Complex sex chromosome system in Eigenmannia sp. (Pisces, Gymnotiformes)

Chromosomes of Eigenmannia sp. (7 males and 15 females) collected from the Tietê River in Botucatu (SP, Brazil) were examined from gill, kidney and testicular cells. The diploid chromosome number in males was 2n=31 and in females, 2n=32. In both sexes the number of chromosomal arms was 40. The difference in diploid number was due to the fusion of two acrocentrics. Mitotic and meiotic studies suggested that one of the fused acrocentrics was the Y chromosome. The sex-determining mechanism in Eigenmannia sp. could therefore be XX, AA in the female and X, \-YA A in the males. One of the males presented 2n=30 chromosomes due to the occurrence of another fusion of acrocentrics. C-banding analysis of the mitotic chromosomes revealed constitutive heterochromatin in the centromeric regions of all acrocentrics. However, small metacentrics were C-band negative. The YA chromosome is C-band negative except for a small amount of heterochromatin in the centromeric region. The nucleolar organizer region as identified by Ag-staining is present in the interstitial region of chromosome pair No. 10. © 1984 Dr W. Junk Publishers.

Single loci with major effects in fire ants : the effects of the complementary sex-determining locus and the green beard supergene on gene expression

Etant données la complexité et la redondance des réseaux de gènes influençant de nombreux phénotypes, l'étude des rares cas d'un locus unique ayant des effets importants sur de nombreux phénotypes peut fournir des informations cruciales sur l'évolution des traits complexes. Nous avons séquencé le génome de la fourmi de feu Solenopsis invicta pour étudier comment l'expression des gènes détermine les effets majeurs et étendus de deux loci uniques sur le phénotype. Le premier locus concerne la détermination du sexe par le modèle des allèles complémentaires. Ce locus est connu pour déterminer le sexe chez tous les hyménoptères mais n'a été caractérisé que chez les abeilles. Les hétérozygotes pour ce locus se développent en reines diploïdes (ou ouvrières stériles) alors que les homozygotes se développent en mâles diploïdes incapables de produire du sperme et les hémizygotes en mâles haploïdes fertiles. Nous avons comparé l'expression des gènes entre les reines et les deux types de mâles au stade pupe, ainsi que 1 et 11 jours après l'émergence. Nous avons trouvé un changement prononcé de l'expression des gènes chez les mâles diploïdes, passant de très proche de celle des reines au stade pupe à identique aux mâles haploïdes 11 jours après l'émergence. Cela signifie que les mâles diploïdes sont condamnés à être stériles parce que les effets après émergence du locus de détermination du sexe ne per¬mettent pas d'effacer les effets de la ploïdie sur l'expression des gènes pendant le stade pupe, quand la spermatogénèse prend place. Le second locus aux effets majeurs que nous avons étudié est le supergène dit "green beard", qui consiste en 616 gènes couvrant 55% d'un chromosome (13 Mb) et est caractérisé par une absence de recombinaison entre les deux variants du supergène : "Social B" et "Social b" (SB et Sb). Au travers de l'effet "green beard", par lequel les ouvrières avec le supergène Sb discriminent favorablement les reines qui partagent ce supergène de façon perceptible, le génotype des reines fondatrices au niveau de ce supergène détermine l'organisation de la colonie : soit elle contient une seule reine SB/SB, soit plusieurs reines SB/Sb. Nous avons montré que le chromosome Sb a évolué comme le chromosome Y, accumulant probablement des allèles favorables