Banding Pattern of Human Chromosomes

STUDIES of mammalian chromosomes have advanced rapidly since the introduction of the quinacrine fluorescence technique¹ and other procedures which are believed to locate repetitive DNA sequences in cytological chromosome preparations^2,3. To improve these techniques, a method has been developed which can provide further information about chromosome organization.     

Banding Human Chromosomes Using a Combined C-Banding-Fluorochrome Staining Technique

Slides pretreated for C-banding and stained with DAPI or CMA3 show different banding patterns in human metaphase chromosomes compared to those obtained with either standard Giemsa C-banding or fluorochrome staining alone. Human chromosomes show C-plus DA-DAPI banding after C-banding plus DAPI and enhanced R-banding after C-banding plus Chromomycin A3 staining. If C-banding preferentially removes certain classes of DNA and proteins from different chromosome domains, C-banding pre-treatment may cause a differential DNA extraction from G- and R-bands in human chromosomes, resulting in a preferential extraction of DNA included in G-bands. This hypothesis is partially supported by the selective cleavage and removal of DNA from R-bands of restriction endonuclease HaeIII with C-banding combined with DAPI or Chromomycin A3 staining. Structural factors relating to regional differences in DNA and/or proteins could also explain these results.     

The Relationship of Slide Maturity and Trypsin Exposure Time in the Differential Giemsa Banding of Chromosomes

Difficulty has been reported in attaining consistent chromosome banding using trypsin-Giemsa techniques. When trypsin concentration, incubation temperature, and the duration of staining are held constant, a logarithmic relationship has been found to exist between the length of time since a slide was made and the length of trypsin treatment necessary to produce optimal banding when it is stained. Under the conditions specified in this report, ten seconds digestion was optimal for ten-day-old slides. This time approximately doubled for each additional eight days that slides were allowed to age. From a plot of the experimental results, treatment times appropriate for slides of any age from 10 to 60 days were obtained. Use of these times produced satisfactory banding in 85% of mitoses.     

Trypsin-Orcein Banding in Plant Chromosomes

A convenient and quick method using trypsin-orcein for handing plant chromosomes (O-banding) is suggested. The technique is directly applicable to meristematic tissues (e.g. root tip) and involves the treatment of root tip with 1-2% solution of trypsin either in buffer or in 05 N HCI for 5-10 minutes at 37 C or for 30-60 minuta near 0 C followed by staining with 1.5% acetic orcein: 1 N HCI (19:1). Dark staining bands are reproducible and species specific. These bands possibly represent specific DNA-protein-dye interaction.     

鱼类染色体的荧光显带研究

应用GC碱基特异性荧光染料色霉素A,辅以AT减基特异性荧光染料Hoechst33258,DAPI或喹吖因对鲤鱼,鲫鱼,大鳞副泥鳅和的有丝分裂染色体及黄鳝的有丝分裂和减数分裂染色体进行了荧光显带研究,结果发现,色霉素A3可以特异性地显示鱼类有丝分裂及减数分裂各个时期核仁组织区NORS的存在,Hoechst33258,DAPI或喹吖因则使这些区域（NORs)淡染,大鳞副泥鳞的染色体NORs 分布位置具有性别,根据实验结果,对有关鱼类染色体的荧光染色研究及其应用进行了讨论。     

Application of Ethidium Bromide for High-Resolution Banding Analysis of Chromosomes from Human Malignant Cells

Ethidium bromide was added to cultured human leukemic bone marrow and solid tumor cells to evaluate its inhibitory effect on mitotic chromosome condensation and its possible application to high-resolution banding analysis. In most experiments ethidium bromide treatment resulted in a high proportion of mitotic cells having elongated chromosomes, without remarkable reduction in either the mitotic index or quality of metaphase chromosomes. Optimal effect on chromosome length was obtained by adding 10 μg/ml of ethidium bromide during the final 2 hr of culture. Because of the simplicity and reproducibility of the technique involved, ethidium bromide can be used routinely to extend the length of chromosomes for fine-banding analysis of malignant cells.     

Chromosomes in patients treated with Azathioprine

Summary Chromosomes from leukocyte cultures and direct bone marrow preparations from patients under Azathioprine treatment were examined and compared with those from uremic patients and normal controls not taking Azathioprine. The results indicate a toxic effect of the drug, as well as an effect of uremia on the chromosome complement. The significance of these findings is discussed, especially in relation to several reports concerning malignant processes arising during or after Azathioprine intake. According to our findings, a relation between immunosuppression and malignancy seems more likely than a direct cancerogenicity of Azathioprine.

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Zusammenfassung Chromosomen aus peripherem Blut und direkten Knochenmarkspräparaten von Patienten unter Azathioprin-Therapie wurden verglichen mit solchen von gesunden Kontrollpersonen und urämischen Patienten ohne Azathioprin-Einnahme. Die Ergebnisse sprechen für eine toxische Chromosomenschädigung sowohl durch Azathioprin als auch die urämische Stoffwechsellage. Die Bedeutung dieser Befunde, insbesondere im Zusammenhang mit mehrfach berichteten malignen Prozessen nach Azathioprin-Einnahme, wird diskutiert. Unsere Ergebnisse lassen eher einen Zusammenhang zwischen Immunosuppression und malignem Geschehen vermuten als eine direkte cancerogen Wirkung von Azathioprin.

