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Uptake and distribution of fluorescein-labeled D2 dopamine receptor antisense oligodeoxynucleotide in mouse brain

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Abstract

To determine the uptake and distribution of oligodeoxynucleotides in brain, a 20-mer phosphorothioated oligodeoxynucleotide complementary to a portion of the D2 dopamine receptor mRNA was fluorescently labeled with fluorescein isothiocyanate (FITC) and injected into the lateral cerebral ventricles of mice. At various survival times after the injection, the brains were removed, fixed, sectioned, and viewed under a fluorescent microscope. The results showed that the oligodeoxynucleotide was rapidly taken up into the brain. Initially the label was relatively diffusely spread throughout the interstitial spaces of the brain, then became redistributed to the cellular compartments. The signal extended from those forebrain nuclei located immediately in contact with the ventricles, such as the corpus striatum, septum, and hippocampus, to areas further removed from the ventricles, such as the cerebral cortex, nucleus accumbens, and substantia nigra. When the FITC-labeled D2 antisense oligodeoxynucleotide was given once daily for 4 d, the signal intensity seen 24 h after the last injection appeared to be of greater intensity overall compared to that seen after a single injection. At early time-points the oligodeoxynucleotide signals appeared to be punctuated and were found in cell bodies as well as in proximal dendritic processes. However, not all cells were equally labeled, suggesting an uneven uptake and accumulation of the D2 antisense into the various cell types. At later time-points the fluorescent signal appeared granular; at these times the injected material was largely degraded. These studies show that a D2 dopamine receptor antisense oligodeoxynucleotide is rapidly taken up from cerebral ventricles into brain, becomes widely distributed throughout the brain tissue to areas far removed from direct contact with the ventricles, and appears to accumulate to a different extent in the various brain areas and cell types.

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References

  • Adams J. U., Chen X. H., DeRiel J. K., Adler M. W., and Liu-Chen L.-Y. (1994)In vivo treatment with antisense oligodeoxynucleotide to kappa-opioid receptors inhibited kappa-agonist-induced analgesia in rats.Regul. Pept. 54, 1,2.

    Article  CAS  Google Scholar 

  • Agrawal S. and Iyer R. P. (1995) Modified oligonucleotides as therapeutic and diagnostic agents.Curr. Opin. Biotechnol. 6, 12–19.

    Article  PubMed  CAS  Google Scholar 

  • Agrawal S., Temsamani J., Galbraith W., and Tang J. (1995) Pharmacokinetics of antisense oligonucleotides.Clin. Pharmacokinet. 28, 7–16.

    Article  PubMed  CAS  Google Scholar 

  • Agrawal S., Temsamani J., and Tang J. Y. (1991) Pharmacokinetics, biodistribution, and stability of oligodeoxynucleotide phosphorothioates in mice.Proc. Natl. Acad. Sci. USA 88, 7595–7599.

    Article  PubMed  CAS  Google Scholar 

  • Beltinger C., Saragovi H. U., Smith R. M., LeSauteur L., Shah N., DeDionisio L., Christensen L., Raible A., Jarett L., and Gewirtz A. M. (1995) Binding, uptake, and intracellular trafficking of phosphorothioate-modified oligodeoxynucleotides.J. Clin. Invest. 95, 1814–1823.

    Article  PubMed  CAS  Google Scholar 

  • Caceres A. and Kosik K. S. (1990) Inhibition of neurite polarity by tau antisense oligonucleotides in primary cerebellar neurons.Nature 343, 461–463.

    Article  PubMed  CAS  Google Scholar 

  • Crooke S. T. (1992) Therapeutic applications of oligonucleotides.Ann. Rev. Pharmacol. Toxicol. 32, 329–376.

    Article  CAS  Google Scholar 

  • Goodale D. B., Jacobi A. G. M., Winkler J. D., and Weiss B. (1985) Rapid stereotaxic injections into mouse striatum using a plastic mold.Soc. Neurosci. 11, 772.

