Coevolution of male and female reproductive traits in a simultaneously hermaphroditic land snail

Inter- and intraspecific studies in gonochoristic animals reveal a covariation between sperm characteristics and the size of the female reproductive tract, indicating a rapid evolutionary divergence, which is consistent with the theory of post-copulatory sexual selection. Simultaneous hermaphrodites differ from species with separate sexes (gonochorists) in that they possess both functional male and female reproductive organs at the same time. We investigated whether in hermaphroditic animals intraspecific variation in reproductive traits results from divergent coevolution, by quantifying the variation in male and female traits among six natural populations of the snail Arianta arbustorum and examining the covariation in interacting traits. There was a significant among-population variation in spermatophore volume, number of sperm transferred and sperm length, as well as in volume of the sperm storage organ (spermatheca) and number of tubules, but not in spermatheca length. We found a positive association between sperm number transferred and spermatheca volume. This result suggests that the same post-copulatory mechanisms as in gonochorists drive the correlated evolution of reproductive characters in hermaphrodites.     

Effects of Intermating Interval on Spermatophore Size and Sperm Number in the Simultaneously Hermaphroditic Land Snail Arianta arbustorum

We examined the effect of intermating interval on spermatophore size and number of sperm delivered in the simultaneously hermaphroditic land snail, Arianta arbustorum . Snails that remated after 6–7 d transferred smaller spermatophores which contained fewer sperm than snails that remated after eight and more days. This indicates that individuals of A. arbustorum need at least 8 d to completely replenish their sperm reserves after a successful copulation. We suggest that the interval between two copulations is also influenced by the energy costs of the long-lasting courtship behaviour with extensive mucus production.     

The spermatheca in the land snail, Arianta arbustorum (Pulmonata: Stylommatophora): muscle system and potential role in sexual selection

Abstract. Previous studies have shown variable patterns of paternity after multiple mating, and also variation in sperm storage among individuals of Arianta arbustorum , which suggests that the spermatheca may influence paternity in this promiscuous land snail. To identify possible morphological correlates of sperm manipulation, we investigated arrangement and ultrastructure of the muscles of the spermatheca. The musculature surrounding the 2–9 spermathecal tubules is arranged in a complex three dimensional network. In addition, each tubule has a thin sheath in which longitudinally oriented cells make up the innermost layer. Usually, the smooth muscle cells are enclosed by connective tissue. Only occasionally is direct muscle-muscle contact established through dense plaques. The short thick filaments, their small diameter, the relatively weak development of the tubular system and sarcoplasmic reticulum, and the low density of mitochondria indicate that the muscle cells contract relatively fast but with little strength, that they recover slowly, and have low endurance. A single muscle cell may be innervated by several axons and one axon may contact several muscle cells. Combining evidence of the present paper and a foregoing investigation on the spermathecal epithelium, we suggest that the main function of the spermathecal muscles is to expel sperm stored for fertilization, while the ciliation of the common duct is probably responsible for the distribution of sperm among the tubules.     

Expandable spermatheca influences sperm storage in the simultaneously hermaphroditic snail Arianta arbustorum

Summary

In many simultaneously hermaphroditic land snail species, the sperm storage organ (spermatheca) is highly structured, suggesting that the female function might be able to influence offspring paternity. Physical properties of the sperm storage organ, including its initial size and sperm storage capacity, may also affect fertilization patterns in multiply mated snails. We examined the structure, volume and tubule length of empty spermathecae in the land snail, Arianta arbustorum, and assessed differences in spermatheca size following a single copulation. The number of spermathecal tubules ranged from 2–7, but was not correlated with the volume of empty spermathecae. The volume of sperm stored in the spermatheca after a copulation was correlated with neither the number of spermathecal tubules nor copulation duration. Mean spermathecal volume more than doubled between two and thirty-six hours after sperm uptake, but the length of the spermathecal tubules did not change. Interestingly, the volume of sperm stored in the spermatheca seems not to be related to the size of the spermatophore and thus not to the number of sperm received (= allosperm). The amount of allosperm digested in the bursa copulatrix was highly variable and no significant relationship with the size of the spermatophore received was found. These findings suggest that numerical aspects of sperm transfer are less important in influencing fertilization success of sperm in A. arbustorum than properties of the female reproductive tract of the sperm receiver.     

