Ana Luiza de Brito Portela-Castro

New occurrence of microchromosomes B in Moenkhausia sanctaefilomenae (Pisces, Characidae) from the Paraná River of Brazil: analysis of the synaptonemal complex

The Moenkhausia sanctaefilomenae specimens showed a karyotype consisting of 2n = 50 chromosomes with 12 metacentrics, 36 submetacentrics and two subtelocentrics. In addition to the basic karyotype, all the males specimens have cells ranging from zero to two B microchromosomes in mitotic metaphases. These chromosomes were not observed in the female specimens. C-band analysis showed a distribution pattern of characteristic heterochromatin with interstitial and centromeric blocks. However, the B chromosomes were faintly stained with C-banding and were not fluorescent with CMA3 staining. The meiotic studies showed the formation of bivalents in metaphase I and in pachytene under an optical microscope. Through synaptonemal complex analysis with an electron microscope, the pachytene showed 25 bivalents completely paired and a small bivalent corresponding to the B chromosomes. In the same preparation, one of the B chromosomes was observed in a univalent form. On the basis of pairing behavior and morphology it is assumed that B chromosomes of M. sanctaefilomenae show homology between them and their evolutionary aspects are discussed.

Chromosomal Polymorphism in Rineloricaria Lanceolata Gunther, 1868 (Loricariidae: Loricariinae) of the Paraguay Basin (Mato Grosso do Sul, Brazil): Evidence of Fusions and Their Consequences in the Population

Rineloricaria is the most species-rich genus of the Loricariinae (armored catfish) with 65 valid species. However, the karyotype structure is known only for eight species in this group. This study provides cytogenetic data for Rineloricaria lanceolata collected from the upper Paraguay basin (Mato Grosso do Sul). The specimens revealed extensive chromosomal polymorphism constituting 10 karyotypes, which differed in the diploid number (48 to 45 chromosomes) and fundamental number (FN) between 52 and 55. Three types of chromosome variants were observed: a medium-sized submetacentric, a large submetacentric, and a small acrocentric form. Internal telomere sequences were demonstrated by a telomeric (TTAGGG)n probe in submetacentric chromosome variants, suggesting Robertsonian and tandem fusions. Considering the karyotype 2n=48 (4m+2st+42a, FN=54) as the starting point for this polymorphism, these rearrangements contributed to the reduction in diploid number (48-45). Furthermore, a remarkable polymorphism of 18S rDNA resulted in three nucleolus organizer region phenotypes (I, II, and III) with variable frequencies. Interestingly, this polymorphism has remained in the population through interbreeding between specimens, resulting in different viable combinations. The data obtained confirm that diversification/karyotype evolution in Rineloricaria was marked by numerous chromosomal rearrangements which appear to be well tolerated in the panmitic population.

Chromosomal polymorphism and speciation in Laetacara cf. dorsigera (Teleostei, Perciformes, Cichlidae) from the river Paraná PR Brazil

Abstract Four sympatric and syntopic karyotype forms of Laetacara cf. dorsigera, centric fusion-originated were analyzed. Specimens with 2n=46, 2n=45, 2n=44 and 2n=43 chromosomes and differences with regard to the number of metacentric chromosomes, inversely proportional to their diploid number, were reported. Karyotypic formulae of the four cytotypes were: cytotype A=2M + 44A; cytotype B=3M + 42A; cytotype C=4M + 40A and cytotype D=5M + 38A and fundamental number constant (NF=48). Two centric fusion events are suggested: a) Robertsonian trans-location, which originated the first and second metacentric chromosomal pairs; b) isochromosome formed by the fusion of acrocentric homologous chromosomes in cytotypes with odd chromosomes. Genetic balance in the four karyotype forms of Laetacara cf. dorsigera seemed to have been determinant to their reproductive isolation. The presence of individuals with different karyotypic structures living in sympatry and syntopy and the absence of “intermediate individuals” (recombinants), that would have been expected in a panmitic population, permit one to conclude that probably form 4 cryptic species exist in the population.

Cytogenetics of the Thorny Catfish Trachydoras paraguayensis (Eigenmann & Ward, 1907), (Siluriformes, Doradidae): Evidence of Pericentric Inversions and Chromosomal Fusion.

The karyotype and chromosomal characteristics of Trachydorasparaguayensis, a representative of the South American catfish family Doradidae, were analyzed by conventional (Giemsa staining, silver staining, C-banding) and molecular (FISH with rDNA and telomeric probes) cytogenetic techniques. The diploid chromosome number was 2n = 56, with 36 metacentric, 16 submetacentric, and 4 subtelocentric chromosomes in both sexes; however, a remarkable heteromorphism in pair 22 (submetacentric and metacentric elements) was detected in 6 individuals. Compared to other representatives of Doradidae which mostly have 58 chromosomes, the karyotype of T. paraguayensis suggests a reduction in 2n due to chromosomal fusion, as could be deduced from the presence of an interstitial telomere sequence in the submetacentric pair 19. Pale heterochromatic blocks were present in the terminal regions of some chromosomes, very similar to other species of Doradidae. The interstitial position of the NORs observed in the karyotype of T. paraguayensis differs from those reported for most Doradidae species, indicating that it is a derived character. FISH with 5S rDNA revealed 2 interstitial fluorescent signals in the submetacentric pair 22, and the polymorphism of these sites likely resulted from a pericentric inversion.

