Insemination,intrafollicular fertilization and development of the fertilization plug during gestation in Heterandria formosa (Poeciliidae)

The viviparous teleost Heterandria formosa is a remarkable species for its reproductive characters including: (a) the smallest oocyte in viviparous fish species; (b) a high level of matrotrophy with a complex placenta; and (c) the highest level of superfetation. Superfetation involves (d) the continuous development of oocytes and fertilization at the same time with embryos in gestation. The sequential fertilization of oocytes requires (e) storage of spermatozoa in the ovary. Among these characteristics, fertilization is of fundamental interest, specifically the intrafollicular fertilization of poeciliids, species that do not present micropyle, and the consequent formation of the fertilization plug, a structure developed at the periphery of the follicle where the entrance of spermatozoa occurs. Both processes intrafollicular fertilization and formation of the fertilization plug have been rarely described. There is only one study illustrating, the fertilization plug of H. formosa with a drawing. In the context of reproductive aspects of H. formosa, the goal of this study is to describe the morphology of the ovary during insemination, intrafollicular fertilization and development of the fertilization plug. After insemination, spermatozoa enter the ovary and occupy folds of the lamella near follicles of all stages of oogenesis, the delle, where the germinal epithelium establishes contact with the follicular epithelium. The results of the present study provide evidence that both epithelia open at the distal end of the delle, this morphological change allow that the spermatozoa to make contact with the zona pellucida of the oocyte. After fertilization, the delle becomes blocked by proliferation of cells of the germinal epithelium, to form the fertilization plug that persists throughout gestation. Abundant reticular fibers and blood vessels are seen around the fertilization plug. Persistence of the fertilization plug suggests that it could be the site where the juvenile will gain entrance to the ovarian lumen during birth.     

Proliferation of Oogonia and folliculogenesis in the viviparous teleost Ilyodon whitei (Goodeidae)

Oogonial proliferation in fishes is an essential reproductive strategy to generate new ovarian follicles and is the basis for unlimited oogenesis. The reproductive cycle in viviparous teleosts, besides oogenesis, involves development of embryos inside the ovary, that is, intraovarian gestation. Oogonia are located in the germinal epithelium of the ovary. The germinal epithelium is the surface of ovarian lamellae and, therefore, borders the ovarian lumen. However, activity and seasonality of the germinal epithelium have not been described in any viviparous teleost species regarding oogonial proliferation and folliculogenesis. The goal of this study is to identify the histological features of oogonial proliferation and folliculogenesis during the reproductive cycle of the viviparous goodeid Ilyodon whitei. Ovaries during nongestation and early and late gestation were analyzed. Oogonial proliferation and folliculogenesis in I. whitei, where intraovarian gestation follows the maturation and fertilization of oocytes, do not correspond to the late oogenesis, as was observed in oviparous species, but correspond to late gestation. This observation offers an example of ovarian physiology correlated with viviparous reproduction and provides elements for understanding the regulation of the initiation of processes that ultimately result in the origin of the next generation. These processes include oogonia proliferation and development of the next batch of germ cells into the complex process of intraovarian gestation. J. Morphol. 275:1004–1015, 2014. © 2014 Wiley Periodicals, Inc.     

Superfetation in the viviparous fish Heterandria formosa (Poeciliidae)

Heterandria formosa is a viviparous poeciliid native of the southeastern of United States of America. H. formosa exhibits unique reproductive features as: (a) production of extremely small eggs with scarce quantity of yolk (microlecithal eggs), (b) consequently, a high level of matrotrophy and development of a complex follicular placenta, (c) ovarian sperm storage that allows the continuous fertilization of oocytes and production of offspring and (d) development of high degree of superfetation. The degree of superfetation refers to the number of broods in different simultaneous stages of gestation. Morphological evidence of the degree of superfetation in H. formosa has not been documented. Therefore, and because of the general interest in the complex process of superfetation, the goal of this study is to morphologically define the degree of superfetation of H. formosa through two procedures: (a) histological analysis of entire ovaries in gestation and (b) dissection of visible embryos and the histological analysis of the remaining ovarian tissue. Results indicate that H. formosa can gestate up to seven broods at the same time.     