dans des colonies avec plusieurs reines mais défavorables dans des colonies avec une seule reine (cf. gènes sexuellement antagonistes), ainsi que des transposons et des séquences répéti¬tives. Nous avons également montré que le polymorphisme du supergène cause de grandes différences d'expression chez les ouvrières et particulièrement les reines mais pas chez les mâles. Pour comprendre comment le polymorphisme du supergène chez les reines peut affecter l'organisation de la colonie, nous avons comparé l'expression entre les génotypes SB/SB et SB/Sb chez des reines vierges (1 et 11 jours) et des reines matures. Nous avons montré que les reines SB/SB sur-régulent des gènes impliqués dans la reproduction, expli-quant pourquoi elle grandissent plus rapidement et peuvent fonder des colonies de façon indépendante, tandis que les reines SB/Sb (qui ne peuvent fonder une nouvelle colonie) sur-régulent des gènes de signalement chimique qui affectent l'organisation des colonies par l'effet "green beard". - Given the complexity and redundancy of the gene networks that underlie many pheno- types, the study of rare cases of a single locus having major effects on many phenotypes can give powerful insights into the evolution of complex traits. We sequenced the genome of Solenopsis invicta fire ants to study how gene expression mediates the widespread major effects of two single loci on phenotype. The first is the complementary sex-determining locus, which is known to exist in most Hymenoptera despite being characterized only for honeybees. Heterozygotes at this locus become diploid queens (or sterile workers), homozy¬gotes become aspermic diploid males, and hemizygotes become fertile haploid males. We compared gene expression between queens and both types of males in pupae and 1 and 11 days after eclosion. We found a pronounced shift in gene expression in diploid males, from being nearly identical to queens as pupae to identical to haploid males 11 days after eclosion. This means that diploid males are condemned to sterility because the overriding effects of the sex locus after eclosion cannot undo the ploidy effects on expression during the pupal stage, when spermatogenesis must be completed. The second locus with major ef¬fects that we studied was the so-called "green beard" supergene, which consists of 616 genes encompassing 55% of one chromosome (13 Mb), without recombination between the two variants "Social B" and "Social b" (SB and Sb) supergene. Through the green beard effect, i.e. workers with the Sb supergene discriminating in favor of queens who perceptibly share this supergene, the founding queen's genotype at the supergene determines colony organi¬zation: either headed by a single SB/SB queen or many SB/Sb queens. We show that the Sb chromosome evolved like a Y-chromosome, probably accumulating alleles beneficial in multi-queen colonies but disadvantageous in single-queen colonies (cf. sexually antagonistic genes), as well as transposons and repetitive sequences. We also show that the polymor¬phism of the supergene causes widespread expression differences in workers and especially queens but not in males. To understand how the polymorphism at the supergene in queen can transform colony organization, we compared the expression between SB/SB and SB/Sb virgin queens (1 and 11 days) and mother queens. We show that SB/SB queens up-regulate genes involved in reproduction, explaining why they mature faster and can found colonies independently, while SB/Sb queens (which cannot found colonies) up-regulate chemical signaling genes that can transform colonies through the green beard effect.

