Chromosome organization in meiosis revealed by light microscope analysis of silver-stained cores

Rufas, Julio S.; Giménez-Abián, Juan F.; Suja, José Á.; Vega, Carlos García de la

doi:10.1139/g87-121

Cited by 53 publications

(43 citation statements)

References 16 publications

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“…For DAPI/DA staining, the slides were treated for 20 min with DAPI and 40 min with DA. Silver nitrate impregnation was performed as described by Rufas et al (1987). In brief, the slides were pre-treated with 2X SSC at 60°C for 10 min and stained with silver nitrate (1 g/mL) at 70°-80°C.…”

Section: Methodsmentioning

confidence: 99%

Cytogenetics of the darkling beetles Zophobas aff. confusus and Nyctobates gigas (Coleoptera, Tenebrionidae)

Lira-Neto

Silva²,

Moura³

et al. 2012

Genet. Mol. Res.

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ABSTRACT. Males of Zophobas aff. confusus and Nyctobates gigas (Tenebrionidae) collected in the State of Pernambuco, Brazil, were studied through conventional staining, C-banding, silver nitrate impregnation (AgNO 3 ), and the base specific fluorochromes CMA 3 and DAPI. Z. aff. confusus was found to have 2n = 20 (9+Xyp) while N. gigas exhibited 2n = 18 (8+neoXY). Large pericentromeric blocks of constitutive heterochromatin (CH) were detected throughout the autosomal complement of the two species, except in one autosomal pair of N. gigas in which no heterochromatic block was observed. The sex chromosomes of both species were almost totally heterochromatic. Double staining with CMA 3 /DA (distamycin) and DAPI/DA marked CH in Z. aff. confusus. However, DAPI staining was more intense. N. gigas was found to possess blocks of CH-positive CMA 3 and homogeneous DAPI. AgNO 3 staining also revealed differences between the two species. In Z. confusus an NOR was observed in the sexual bivalent Xyp and N. gigas was found to have an autosomal NOR.

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Section: Methodsmentioning

confidence: 99%

Cytogenetics of the darkling beetles Zophobas aff. confusus and Nyctobates gigas (Coleoptera, Tenebrionidae)

Lira-Neto

Silva²,

Moura³

et al. 2012

Genet. Mol. Res.

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“…Slides used for C-banding, silver nitrate staining and FISH analysis were prepared in 45% acetic acid and coverslips were removed after freezing the preparations by immersing in liquid nitrogen for a few seconds. C-banding was performed according to the method described by Sumner (1972) and silver nitrate staining was conducted according to Rufas et al (1987).…”

Section: Methodsmentioning

confidence: 99%

Chromosomal mapping of repetitive DNAs in the beetle Dichotomius geminatus provides the first evidence for an association of 5S rRNA and histone H3 genes in insects, and repetitive DNA similarity between the B chromosome and A complement

2009

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Chromosomal banding techniques and repetitive DNA mapping are useful tools in comparative analysis and in the elucidation of genome organization of several groups of eukaryotes. In this study, we contributed to the knowledge of Coleoptera genomes by reporting the chromosomal organization of repetitive DNA sequences, as well as the presence and characteristics of a B chromosome in two natural populations of Dichotomius geminatus (Coleoptera; Scarabaeidae) using classical, chromosomal banding and molecular cytogenetic techniques. As in other coleopteran species, the heterochromatin was mainly concentrated in pericentromeric regions and the B chromosome was composed almost entirely of heterochromatin. Physical mapping using double fluorescent in situ hybridization was performed for the first time in Coleoptera; using DNA probes for 5S and 18S ribosomal RNA (rRNA) and histone H3 genes, we showed that ribosomal 18S rDNAs are located in chromosomes 3 and 4, whereas 5S rRNA and histone H3 genes are colocalized in chromosomal pair 2 and show an apparently interspersed organization. Moreover, these genes are not present in the B chromosome, suggesting that the B chromosome did not originate from chromosomal pairs 2, 3 or 4. On the other hand, mapping of the C 0 t-1 DNA fraction showed that the B chromosome is enriched in repetitive DNA elements, also present in the standard complement, indicating an intraspecific origin of this element in D. geminatus. These results will contribute to our understanding of genome organization and evolution of repetitive elements in Coleoptera and other insects regarding both A and B chromosomes.

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“…• C. Triple combination of the fluorochromes CMA 3 /DA/DAPI and silver staining were performed using the protocols of Schweizer (1976) and Rufas et al (1987) respectively, and FISH followed the procedure of Moscone et al (1996), using a probe of 45S rDNA isolated from Arabidopsis thaliana. The images were captured in Leica DM LB microscope (Leica Microsystems, Wetzlar, Germany) and the figures were organized using the Corel Photo-Paint 12 software (Corel corporation 2003, Ottawa, Canada).…”

Section: Methodsmentioning

confidence: 99%

Cytogenetic characterization of Eurysternus caribaeus (Coleoptera: Scarabaeidae): evidence of sex-autosome fusion and diploid number reduction prior to species dispersion

Arcanjo

Cabral-de-Mello

Silva

et al. 2009

J Genet

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Mitotic and meiotic chromosomes of several populations of Eurysternus caribaeus (Coleoptera: Scarabaeidae) were analysed through conventional staining, C-banding, base-specific fluorochromes, silver nitrate staining and fluorescent in situ hybridization (FISH). All specimens showed 2n = 8 in their karyotypes, with a neo-XY sex system (Y is a submetacentric and X a metacentric) and three pairs of submetacentric autosomes. The analysis of constitutive heterochromatin (CH) revealed small blocks located in the centromeric region of all chromosomes which do not present positive staining under the fluorochromes CMA 3 and DAPI. Silver nitrate staining revealed that the nucleolar organizer region (NORs) is associated with the sex chromosomes. The FISH technique revealed that rDNA sites in the X and Y are different in size. Data from different populations indicate that the diploid number reduction (2n = 8) observed in E. caribaeus is established and presumably has preceded the dispersion of this species. Moreover, this reduction occasioned the translocation of rDNA sites to the sex chromosomes, X and Y, an uncommon pattern in Scarabaeidae that was observed for the first time by the FISH in this work.

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Chromosome organization in meiosis revealed by light microscope analysis of silver-stained cores

Cited by 53 publications

References 16 publications

Cytogenetics of the darkling beetles Zophobas aff. confusus and Nyctobates gigas (Coleoptera, Tenebrionidae)

Cytogenetics of the darkling beetles Zophobas aff. confusus and Nyctobates gigas (Coleoptera, Tenebrionidae)

Chromosomal mapping of repetitive DNAs in the beetle Dichotomius geminatus provides the first evidence for an association of 5S rRNA and histone H3 genes in insects, and repetitive DNA similarity between the B chromosome and A complement

Cytogenetic characterization of Eurysternus caribaeus (Coleoptera: Scarabaeidae): evidence of sex-autosome fusion and diploid number reduction prior to species dispersion

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