     

Giemsa Banding in Chromosomes of Douglas Fir Seedlings and Plantlets

Douglas fir plantlets have been produced by tissue culture.Karyotypes of seedlings and plantlets were prepared from roottip squash preparations using standard histological procedures.Adiploidchromosome number of 26 was common to both. The relative lengths of seedling and plantlet chromosomes werefound to be similar. The frequency of occurrences of secondaryconstrictions was found to be high in chromosomes three andten. Giemsa staining was successfully used to distinguish uniquechromosomal banding patterns in seedlings and plantlets. Pseradotsuga menziesii, Douglas fir, chromosomes, giemsa staining     

Specific Heterochromatic Banding of Metaphase Chromosomes Using Nuclear Yellow

The bis-benzimidazole compound nuclear yellow (NY) belongs to the same chemical family as the DNA binding fluorochromes Hoechst 33258 and Hoechst 33342. Spectroscopic studies of NY alone and in the presence of calf thymus DNA show high DNA binding affinity and behavior similar to the Hoechst fluorochromes above. Mitotic metaphase chromosomes from Balb/c mice stained with NY show C-banding and weak G/Q-banding, both of them disappearing after distamycin A (DA) or methyl green (MG) counterstaining. The same staining of human metaphase chromosomes from lymphocyte cultures, however, reveal only faint G/Q-banding (NY) and a characteristic DA-DAPI-like banding (NY-DA, NY-MG). Image analysis of NY stained human chromosomes, confirms that NY is suitable for studying polymorphisms affecting size in the pericentromeric hete-rochromatin of pairs 1, 9 and 16, and shows significant enhancement of NY fluorescence induced by DA in DA-DAPI heterochromatin. Our spectroscopic and cytological results show that NY, either alone or counterstained with DA or MG, can be used for DNA cytochemistry and chromosome banding. Possible mechanisms for the banding patterns induced by NY are discussed.     

Specific Heterochromatic Banding of Metaphase Chromosomes Using Nuclear Yellow

The bis-benzimidazole compound nuclear yellow (NY) belongs to the same chemical family as the DNA binding fluorochromes Hoechst 33258 and Hoechst 33342. Spectroscopic studies of NY alone and in the presence of calf thymus DNA show high DNA binding affinity and behavior similar to the Hoechst fluorochromes above. Mitotic metaphase chromosomes from Balb/c mice stained with NY show C-banding and weak G/Q-banding, both of them disappearing after distamycin A (DA) or methyl green (MG) counterstaining. The same staining of human metaphase chromosomes from lymphocyte cultures, however, reveal only faint G/Q-banding (NY) and a characteristic DA-DAPI-like banding (NY-DA, NY-MG). Image analysis of NY stained human chromosomes, confirms that NY is suitable for studying polymorphisms affecting size in the pericentromeric hete-rochromatin of pairs 1, 9 and 16, and shows significant enhancement of NY fluorescence induced by DA in DA-DAPI heterochromatin. Our spectroscopic and cytological results show that NY, either alone or counterstained with DA or MG, can be used for DNA cytochemistry and chromosome banding. Possible mechanisms for the banding patterns induced by NY are discussed.     

Chromosomal Banding Patterns Produced by Methyl Green-Pyronin Staining After Trypsin Treatment

A method is described for producing banding pattern with methyl green-pyronin (MGP) stain in chromosomes of fibrosarcoma cells. 1) The stain was made by mixing equal volumes of 2% aqueous pyronin G, 2% aqueous methyl green, distilled water, and 0.1 M acetate Mer (pH 5.7). 2) Treatment with colcemide and hypotonic KCI (0.075 M) was performed u usual. 3) Metaphase chromosomes were prepared using the flame-drying technique and treated with 0.25% trypsin at 37 C for 45 to 90 seconda. Before staining, the slides were rid in PBS, in distilled water, and then were dipped in 0.05 M acetate buffer. 4) Chromosomes were stained for more than 20 minuta, rinsed in distilled water, and hot-air dried. satisfactory results were obtained in uncontracted metaphase chromosomes. MCP stain hm the advantage of permitting much longer trypsin treatment and staining time than the trypsin-Giemsa method while providing satisfactory banding pattern.     

鱼类基因组结构研究——Ⅰ.染色体的限制性内切酶显带

本文报道了1种新的鱼类染色体研究方法——限制性内切酶显带技术。我们应用限制性内切酶Bsp631等分别对黄鳝和长春鳊的染色体进行显带处理,结果表明,Bsp631能使这两种鱼的染色体发生稳定的带纹分化:适度的处理产生多重的G-带状带型,而过度的处理则产生特殊的限制性内切酶抗性带型。根据显带结果分析,我们对鱼类染色体限制性内切酶显带的可行性和实用价值进行了讨论。     

Banding of Allium Chromosomes Protected Against Dnase Digestion by Dna-Binding Drugs

Metaphase chromosome bands were induced in Allium flavum (Liliaceae) by protecting the chromosomal DN A with DN A-binding compounds of different base specificities against DNase digestion. The bands obtained represented different subsets of C-band heterochromatin.