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Molecular Neurobiology

, Volume 56, Issue 10, pp 6770–6776 | Cite as

Brain Metabolic DNA Is Reverse Transcribed in Cytoplasm: Evidence by Immunofluorescence Analysis

  • Marina Prisco
  • Joyce Casalino
  • Carolina Cefaliello
  • Antonio GiudittaEmail author
Article
  • 98 Downloads

Abstract

In a previous study (Mol Neurobiol 55:7476–7486, 2017), newly synthesized brain metabolic DNA (BMD) from rat subcellular fractions has been shown to behave as a DNA-RNA hybrid when analyzed in cesium gradients at early [3H] thymidine incorporation times but to assume the double-stranded configuration at later times. Conversely, BMD from purified nuclei displayed the dsDNA configuration even at early incorporation times. The results were interpreted to support the BMD origin by reverse transcription in the cytoplasm and its later acquisition of the double-stranded configuration before the partial transfer to the nuclei. This interpretation has now been confirmed by immunofluorescence analyses of newly synthesized BrdU-labeled BMD from the mouse brain that demonstrates its cytoplasmic localization and colocalization with DNA-RNA hybrids. In addition, BrdU-labeled BMD has been shown to colocalize with astroglial anti-GFAP antibodies and with presynaptic anti-synaptophysin antibodies.

Keywords

Brain metabolic DNA (BMD) DNA synthesis Reverse transcription Astroglia Synaptosomes 

Notes

Acknowledgements

We warmly thank Prof. M. D’Amico and. S. Maione for their generous permission to use the facilities of the Second University of Naples Medical School.

