The first cytogenetic report of fireflies (Coleoptera, Lampyridae) from Brazilian fauna

The first cytogenetic analysis of fireflies from Brazilian fauna was carried out in this work. The investigation of two species of the subfamily Lampyrinae, Aspisoma maculatum and Photinus sp. (aff. pyralis), showed the diploid number 2n = 19 and an X0 sex determination system in males. These observations are similar to those already described for all the Lampyrinae species previously studied. In contrast, Bicellonycha lividipennis (Photurinae) revealed the karyotype 2n = 16 + neoXY, which has not yet been registered for any firefly species. The neoXY sex determination system encountered in this species probably arose through fusion between an ancestral X sex chromosome, belonging to the X0 system, and an autosomal element. This event also reduced the diploid number from 2n = 19, which is more frequent in the family Lampyridae, to 2n = 18 in B. lividipennis. The analysis of meiotic cells showed that the neoXY sexual bivalent of B. lividipennis exhibited a prominent terminal chiasma, indicating that the sex chromosomes are not wholly differentiated and still retain a region of homology. A review of the cytogenetic data known for the family Lampyridae was also documented in this work, as well as a discussion on the main trends of chromosomal evolution that seem to have occurred in this group.

Sex determination studies in two species of teleost fish, Oreochromis niloticus and Leporinus elongatus

Genetic analyses of sex determination have identified sex chromosomes in many teleost fish species. However, there are several cases for which sex ratios do not fit perfectly with the expectations of heterogametic systems, suggesting the influence of either minor sex determining genes or environmental influences on the process of sex differentiation. The frequent absence of sex chromosome markers makes the identification of minor sex-determining genes very difficult. It is easier to test first the hypothesis of environmental sex determination (ESD) by studying the temperature effect, since temperature-dependent sex determination has been demonstrated to occur in several vertebrate groups including 1 fish species. To contribute to a better understanding of fish sex determination, we have tested the effects of high temperatures on sex ratios of Oreochromis niloticus, and have attempted to isolate sex chromosome molecular markers in Leporinus elongatus. Treatments of O. niloticus fry at 36 degrees C applied for 10 days and more, and starting 1 week after fertilization markedly increased the proportion of males, and progeny-testing these males confirmed that some of them are sex-reversed genetic females. Two non-coding sequences of L. elongatus Z and W chromosomes were cloned by genomic subtraction. They cross-hybridized with the genome of a close species without providing sex-specific patterns. A collection of L. elongates individuals was subjected to gonadal and chromosomal sexing, and DNA hybridization with both sequences. These analyses revealed 3 individuals having atypical W chromosomes. Interestingly, 2 of these were males having a ZW karyotype. We assume that these atypical sex chromosome arise by exchanges between Z and W chromosomes, and that a transition between female and male heterogamety is underway in this species.

Chromosomal evolution in Gekkonidae. I. Chromosome painting between Gekko and Hemidactylus species reveals phylogenetic relationships within the group

Geckos are a large group of lizards characterized by a rich variety of species, different modes of sex determination and diverse karyotypes. In spite of many unresolved questions on lizards' phylogeny and taxonomy, the karyotypes of most geckos have been studied by conventional cytogenetic methods only. We used flow-sorted chromosome-specific painting probes of Japanese gecko (Gekko japonicus), Mediterranean house gecko (Hemidactylus turcicus) and flat-tailed house gecko (Hemidactylus platyurus) to reveal homologous regions and to study karyotype evolution in seven gecko species (Gekko gecko, G. japonicus, G. ulikovskii, G. vittatus, Hemidactylus frenatus, H. platyurus and H. turcicus). Generally, the karyotypes of geckos were found to be conserved, but we revealed some characteristic rearrangements including both fissions and fusions in Hemidactylus. The karyotype of H. platyurus contained a heteromorphic pair in all female individuals, where one of the homologues had a terminal DAPI-negative and C-positive heterochromatic block that might indicate a putative sex chromosome. Among two male individuals studied, only one carried such a polymorphism, and the second one had none, suggesting a possible ZZ/ZW sex determination in some populations of this species. We found that all Gekko species have retained the putative ancestral karyotype, whilst the fission of the largest ancestral chromosome occurred in the ancestor of modern Hemidactylus species. Three common fissions occurred in the ancestor of Mediterranean house and flat-tailed house geckos, suggesting their sister group relationships. PCR-assisted mapping on flow-sorted chromosome libraries with conserved DMRT1 gene primers in G. japonicus indicates the localization of DMRT1 gene on chromosome 6.

Chromosome banding patterns of four species of bats, with special reference to a case of X-autosome translocation

The karyotypes of 4 species of bats, Artibeus lituratus (Phyllostomatidae), Pipistrellus pipistrellus (Vespertilionidae), Pteropus alecto and P. giganteus (Pteropodidae), were studied after several banding techniques. For A. lituratus, in which an X-autosome translocation was observed, an analysis of the replication pattern in the rearranged chromosome was also made after BrdU incorporation.

