Classical and molecular cytogenetic analysis in head and neck squamous cell carcinomas

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

New occurrence of microchromosomes B in Moenkhausia sanctaefilomenae (Pisces, Characidae) from the Parana 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 CMA(3) 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.

Organization of Repeated DNA Elements in the Genome of the Cichlid Fish Cichla kelberi and Its Contributions to the Knowledge of Fish Genomes

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

Phosphorylation of Histone H3S10 in Animal Chromosomes: Is There a Uniform Pattern?

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Karyotypic similarity among Barycholos temetzi and five species of the genus Eleutherodactylus from southeastern Brazil (Anura, Brachycephalidae)

Comparative cytogenetic analyses were carried out in six species of Brachycephalidae from southeastern Brazil. Barycholos ternetzi, Eleutherodactylus binotatus, Eleutherodactylus guentheri, Eleutherodactylus juipoca, Eleutherodactylus parvus and Eleutherodactylus sp. have 2n = 22 karyotypes with a marked variation in the morphology of chromosome pairs 8, 10 and 11, which are of telocentric or metacentric types, resulting in FN = 38, 40 and 44. Eleutherodactylus have a single chromosome pair bearing Ag-NOR, i.e. pair 1 in E. binotatus, pair 6 in E. guentheri and E. parvus, and pair 11 in E. juipoca and Eleutherodactylus sp. In contrast, B. ternetzi showed Ag-positive sites in the chromosome pairs 1, 4, 5, 9 and 11, and only one to three labelings per metdphase in each individual. Nevertheless, the main chromosome pair with Ag-NOR in the species seems to be the 11th, like in E. juipoca and Eleutherodactylus sp. The NOR site was confirmed by fluorescence in situ hybridization (FISH) technique in E. binotatus and in B. ternetzi, bearing 1p1p and 9p11p11p Ag-NOR pattern, respectively. All the species exhibited predominantly centromeric C-banding pattern, but interstitial bands have also been observed in some cases. In E. binotatus, there is an indication of geographical difference in the distribution of the interstitial C-bands. The fluorochromes GC-specific chromomycin A(3) (CMA(3)) and AT-specific 4',6-diamidino-2-phenylindole (DAPI), with distamycin A (DA) counterstaining, provided the molecular content of some repetitive regions in the karyotypes of the species. One male of E. binotatus presented an extensive heteromorphism, involving at least five different pairs, probably as a consequence of multiple reciprocal translocations. Such rearrangements might be responsible for the multivalent chain seen in the meiosis of this specimen, as well as in another male, although not exhibiting chromosome heteromorphism. The remaining males and those belonging to the other species have always shown 11 bivalents in diplotene and metaphase I cells. In all male specimens, metaphases II presented 11 chromosomes. Despite the observed discrepancies, the five species of Eleutherodactylus have a great uniformity in the 2n = 22 karyotypes, suggesting an assemblage of species from southeastern and southern Brazil, in contrast to northern and northeastern assemblage which is characterized by higher diploid numbers. Undoubtedly, B. ternetzi could be included in that proposed assemblage, due to its karyotypic similarity with the Eleutherodactylus species, as evidenced in the present study. This fact strongly supports the close relationships of both genera, previously inferred on the basis of several characters shared by their species. (C) 2006 Elsevier Ltd. All rights reserved.

Comparative karyotype analysis and chromosome evolution in the genus Aplastodiscus (Cophomantini, Hylinae, Hylidae)

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

Cytogenetic analyses of eight species in the genus Leptodactylus Fitzinger, 1843 (Amphibia, Anura, Leptodactylidae), including a new diploid number and a karyotype with multiple translocations

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Assignment of chromosome rearrangements between X chromosomes of human and cattle by laser microdissection and Zoo-FISH