References

  1. 1.
    Perrone Capano C, D'Onofrio G, Giuditta A (1982) DNA turnover in rat cerebral cortex. J Neurochem 38:52–56CrossRefGoogle Scholar
  2. 2.
    Reinis S (1972) Autoradiographic study of 3H-thymidine incorporation into brain DNA during learning. Physiol Chem Phys 4:391–397Google Scholar
  3. 3.
    Reinis S, Lamble RW (1972) Labeling of brain DNA by 3H-thymidine during learning. Physiol Chem Phys 4:335–338Google Scholar
  4. 4.
    Ashapkin VV, Romanov GA, Tushmalova NA, Vanyushin BF (1983) Selective DNA synthesis in the rat brain induced by learning. Biokhimija 48:355–362Google Scholar
  5. 5.
    Scaroni R, Ambrosini MV, Principato GB, Federici F, Ambrosi G, Giuditta A (1983) Synthesis of brain DNA during acquisition of an active avoidance task. Physiol Behav 30:577–582CrossRefGoogle Scholar
  6. 6.
    Papa M, Pellicano MP, Cerbone A, Lamberti-D'Mello C, Menna T, Buono C, Giuditta A, Welzl H et al (1995) Immediate early genes and brain DNA remodeling in the naples high and low-excitability rat lines following exposure to a spatial novelty. Brain Res Bull 37:111–118CrossRefGoogle Scholar
  7. 7.
    Giuditta A et al. (1986) Synthesis of rat brain DNA during acquisition of an appetitive task. Pharmacol Biochem Behav25:651–658Google Scholar
  8. 8.
    Ivashkina OI, Zots MA, Bezriadnov DV, Anokhin KV (2012) Increased 5′-bromo-2′-deoxyuridine incorporation in various brain structures following passive avoidance training in mice. Bull Exp Biol Med 154:171–173CrossRefGoogle Scholar
  9. 9.
    Giuditta A, Ambrosini MV, Scaroni R, Chiurulla C, Sadile A (1985) Effect of sleep on cerebral DNA synthesized during shuttle-box avoidance training. Physiol Behav 34:769–778CrossRefGoogle Scholar
  10. 10.
    Langella M, Colarieti L, Ambrosini MV, Giuditta A (1992) The sequential hypothesis of sleep function. IV. A correlative analysis of sleep variables in learning and non learning rats. Physiol Behav 51:227–238CrossRefGoogle Scholar
  11. 11.
    Grassi Zucconi G, Menichini E, Castigli E, Belia S, Giuditta A (1988a) Circadian oscillations of DNA synthesis in rat brain. Brain Res 447:253–261CrossRefGoogle Scholar
  12. 12.
    Grassi Zucconi G, Carandente ME, Belia S, Giuditta A (1988b) Circadian rhythms of DNA content in brain and kidney: effects of environmental stimulation. Chronobiol 15:195–204Google Scholar
  13. 13.
    Grassi Zucconi G, Crognale MC, Bassetti MA, Giuditta (1990) Environmental stimuli modulate the circadian rhythm of (3H-methyl) thymidine incorporation into brain a DNA of male rats. Behav Brain Res 41:103–110Google Scholar
  14. 14.
    Giuditta A, Grassi-Zucconi G, Sadile A (2017) Brain metabolic DNA in memory processing and genome turnover. Rev Neurosci 28:21–30CrossRefGoogle Scholar
  15. 15.
    Pelc SR (1964) Labelling of DNA and cell division in so called non-dividing tissues. J Cell Biol 22:21–28CrossRefGoogle Scholar
  16. 16.
    Pelc SR (I968a) Turnover of DNA and function. Nature 219:162–l63Google Scholar
  17. 17.
    Pelc SR (1968b) Biological implications of DNA-turnover in higher organisms. Acta Histochem Suppl 8:441–452Google Scholar
  18. 18.
    Pelc SR (1972) Metabolic DNA in ciliated protozoa, salivary gland chromosomes, and mammalian cells. Int Rev Cytol 32:327–355CrossRefGoogle Scholar
  19. 19.
    Roels H (1966) Metabolic’ DNA: a cytochemical study. Int Rev Cytol 19:1–34CrossRefGoogle Scholar
  20. 20.
    Stroun M, Charles P, Anker P, Pelc SR (1967) Metabolic DNA in heart and skeletal muscle and in the intestine of mice. Nature 216:716–717CrossRefGoogle Scholar
  21. 21.
    Gahan PB, Anker P, Stroun M (2008) Metabolic DNA as the origin of spontaneously released DNA? Ann N Y Acad Sci 1137:7–17CrossRefGoogle Scholar
  22. 22.
    Rutigliano B, Giuditta A (2015) The unexpected recovery of misplaced data on brain metabolic DNA. Rend Acc Sci Fis Mat LXXXII:99–106Google Scholar
  23. 23.
    Giuditta A, Rutigliano B (2017) Brain metabolic DNA in rat cytoplasm. Mol Neurobiol 55:7476–7486.  https://doi.org/10.1007/s12035-018-0932-0 CrossRefGoogle Scholar
  24. 24.
    Salganik RI, Parvez H, Tomson VP, Shumskaya IA (1983) Probable role of reverse transcription in learning: correlation between hippocampal RNA-dependent DNA synthesis and learning ability in rats. Neurosci Lett 36:317–322CrossRefGoogle Scholar
  25. 25.
    Prisco M, Casalino J, Cefaliello C, Giuditta A(2015) DNA synthesis in mouse brain cytoplasm. Rend Acc Sc fis mat Napoli LXXXII 149-152Google Scholar
  26. 26.
    Cefaliello C, Prisco M, Crispino M, Giuditta A (2018) DNA in squid synaptosomes. Mol Neurobiol 56:56–60.  https://doi.org/10.1007/s12035-018-1071-3 CrossRefGoogle Scholar
  27. 27.
    Cefaliello C, Prisco M, Crispino M, Giuditta A (2015) Newly synthesized DNA in squid nerve terminals. Rend Acc Sci fis mat LXXXII 61-64Google Scholar
  28. 28.
    Eyman M, Cefaliello C, Ferrara E, de Stefano R, Lavina ZS, Crispino M, Squillace A, van Minnen J et al (2007) Local synthesis of axonal and presynaptic RNA in squid model systems. Eur J Neurosci 25:341–350CrossRefGoogle Scholar
  29. 29.
    Crispino M, Chun JT, Cefaliello C, Perrone Capano C, Giuditta A (2014) Local gene expression in nerve endings. Dev Neurobiol 74:279–291CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Biology DepartmentUniversity of Naples Federico IINaplesItaly
  2. 2.Department of NeurologyUniversity of Massachusetts Medical SchoolWorcesterUSA
  3. 3.Accademia di Scienze Fisiche e MatematicheNaplesItaly

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