Functional morphology of the gonoduct of the viviparous teleost Poeciliopsis gracilis (Heckel, 1848) (Poeciliidae)

Female teleosts do not develop Müllerian ducts; consequently, the ovary of teleosts contains two zones: germinal and gonoduct. The gonoduct lacks germinal cells, but has relevant functions in the reproductive process. We describe the functional morphology of the gonoduct in the viviparous teleost Poeciliopsis gracilis during nongestation and gestation stages. This study tests the hypothesis that the gonoduct functions as a barrier between the germinal zone and the exterior. By providing information about morphology and function of the gonoduct we show that this part of the ovary has an essential role in the reproduction of teleosts. The ovaries were processed by histological technique and stained with hematoxylin‐eosin (H‐E), Masson's trichrome, toluidine blue and periodic acid‐Schiff (PAS). The gonoduct is divided into three regions: cephalic, middle, and caudal. In the cephalic and middle regions there are mucosal folds that extend into the gonoductal lumen, forming structures similar to a cervix. The caudal region has two portions: the anterior contains a dorsal invagination and exocrine glands among columnar cells; the posterior has a ventral flexion and stratified epithelium with apical secretory cells. The morphology of this epithelium indicates two functions: (a) secretory by the apical columnar cells, and (b) protection through the stratification. Another peculiarity of the caudal region is that both ducts, reproductive and digestive, converge in a common cavity at their caudal ends, forming a cloacal region. The histology of the gonoduct indicates relevant functions including: (1) the control of the luminal diameter by the muscle and the presence of mucosal folds, like a cervix; (2) the relationship with the spermatozoa during insemination and storing them in mucosal folds; (3) the support of immunological processes; (4) secretory activities; (5) forming the duct during birth; and (6) possibly, acts as a barrier against parasite infestations.     

Oogenesis of microlecithal oocytes in the viviparous teleost Heterandria formosa

Viviparous teleosts exhibit two patterns of embryonic nutrition: lecithotrophy (when nutrients are derived from yolk that is deposited in the oocyte during oogenesis) and matrotrophy (when nutrients are derived from the maternal blood stream during gestation). Nutrients contained in oocytes of matrotrophic species are not sufficient to support embryonic development until term. The smallest oocytes formed among the viviparous poeciliid fish occur in the least killifish, Heterandria formosa, these having diameters of only 400 μm. Accordingly, H. formosa presents the highest level of matrotrophy among poeciliids. This study provides histological details occurring during development of its microlecithal oocytes. Five stages occur during oogenesis: oogonial proliferation, chromatin nucleolus, primary growth (previtellogenesis), secondary growth (vitellogenesis), and oocyte maturation. H. formosa, as in all viviparous poeciliids, has intrafollicular fertilization and gestation. Therefore, there is no ovulation stage. The full‐grown oocyte of H. formosa contains a large oil globule, which occupies most of the cell volume. The oocyte periphery contains the germinal vesicle, and ooplasm that includes cortical alveoli, small oil droplets and only a few yolk globules. The follicular cell layer is initially composed of a single layer of squamous cells during early previtellogenesis, but these become columnar during early vitellogenesis. They are pseudostratified during late vitellogenesis and reduce their height becoming almost squamous in full‐grown oocytes. The microlecithal oocytes of H. formosa represent an extreme in fish oogenesis typified by scarce yolk deposition, a characteristic directly related to matrotrophy. J. Morphol., 2011. © 2010 Wiley‐Liss, Inc.     