Sex allocation conflict in ants: when the queen rules.

Insect societies are paramount examples of cooperation, yet they also harbor internal conflicts whose resolution depends on the power of the opponents. The male-haploid, female-diploid sex-determining system of ants causes workers to be more related to sisters than to brothers, whereas queens are equally related to daughters and sons. Workers should thus allocate more resources to females than to males, while queens should favor an equal investment in each sex. Female-biased sex allocation and manipulation of the sex ratio during brood development suggest that workers prevail in many ant species. Here, we show that queens of Formica selysi strongly influenced colony sex allocation by biasing the sex ratio of their eggs. Most colonies specialized in the production of a single sex. Queens in female-specialist colonies laid a high proportion of diploid eggs, whereas queens in male-specialist colonies laid almost exclusively haploid eggs, which constrains worker manipulation. However, the change in sex ratio between the egg and pupae stages suggests that workers eliminated some male brood, and the population sex-investment ratio was between the queens' and workers' equilibria. Altogether, these data provide evidence for an ongoing conflict between queens and workers, with a prominent influence of queens as a result of their control of egg sex ratio.

Sex-chromosome evolution of Palearctic tree frogs in space and time

Sexual reproduction is nearly universal in eukaryotes and genetic determination of sex prevails among animals. The astonishing diversity of sex-determining systems and sex chromosomes is yet bewildering. Some taxonomic groups possess conserved and dimorphic sex chromosomes, involving a functional copy (e.g. mammals' X, birds' Z) and a degenerated copy (mammals' Y, birds' W), implying that sex- chromosomes are expected to decay. In contrast, others like amphibians, reptiles and fishes yet maintained undifferentiated sex chromosomes. Why such different evolutionary trajectories? In this thesis, we empirically test and characterize the main hypotheses proposed to prevent the genetic decay of sex chromosomes, namely occasional X-Y recombination and frequent sex-chromosome transitions, using the Palearctic radiation of Hyla tree frogs as a model system. We take a phylogeographic and phylogenetic approach to relate sex-chromosome recombination, differentiation, and transitions in a spatial and temporal framework. By reconstructing the recent evolutionary history of the widespread European tree frog H. arborea, we showed that sex chromosomes can recombine in males, preventing their differentiation, a situation that potentially evolves rapidly. At the scale of the entire radiation, X-Y recombination combines with frequent transitions to prevent sex-chromosome degeneration in Hyla: we traced several turnovers of sex-determining system within the last 10My. These rapid changes seem less random than usually assumed: we gathered evidences that one chromosome pair is a sex expert, carrying genes with key role in animal sex determination, and which probably specialized through frequent reuse as a sex chromosome in Hyla and other amphibians. Finally, we took advantage of secondary contact zones between closely-related Hyla lineages to evaluate the consequences of sex chromosome homomorphy on the genetics of speciation. In comparison with other systems, the evolution of sex chromosomes in Hyla emphasized the existence of consistent evolutionary patterns within the chaotic diversity of flexibility of cold-blooded vertebrates' sex-determining systems, and provides insights into the evolution of recombination. Beyond sex-chromosome evolution, this work also significantly contributed to speciation, phylogeography and applied conservation research. -- La reproduction sexuée est quasi-universelle chez les eucaryotes et le sexe est le plus souvent déterminé génétiquement au sein du règne animal. L'incroyable diversité des systèmes de reproduction et des chromosomes sexuels est particulièrement étonnante. Certains groupes taxonomiques possèdent des chromosomes sexuels dimorphiques et très conservés, avec une copie entièrement fonctionnelle (ex : le X des mammifères, le Z des oiseaux) et une copie dégénérée (ex : le Y des mammifères, le W des oiseaux), suggérant que les chromosomes sexuels sont voués à se détériorer. Cependant les chromosomes sexuels d'autres groupes tels que les amphibiens, les reptiles et les poissons sont pour la plupart indifférenciés. Comment expliquer des trajectoires évolutives si différentes? Au cours de cette thèse, nous avons étudié empiriquement les processus évolutifs pouvant maintenir les chromosomes sexuels intacts, à savoir la recombinaison X-Y occasionnel ainsi que les substitutions fréquentes de chromosomes sexuels, en utilisant les rainettes Paléarctiques du genre Hyla comme modèle d'étude. Nous avons adopté une approche phylogéographique et phylogénétique pour appréhender les événements de recombinaison, de différenciation et de transitions de chromosomes sexuels dans un contexte spatio-temporel. En retraçant l'histoire évolutive récente de la rainette verte H. arborea, nous avons mis en évidence que les chromosomes sexuels pouvaient recombiner chez les mâles, empêchant ainsi leur différenciation, et que ce processus avait le potentiel d'évoluer très rapidement. A l'échelle plus globale de la radiation, il apparait que les phénomènes de recombinaison X-Y soient également accompagnés de substitutions de chromosomes sexuels, et participent de concert au maintien de chromosomes sexuels intacts dans les populations: le système de détermination du sexe des rainettes a changé plusieurs fois au cours des 10 derniers millions d'années. Ces transitions fréquentes ne semblent pas aléatoires: nous avons identifié une paire de chromosomes qui présente des caractéristiques présageant d'une spécialisation dans le déterminisme du sexe (notamment car elle possède des gènes importants pour cette fonction), et qui a été réutilisée plusieurs fois comme tel chez les rainettes ainsi que d'autres amphibiens. Enfin, nous avons étudié l'hybridation entre différentes espèces dans leurs zones de contact, afin d'évaluer si l'absence de différenciation entre X et Y jouaient un rôle dans les processus génétiques de spéciation. Outre son intérêt pour la compréhension de l'évolution des chromosomes sexuels, ce travail contribue de manière significative à d'autres domaines de recherche tels que la spéciation, la phylogéographie, ainsi que la biologie de la conservation.

