Cycloactive, aneugenic and clastogenic effects of Paracoccidioides brasiliensis exoantigen in human lymphocyte cultures

The in vitro effect of Paracoccidioides brasiliensis exoantigen on the human lymphocytes cell cycle and chromosomes was studied. Human peripheral blood lymphocyte cultures from ten healthy, white, non-smoking, non-related adult males (mean age 31·3 ± 8·2 years) were studied. Blood cultures were treated with three exoantigen concentrations (0·25, 2·50 and 10·00 μg ml -1). At least 1000 metaphases were analysed at each concentration, for evaluation of numerical and structural chromosome aberrations (cA) and 30 000 for mitotic index (MI). Among the treated cultures, statistically significant differences in the frequencies of MI and cA were not observed. Nevertheless, when compared with control cultures, they all showed a significantly lower frequency of MI and higher frequency of cA. It is suggested that the detected alterations were caused by the exoantigen, its fractions or its metabolites. © 1996 Informa UK Ltd All rights reserved.

Synapsis in supernumerary chromosomes of Prochilodus lineatus (Teleostei: Prochilodontidae)

Meiotic analysis performed on a sample of 10 specimens of Prochilodus lineatus revealed continuous filaments of different sizes stained by AgNO3, corresponding to the bivalent of the normal complement. Small supernumerary chromosomes were observed as isolated and well stained bodies, scattered among the other elements. Synaptonemal complex studies have shown that the beginning of chromosome pairing process in P. lineatus usually occurs from the telomeres to the pericentromeric region. At the end of the pachytene 27 bivalents are perfectly paired and the small supernumerary chromosomes of this species are seen as bivalents, trivalents, or tetravalents. The central region of these small chromosomes show a trick staining when they formed bivalents or tetravalents. This portion seems to correspond to the pericentromeric region of the regular chromosomes with the heterochromatic characteristic of B chromosomes.

Nucleotide sequence of 5S rDNA and localization of the ribosomal RNA genes to metaphase chromosomes of the Tilapiine cichlid fish, Oreochromis niloticus

In this study, we report the cloning and nucleotide sequence of PCR-generated 5S rDNA from the Tilapiine cichlid fish, Oreochromis niloticus. Two types of 5S rDNA were detected that differed by insertions and/or deletions and base substitutions within the non-transcribed spacer (NTS). Two 5S rDNA loci were observed by fluorescent in situ hybridization (FISH) in metaphase spreads of tilapia chromosomes. FISH using an 18S rDNA probe and silver nitrate sequential staining of 5S-FISH slides showed three 18S rDNA loci that are not syntenic to the 5S rDNA loci.

Cytogenetic analysis of Epicauta atomaria (Meloidae) and Palembus dermestoides (Tenebrionidae) with Xyp sex determination system using standard staining, C-bands, NOR and synaptonemal complex microspreading techniques

The mitotic and meiotic chromosomes of the beetles Epicauta atomaria (Meloidae) and Palembus dermestoides (Tenebrionidae) were analysed using standard staining, C-banding and silver impregnation techniques. We determine the diploid and haploid chromosome numbers, the sex determination system and describe the chromosomal morphology, the C-banding pattern and the chromosome(s) bearing NORs (nucleolar organizer regions). Both species shown 2n = 20 chromosomes, the chromosomal meioformula 9 + Xyp, and regular chromosome segregation during anaphases I and II. The chromosomes of E. atomaria are basically metacentric or submetacentric and P. dermestoides chromosomes are submetacentric or subtelocentric. In both beetles the constitutive heterochromatin is located in the pericentromeric region in all autosomes and in the Xp chromosome; additional C-bands were observed in telomeric region of the short arm in some autosomes in P. dermestoides. The yp chromosome did not show typical C-bands in these species. As for the synaptonemal complex, the nucleolar material is associated to the 7th bivalent in E. atomaria and 3rd and 7th bivalents in P. dermestoides. Strong silver impregnated material was observed in association with Xyp in light and electron microscopy preparations in these species and this material was interpreted to be related to nucleolar material.

Morphologically differentiated sex chromosomes in neotropical freshwater fish

A general survey of the occurrence of morphologically differentiated sex chromosomes in the neotropical freshwater fishes is presented. The total number of 32 occurrences involving simple XX-XY and ZZ-ZW, and multiple X1X2Y, XY1Y2 and ZW1W2 sex chromosome systems is described, with comments on the aspects of sex chromosome evolution in this fish fauna. The occurrence of different sex chromosome systems in related species of the same genus, or in different populations of the same nominal species, involving male and sometimes female heterogamety, and differences in the molecular composition of sex-linked heterochromatin, are considered as indicative of the early stage of sex chromosomes evolution in fish.