Ovarian structure and oogenesis of the oviparous goodeids Crenichthys baileyi (Gilbert, 1893) and Empetrichthys latos Miller, 1948 (teleostei,Cyprinodontiformes)

The cyprinodontiform family Goodeidae comprises two biogeographically disjunct subfamilies: the viviparous Goodeinae endemic to the Mexican Plateau, and the oviparous Empetrichthyinae, known only from relict taxa in Nevada and California. Ovarian characteristics of two oviparous species of goodeid, Crenichthys baileyi and Empetrichthys latos, studied using museum collections, are compared with those of viviparous species of goodeids. Both subfamilies have a single, cystovarian ovary. The ovary in the viviparous Goodeinae has an internal septum that divides the ovarian lumen into two compartments, and it may possess oogonia. There is no ovarian septum in the oviparous C. baileyi and E. latos. Oogenesis is similar in both subfamilies with regard to the proliferation of oogonia, initiation of meiosis, primary growth and development of an oocyte during secondary growth in which fluid yolk progressively fuses into a single globule. Notably, eggs of C. baileyi and E. latos are approximately double the size of those of the viviparous Goodeinae in which embryos develop inside the ovarian lumen and are nourished, in part, by nutrients transferred from the maternal tissues, a mode of embryo development called matrotrophy. Egg envelopes of the two subfamilies differ in that those of C. baileyi and E. latos have a relatively thick zona pellucida, attachment fibrils or filaments that develop between the follicle cells during oogenesis, and a micropyle observed only in E. latos. In contrast, viviparous goodeid eggs have a relatively thin zona pellucida, but lack adhesive fibrils, and a micropyle was not observed. These reproductive characters are compared with those of species of the eastern North American Fundulus, a representative oviparous cyprinodontiform. One newlyrecognized shared, derived character, a single, median ovoid ovary with no obvious external evidence of fusion, supports monophyly of the Goodeidae. Differences among the goodeid subfamilies and Fundulus are interpreted relative to the oviparous versus viviparous modes of reproduction. J. Morphol., 2012. © 2011 Wiley Periodicals, Inc.     

Bioenergetic components of reproductive effort in viviparous snakes: costs of vitellogenesis exceed costs of pregnancy

Reproductive effort has been defined as the proportion of an organism's energy budget that is allocated to reproduction over a biologically meaningful time period. Historically, studies of reproductive bioenergetics considered energy content of gametes, but not costs of gamete production. Although metabolic costs of vitellogenesis (MCV) fundamentally reflect the primary bioenergetic cost of reproductive allocation in female reptiles, the few investigations that have considered costs of reproductive allocation have focused on metabolic costs of pregnancy (MCP) in viviparous species. We define MCP as energetic costs incurred by pregnant females, including all costs of maintaining gestation conditions necessary for embryogenesis. MCP by our definition do not include fetal costs of embryogenesis. We measured metabolic rates in five species of viviparous snakes (Agkistrodon contortrix, Boa constrictor, Eryx colubrinus, Nerodia sipedon, and Thamnophis sirtalis) during vitellogenesis and pregnancy in order to estimate MCV and MCP. Across all species, MCV were responsible for 30% increases in maternal metabolism. Phylogenetically-independent contrasts showed that MCV were significantly greater in B. constrictor than in other species, likely because B. constrictor yolk energy content was greater than that of other species. Estimates of MCP were not significantly different from zero in any species. In viviparous snakes, MCV appear to represent significant bioenergetic expenditures, while MCP do not. We suggest that MCV, together with yolk energy content, represent the most significant component of reptilian reproductive effort, and therefore deserve greater attention than MCP in studies of reptilian reproductive bioenergetics.     