Cross-species fluorescence in-situ hybridization (Zoo-FISH) was performed on cattle metaphase spreads using Homo sapiens X chromosome (HSAX) painting probes specific for the p- and q-arms to identify the cytogenetic location of a chromosome breakpoint between HSAX and the Bos taurus X chromosome (BTAX). The existence of a breakpoint is strongly suggested by recent radiation hybrid and FISH mapping results. Hybridization probes were generated by microdissection of HSAX p- and q-arms using the contact-free technology of Laser Microdissection and Pressure Catapulting (LMPC), amplification of the isolated chromosome material by DOP-PCR, and labelling of the PCR products with digoxigenin in a secondary PCR. Independent Zoo-FISH of the two painting probes on bovine metaphase chromosomes (detected by antidigoxigenin-fluorescein) resulted in clear hybridization signals on BTAX. A breakpoint was identified between HSAXp and HSAXq on BTAX, and narrowed down between the G-bands BTAXq25 and BTAXq26. The assumed centromere transposition between HSAX and BTAX associated with the rearranged chromosome segments is supported by cytogenetic assignments of the genes BGN and G6PD to BTAX.

Meiosis aspects and nucleolar activity in Triatoma vitticeps (Triatominae, Heteroptera)

Some aspects of both the nucleolar organizer activity and meiosis were studied in the testes of Triatoma vitticeps (Heteroptera, Triatominae). The techniques used included squashing followed by lacto-acetic orcein staining, silver-ion impregnation, fluorescent banding (CMA(3), Quinacrine mustard and DAPI) and fluorescent in situ hybridization (FISH). A close relationship between heterochromatin and nucleolus in testicular cells was observed. During meiosis, the silver-ion impregnation pattern varied. At metaphase plate, a small body appeared apart from the chromosomes. In the spermatids this small body was seen in preparations stained with orcein and silver- ion impregnation but not with fluorochromes or FISH. These characteristics combined suggest that these corpuscles represent a source of ribonucleoproteins (RNP) - RNA and specific nucleolar proteins. Silver-ion impregnation and (FISH) revealed nucleolar organizer activity in two metaphase sex chromosomes (X). These results indicate that, in these species, nucleolar organizer regions (NORs) are located in the sex chromosomes, X chromosomes were were CMA(3)(+) and Y chromosome was DAPI(+).

Spermatogenesis and nucleolar activity in Triatoma klugi (Triatomine, Heteroptera)

Triatoma klugi is a Chagas disease vector in the Rio Grande do Sul State. Triatominae chromosomes are holocentric and sex chromosomes segregation is post-reductional. In this paper we describe the karyotype of male T. klugi and a meiotic analysis including the nucleolar behavior during spermatogenesis. Testis cells were analyzed after lacto-acetic orcein and silver nitrate staining. Two autosomes and the heterochromosomes presented nucleolar activity (Ag-NORs) during diplotene-diakinesis. The analysis of metaphase I and II revealed a karyotype with 2n = 20+XY. In metaphase I a prominent nucleolar mass was observed in the cell periphery and small silver grains were detected in metaphase II. During anaphase, the chromosomes segregated in parallel and a typical holocentric late migration behavior was observed. The restoration of the nucleolus was an important feature in this phase. During telophase nucleolar masses persisted and in early spermiogenesis the spermatids presented a small peripheral mass until elongation. The present study is a contribution to the study of chromatin behavior and nucleolar persistence in meiosis.

Numerical chromosome polymorphism in Mikania cordifolia Willd. (Asteraceae)

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

Pattern of nucleolar activity during spermiogenesis in triatomines (Heteroptera, Reduviidae) as analysed by silver staining

Nucleoli are the sites of biosynthesis of ribosomal precursors. In this work the nucleolar activity at interphase and the meiotic cells of the testis in five species of triatomines were analysed by means of silver staining. Several nucleolar blocks in the polyploid nuclei of testicular tubules were observed, whereas only one nucleolar body could be seen in the spermatogonial nuclei of all five species. A single nucleolar body was evident in the 'confused stage' of Triatoma brasiliensis, T. delpontei, T. lecticularia and T. rubrovaria, while T. sordida presented two nucleolar dots. The existence of small, silver-stained dots in some metaphase I chromosomes of T. brasiliensis and T. sordida is reported. The number of nucleolar dots present in spermatids of each species varied within and among species. It is suggested that in addition to providing information on rRNA biosynthesis, studies of nucleolar organizing activity can also be important sources of data on differentiation patterns and species development.