日本沼虾输精管的结构及其在精荚形成中作用的研究

应用光镜和透射电镜技术研究了日本沼虾输精管的结构及其在精荚形成中的作用。结果表明,日本沼虾输精管从形态结构上可分为近端输精管、卷曲输精管、远端输精管和膨大的远端输精管四部分。各部分的管壁皆由分泌上皮、基膜、肌肉层和结缔组织构成,其中分泌上皮包括高度明显不同的低柱状上皮和高拄状上皮两部分。输精管各部分管腔内含有处于不同形成阶段的精荚。进入近端输精管内的精子被支撑在一种嗜酸性基质中。近端输精管的分泌物主要帮助形成精子团,同时形成精荚壁的极小部分。卷曲和远端输精管分泌形成精荚壁的绝大部分,其分泌物由细胞顶端通过外排作用和顶泌机制分泌产生。膨大的远端输精管具有贮存精荚的作用,其分泌上皮也通过外排作用和顶泌机制产生分泌物包裹在已基本形成的精荚外侧,管壁肌肉层在雌雄交配时将管腔内的精荚切割成适宜长度并排出体外。       

Ejaculate production schedule and the degree of protandry in fishflies (Megaloptera: Corydalidae)

1. To examine the relationship between male–female emergence patterns and ejaculate dynamics, patterns producing sperm and seminal fluids in male internal reproductive organs, the size of a spermatophore transferred at mating and the fate of spermatophore contents moved into female storage organs were compared among the fishflies (Megaloptera: Corydalidae: Chauliodinae), Parachauliodes continentalis, P. japonicus and Neochauliodes sinensis .
2. Spermatophore contents moved into female storage organs decreased rapidly in P. japonicus and N. sinensis , but hardly at all in P. continentalis . This suggests that the females of the former two species may remate sooner than the latter species as it is known in insects that material remaining in the storage organs mechanically inhibits receptivity to mating.
3. Male P. japonicus and N. sinensis increased in internal reproductive organ mass continuously after adult eclosion, and the spermatophore size produced at the first mating increased with male age. In contrast, the internal reproductive organs of P. continentalis were relatively small and did not increase in mass after emergence. P. continentalis transferred a constantly smaller spermatophore at any copulation than the former two species.
4. Males of P. japonicus and N. sinensis emerged earlier than females, while P. continentalis showed a nearly simultaneous emergence pattern between the sexes. It seems that males of P. japonicus and N. sinensis (more polyandrous than P. continentalis ) inhibit female receptivity for a longer time period by emerging earlier and transferring larger ejaculates. In these two species, the number of sperm ejaculated at the first mating also increased with male age. The increase in sperm number by emerging earlier may be adaptive for the males owing to numerical sperm competition when the female remates.     

Sperm storage by spermatodoses in the spermatheca of Trioza alacris (Flor, 1861) hemiptera,psylloidea, triozidae: A structural and ultrastructural study

Female insects generally store sperm received during mating in specific organs of their reproductive tract, i.e., the spermathecae, which keep the sperm alive for a long time until fertilization occurs. We investigated spermatheca morphology and ultrastructure in the psylloidean insect Trioza alacris (Flor, 1861 ) in which spheroidal sperm packets that we refer to as ‘spermatodoses’ are found after mating. The ectoderm‐derived epithelium of the sac‐shaped spermatheca that has a proximal neck, consists of large secretory and flat cuticle‐forming cells. Secretory cells are characterized by a wide extracellular cavity, bordered by microvilli, in which electron‐dense secretion accumulates before discharge into the spermathecal lumen. The cuticle‐forming cells produce the cuticular intima of the organ and a peculiar specialized apical structure, through which secretion flows into the lumen. At mating, the male transfers bundles of sperm cells embedded in seminal fluid into the spermathecal neck. Sperm cells proceed towards the spermathecal sac lumen, where they are progressively compacted and surrounded with an envelope that also encloses secretions of both male and female origin. We describe the formation of these sperm containing structures and document the contribution of the female secretion to spermatodose or female‐determined spermatophore construction. We also discuss the choice of the term ‘spermatodose’ for T. alacris and suggest it be used to refer to sperm masses constructed in the female reproductive organs, at least when they involve the contribution of female secretion. © 2011 Wiley Periodicals, Inc.     