Development of the uterine shell glands during the preovulatory and early gestation periods in oviparous and viviparous Lacerta vivipara

The evolutionary process leading to the emergence of viviparity in Squamata consists of lengthening the period of egg retention in utero coupled with marked reduction in the thickness of the eggshell. We used light microscopy and scanning electron microscopy to study uterine structure during the reproductive cycle of oviparous and viviparous females of the reproductively bimodal Lacerta vivipara. We compared the structure of the uterine shell glands, which secrete components of the eggshell, during preovulatory and early gestation phases of the reproductive cycle and also compared histochemistry of the eggshells. The uterine glands of both reproductive forms undergo considerable growth within a period of a few weeks during folliculogenesis and vitellogenesis preceding ovulation. The majority of the proteinaceous fibers of the shell membrane are secreted early in embryonic development and the uterine glands regress shortly thereafter. This supports previous observations indicating that, in Squamata, secretion of the shell membrane occurs very rapidly after ovulation. The most striking differences between reproductive modes were larger uterine glands at late vitellogenesis in oviparous females, 101 microm compared to 60 microm in viviparous females, and greater thickness of the shell membrane during early gestation in oviparous females (52-73 microm) compared to viviparous females (4-8 microm). Our intraspecific comparison supports the conclusions of previous studies that, prior to ovulation, the uterine glandular layer is less developed in viviparous than in oviparous species, and that this is the main factor accounting for differences in the thickness of the shell membrane of the two reproductive forms of squamates.     

Morphology of the male reproductive system of freshwater prawn Macrobrachium carcinus (Decapoda,Caridea): Functional and comparative aspects

Testes and vasa deferentia are parts of the male reproductive system of decapod crustaceans. Both organs show morphological differences among decapod species in terms of anatomical and histological patterns reflecting the diversity of this group. Describing these features may assist in systematics, phylogenetics, and studies of reproductive behavior, especially for species of commercial interest, such as Macrobrachium carcinus, a native American species that, unusually for this genus, has no precopulation courting behavior. This study aims to describe the reproductive morphology and spermatogenesis of the male freshwater prawn M. carcinus. The male reproductive system of this species consisted of lobed testes connected to the vasa deferentia. The testis of M. carcinus was divided into several lobules. Each lobule was formed by a cluster of germ cells surrounded by connective tissue and nurse cells. This microscopic anatomy and histology of the testicular histoarchitecture has been described for many species of Decapoda and may represent a derived design of the testes. Unlike that in other decapod species, spermatogenesis proceeds in short transitory phases that produce spermatozoa at high concentrations and frequencies, corroborating the uncommon male reproductive behavior of this species. In the spermatic pathway, the lobules develop and fuse before releasing spermatozoa from the testes; however, this process has not been observed in decapods, yet. The neutral compounds secreted by the vas deferens are important for sperm nutrition as females secrete a substance for spermatophore adhesion during reproduction. This study presents different features and dynamics of the spermatogenic process in the male reproductive system of M. carcinus that have not yet been presented in the literature for decapods.     

Endocytosis-mediated vitellogenin absorption and lipid metabolism in the hindgut-derived placenta of the viviparous teleost Xenotoca eiseni

Certain viviparous animals possess mechanisms for mother-to-embryo nutrient transport during gestation. Xenotoca eiseni is one such viviparous teleost species in which the mother supplies proteins and other components to the offspring developing in the ovary. The embryo possesses trophotaenia, hindgut-derived placental structure, to receive the maternal supplement. However, research on the molecular mechanisms underlying viviparous species is scarce in non-mammalian vertebrates, including teleosts. Thus, we conducted this study to investigate the mechanism for nutrient absorption and degradation in trophotaeniae of X. eiseni. A tracer assay indicated that a lipid transfer protein, vitellogenin (Vtg), was absorbed into the epithelial layer cells of the trophotaeniae. Vtg uptake was significantly suppressed by Pitstop-2, an inhibitor of clathrin-mediated endocytosis. Gene expression analysis indicated that the genes involved in endocytosis-mediated lipolysis and lysosomal cholesterol transport were expressed in the trophotaeniae. In contrast, plasma membrane transporters expressed in the intestinal tract were not functional in the trophotaeniae. Our results suggested that endocytosis-mediated lysosomal lipolysis is one of the mechanisms underlying maternal component metabolism. Thus, our study demonstrated how viviparous teleost species have acquired a unique developmental system that is based on the hindgut-derived placenta.     

