The Structure of the Dna-Acridine Complex

Lerman, Leonard S.

doi:10.1073/pnas.49.1.94

Cited by 652 publications

(185 citation statements)

References 9 publications

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“…The differential staining of double-stranded DNA versus single-stranded nucleic acids (DNA or RNA) is due to the metachromatic properties of AO: the dye fluoresces green (F530) when intercalated into doublestranded nucleic acids (31) and exhibits red (F600) fluorescence when bound to single-stranded polymers (3, 25,26). A 0 stained normal, mature sperm fluoresce primarily green but at about one-tenth the level of so-matic cells due to their haploid DNA content and a high level of chromatin condensation that restricts the dye binding to DNA (9,171.…”

Section: Sample Preparationmentioning

confidence: 99%

Flow cytometric analysis of mouse spermatogenic function following exposure to ethylnitrosourea

et al. 1985

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The effects of the mutagenic agent ethylnitrosourea ( E m on spermatogenic function and sperm chromatin structure were studied by flow cytometry and the results compared with sperm head morphology measurements. Groups of mice received daily exposures ranging from 0 to 75 mgkg body weight x 5 days and were sacrificed 28 days later. Fresh testicular cell suspensions and epididymal sperm were stained with acridine orange (AO) and measured by flow cytometry. Sperm nuclei were isolated, fixed, rehydrated, and then either subjected to thermal stress or not prior to staining with AO. Body weights were unaffected by the chemical exposure while the testicular weights were reduced by about 50%. Two-parameter (DNA, RNA) flow cytometry measurements showed a dose-response relationship in the loss of certain cell types, particularly the elongated spermatids, from the testes of treated animals. Flow cytometric analysis of both heat-stressed and non-heatstressed nuclei showed a relationship between dosage and the coefficient of variation of at {redked + green fluorescence)] measurements of A0 stained nuclei, thereby demonstrating that alterations of chromatin structure occurred in response to ENU. Enzymatic digestions with RNAse, DNAse, and nuclease S1 suggest that the increase in red fluorescence is due to an increase of singlestranded DNA induced by heat or acid treatment of chemically altered chromatin structure. The lowest daily dosage used (5 mgkg) caused no significant changes in ratios of testicular cell types, a questionable increase in abnormal sperm head morphology and a detectable change in chromatin structure expressed as at. This report shows that our technique for assaying sperm nuclear chromatin structure appears to have the same level of sensitivity to ENU induced nuclear alterations as the sperm head morphology test.Key terms: Flow cytometry, sperm head morphology, toxicology, testis, chromatin structureThe production of sperm represents one of the most dramatic examples of cellular proliferation and differentiation in any mammalian organ system. Although many biological parameters have been related to male fertility, sperm morphology is frequently used to assay testicular function. Sperm head morphology, consistent with nuclear morphology, is genotype specific (28), and is directed by genes on both the autosomes and sex chromosomes (2,411. Heritable changes in sperm head morphology can be induced by radiation and alkylating agents and followed quantitatively through successive generations (22,38,40). Wyrobek and Bruce (42,43) developed a mouse model assay system to study the mutagenic andor carcinogenic potential of chemicals using sperm head morphology as an end point. This test is relatively rapid and is considered very useful in the assessment of potentially toxic chemicals, since 100% of known mutagens tested were correctly identified as positives (44). Thus, demonstration of elevated levels of sperm abnormalities following exposure to a physical or chemical agent raises the possibility that the agent is i...

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Section: Sample Preparationmentioning

confidence: 99%

Flow cytometric analysis of mouse spermatogenic function following exposure to ethylnitrosourea

et al. 1985

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“…It has been proposed that aminoacridines intercalate between normally neighboring DNA base pairs in a plane perpendicular to the helix axis (27); therefore, 9-AMA may be assumed to bind indiscriminately to DNA against the base pairs. This characteristic of 9-AMA should be regarded as an advantage over bromodeoxyuridine, which is often used to suppress chromosome condensation because it is strictly AT-specific and, hence, its influence on the chromosome banding pattern differs from than in AT/GC-enriched regions.…”

Section: Discussionmentioning

confidence: 99%

9‐Aminoacridine: An efficient reagent to improve human and plant chromosome banding patterns and to standardize chromosome image analysis

et al. 2002

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BackgroundSuccessful automated chromosome analysis requires the development of new techniques to increase and standardize chromosome length and improve banding patterns.MethodsHuman and plant cells were pretreated with the DNA intercalator 9‐aminoacridine (9‐AMA), and chromosomes were stained with GTG and aceto‐orcein banding techniques and investigated by an image analysis system.ResultsThe human optimal chromosome spreads with the 850 G‐band resolution level, suitable for image analysis, were obtained by 9‐AMA pretreatment for 1 h at a final concentration of 0.5–1 μg/ml, as compared with 600–700 bands after ethidium bromide treatment and about 400 bands without pretreatment. The best results for plant chromosomes were obtained after pretreatment with 1–2 μg/ml of 9‐AMA for 12–24 h. The chromosomes elongated approximately 1.5‐fold, and the resolution of chromosome banding patterns increased, reaching approximately 140 bands per haploid set in the case of camomile.Conclusions9‐AMA is an efficient reagent for the standardization and increasing the resolution of chromosome banding patterns in human and plant chromosomes. It is extremely important for chromosome investigation in small plants. Cytometry Part A 51A:52–57, 2003. © 2002 Wiley‐Liss, Inc.

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“…In vitro experiments have shown that acridines are intercalated between specific adjacent base pairs in DNA (Lerman, 1963) and inhibit synthesis of DNA (Hurwitz, Furth, Malamy &Alexander, 1962). The base specificity of the affinity has been demonstrated to differ according to the kinds of acridines (Tubbs, Eitmars & van Winkle, 1964;Weisblum & de Haseth, 1972).…”

Section: Eflect Of Fatty Acids On Curingmentioning

confidence: 99%

Plasmid-instability in acrA Mutants of Escherichia coli K12

Nakamura

1974

Journal of General Microbiology

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The Structure of the Dna-Acridine Complex

Cited by 652 publications

References 9 publications

Flow cytometric analysis of mouse spermatogenic function following exposure to ethylnitrosourea

Flow cytometric analysis of mouse spermatogenic function following exposure to ethylnitrosourea

9‐Aminoacridine: An efficient reagent to improve human and plant chromosome banding patterns and to standardize chromosome image analysis

Plasmid-instability in acrA Mutants of Escherichia coli K12

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