An examination of surface epithelium structures of the embryo across the genus Poeciliopsis (Poeciliidae).

In viviparous, teleost fish, with postfertilization maternal nutrient provisioning, embryonic structures that facilitate maternal-fetal nutrient transfer are predicted to be present. For the family Poeciliidae, only a handful of morphological studies have explored these embryonic specializations. Here, we present a comparative morphological study in the viviparous poeciliid genus, Poeciliopsis. Using microscopy techniques, we examine the embryonic surface epidermis of Poeciliopsis species that vary in their level of postfertilization maternal nutrient provisioning and placentation across two phylogenetic clades and three independent evolutionary origins of placentation. We focus on surface features of the embryo that may facilitate maternal-fetal nutrient transfer. Specifically, we studied cell apical-surface morphology associated with the superficial epithelium that covers the body and sac (yolk and pericardial) of embryos at different developmental stages. Scanning electron microscopy revealed common surface epithelial cells across species, including pavement cells with apical-surface microridges or microvilli and presumed ionocytes and/or mucus-secreting cells. For three species, in the mid-stage embryos, the surface of the body and sac were covered in microvillus epithelium. The remaining species did not display microvillus epithelium at any of the stages examined. Instead, their epithelium of the body and sac were composed of cells with apical-surface microridges. For all species, in the late stage embryos, the surface of the body proper was composed of apical-surface microridges in a "fingerprint-like arrangement." Despite the differences in the surface epithelium of embryos across Poeciliopsis species and embryonic developmental stages, this variation was not associated with the level of postfertilization maternal nutrient provisioning. We discuss these results in light of previous morphological studies of matrotrophic, teleost fish, phylogenetic relationships of Poeciliopsis species, and our earlier comparative microscopy work on the maternal tissue of the Poeciliopsis placenta.

Adenylate cyclase 5 is required for melanophore and male pattern development in the guppy (Poecilia reticulata).

Guppies (Poecilia reticulata) are colorful fish that have attracted the attention of pigmentation researchers for almost a century. Here, we report that the blond phenotype of the guppy is caused by a spontaneous mutation in the guppy ortholog of adenylate cyclase 5 (adcy5). Using double digest restriction site-associated DNA sequencing (ddRADseq) and quantitative trait locus (QTL) mapping, we linked the blond phenotype to a candidate region of 118 kb, in which we subsequently identified a 2-bp deletion in adcy5 that alters splicing and leads to a premature stop codon. We show that adcy5, which affects life span and melanoma growth in mouse, is required for melanophore development and formation of male orange pigmentation traits in the guppy. We find that some components of the male orange pattern are particularly sensitive to loss of Adcy5 function. Our work thus reveals a function for Adcy5 in patterning of fish color ornaments.

Alternative splicing of Wilms tumor suppressor 1 (Wt1) exon 4 results in protein isoforms with different functions.

The Wilms tumor suppressor gene Wt1 encodes a zinc finger transcription factor that is essential for development of multiple organs including kidneys, gonads, spleen and heart. In mammals Wt1 comprises 10 exons with two characteristic splicing events: inclusion or skipping of exon 5 and alternative usage of two splice donor sites between exons 9 and 10. Most fish including zebrafish and medaka possess two wt1 paralogs, wt1a and wt1b, both lacking exon 5. Here we have characterized wt1 in guppy, platyfish and the short-lived African killifish Nothobranchius furzeri. All fish except zebrafish show alternative splicing of exon 4 of wt1a but not of wt1b with the wt1a(-exon 4) isoform being the predominant splice variant. With regard to function, Wt1a(+exon 4) showed less dimerization but stimulated transcription more effectively than the Wt1a(-exon 4) isoform. A specific knockdown of wt1a exon 4 in zebrafish was associated with anomalies in kidney development demonstrating a physiological function for Wt1a exon 4. Interestingly, alternative splicing of exon 4 seems to be an early evolutionary event as it is observed in the single wt1 gene of the sturgeon, a species that has not gone through teleost-specific genome duplication.

Shared and unique patterns of embryo development in extremophile poeciliids.