X chromosome heterochromatic pattern and nucleolar synthesis in pupal ovaries of Aedes aegypti

C-banding and silver-staining techniques were used to examine pupal ovaries of Aedes aegypti from Sao Jose do Rio Preto (Brazil). Silver staining in ovary cystocytes showed two basic patterns relative to the nucleolar morphology: viz (1) a single, compact small body; and (2) multiple bodies encompassing large nuclear areas. These two types of cystocytes were present in the ratio of 7:1, which is the same as the number of nurse cells and oocytes, respectively, in each follicle. This suggests the possibility of eventually using such a nucleolar morphological difference to recognize both cell types in developmental stages before emergence. Silver nitrate staining in metaphase chromosomes revealed centromeric bands on all six chromosomes. The C-banding pattern in metaphase chromosomes showed an intercalary band in one of the X arms, as described previously in other populations. In ovary cystocytes (pachytene stage) this C-positive band seemed to consist of two chromomeres. Phase contrast microscopy showed that the nucleolus was associated with the distal chromomere of this intercalary C-band, indicating that the nucleolus organizer region was located in that part of the heterochromatic band.

Recombination nodules in coleopteran species: Palembus dermestoides(Tenebrionidae) and Epicauta atomaria (Meloidae)

This work describes the first report about the occurrence of recombination nodules (RNs) in spread pachytene cells of two species of Coleoptera: Palembus dermestoides (Tenebrionidae) and Epicauta atomaria (Meloidae). The RNs were observed in preparations contrasted with phosphotungstic acid. Considering RN morphology and its occurrence in pachytene bivalents (one per autosome bivalent) these structures were interpreted to be late RNs. P. dermestoides and E. atolraria have 2n = 20 chromosomes including an Xyp sex determination system. In spite of most frequently subtelocentric morphology observed in the autosomes of both species, the occurrence of RNs is limited only to the synaptonemal complex (SC) structure of the long arms. These findings are in agreement with those obtained using light microscopy analysis in which only one chiasma or terminalization event is observed per autosomal bivalent in early or late metaphase I cells. The RNs have the same average width of the SC of each analyzed species, a circular shape, strong electron density, and are observed mainly between the lateral elements of the SC. The RNs of P. dermestoides and E. atomaria have approximately the same average size (width), 180 +/- 20 nm and 160 +/- 80 nm, respectively. The absence of RNs in the short arms and its occurrence in the long arms are discussed considering the short arm pericentromeric and pro-centric heterochromatin. Copyright (C) 2003 S. Karger AG, Basel

POPULATIONAL STUDY OF CHROMOSOME POLYMORPHISM IN ZONOTRICHIA-CAPENSIS (EMBERIZIDAE, PASSERIFORMES, AVES)

The karyotypes of 328 specimens of Zonotrichia capensis from four regions, three in the state of Parana and one in the state of São Paulo (Brazil), were studied. This species presents a chromosomal polymorphism due to pericentric inversion, involving the 3rd and 5th chromosome pairs. Chromosome 3 exists in the 3st and 3sm conditions and chromosome 5, in 5st and 5m conditions. Mitotic metaphase spreads were obtained from bone-marrow material of colchicine-treated animals.At all collection sites the frequency of chromosome 5m was considerably lower than that of the inverted chromosome 3sm.The present work tried to establish a correlation between the frequencies of the 3rd and 5th chromosome pairs with the climatical and geographical parameters at the collecting regions.Data show that the frequency of chromosome 5m is higher in regions with low temperature and high altitude.