Quantitative determination of single-stranded sections in DNA using the fluorescent probe acridine orange

Ichimura, Sachiko; Zama, Mitsuo; Fujita, Hitoshi

doi:10.1016/0005-2787(71)90705-2

Cited by 59 publications

(33 citation statements)

References 14 publications

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“…Acridine orange has been used to label nucleic acids in solution and intact cells. [59][60][61][62] Acridine orange intercalates into double-stranded DNA as a monomer, whereas it binds to single-stranded DNA as an aggregate. Upon excitation at 470-490 nm, the monomeric acridine orange bound to double-stranded DNA fluoresces green, with an emission maximum at 530 nm.…”

Section: Surfactant-dna Gel Particlesmentioning

confidence: 99%

Surfactant−DNA Gel Particles: Formation and Release Characteristics

Morán

Miguel

Lindman³

2007

Biomacromolecules

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Aqueous mixtures of oppositely charged polyelectrolytes undergo associative phase separation, resulting in coacervation, gelation, or precipitation. This phenomenon has been exploited here to form DNA gel particles by interfacial diffusion. We report on the formation of DNA gel particles by mixing solutions of DNA (either singlestranded (ssDNA) or double-stranded (dsDNA)) with solutions of cationic surfactant cetyltrimetrylammonium bromide (CTAB). By using CTAB, the formation of DNA reservoir gel particles, without adding any kind of cross-linker or organic solvent, has been demonstrated. Particles have been characterized with respect to the degree of DNA entrapment, surface morphology, and secondary structure of DNA in the particles. The swelling/ deswelling behavior and the DNA release have been investigated in response to salt additions. Analysis of the data has suggested a higher degree of interaction between ssDNA and the cationic surfactant, confirming the stronger amphiphilic character of the denatured DNA. Fluorescence microscopy studies have suggested that the formation of these particles is associated with a conservation of the secondary structure of DNA.

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Section: Surfactant-dna Gel Particlesmentioning

confidence: 99%

Surfactant−DNA Gel Particles: Formation and Release Characteristics

Morán

Miguel

Lindman³

2007

Biomacromolecules

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“…Acridine orange intercalates into double-stranded DNA as a monomer, whereas it binds to single-stranded DNA as an aggregate. Upon excitation at 470-490 nm, the monomeric acridine orange bound to double-stranded DNA fluoresces green with an emission maximum at 530 nm, and the aggregated acridine orange on single-stranded DNA fluoresces red with an emission maximum at about 640 nm (Ichimura et al, 1971;Peacocke, 1973). Kosower et al (1992) compared the colour of acridine orange fluorescence in sperm nuclei after acid treatment with the 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 252 S. Said et al…”

Section: Discussionmentioning

confidence: 99%

“…Sperm DNA with disulphide poor protamines appears to be denatured following treatment with acid, whereas the DNA of disulphide-rich sperm nuclei is resistant to denaturation even after exposure to acid (Kosower et al, 1992). Binding of acridine orange to denatured DNA emits red fluorescence, whereas binding to native DNA emits green fluorescence (Ichimura et al, 1971;Kosower et al, 1992). A previous observation using mBBr has demonstrated that the reactive thiols of rat spermatozoa decrease markedly between the corpus and the cauda of the epididymis (Shalgi et al, 1989).…”

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confidence: 99%

DNA stability and thiol-disulphide status of rat sperm nuclei during epididymal maturation and penetration of oocytes

Said

Funahashi

Niwa

1999

Zygote

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DNA stability and thiol-disulphide status of rat sperm nuclei was observed in vivo during maturation in the epididymis and penetration of oocytes. When spermatids and spermatozoa were stained with acridine orange after fixation with acetic alcohol, the red/green fluorescence ratio observed under a confocal microscope was not different between spermatids (3.81 ± 0.16) and testicular spermatozoa (4.03 ± 0.34), and then decreased sharply (p < 0.01) as the spermatozoa descended the epidymis to the caput epididymis (1.13 ± 0.03). However, the ratio was not different among corpus (0.69 ± 0.01), cauda epididymis (0.68 ± 0.11) and ejaculated spermatozoa (0.63 ± 0.01). On the other hand, when spermatozoa were labelled with monobromobimane (mBBr), the fluorescence intensities gradually decreased (p < 0.01) during passage of spermatozoa from testis (4.74 ± 0.16) through epididymis (caput, 2.72 ± 0.08; corpus, 1.07 ± 0.03; cauda, −0.05 ± 0.05; ejaculated, 0.08 ± 0.03). The acridine orange red/green fluorescence ratio increased (p < 0.01) during zona penetration (binding sperm, 0.52 ± 0.09; perivitelline sperm, 0.64 ± 0.16) and sperm decondensation (decondensed sperm, 0.69 ± 0.12). When spermatozoa in the perivitelline space were labelled with mBBr, the fluorescence was detected. These results demonstrate that DNA stability against acid appears to be ahead of the oxidation of protamine during sperm maturation in the epididymis and is an initial event of the unpackaging process in rat genome occurring during or just after zona penetration but before ooplasm penetration.

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“…has been used to label nucleic acids in solution and in intact cells [78][79][80][81]. In the case of AO-dsDNA, the fluorescence emission shows a maximum around 530 nm, in the green spectra.…”

Section: Morphological Characterization Of the Dna Gel Particlesmentioning

confidence: 99%

DNA gel particles: An overview

Morán

Vinardell

Infante

et al. 2014

Advances in Colloid and Interface Science

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A general understanding of interactions between DNA and oppositely charged compounds forms the basis for developing novel DNA-based materials, including gel particles. The association strength, which is altered by varying the chemical structure of the cationic cosolute, determines the spatial homogeneity of the gelation process, creating DNA reservoir devices and DNA matrix devices that can be designed to release either single-(ssDNA) or double-stranded (dsDNA). This review covers recent developments on the topic of DNA gel particles formed in water-water emulsion-type interfaces. The degree of DNA entrapment, particle morphology, swelling/dissolution behaviour and DNA release responses are discussed as a function of the nature of the cationic agent used. On the basis of designing DNA gel particles for therapeutic purposes, recent studies on the determination of the surface hydrophobicity, the haemolytic and the cytotoxic assessments of the obtained DNA gel particles have been also reported.

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Quantitative determination of single-stranded sections in DNA using the fluorescent probe acridine orange

Cited by 59 publications

References 14 publications

Surfactant−DNA Gel Particles: Formation and Release Characteristics

Surfactant−DNA Gel Particles: Formation and Release Characteristics

DNA stability and thiol-disulphide status of rat sperm nuclei during epididymal maturation and penetration of oocytes

DNA gel particles: An overview

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