BACKGROUND: Closely related lineages of livebearing fishes have independently adapted to two extreme environmental factors: toxic hydrogen sulphide (H(2)S) and perpetual darkness. Previous work has demonstrated in adult specimens that fish from these extreme habitats convergently evolved drastically increased head and offspring size, while cave fish are further characterized by reduced pigmentation and eye size. Here, we traced the development of these (and other) divergent traits in embryos of Poecilia mexicana from benign surface habitats ("surface mollies") and a sulphidic cave ("cave mollies"), as well as in embryos of the sister taxon, Poecilia sulphuraria from a sulphidic surface spring ("sulphur mollies"). We asked at which points during development changes in the timing of the involved processes (i.e., heterochrony) would be detectible. METHODS AND RESULTS: Data were extracted from digital photographs taken of representative embryos for each stage of development and each type of molly. Embryo mass decreased in convergent fashion, but we found patterns of embryonic fat content and ovum/embryo diameter to be divergent among all three types of mollies. The intensity of yellow colouration of the yolk (a proxy for carotenoid content) was significantly lower in cave mollies throughout development. Moreover, while relative head size decreased through development in surface mollies, it increased in both types of extremophile mollies, and eye growth was arrested in mid-stage embryos of cave mollies but not in surface or sulphur mollies. CONCLUSION: Our results clearly demonstrate that even among sister taxa convergence in phenotypic traits is not always achieved by the same processes during embryo development. Furthermore, teleost development is crucially dependent on sufficient carotenoid stores in the yolk, and so we discuss how the apparent ability of cave mollies to overcome this carotenoid-dependency may represent another potential mechanism explaining the lack of gene flow between surface and cave mollies.

Do embryos influence maternal investment? Evaluating maternal-fetal coadaptation and the potential for parent-offspring conflict in a placental fish.

The evolution of matrotrophy introduces the potential for genomic conflicts between mothers and embryos. These conflicts are hypothesized to accelerate the evolution of reproductive isolation and to influence the evolution of life-history traits, reproductive structures, and genomic imprinting. These hypotheses assume offspring can influence the amount of maternal investment they receive and that there is a trade-off between maternal investment into individual offspring and maternal survival or fecundity. We used field data and laboratory crosses to test whether these assumptions are met in the matrotrophic poeciliid fish Heterandria formosa. Comparisons of life histories between two natural populations demonstrated a trade-off between the level of maternal investment into individual embryos and maternal fecundity. Laboratory crosses between individuals from these populations revealed that offspring genotype exerts an influence on the level of maternal investment and affects maternal fecundity through higher rates of embryo abortion and lower numbers of full-term offspring. Our results show that the prerequisites for parent-offspring conflict to be a potent evolutionary force in poeciliid fish are present in H. formosa. However, determining whether this conflict has shaped maternal investment in nature will require disentangling any effects of conflict from those of several ecological factors that are themselves correlated with the expected intensity of conflict.

Ecology of Phallotorynus pankalos (Cyprinodontiformes: Poeciliidae) in a first-order stream of the upper Paraná Basin

Some aspects of the population ecology of Phallotorynus pankalos in a first-order stream of the Iguatemi River Basin are described based on samples taken from March/2007 to February/2008. A total of 2680 individuals, including 948 males and 1732 females was collected. Adult females are larger than males; theirs mean fecundity was estimated as 6.5 embryos/female. There was a strong correlation between standard length and ovary weight, relative ovary weight, and number of embryos. The size of the first maturation of 50 percent of female population was estimated as 18.24 mm of standard lenght. High female mortality was observed after the first reproduction and sex ratio presents seasonal variation with higher female proportion in the winter.(AU)

Para descrever alguns aspectos da ecologia populacional de Phallotorynus pankalos em um riacho de primeira ordem da bacia do rio Iguatemi foram realizadas amostragens de março/2007 a fevereiro/2008. Um total de 2680 indivíduos, distribuídos em 948 machos e 1732 fêmeas, foi coletado. Fêmeas adultas foram maiores que os machos e sua fecundidade média foi estimada em 6,5 embriões/fêmea. Foi observada forte correlação entre o comprimento padrão e o peso dos ovários, peso relativo dos ovários e número de embriões. O tamanho da primeira maturação de 50 por cento da população de fêmeas foi estimado em 18,24 mm de comprimento padrão. Observamos elevada mortalidade de fêmeas após a primeira reprodução e variação sazonal na proporção sexual, com maior proporção de fêmeas no inverno.(AU)

In vitro culture of embryos of the guppy, Poecilia reticulata.