The outcome of sperm competition is affected by behavioural and anatomical reproductive traits in a simultaneously hermaphroditic land snail

Sperm competition is important in species with reproductive strategies that involve multiple mating and prolonged sperm storage such as the simultaneously hermaphroditic land snail Cornu aspersum. Double mating trials in this species have revealed that mating order and courtship behaviour affect paternity success. We investigated the effect of behavioural and anatomical reproductive traits on paternity success from triple mating trials. Triple mating resulted in triple fertilization in 58% of the cases whereas zero paternity was observed in 16% of sperm donors. Third sperm donors achieved higher paternity followed by first and second sperm donors. Snails with a longer epiphallus, the spermatophore forming organ, sired more offspring regardless of their mating order. Genetic compatibility between sperm donor and recipient did not influence paternity success. The results of the present study identified mating order and epiphallus length, as traits affecting the outcome of sperm competition in this species.     

Ultrastructural analysis of the morphology and function of the spermatheca of the pulmonate slug Arion subfuscus

A study of the histology, histochemistry and ultrastructure of the spermatheca of Arion subfuscus in different stages of its life cycle shows that the spermatheca, an accessory organ of the femal genital tract of pulmonates, is a spherical-shaped organ lined with a columnar and non-ciliated epithelium surrounded by a thin network of connective tissue with some muscle fibres. The narrow epithelial cells possess numerous microvilli that, excepting young specimens, are provided with long, thin and generally curved, membranous process. Golgi apparatus, RER and mitochondria are abundant. Basal infolds are not very deep. In specimens killed two days after-copula, the spermatheca is swollen and its lumen is full of degenerating spermatozoa, mucus masses and spermatophore fragments. The apex of the cell is rich in granules with varied content, including multivesicular bodies. In specimens killed two weeks after-copula there are numerous endocytic vesicles in the apex and big vacuoles containing lipid. It is considered that after mating the excess of exogenous spermatozoa and copulatory seminal fluids are digested in the spermatheca. First, there is extracellular digestion that may be carried out by the enzymes contained in the multivesicular bodies exocytosed to the lumen, as well as by the enzymes secreted in apocrine vesicles. The partially digested materials would then be absorbed by endocytosis and further digested intracellularly. The great accumulation of lipid in the epithelial cells two weeks after mating suggest that the spermatheca could be involved in lipid synthesis, acting as a reserve organ.     

SPERMATOPHORE SIZE IN BUSHCRICKETS: COMPARATIVE EVIDENCE FOR NUPTIAL GIFTS AS A SPERM PROTECTION DEVICE

During courtship and copulation, males of many insect species provide the female with a nuptial gift of a prey item or synthesized material. These gifts may be explained as a form of paternal investment by increasing female reproductive output, or in terms of mating effort by increasing male fertilization success. These explanations, while not mutually exclusive, are controversial. While experimental studies examine the maintenance of nuptial gifts in single species, comparative studies are required to indicate more general evolutionary trends. Male bushcrickets provide females with a nuptial gift, a spermatophylax, which is transferred to females at mating along with the sperm-containing ampulla. Analysis of comparative data of 28 species of bushcrickets (Orthoptera: Tettigoniidae), reveals that male spermatophore size (spermatophylax and ampulla weight) is positively correlated with female refractory period, which, in turn, correlates with male fertilization success. Moreover, gift size (the spermatophylax) covaries with ejaculate size (the ampulla), which is consistent with the hypothesis that it serves as a sperm protection device. In contrast, there is no significant correlation between any measure of female fecundity and male spermatophylax size. This indicates that the variation in spermatophore size among bushcrickets is better explained by a mating-effort function than a paternal investment function.     