Independent fusions and recent origins of sex chromosomes in the evolution and diversification of glass knife fishes (Eigenmannia)

The genus Eigenmannia comprises several species groups that display a surprising variety of diploid chromosome numbers and sex-determining systems. In this study, hypotheses regarding phylogenetic relationships and karyotype evolution were investigated using a combination of molecular and cytogenetic methods. Phylogenetic relationships were analyzed for 11 cytotypes based on sequences from five mitochondrial DNA regions. Parsimony-based character mapping of sex chromosomes confirms previous suggestions of multiple origins of sex chromosomes. Molecular cytogenetic analyses involved chromosome painting using probes derived from whole sex chromosomes from two taxa that were hybridized to metaphases of their respective sister cytotypes. These analyses showed that a multiple XY system evolved recently (<7 mya) by fusion. Furthermore, one of the chromosomes that fused to form the neo-Y chromosome is fused independently to another chromosome in the sister cytotype. This may constitute an efficient post-mating barrier and might imply a direct function of sex chromosomes in the speciation processes in Eigenmannia. The other chromosomal sex-determination system investigated is shown to have differentiated by an accumulation of heterochromatin on the X chromosome. This has occurred in the past 0.6 my, and is the most recent chromosomal sex-determining system described to date. These results show that the evolution of sex-determining systems can proceed very rapidly. Heredity (2011) 106, 391-400; doi:10.1038/hdy.2010.82; published online 23 June 2010

Origin and differentiation of a sex chromosome system in Parodon hilarii (Pisces, Parodontidae). Satellite DNA, G- and C-banding

A satellite DNA sequence of Parodon hilarii ( named pPh2004) was isolated, cloned and sequenced. This satellite DNA is composed of 200 bp, 60% AT rich. In situ hybridization ( FISH) results revealed that the satellite DNA pPh2004 is located in the terminal regions of several chromosomes, forming highly evident blocks in some and punctual marks in others. The comparison between the FISH and C-banding results showed that the location of this satellite DNA coincides with that of most terminal heterochromatins. However, some regions are only marked by FISH whereas other regions are only marked by C-banding. The possible existence of more than one satellite DNA family could explain these partial differences. The in situ hybridization with the satellite DNA and the G- and C-bandings confirmed the presence of a sex chromosome system of the ZZ/ZW type in P. hilarii, as well as the correct identification of the Z chromosome in the karyotype. This chromosome displays a segment of terminal heterochromatin in the long arm, similar to the segment observed in the short arm of the W chromosome, also showing a G- banding pattern similar to that of the short arm and part of the long arm of the W chromosome. A hypothesis on the origin of the W chromosome from an ancestral chromosome similar to the Z chromosome is presented.

Unusual occurrence of a ZZ/ZW sex-chromosome system and supernumerary chromosomes in Characidium cf. fasciatum (Pisces, Characiformes, Characidiinae)

Individuals of two populations of the fish Characidium cf. fasciatum were cytogenetically studied and showed a basic diploid number of 50 chromosomes. Some fishes were found to have 51 to 54 chromosomes due to the presence of one to four small subtelocentric/acrocentric supernumerary chromosomes. When analyzed by conventional Giemsa staining, male and female specimens of C. cf. fasciatum from the Quinta stream and Pardo River presented the same basic karyotypic macro- and microstructure, consisting of 32 metacentric and 18 submetacentric chromosomes.Ag-NORs were terminally located on the long arms of two submetacentric chromosome pairs. Constitutive heterochromatin was identified by C-banding as small pericentromeric blocks in the majority of the chromosomes, and B-chromosomes were found to be heterochromatic. The occurrence of one totally heterochromatic submetacentric chromosome restricted to females and considered as an unusual feature in fish karyotypes led to the identification of a ZZ/ZW sex-chromosome system. The implications of chromosomic differentiation observed in the genus Characidium are discussed.