Chromosomal characteristics of rDNA in a conserved karyotype of two Sternopygus macrurus (Gymnotiformes: Sternopygidae) populations from upper Paraná River basin

Abstract Karyotype and chromosomal characteristics of both minor and major rDNA of Sternopygus macrurus, a weakly electric South American fish, from two populations of the upper Paraná River basin, were investigated using conventional (Giemsa staining, silver staining, C-banding and base-specific fluorochromes) and molecular (fluorescent in situ hybridization (FISH) with 5S and 18S rDNA probes) cytogenetic techniques. Diploid chromosome number was invariably 2n = 46 and karyotype composed of 23 pairs of biarmed chromosomes (28m+18sm). The nucleolus organizer regions (NORs) were located in the secondary constriction of the p arm of pair No. 2; this site corresponded with CMA3 positive as well as with 18S rDNA signals, respectively. This 18S rDNA cluster was not syntenic to the 5S rDNA sites located at pairs Nos. 1, 5 and 15. The karyotypes and other chromosomal characteristics of individuals from the two populations in the upper Paraná River basin were identical. The karyotype differences among individuals identified as S. macrurus from Paraná River and the São Francisco and Amazon River basins, respectively, may indicate that these taxa might represent distinct species.

Histological and electron microscopy observations on the testis and spermatogenesis of the butterfly Dione juno (Cramer, 1779) and Agraulis vanillae (Linnaeus, 1758) (Lepidoptera: Nymphalidae)

Lepidopteran species present an interesting case of sperm polymorphism and testicular fusion. The study of these features are of great importance in understanding the reproductive biology of these insects, especially in the case of those considered pests. Dione juno and Agraulis vanillae stand out as the most important pests of passion fruit (Passiflora sp.) crops in Brazil. Therefore, the objective of the present study was to characterize the testes and germ cells of Dione juno and Agraulis vanillae at different life stages, using light microscopy and scanning and transmission electron microscopy, to understand the maturation mechanisms of the male gametes in these species. The study showed that the larvae of both species have a pair of brown kidney-shaped testes, covered by epithelial cells which divide the organ into four follicles. The testes are full of spermatogonia which begin to differentiate in the third larval instar. In the fifth larval instar, spermatozoa can be observed. When they enter the prepupal stage the testes begin a fusion process that is completed in the adult insects, where they present as spherical organs divided into eight follicles, containing all the cells of the germ line. Spermatogenesis occurs centripetally, and in both species, sperm dimorphism is observed, where two different types of spermatozoa are formed, eupyrene (nucleated) and apyrene (anucleate), which differ in morphology and function. Apart from contributing to scientific basic research on the reproductive biology of these insects, the present study provides important data that can aid in research on the physiology, systematics, and control of these species.

Hidden Diversity in the Populations of the Armored Catfish Ancistrus Kner, 1854 (Loricariidae, Hypostominae) from the Paraná River Basin Revealed by Molecular and Cytogenetic Data

Only one species of armored catfish, Ancistrus cirrhosus Valenciennes 1836, has been historically described in the basin of the Paraná River, from Misiones (Argentina). However, the ample variation found in the morphology and coloration of the populations sampled in the tributaries of the Brazilian state of Paraná makes it difficult to establish the real taxonomic status and evolutionary history of the Ancistrus specimens, suggesting that A. cirrhosus is not the only species found in this basin. By combining data on mitochondrial DNA (COI gene) and chromosomal markers from different Ancistrus populations, totaling 144 specimens, in the tributaries of the Paraná, and specimens from Misiones (type-locality of A. cirrhosus), we detected five distinct evolutionary lineages. All the specimens were 2n = 50, but had four distinct karyotype formulae. The results of the Generalized Mixed Yule Coalescent (GYMC) and the genetic distances (uncorrected P-values) between lineages ranged from 3 to 5%. Clusters of 18S rDNA were observed in a single chromosome pair in seven populations of Ancistrus, but at different positions, in some cases, in synteny with the 5S rDNA sites. Multiple 5S sites were observed in all populations. Overall, the cytogenetic data reinforce the genetic evidence of the diversification of lineages, and indicate the existence of candidate species in the study region. The evidence indicates that at least four candidate species of the Ancistrus may coexist in the Paraná basin besides A. cirrhosus. Overall, our results provide a comprehensive scenario for the genetic variation among Ancistrus populations and reinforce the conclusion that the true diversity of the freshwater fish of the Neotropical regions has been underestimated.