Effects of luteectomy in early pregnancy on the maintenance of gestation and plasma progesterone concentrations in the viviparous temperate lizard <Emphasis Type="Italic">Barisia imbricata imbricata</Emphasis>

Background  

Several studies have shown that the corpus luteum is the principal source of progesterone during the gravidity period in reptiles; however, its participation in the maintenance of gestation in the viviparous squamata is in dispute. The effects of ovariectomy or luteectomy vary according to the species and the time at which the procedure is performed. In this paper, we describe the effects of luteectomy during early pregnancy on the maintenance of gestation and progesterone concentrations in the temperate Mexican viviparous lizard Barisia imbricata imbricata.     

Specialized basophilic cells in the ventral lobe of the pituitary of the smooth dogfish,Mustelus canis

The ventral lobe of the adenohypophysis of the smooth dogfish, Mustelus canis, a viviparous elasmobranch, has been found to possess distinctive cells identified as basophils on the basis of staining properties. At maximum size, such a cell consists of a distended vesicle containing PAS-positive, AF-negative material surrounded by a thin envelope of cytoplasm and an eccentric nucleus. In earlier stages of these cells, vesicles are small or absent and granules in the more abundant cytoplasms are AF or Alcian-positive. Basophil numbers are high in pre-ovulation and mid-ovulation females, decrease markedly after the end of ovulation until embryos are about 1 cm long then increase greatly during August and September while embryos grow to 8 cm in length. Early high counts, if these basophils are gonadotropes, may be correlated with stimulation of the ovary and ovulation; reduced numbers suggest inhibition, possibly by ovarian hormones for a period, while subsequent increase may indicate indirect involvement in uterine conditions in this viviparous species. Conclusion are, admittedly, tentative as specimens were available during only a fraction of the ten month gestation period.     

Reproductive histology of Tomeurus gracilis Eigenmann, 1909 (Teleostei: Atherinomorpha: Poeciliidae) with comments on evolution of viviparity in atherinomorph fishes

Tomeurus gracilis is a species long considered pivotal in understanding the evolution of livebearing in atherinomorph fishes. Tomeurus gracilis is a zygoparous or embryoparous poeciliid: internal fertilization is followed by females laying fertilized eggs singly or retaining fertilized eggs until or near hatching. Tomeurus was hypothesized as the sister group of the viviparous poeciliids until it was proposed as a close relative of a derived viviparous poeciliid, Cnesterodon, hence nested among viviparous taxa rather than near the root of the tree. Here, we describe and compare reproductive morphological characters of the little‐known Tomeurus with those of representative atherinomorphs. In Tomeurus and Cnesterodon, sperm are packaged in naked sperm bundles, or spermatozeugmata, in a configuration considered here diagnostic of viviparous poeciliids. Testes are single and free sperm are stored in the ovary in both taxa in contrast to oviparous atherinomorphs in which testes are paired and sperm are not packaged and not stored in the ovary. Efferent ducts in Cnesterodon testes and other viviparous poeciliids have a PAS‐positive secretion demonstrating presence of a glycoprotein that inactivates sperm or prevents final sperm maturation. No PAS‐positive staining secretion was observed in Tomeurus or oviparous atherinomorphs. Tomeurus shares apomorphic reproductive characters, such as sperm bundle and testis morphology and a gonopodium, with viviparous poeciliids and plesiomorphic characters, such as a thick zona pellucida with filaments, with oviparous taxa. We do not postulate loss or reversal of viviparity in Tomeurus, and we corroborate its phylogenetic position as sister to the viviparous poeciliids. J. Morphol., 2010. © 2010 Wiley‐Liss, Inc.     