Karyotypic relationships among the tribes of Hypostominae (Siluriformes : Loricariidae) with description of XO sex chromosome system in a neotropical fish species

The family Loricariidae, with about 683 species, is one the largest fish families in the world. The subfamily Hypostominae was recently reviewed and is now divided in five tribes. With the main objective of contributing to a better understanding of the relationships of the members of the subfamily Hypostominae, cytogenetic analyses were conducted in seven species (three Hypostomini, three Pterygoplichthini and two Ancistrini) from Brazil and Venezuela. In Pterygoplichthini, all species show 2n = 52 chromosomes. In Hypostomini Hypostomus ancistroides has 2n = 68, H. regani 2n = 72 and Hypostomus goyazensis 2n = 72 chromosomes. In Ancistrini Ancistrus n. sp. 1 has 2n = 39/40 with a sex chromosome system of the type XX/X0, which is a novelty for neotropical fishes, and Ancistrus n. sp. 2 has 2n = 52 chromosomes. Six species have single Ag-NORs and two multiple Ag-NORs. The possible cytogenetic relationships among the species of Hypostominae are discussed.

XX : XY sex chromosome system with X heterochromatinization: an early stage of sex chromosome differentiation in the Neotropic electric eel Eigenmannia virescens

An early stage of sex chromosome differentiation is reported to occur in the electric eel Eigenmannia virescens (Pisces, Sternopygidae) from populations of two tributaries of the Parana river system (Brazil). Cytogenetic studies carried out in the two populations showed that the Mogi-Guacu population is characterized by 2n = 38 chromosomes and undifferentiated sex chromosomes and the Tiete population presents 2n = 38 both for males and females and an XX:XY sex chromosome system. The X-chromosome is acrocentric, easily recognized by the presence of a conspicuous heterochromatin block in its distal portion; the Y-chromosome is probably one of the medium sized acrocentrics present in the male karyotype. BrdU induced R-bands of the two populations did not reveal any difference in the euchromatic regions of the chromosomes. AluI and HaeIII restriction enzyme digestion patterns and chromomycin A3 staining of the X-chromosome are presented. The possible role of heterochromatinization in the evolution of sex chromosomes in fish is discussed.

A novel ZZ/ZW sex chromosome system for the genus Leporinus (Pisces, Anostomidae, Characiformes)

A wide range of sex chromosome mechanisms, including simple and multiple chromosome systems is characteristic of fishes. The Leporinus genus represent a good model to study sex chromosome mechanisms, because an unambiguous ZZ/ZW sex chromosome system was previously described for seven species, while the remaining studied species of the genus do not show differentiated sex chromosomes. The occurrence of sex chromosomes in Leporinus trifasciatus and Leporinus sp2 from the Araguaia river, Amazon basin, Brazil, was here investigated. ZZ/ ZW sex chromosomes were detected for both species. The Z and W chromosome morphology of L. trifasciatus is the same as described for other species of the genus Leporinus. However, the Z and W chromosomes of L. sp2 were quite different in their morphology and banding pattern suggesting that the ZW system of this species have originated independently from the ZW system previously described for other Leporinus.

The chromosomes and the sex determining mechanism of Scaphura nigra (Orthoptera, Ensifera, Tettigoniidae, Phaneropterinae).

Scaphum nigra has a uniquechromosomecomplement among approximately 100 species studied so far belonging to the subfamily Phaneropterinae. It is formed by 2n ([male]) = 26 and a FN = 29 and derived from the ancestral karyotype of the group 2n ([male]) = 31, FN = 31, by means of two centric fusions and one tandem fusion. The first between the X chromosome and a medium-sized autosome giving rise to a neo-XY sex chromosome mechanism of recent origin, and the second between two acrocentric ones, the bigger and a medium size, that gave rise to a large submetacentric element whose length is very uncommon in the subfamily. This process has created a bimodal karyotype that contrasts with the majority in this group, whose chromosomes usually can be arranged in a decreasing order of size. A third rearrangement incorporating the chromatin of a medium-sized autosome to the bigger one, explains the reduction observed in the number of chromosomes and the enlarged size of the submetacentric elements. These features demonstrate the effectiveness of chromosome number, their morphology and the change of the sex mechanism as useful tools for taxonomy.

Karyotype description and evidence of multiple sex chromosome system X1X1X2X2/X1X2Y in Potamotrygon aff. motoro and P. falkneri (Chondrichthyes: Potamotrygonidae) in the upper Paraná River basin, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Description of a ZZ/ZW sex chromosome system in Thoracocharax cf. stellatus (Teleostei, Characiformes, Gasteropelecidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Evidence of an XX/XY sex chromosome system in the fish Dormitator maculatus (Teleostei, Eleotrididae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)