Delayed spermatophore removal in the land snail Euhadra peliomphala

Although many studies have demonstrated that variation in male genital morphology can influence male fertilization success, it remains unclear in many cases which mechanism(s) of sexual selection and sexual conflict produce the morphological divergence of genitalia. Distinguishing between such mechanisms requires, at the very least, information on the process by which genital morphology influences fertilization success. The length of the spermatophore‐producing organ (SPO) of the stylommatophoran land snails is a case in point. The length of the spermatophore may have an important influence on the transfer of the spermatophore to the recipient's genital tract. To test this hypothesis, we investigated in Euhadra peliomphala (Pfeiffer, 1850) the relationship between the length of the SPO in sperm donors and the position of their spermatophore in sperm recipients at 72, 84, and 96 h after mating. The results showed that longer spermatophores required longer times to reach a gametolytic organ and to begin to be digested. These findings suggest that this delay in spermatophore digestion explains the mechanism for the high fertilization success shown by snails with a long SPO. © 2013 The Linnean Society of London     

Social group size, potential sperm competition and reproductive investment in a hermaphroditic leech, Helobdella papillornata (Euhirudinea: Glossiphoniidae)

Social group size may affect the potential for sperm competition, and this in turn may favour ontogenetic adjustments in testicular mass according to the likely requirements for sperm and spermatophore production. In a number of comparative analyses of testis mass among vertebrate species that differ in mating system or social organization, increasing potential for sperm competition is associated with larger testis size. Intraspecific phenotypic plasticity should be able to produce the same pattern if social group size is heterogenous and reflects differing degrees of average sperm competition, but this intraspecific effect is less well studied. We tested the effect of social groups on both male and female investment in the simultaneously hermaphroditic leech, Helobdella papillornata. Leeches were placed in groups of one, two, four or eight. Sexual investment at the onset of reproductive maturity was quantified as the total testisac volume for male function and total egg volume for female function. We found that testisac volume (statistically adjusted for body size) showed a significant increase with increasing group size. Total egg volume (also adjusted for body size) was unaffected by group size. Our findings indicate adaptive developmental plasticity in male gonad investment in response to the potential for sperm competition.     

Effects of male mating interval on spermatophore formation,transfer, and subsequent female receptivity and fecundity in the sorghum plant bug,Stenotus rubrovittatus

Males of the sorghum plant bug, Stenotus rubrovittatus (Matsumura) (Heteroptera: Miridae), transfer a spermatophore to females during copulation. After a 1‐day interval between the first and second copulation, males transferred both sperm and a spermatophore to females during the second copulation. However, when male mating interval was <1 h, they transferred sperm but no spermatophores to females during the second copulation. Therefore, the male mating interval probably produces two types of mated females, those with and those without a spermatophore. Mated females of S. rubrovittatus do not remate for at least 3 days after mating, even when courted, and lay more eggs than virgin females at the beginning of the oviposition period. The effects of spermatophores on female sexual receptivity and fecundity were examined using mated females with or without a spermatophore. Only one of the 40 (2.5%) mated females with a spermatophore remated, whereas 10 of the 26 (38.5%) without a spermatophore remated. Furthermore, mated females with a spermatophore laid more eggs than those without a spermatophore. These results suggest that spermatophores participate in reducing female sexual receptivity and enhancing female fecundity in S. rubrovittatus.     

Impact of male condition on his spermatophore and consequences for female reproductive performance in the Glanville fritillary butterfly

In butterflies, male reproductive success is highly related to the quality and the size of the spermatophore transferred to the female. The spermatophore is a capsule produced by the male during copulation, which in many species contains sperm in addition to a nuptial gift, and which is digested by the female after copulation. The nuptial gift may contribute to egg production and offspring quality, and in some cases also to female body maintenance. The production of the spermatophore, however, represents a cost for the male and, in polyandrous species, ejaculates are sometimes allocated adaptively across matings. Nonetheless, although the ecological factors affecting the reproductive success of female butterflies have been the topic of numerous studies, little information exists on the factors affecting males’ contribution to reproduction, and the indirect impacts on female fecundity and fitness. We used the Glanville fritillary butterfly, Melitaea cinxia (Linnaeus, 1758) (Nymphalidae), in order to assess variation in male allocation to matings. In this species, smaller males produce smaller spermatophores, but variation in spermatophore size is not correlated with female reproductive success. We show that spermatophore size increases with male age at first mating, decreases with mating frequency and adult food‐deprivation, and is not influenced by developmental food‐limitation. The length of copulation period does not influence the spermatophore size nor influences the polyandrous mating behavior in this species. Male contribution to his spermatophore size is clearly influenced by his condition and adult‐resource at the time of mating. Despite this variation, spermatophore size does not seem to have a direct impact on female reproductive output or mating behavior.     