Genitalia, acoustics and chromosomes of Urogryllus toledopizai (Orthoptera: Gryllidae)

Morphological structures of male and female genitalia and the pars stridens of a population of Urogryllus toledopizai Mello (1988) as well as the karyotype of the species are illustrated. The characteristics of song signals and associated behavior are described and information on the seasonality of the species is provided. Disagreements with the results published by other authors are discussed.

Molecular characterization and chromosomal localization of two families of satellite DNA in Prochilodus lineatus (Pisces, Prochilodontidae), a species with B chromosomes

Prochilodus lineatus, an abundant species in the Mogi-Guaçu river basin, represents a large part of the region's fishing potential. Karyotypic analyses based on classic cytogenetic techniques have revealed the presence of 54 metasubmetacentric type chromosomes, together with the occurrence of small supernumerary chromosomes with intra and interindividual variations. This paper describes the genomic organization of two families of satellite DNA in the P. lineatus genome. The chromosomal localization these two repetitive DNA families through fluorescence in situ hybridization (FISH) demonstrated that the SATH1 satellite DNA family, composed of approximately 900 bp, was located in the pericentromeric region of a group of chromosomes of the standard complement, as well as on all the B chromosomes. The SATH2 satellite family has a monomeric unit of 441 bp and was located in the pericentromeric regions of some chromosomes of the standard complement, but was absent in the B chromosomes. Double FISH analyses showed that these two families participate jointly in the pericentromeric organization of several chromosomes of this species. The data obtained in this study support the hypothesis that the B chromosomes derive from chromosomes of the standard complement, which are carriers of the SATH1 satellite DNA.

Cytogenetic study of holocentric chromosomes in three species of triatomines (Heteroptera, Reduvidae)

In the present work, spermatogenesis was analyzed in 3 species of the genus Triatoma (T. platensis, T. proctata, T. tibiamaculata). Lacto-acetic orcein staining was used in order to investigate chromosomal meiotic behavior of these species. It allowed the identification of the T. tibiamaculata karyotype (20, X 1X 2Y), the observation that in T. protacta doesn't occur late migration of sexual chromosomes and corroborated knowledgments about holocentric chromosome nature.

Cytogenetic analysis of A-, B-chromosomes and ZZ/ZW sex chromosomes of Characidium gomesi (Teleostei, Characiformes, Crenuchidae)

Different cytogenetic techniques were used to analyze the chromosomes of Characidium gomesi with the main objective of comparing the base composition of ZZ/ZW sex-chromosomes, B-chromosomes and the heterochromatin of A-chromosomes. The results of digestion of chromosomes with AluI restriction endonuclease (RE), silver and CMA3 staining, C-banding and fluorescence in situ hybridization (FISH) with the 18S rDNA probe suggested the existence of compositional differences between the heterochromatin of ZZ/ZW sex-chromosomes, A- and B-chromosomes, and indicated the presence of different numbers and morphology of B-chromosomes in the samples of this population.

Evaluating the karyotypic diversity in species of Hyla (Anura, Hylidae) with 2n = 30 chromosomes based on the analysis of ten species

Ten species of Hyla with 2n = 30 from Brazilian fauna were analysed cytogenetically. Hyla minuta is the unique presenting all bi-armed metacentric or submetacentric chromosomes in the karyotype, therefore, with the highest FN = 60. The remaining species have a variable number of uni-armed telocentric or subtelocentric chromosomes: H. cruzi, H. elianeae, and H. rubicundula with three pairs (FN = 54), H. berthalutzae, H. elegans, H. microps, and H. nana with four pairs (FN = 52), and H. nahdereri and H. sanborni with five pairs (FN = 50). The uni-armed elements are among pairs 5, 6, 7, 11, 14, and 15, which also appeared with metacentric or submetacentric morphology. The remaining chromosome pairs 1, 2, 3, 4, 8, 9, 10, 12, and 13 were never found to be telocentric or subtelocentric. AgNOR patterns are species-specific, the majority of the species exhibiting a single pair with AgNORs, with the exception of H. elegans and H. nana with more than one chromosome pair bearing this cytological marker. C banding was obtained in H. berthalutzae, H. cruzi, H. elegans, H. elianeae, H. microps, H. minuta, H. nahdereri, and H. nana, which showed positively stained centromeric heterochromatin. Our analysis confirms the great karyotypic diversity in the species of Hyla with 2n = 30, with no species sharing identical karyotypes.

Localization of rDNA sites in holocentric chromosomes of three species of triatomines (Heteroptera, Triatominae).