Reproduction and development of the South American freshwater stingrays,Potamotrygon circularis and P. motoro

Synopsis Observations of reproductive features and body measurements were made on wild-caught, freshwater stingrays, Potamotrygon circularis and P. motoro, from the Amazon drainage of western Brazil and southern Colombia. Further observations were made in Detroit's Belle Isle Aquarium on a captive pair of P. motoro and their descendants, which constitute the first known captive breeding colony of potamotrygonids. The gross structure and function of female and male reproductive systems are described. There is no obvious difference between those of the two species. They are aplacentally viviparous, the young being nourished in advanced stages by uterine milk secreted by trophonemata. Size at onset and completion of sexual maturation, breeding season and behavior, gestation period, litter size and sex ratios are discussed. Up to 21 proportional measurements were made on several fetal and postnatal stages of both species. Several proportional changes occur in very early fetal life, but most body proportions undergo only minor changes from advanced fetal through adult stages. A growth curve is proposed for P. motoro based on observations of the captive colony.     

Morphological structures for potential sperm storage in poeciliid fishes. Does superfetation matter?

Sperm storage within the female reproductive tract has been reported as a reproductive strategy in several species of vertebrates and invertebrates. However, the morphological structures that allow for sperm to be stored and kept viable for long periods are relatively unknown in osteichthyes. We use histological and stereological tools to identify and quantify sperm storage structures (spermathecae) in 12 species of viviparous Poeciliidae. We found spermathecae in nine species, six of which exhibit superfetation (the ability of females to simultaneously carry within the ovary two or more broods of embryos at different stages of development). These spermathecae are folds of ovarian tissue that close around spermatozoa. We compared the number and size (volume) of spermathecae between species with and without superfetation. Species that exhibit superfetation had a significantly higher number of spermathecae than species that do not exhibit this reproductive strategy. In addition, we found that the mean volume of spermathecae and total volume of spermathecae present in the ovary are marginally higher in species with superfetation. Our results contribute to the understanding of the morphological structures that allow for sperm storage in viviparous osteichthyes and suggest a positive relationship between superfetation and the capacity of females to store sperm.     

Energy consumption by embryos of a viviparous lizard, Eulamprus tympanum, during development

Energy consumption during development has been measured in many oviparous lizards, but not in viviparous lizards in utero. It has always been assumed that energy consumption by embryos of viviparous lizards during development is similar to that of oviparous species. Estimation of energy consumption of viviparous lizards in vivo are confounded by the possible influence of pregnancy on maternal metabolism. Here we separated maternal and embryonic metabolism in measurements of pregnant Eulamprus tympanum throughout pregnancy. Our data support the hypothesis that the energetic cost of development in viviparous lizards (19.8 kJ g⁻¹) is similar to that in oviparous lizards (mean 16.2 kJ g⁻¹), at least for a species with a simple placenta. An increase in maternal metabolism of 29% above that for non-pregnant E. tympanum goes to maintain pregnancy, and represents an important component of the reproductive effort in E. tympanum.     

Structure of the female gonoduct of the viviparous teleost Poecilia reticulata (Poeciliidae) during nongestation and gestation stages