中华绒螯蟹(Eriocheir sinensis)雄性生殖系统的组织学研究

中华绒螯蟹雄性生殖系统的组织学研究表明:生精小管一侧为管壁上皮,另一侧为生发区,生殖细胞由生发区基底部同管腔增殖。输精管分为输精细管和贮精囊,管壁上皮具分泌功能,贮精囊有肌肉层。射精管壁肌肉层较厚,粘膜形成纵行皱襞。副性腺内壁为单层立方上皮。生殖系统发育有明显的季节性,8月开始发育加速,10月进入高峰,4月开始发育停滞。     

Morphology and function of reproductive organs in Neodasys chaetonotoideus (Gastrotricha: Neodasys) with a phylogenetic assessment of the reproductive system in Gastrotricha

The reproductive system of the important basal gastrotrich Neodasys chaetonotoideus is described and reconstructed on the basis of light microscopy, serial ultrathin sections (ultrastructure) and scanning electron microscopy. Starting frontally, the hermaphroditic reproductive system consists of paired and tube shaped lateral testes that do not possess elongated seminal ducts but most likely open directly via paired ventral pores. The unpaired, medio‐dorsal ovary region contains early oogenic stages that mature caudally towards the uterus region, where the most mature egg is positioned laterally to the midgut. The ovary region is not covered with an epithelial lining whereas the uterus region possesses a distinct epithelial wall. Between ovary and uterus region, we have detected a conspicuous section of the female gonad, the vitellogenic oviduct that consists of a thick epithelial wall which forms cellular protuberances into the developing oocytes passing the oviduct. We interpret this as a special, hitherto undescribed mode of vitellogenesis in Gastrotricha. Further caudally, the uterus continues with the fronto‐caudal organ, a complex of two substructures that are apparently homologous to the frontal organ and the caudal organ of many species of the Gastrotricha Macrodasyida. Neodasys chaetonotoideus obviously engages in spermatophore formation and transfer. In this study we develop a morpho‐functional scenario for the gonads and accessory organs in terms of spermatophore production, exchange and oviposition. We compare our newly obtained data with already published results on the reproductive organs of several species of Gastrotricha by means of a species‐character matrix and provide a computer aided evaluation by a parsimonious character optimization. A reconstruction of the reproductive system of the stem species of Gastrotricha on the basis of three recent phylogenetic analyses is presented. These reconstructions give support for a Neodasys‐like reproductive system in the ground pattern of Gastrotricha with slight morphological differences and direct transfer of spermatozoa rather than spermatophore transfer. The evolution of selected characters is traced thus revealing some incidents of convergent evolution as well as the evolutionary replacement of the ancestral frontal organ by the derived frontal sac in at least two separated lineages.     

Prenuptial agreements: mating frequency predicts gift-giving in Heliconius species

Theory predicts that when males provision females with nuptial gifts that include nutrients, the degree of polyandry should be positively correlated with the size or quality of the gift. This is because larger and more nutritious gifts tend to increase female refractory period, reducing the chances the female will remate soon. This decreases the likelihood of sperm competiton and consequently increases the donor male fitness. Butterflies in the genus Heliconius Kluk (Lepidoptera: Nymphalidae: Heliconini) exhibit variable mating systems that include monandry and polyandry. In addition to protein in the spermatophore, males increase gift quality by providing females with cyanide, which may contribute to protection of the female or her eggs. We tested whether degree of polyandry and gift quality (spermatophore weight and cyanide content) were correlated in nine Heliconius species from greenhouse populations. As predicted, both spermatophore weight and cyanide content were correlated with mating frequency. This is the first report to show that degree of polyandry correlates with allocation of defensive chemical as part of a nuptial gift.     