Chromatin organization in the holocentric chromosomes of three triatomines species was cytologically studied by fluorescent in situ hybridization with a 45S rDNA probe of Drosophila melanogaster to localize ribosomal genes. In Triatoma tibiamaculata, metaphases I showed telomeric highlights in a single, larger bivalent. In T. protacta, hybridization was detected in one of the telomeres of an autosomal chromosome. In T. platensis, there were highlights in a single, smaller chromosome (X chromosome). The results obtained did not agree with the expected localization of rDNA genes in the sex chromosomes of triatomines, as demonstrated by silver impregnation, and suggest that the chromosome reorganization that occurred in this group during evolution may be a more important mechanism involved in rDNA distribution.

Chromosomes of Crossopriza lyoni (Blackwall 1867), intraindividual numerical chromosome variation in Physocyclus globosus (Taczanowski 1874), and the distribution pattern of NORs (Araneomorphae, Haplogynae, Pholcidae)

Pholcidae (Haplogynae) encompasses 967 described species, of which only 14 have been cytogenetic analyzed. Several chromosomal features have already been described including presence of meta- and sub-metacentric chromosomes and sex determination chromosome system (SDCS) of the X, X1X2Y, and X1X2 types, which contrast with the telo- and acrocentric chromosomes and SDCS of the X1X2 type typical of entelegyne spiders. To obtain further cytogenetic information for the family, we examined two pholcid species, Crossopriza lyoni (Blackwall 1867) and Physocyclus globosus (Taczanowski 1874) using both conventional staining and silver staining techniques. Crossopriza lyoni exhibited 2n = 23 = 22 + X in males and 2n = 24 = 22 + XX in females, while P. globosus showed 2n = 15 = 14 + X and 4n = 30 = 28 + 2X, both in male adults, 2n = 16 = 14 + XX in female adults and embryos, and 2n = 15 = 14 + X in male embryos. Both species revealed predominately metacentric and submetacentric chromosomes and a SDCS of the X/XX type. The cytogenetic data obtained in this work and those already recorded for C. lyoni indicate interpopulational and intraspecific numerical chromosome variation, suggesting the presence of chromosomal races or cytotypes in this species. The intraindividual numerical chromosome variation observed in male adult specimens of P. globosus may be explained by the presence of cytoplasmatic bridges between germ cells. The use of the silver staining technique to reveal the nucleolar organizer region (NOR) showed that chromosome pairs 4 and 6 and the X chromosome in C. lyoni are telomeric NOR-bearers, and that the chromosome pair 2 in P. globosus possesses a proximal NOR in the long arm.

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.

Differential characterization of holocentric chromosomes in triatomines (Heteroptera, Triatominae) using different staining techniques and fluorescent in situ hybridization

A comparative study of holocentric chromosomes in the triatomine species Panstrongylus megistus, Rhodnius pallescens and Triatoma infestans was carried out in order to characterize heterochromatin, rDNA active sites and nucleolar proteins. Cytological preparations of seminiferous tubules were stained by silver impregnation, C banding, fluorochromes CMA 3/DA and DAPI/DA, and fluorescent in situ hybridization (FISH) with Drosophila melanogaster 28S rDNA probe. Our results showed interesting aspects of the organization of chromatin and chromosomes in the meiotic cells of these insects. In R. pallescens, sex chromosomes (X, Y) were distinct from autosomes, when submitted to silver impregnation, C banding, CMA 3 staining, and FISH, confirming that these chromosomes bear nucleolar organizer regions (NORs). In P. megistus, two of the three sex chromosomes were CMA 3/DAPI-; at early meiotic prophase and at diakinesis, silver impregnation corresponded with FISH signals, indicating that in this species, two chromosomes (probably a sex chromosome and an autosome) bear NORs. In T. infestans, silver nitrate and FISH also stained corresponding areas on meiotic chromosomes. Our data suggest that in triatomines, in general, the number and location of NORs are species-specific. These regions may be considered important chromosome markers for comparative studies to improve the understanding of evolutionary mechanisms in these hematophagous insects. ©FUNPEC-RP.

Induction of mitotic crossing-over in diploid strains of Aspergillus nidulans using low-dose X-rays

As a contribution towards detecting the genetic effects of low doses of genotoxic physical agents, this paper deals with the consequences of low-dose X-rays in the Aspergillus nidulans genome. The irradiation doses studied were those commonly used in dental clinics (1-5 cGy). Even very low doses promoted increased mitotic crossing-over frequencies in diploid strains heterozygous for several genetic markers including the ones involved in DNA repair and recombination mechanisms. Genetic markers of several heterozygous strains were individually analyzed disclosing that some markers were especially sensitive to the treatments. These markers should be chosen as bio-indicators in the homozygotization index assay to better detect the recombinogenic/ carcinogenic genomic effects of low-dose X-rays. ©FUNPEC-RP.