Female teleosts do not have oviducts because Müllerian ducts do not develop. Instead, the caudal region of the ovary, the gonoduct, connects to the exterior. Because of the lack of oviducts in viviparous teleosts, the embryos develop in the ovary, as an intraovarian gestation, unique in vertebrates. This is the first study to address the histology of the gonoduct in a viviparous teleost. The gonoduct of Poecilia reticulata was analyzed during previtellogenesis, vitellogenesis, and gestation. The gonoduct lacks germinal cells. From deep to superficial, the wall has simple cuboidal or columnar epithelium, loose connective tissue, longitudinal layer of smooth muscle, and visceral peritoneum. Cells of the immune system occur in the lumen and in the mucosa. The gonoduct was divided in three regions: 1) cephalic, 2) middle, and 3) caudal. At the initial part of each region, thin mucosal folds extend into the lumen. The cephalic region forms a tubular structure with light and irregular folds. The middle region has a wider lumen and is more irregular due to ventral invaginations and irregular and short mucosal folds; beneath the epithelium there are melano‐macrophage centers. The caudal region is delimited from the middle region by folds; however, they are thinner than these of the other regions. Ventral invaginations form exocrine glands, and the smooth muscle is thicker than in the other regions. During gestation, cells of the immune system are abundant; melano‐macrophage centers become larger and the glands exhibit desquamated cells. These observations suggest roles of the gonoduct in reducing the diameter of the lumen; receiving sperm during vitellogenesis; producing secretions, more abundant during vitellogenesis; and in immunological activity throughout the reproductive cycle. The ciliated epithelium and the thick muscle of the caudal region may be involved during birth. J. Morphol. 275:247–257, 2014. © 2013 Wiley Periodicals, Inc.     

Behavioural and life‐history regulation in a unisexual/bisexual mating system: does male mate choice affect female reproductive life histories?

In theory, unisexual taxa have an advantage over ecologically similar bisexual species because unisexuals produce twice as many daughters and, thus, should quickly outcompete coexisting bisexuals in any given population. For sperm‐dependent unisexual (gynogenetic) species, stable coexistence with their bisexual sperm donors can be postulated if male mate choice puts unisexual females at a disadvantage through sperm limitation, thus halving their reproductive output compared to bisexuals (‘behavioural regulation hypothesis’). We tested for a potential life‐history signature of male mate choice in a system of coexisting bisexual sailfin mollies (Poecilia latipinna) and gynogenetic Amazon mollies (Poecilia formosa). Specifically, we gave P. latipinna males an opportunity to freely interact (and mate) with both types of females and, after 25 days, quantified the proportion of (1) females with sperm in their genital tract and (2) pregnant females. A higher proportion of P. latipinna females (53.7%) had sperm in their genital tract (compared to only 25.9% in P. formosa), corroborating a previous study on wild‐caught fish. This translated into a higher frequency (42.6%) of P. latipinna females being pregnant (compared to 29.6% in P. formosa); however, among pregnant females, no significant differences between species in reproductive life‐history traits (such as offspring number or size) were uncovered. Hence, although the findings of the present study confirm that male discrimination against unisexual females leads to reduced reproductive output in unisexuals, the observed magnitude of differences in targeted life histories between the two types of females is unlikely to be the sole factor regulating stable coexistence in this system. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 106 , 598–606.     

Do density‐driven mating system differences explain reproductive incompatibilities between populations of a placental fish?

Matrotrophy, the provisioning of embryos between fertilization and birth, creates the potential for conflict between mothers and embryos over the level of maternal investment. This conflict is predicted to drive the evolution of reproductive isolation between populations with different mating systems. In this study, we examine whether density‐driven mating system differences explain the patterns of asymmetric reproductive isolation observed in previous studies involving four populations of the matrotrophic least killifish, Heterandria formosa. Minimum sire number reconstructions suggested that two populations characterized by low densities had lower levels of concurrent multiple paternity than two populations characterized by high densities. However, low levels of genetic variation in the low‐density populations greatly reduced our probability of detecting multiple mating in them. Once we took the lower level of genetic variation into account in our estimations, high levels of multiple paternity appeared the rule in all four populations. In the population where we had the greatest power of detecting multiple mating, we found that multiple paternity in H. formosa typically involves multiple sires contributing to offspring within the same brood instead of different fathers contributing to distinct, simultaneously provisioned broods. Paternity was often skewed towards one sire. Our results suggest that differences between H. formosa populations in the levels of multiple paternity are not sufficient to explain the reproductive isolation seen in previous studies. We suggest that other influences on maternal–foetal conflict may contribute to the pattern of reproductive isolation observed previously. Alternatively, the asymmetric reproductive isolation seen in previous studies might reflect the disruption of maternal–foetal coadaptation.