Cytogenetic studies in Diplopoda

The Diplopoda have received little attention from cytogeneticists owing mostly to technical difficulties in obtaining mitotic chromosomes, restricting the studies to meiosis and eventual spermatogonial metaphases, which limits the use of modern cytogenetical techniques. A literature search shows that only about 0.1% of all known species have been cytogenetically studied. There are 80,000 species estimated for this group, making it the 3rd. larger class in Arthropoda, after Insecta and Arachnida. The diploid chromosomal number in diplopods varies from 2n=8 to 2n=30 and the sex determination mechanism commonly found is XY/XX. In meiotic prophase, the bouquet formation and the diffuse state in pachytene are typical events. The few works performed on Brazilian fauna add up to 16 species, out of an estimated number of 2000 to 3000 species. The present review reports all the species of diplopods that have been cytogenetically studied so far, each with its chromosome number and sex determination system.

Cytogenetical aspects of testicular cells in economically important species of coreidae family (Heteroptera)

Some cytogenetical aspects of spermatozoa formation were studied in 9 Coreidae Brazilian species: Anasa bellator, Athaumastus haematicus, Chariesterus armatus, Dallacoris obscura, Dallacoris pictus, Leptoglossus gonagra, Leptoglossus zonatus, Sphictyrtus fasciatus, and Zicca annulata. Similarly to the other species described to date, all the species studied herein showed cystic spermatogenesis, a reddish membrane covering the testes, a X0 sex determining system, a pair of m-chromosomes, intersticial chiasmata in most autosomes, and autosomes dividing reductionally at first meiotic division and equationally in the second 1 while sex chromosomes, divide equationally and reductionally at first and second meiotic division, respectively. In addition, it was observed that the sex chromosome is heteropycnotic at prophase and that heteropycnotic chromosomal material is found in the nuclei at spermiogenesis. In the species studied, the diploid chromosome number ranged from 19 to 25. It was 19 in S. fasciatus (16A+2m+X0); 21 in A. bellator, A. haematicus, D. obscura, D. pictus, L. gonagra, and L. zonatus (18A+2m+X0); 23 in Z. annulata (20A+2m+X0); and 25 in C. armatus (22A+2m+X0). © 2007 The Japan Mendel Society.

Chromosomal similarities and differences among four Neotropical Elateridae (Conoderini and Pyrophorini) and other related species, with comments on the NOR patterns in Coleoptera

This work deals with the comparative cytogenetic analysis of four Neotropical Elateridae species and reviews the nucleolar organizer region (NOR) patterns on Coleoptera chromosomes, for the first time. The cytogenetic characterization of Conoderus malleatus (Conoderini), Pyrearinus candelarius, Pyrophorus divergens and Pyrophorus punctatissimus (Pyrophorini) was accomplished through the study of mitotic and meiotic cells submitted to standard (Giemsa) and differential staining [silver impregnation and GC-specific chromomycin A 3 (CMA 3) plus AT-specific 4′-6-diamidino-2-phenylindole (DAPI) fluorochromes]. The analysis of spermatogonial cells revealed the diploid numbers: 2n = 17 in C. malleatus and 2n = 15 in P. candelarius, P. divergens and P. punctatissimus. In these species, the X0 type sex-determination system and the acrocentric morphology of almost all chromosomes were observed. The study of meiotic cells of the four species revealed the occurrence of total synapsis between the autosomes, the presence of one terminal or interstitial chiasma in the majority of the bivalents, and the reductional behaviour and regular segregation of all chromosomes. Although the three Pyrophorini species demonstrated many similar karyotypical characteristics, there was one discrepancy, which was noted in the diplotene cells and concerns the number of bivalents with two chiasmata; P. candelarius only presented one bivalent, P. divergens showed two bivalents and P. punctatissimus exhibited up to four bivalents with two chiasmata. Testicular cells impregnated with silver nitrate demonstrated two terminal NORs located on the fourth autosomal pair of the Conoderini species and on the second autosomal pair of the three Pyrophorini representatives. Use of CMA 3/distamycin A (DA)/DAPI staining on the P. candelarius and P. punctatissimus chromosomes revealed that the CMA 3 labelled regions were coincident with the NORs. The main strategies of karyotypical differentiation that have occurred among the four Elateridae species and other related species, and the general trends of the NOR shifts during Coleoptera chromosomal evolution are discussed in this work. © 2007 The Authors.

Chromosomal Mapping of Repetitive DNAs in the Grasshopper Abracris flavolineata Reveal Possible Ancestry of the B Chromosome and H3 Histone Spreading

Supernumerary chromosomes (B chromosomes) occur in approximately 15% of eukaryote species. Although these chromosomes have been extensively studied, knowledge concerning their specific molecular composition is lacking in most cases. The accumulation of repetitive DNAs is one remarkable characteristic of B chromosomes, and the occurrence of distinct types of multigene families, satellite DNAs and some transposable elements have been reported. Here, we describe the organization of repetitive DNAs in the A complement and B chromosome system in the grasshopper species Abracris flavolineata using classical cytogenetic techniques and FISH analysis using probes for five multigene families, telomeric repeats and repetitive C0t-1 DNA fractions. The 18S rRNA and H3 histone multigene families are highly variable and well distributed in A. flavolineata chromosomes, which contrasts with the conservation of U snRNA genes and less variable distribution of 5S rDNA sequences. The H3 histone gene was an extensively distributed with clusters occurring in all chromosomes. Repetitive DNAs were concentrated in C-positive regions, including the pericentromeric region and small chromosomal arms, with some occurrence in C-negative regions, but abundance was low in the B chromosome. Finally, the first demonstration of the U2 snRNA gene in B chromosomes in A. flavolineata may shed light on its possible origin. These results provide new information regarding chromosomal variability for repetitive DNAs in grasshoppers and the specific molecular composition of B chromosomes. © 2013 Bueno et al.

