ejaculation is the cauda epididymidis, with a smaller contribution arising from the vas deferens.     

Variation in sperm precedence in Chorthippus grasshoppers (Caelifera: Gomphocerinae)

Variation in the proportions of offspring fathered by a second male to mate (the P₂ value) has been studied in two species of grasshoppers, Chorthippus parallelus (Zetterstedt) and Ch. biguttulus (Linnaeus), by means of the sterile‐male technique. In both species the P₂‐values of the first egg pods laid were in the range of 50–100%, and the temporal variation of P₂‐values appeared to follow two modes. In one, the P₂‐value steadily declined with time, in the other it remained constant at a high level. It is concluded that sperm is passively lost between two matings. The remaining sperm then either mixes within the spermatheca or is stratified. Further variability in sperm precedence can possibly be explained by spermatophore ejection or differential use of stored sperm.     

Sperm allocation in the three-spined stickleback

Male three-spined stickleback Gasterosteus aculeatus have a fixed amount of sperm during the breeding season because spermatogenesis is inhibited at this time. A method was developed to estimate ejaculate size in situ by removing the sperm from the male's nest. The reliability of the method was tested using known numbers of sperm. In their first mating, males ejaculated 11·64 × 10⁶ sperm (median), representing c. 5% of the male's sperm store (median 27·88 × 10⁷ sperm). The amount of sperm in the testes was significantly reduced in males that had mated several times (median 8·09 × 10⁷). Additionally, ejaculate size was smaller in these experienced males (median 8·79 × 10⁵). Heavier and larger fish invested absolutely and relatively more sperm in a mating than did lighter and smaller fish. Ejaculate size did not correlate with the mass of the egg clutch.     

Larval settlement of the nemertean egg predator Carcinonemertes errans on the Dungeness crab,Metacarcinus magister

Due to their habitat specificity, marine parasites present excellent systems for studying the processes and patterns of larval settlement. Settlement of Carcinonermertes errans, an egg predator of the Dungeness crab, is described here for the first time. Upon contact with a host individual, competent larvae of C. errans settled on the crab's exoskeleton and migrated under the abdominal flap within 24 h. When removed from the host, recently settled worms retained their larval characteristics. After 48 h on the host, however, metamorphosis proceeded and larvae became juvenile worms. Additional field studies showed that competent larvae were present in the waters of the Coos Bay Estuary during the months of August through early November, could infect crab hosts directly from the water column, and exhibited density‐dependent gregarious settlement.