The rich variation in adult color patterns of male guppies (Poecilia reticulata) has attracted the attention of geneticists and ecologists for almost a century. Studies on their embryogenesis, however, have been limited by the fact that guppies are live bearers. We have observed normal development after explantation of guppy embryos from the ovary of pregnant females at various times after last parturition, and found that development of each batch of eggs is slightly asynchronous, most likely due to asynchronous fertilization. We have cultured explanted embryos in vitro and continuously observed their development. Although embryos explanted a few days after fertilization survived up to 4 weeks in culture, they did not complete their development. In contrast, embryos explanted at late stages of gestation could hatch and develop to fertile adults. Our embryo culture techniques overcome some of the limitations of using livebearers as study objects, and they allow continuous observation of and accessibility to live embryos at all stages.

Fecundidad, fertilidad e índice gonadosomático de Poecilia reticulata (Pisces: Poeciliidae)en Heredia, Costa Rica / Fertility, fecundity and gonadosomatic index of Poecilia reticulata (Pisces: Poeciliidae) in Heredia, Costa Rica

Entre setiembre y noviembre de 1998 se determinó la fecundidad,fertilidad e índice gonadosomático de Poecilia reticulata (Peters 1859),a partir de especímenes recolectados en un estanque en Santo Domingo de Heredia,Costa Rica.Las hembras presentaron una longitud total promedio de 34.43 ± 7.26 mm con un ámbito entre 17.80 y 51.50 mm y un peso total promedio de 0.69 ± 0.48 g,con un ámbito entre 0.06 y 2.32 g.Se determinó que esta especie presenta superfetación,matrotrofía y se mantuvo sexualmente activa durante todo el período de estudio.La fertilidad se rigió por la ecuación F =0.527e0.1171Lt (r2 =0.9404)y con una fertilidad promedio para la población de 56.1 ± 43.6,con un ámbito entre 7.98 y 197.58 (huevos +embriones).La fecundidad se rigió por la ecuación Fo = 0.695e0.1076 Lt (r2 =0.9376)con una fecundidad promedio para la población de 48.95 ± 35.99, con un ámbito entre 6.98 y 178.99 (ovocitos +óvulos). El índice gonadosomático en hembras se rigió por la ecuación IGS =0.0014e 0.134 Lt (r 2 =0.8581)y se encontró un índice gonadosomático promedio de 0.25 ± 0.27% con un ámbito entre 0.02 y 1.31%

Between September and November of 1998,1 432 females of Poecilia reticulata were collected in a pond in Santo Domingo,Heredia,Costa Rica.The average total length of females was 34.43 ± 7.26 mm (range 17.80 and 51.50 mm)and the average total weight of 0.69 ± 0.48 g (range 0.06 and 2.32 g). This species is a viviparous matrotrofic and the specimens were sexually mature.The total length-fertility relationship was F =0.527e 0.1171Lt (r2 =0.9404).The average fertility was 56.1 ± 43.6 (eggs +embryos)(range 8.0 and 197.6).The total length-fecundity relationship was Fo =0.695e0.1076 Lt (r2 =0.9376).The average fecundity was of 49.0 ± 36.0 (oocytes +ova)(range 7.0 and 179.0).The total length-gonadosomatic index relationship was IGS =0.0014 e0.134 Lt (r2 =0.8581).The average gonadosomatic index was 0.25 ± 0.27%(range 0.02 and 1.31%)

[Fertility, fecundity and gonadosomatic index of Poecilia reticulata (Pisces: Poeciliidae) in Heredia, Costa Rica]. / Fecundidad, fertilidad e índice gonadosomático de Poecilia reticulata (Pisces: Poeciliidae) en Heredia, Costa Rica.

Between September and November of 1998, 1 432 females of Poecilia reticulata were collected in a pond in Santo Domingo, Heredia, Costa Rica. The average total length of females was 34.43 +/- 7.26 mm (range 17.80 and 51.50 mm) and the average total weight of 0.69 +/- 0.48 g (range 0.06 and 2.32 g). This species is a viviparous matrotrofic and the specimens were sexually mature. The total length-fertility relationship was F = 0.527e 0.1171Lt (r2 = 0.9404). The average fertility was 56.1 +/- 43.6 (eggs + embryos) (range 8.0 and 197.6). The total length-fecundity relationship was Fo = 0.695e(0.1076Lt (r2 = 0.9376). The average fecundity was of 49.0 +/- 36.0 (oocytes + ova) (range 7.0 and 179.0). The total length-gonadosomatic index relationship was IGS = 0.0014 e0.134 Lt (r2 = 0.8581). The average gonadosomatic index was 0.25 +/- 0.27% (range 0.02 and 1.31%).