Estudos citogenéticos em raias do gênero Patamotrygon (Chondrichthyes: Myliobatiformes: Potamotrygonidae) na bacia superior do rio Paraná

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

The karyotype of Alouatta fusca clamitans from Rio de Janeiro, Brazil: Evidence for a y-autosome translocation

Os cariótipos referentes a quatro machos de Alouatta fusca clamitans oriundos do Rio de Janeiro foram analisados através de técnicas de bandamento G, C e NOR. O número diplóide em todos os espécimes foi igual a 49, com a presença de três cromossomos não pareados. A comparação dos padrões de bandamento G com espécimes previamente descritos com 2n = 50 revelou a ocorrência de uma translocação do tipo Y-autossomo, modificando o sistema cromossômico de determinação sexual para o tipo múltiplo, X1X2Y/X1X1 X2X2. Os blocos de heterocromatina constitutiva se distribuíram na região pericentromérica de todos os cromossomos; segmentos intercalares e teloméricos foram visualizados em um par acrocêntrico e em outro submetacêntrico, respectivamente. As regiões organizadoras de nucléolo se localizaram no braço longo de dois pares de pequenos acrocêntricos.

Análise dos cromossomos sexuais de Characidium (Characiformes, Crenuchidae) por microdissecção e pintura cromossômica

Pós-graduação em Ciências Biológicas (Zoologia) - IBB

Identificação e caracterização de elementos retrotransponíveis da família Rex do genoma de espécies do genêro Leporinus SPIX, 1829 (Teleostei: Anostomidae)

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

High Diversity in CMA(3)/DAPI-Banding Patterns in Heteropterans

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Distribution of constitutive heterochromatin in species of triatomines with fragmentation of sex chromosomes X

Cytogenetic analyses of triatomines are considered to be important taxonomic tools. Thus, we analyzed the pattern of constitutive heterochromatin in 7 species of triatomine with fragmentation of the sex chromosome X, focusing on the cytotaxonomy of these triatomines. The species analyzed included Triatoma vitticeps, Triatoma melanocephala, Triatoma tibiamaculata, Triatoma protracta, Meccus pallidipennis, Panstrongylus megistus, and Panstrongylus lignarius. The seminiferous tubules of the adult males were subjected to C-banding. P. megistus and P. lignarius showed differences in chromosome number and disposition of constitutive heterochromatin, as only P. lignarius showed C-blocks in autosomes. C-banding can differentiate these species, since one of the sex chromosome (X) is heterochromatic in T. vitticeps. T. protracta showed C-blocks in both ends of all autosomes, T. tibiamaculata showed terminal C-dots in some autosomal pairs and M. pallidipennis did not show constitutive heterochromatin in autosomes. Thus, we confirmed the heterochromatic pattern of 7 species of insects and emphasized the importance of cytogenetic techniques for C-banding for taxonomy studies of the triatomines, which are important vectors of Chagas disease.

A step to the gigantic genome of the desert locust: chromosome sizes and repeated DNAs

The desert locust (Schistocerca gregaria) has been used as material for numerous cytogenetic studies. Its genome size is estimated to be 8.55 Gb of DNA comprised in 11 autosomes and the X chromosome. Its X0/XX sex chromosome determinism therefore results in females having 24 chromosomes whereas males have 23. Surprisingly, little is known about the DNA content of this locust's huge chromosomes. Here, we use the Feulgen Image Analysis Densitometry and C-banding techniques to respectively estimate the DNA quantity and heterochromatin content of each chromosome. We also identify three satellite DNAs using both restriction endonucleases and next-generation sequencing. We then use fluorescent in situ hybridization to determine the chromosomal location of these satellite DNAs as well as that of six tandem repeat DNA gene families. The combination of the results obtained in this work allows distinguishing between the different chromosomes not only by size, but also by the kind of repetitive DNAs that they contain. The recent publication of the draft genome of the migratory locust (Locusta migratoria), the largest animal genome hitherto sequenced, invites for sequencing even larger genomes. S. gregaria is a pest that causes high economic losses. It is thus among the primary candidates for genome sequencing. But this species genome is about 50 % larger than that of L. migratoria, and although next-generation sequencing currently allows sequencing large genomes, sequencing it would mean a greater challenge. The chromosome sizes and markers provided here should not only help planning the sequencing project and guide the assembly but would also facilitate assigning assembled linkage groups to actual chromosomes.