Mating and the inferred function of the genital system of the nudibranch, Aeolidiella glauca (Gastropoda: Opisthobranchia: Aeolidioidea)

Abstract. We describe the genital system of the aeolid nudibranch gastropod Aeolidiella glauca as a basis for our ongoing analysis of the mating system of this hermaphroditic species. In addition we give a short account of its mating behavior. A. glauca has an androdiaulic genital system with a proximally situated semiserial seminal receptacle. There is no bursa copulatrix. After fertilization, eggs pass through six glands, i.e., the capsule gland and the female gland mass which is comprised of five histologically differentiated parts. The prostate is a long, glandular tube. The everted, unarmed penis is very large and bears a series of 3–4 hook-shaped lobes consisting only of a simple, ciliated epithelium on its ventral side. Their function is unknown. After courtship, which involves moving in circles followed by resting in a head-to-head position, reciprocally touching each other with the tentacles, the slugs glide into a position where the everted genital atria are in contact. The huge penes are protruded simultaneously shortly after this contact occurs. Each animal strokes its partner's back with the penis and deposits a spermatophore of undetermined shape onto the partner's notum. Sperm enter the recipient through histolysis. How the sperm find their way to the seminal receptacle is not known.     

Mating behavior and the function of the male genital spine in the ground beetle Carabus clathratus

The morphologies of male genitalia often appear harmful or aggressive, as if they may inflict physical damage upon females during copulation. Such male genitalia are often thought to function in intra- and intersexual interactions during mating. In the carabid genus Carabus, division Spinulati, males possess a spine (spinula) on the intromittent organ, of which function is unknown. To reveal the function of the spinula, we studied the mating behavior and genital coupling of a Spinulati species, Carabus (Limnocarabus) clathratus. The males positioned the spinula along the inner wall of the vaginal opening throughout copulation. This placement created a small dent and subsequently a melanized patch (wound) on the vaginal wall, but the spinula rarely penetrated the vaginal wall. The spinula did not reach the innermost part of the vagina where the spermatophore is deposited. These results suggest that the spinula is not used for inflicting damage on female genitalia or manipulating spermatophores of rival males. During spermatophore formation, the male partially withdrew the aedeagus, and only the aedeagal tip and endophallus remained within the female. By placing the spinula against the vaginal wall, the male could hold the endophallus within the vaginal chamber in the unstable copulatory posture. Thus, our observations suggest that the spinula primarily functions as an "anchor" to maintain the coupling of the male and female genitalia and thereby ensure insemination.     

Anatomy and ultrastructure of the reproductive organs in Dactylopodola typhle (Gastrotricha: Macrodasyida) and their possible functions in sperm transfer

Abstract. The reproductive anatomy of gastrotrichs is well known for several species, especially for the marine taxon Macrodasyida. However, there is little information on the reproductive organs and the modes of mating and sperm transfer in putative basal taxa, which is necessary for accurate reconstruction of the ground pattern of the Gastrotricha. We present the first detailed morphological investigation of the reproductive system of a putative basal gastrotrich, Dactylopodola typhle , using transmission and scanning electron microscopy, histology, and microscopic observations of living specimens. Dactylopodola typhle is a hermaphrodite that possesses paired female and male gonads, an unpaired uterus with an outlet channel that we call the cervix, and an additional accessory reproductive organ, the so-called caudal organ. We hypothesize that the hollow, secretory caudal organ serves for picking up autospermatozoa (self-sperm), for spermatophore formation, and finally for transferring the autospermatophore to a mating partner. The allospermatophore (foreign spermatophore) is stored within the uterus where fertilization occurs. We think that the mature and fertilized egg is released through the cervix and the dorsolateral female gonopore, and not by rupture of the body wall. Based on the morphology, we provide a plausible hypothesis for spermatophore formation and transfer in D. typhle . Preliminary phylogenetic considerations indicate that the stem species of Macrodasyida, perhaps that of all Gastrotricha, had paired ovaries and paired testes, an unpaired uterus, and only one accessory reproductive organ.