Nonlinkage of major histocompatibility complex class I and class II loci in bony fishes.

In tetrapods, the functional (classical) class I and class II B loci of the major histocompatibility complex (Mhc) are tightly linked in a single chromosomal region. In an earlier study, we demonstrated that in the zebrafish, Danio rerio, order Cypriniformes, the two classes are present on different chromosomes. Here, we show that the situation is similar in the stickleback, Gasterosteus aculeatus, order Gasterosteiformes, the common guppy, Poecilia reticulata, order Cyprinodontiformes, and the cichlid fish Oreochromis niloticus, order Perciformes. These data, together with unpublished results from other laboratories suggest that in all Euteleostei, the classical class I and class II B loci are in separate linkage groups, and that in at least some of these taxa, the class II loci are in two different groups. Since Euteleostei are at least as numerous as tetrapods, in approximately one-half of jawed vertebrates, the class I and class II regions are not linked.

Effects of neurohypophyseal and adenohypophyseal hormones, steroids, eicosanoids, and extrafollicular tissue on ovulation in vitro of guppy (Poecilia reticulata) embryos.

In the viviparous guppy, oocyte maturation is followed by intrafollicular fertilization and gestation. The fully developed embryos are ovulated at term just prior to parturition. Various agents were tested in vitro for their effects on ovulation of embryos in isolated follicles of the guppy. Arachidonic acid (10 and 100 microM), PGE2, PGF2 alpha, and 6-keto-PGF1 alpha (0.1 microgram/ml) induced ovulation, while PGE1, 15-keto-PGF2 alpha, leukotriene B4, 5-, 12-, and 15-HETEs (0.01 to 0.1 microgram/ml), cortisol, 11-deoxycortisol (25 and 250 ng/ml), estradiol-17 beta, testosterone, 17 alpha,20 beta-P, progesterone (5 and 50 ng/ml), isotocin, vasotocin (0.02 to 2 microgram/ml), and guppy pituitary extract (one and two glands per fish) did not. Extrafollicular (EF) ovarian tissue cocultured with isolated follicles induced ovulation, and the medium levels of PGE and PGF in such incubations were higher than those in the control. Indomethacin, the cyclooxygenase inhibitor, did not inhibit ovulation induced by arachidonic acid and EF tissue, although it inhibited PGE and PGF production. NDGA, the lipoxygenase inhibitor, did not inhibit ovulation induced by arachidonic acid or EF tissue. A combination of eicosanoids synthesized by follicles and EF tissue may be involved in the induction of ovulation. Dibutyryl cAMP inhibited ovulation induced by PGE2, PGF2 alpha, 6-keto-PGF1 alpha, and EF tissue suggesting that a low level of cAMP may be associated with ovulation in the guppy.

Development of spontaneous motility in the guppy embryo (Lebistes reticulatus) and the effect of spinal transection.

Guppy (Lebistes reticulatus) embryos pass through a distinct sequence of motor behaviors that leads to swimming capability during the course of their development. We have characterized these activities in order of appearance, with several corresponding morphological features, as belonging to the coil stage, tail-twitch stage, S-movement stage, and swimming stage. A primary feature of development was an increase in the amount of activity per unit of time over these four stages. The developmental pattern of motility was not interrupted by spinal transection until the onset of swimming, implying that supraspinal information is not required for the occurrence of the primitive behaviors that precede swimming. Elimination of swimming by spinal transection did not elicit a reversion to less complex activities, suggesting that once the cerebral control for swimming is developed, it represents a hardwired system not behaviorally reducible to antecedent components.

[Embryonic development of guppies in weightlessness]. / Embrional'noe razvitie guppi v nevesomosti.

The program of the Cosmos-1514 flight included an experiment the purpose of which was to study the effect of weightlessness on the embryonic development of the live-bearing guppy fish: three pregnant females were flown for 5 days. Prelaunch their embryos were at the stage of cerebral vesicle differentiation and somite formation; this implies that the basic stages of organogenesis developed in weightlessness. One female was fixed in Bouin's fluid two days postflight and the second fish was fixed nine days postflight. Fourteen days after flight the third female gave birth to 25 normal fry. Thereafter that fish was mated 6 times more, each time delivering normal offspring. In addition, the offspring of the second generation was normal. Histological analysis of the embryos that were developing in weightlessness revealed no abnormalities. It can be concluded that weightlessness produced no effect on the fish development, beginning with the stage of the axial complex formation.