    Google Scholar 

  • Iversen P. (1991) In vivo studies with phosphorothioate oligonucleotides: pharmacokinetics prologue.Anti-Cancer Drug Design 6, 531–538.

    PubMed  CAS  Google Scholar 

  • Iwanaga T. and Ferriola P. C. (1993) Cellular uptake of phosphorothioate oligodeoxynucleotides is negatively affected by cell density in a transformed rat tracheal epithelial cell line: implication for antisense approaches.Biochem. Biophys. Res. Commun. 191, 1152–1157.

    Article  PubMed  CAS  Google Scholar 

  • Loke S. L., Stein C. A., Zhang X. H., Mori K., Nakanishi M., Subasinghe C., Cohen J. S., and Neckers L. M. (1989) Characterization of oligonucleotide transport into living cells.Proc. Natl. Acad. Sci. USA 86, 3474–3478.

    Article  PubMed  CAS  Google Scholar 

  • Lu X.-M., Fischman A. J., Jyawook S. L., Hendricks K., Tompkins R. G., and Yarmush M. L. (1994) Antisense DNA delivery in vivo: liver targeting by receptor-mediated uptake.J. Nucl. Med. 35, 269–275.

    PubMed  CAS  Google Scholar 

  • McCarthy M. M., Kleopoulos S. P., Mobbs C. V., and Pfaff D. W. (1994a) Infusion of antisense oligodeoxynucleotides to the oxytocin receptor in the ventromedial hypothalamus reduces estrogen-induced sexual receptivity and oxytocin receptor binding in the female rat.Neuroendocrinology 59, 432–440.

    PubMed  CAS  Google Scholar 

  • McCarthy M. M., Masters D. B., Rimvall K., Schwartz-Giblin S., and Pfaff D. W. (1994b) Intracerebral administration of antisense oligodeoxynucleotides to GAD65 and GAD67 mRNAs modulate reproductive behavior in the female rat.Brain Res. 636, 209–220.

    Article  PubMed  CAS  Google Scholar 

  • Meeker R., LeGrand G., Ramirez J., Smith T., and Shih Y.-H. (1995) Antisense vasopressin oligonucleotides: uptake, turnover, distribution, toxicity and behavioral effects.J. Neuroendocrinol. 7, 419–428.

    Article  PubMed  CAS  Google Scholar 

  • Meng H., Wielbo D., Gyurko R., and Phillips M. I. (1994) Antisense oligonucleotide to AT1 receptor mRNA inhibits central angiotensin induced thirst and vasopressin.Regul. Pept. 54, 543–551.

    Article  PubMed  CAS  Google Scholar 

  • Neckers L., Rosolen A., Fahmy B., and Whitesell L. (1992) Specific inhibition of oncogene expressionin vitro andin vivo by antisense oligonucleotides.Ann. NY Acad. Sci. 660, 37–44.

    Article  PubMed  CAS  Google Scholar 

  • Ogawa S., Brown H. E., Okano H. J., and Pfaff D. W. (1995) Cellular uptake of intracerebrally administered oligodeoxynucleotides in mouse brain.Regul. Pept. 59, 143–149.

    Article  PubMed  CAS  Google Scholar 

  • Pilowsky P. M., Suzuki S., and Minson J. B. (1994) Antisense oligonucleotides: a new tool in neuro-science.Clin. Exp. Pharmacol. Physiol. 21, 935–944.

    Article  PubMed  CAS  Google Scholar 

  • Piwnica-Worms D. (1994) Making sense out of antisense: challenges of imaging gene translation with radiolabeled oligonucleotides.J. Nucl. Med. 35, 1064–1066.

    PubMed  CAS  Google Scholar 

  • Sakai R. R., He P. F., Yang X. D., Ma L. Y., Guo Y. F., Reilly J. J., Moga C. N., and Fluharty S. J. (1994) Intracerebroventricular administration of AT1 receptor antisense oligonucleotides inhibits the behavioral actions of angiotensin II.J. Neurochem. 62, 2053–2056.

    Article  PubMed  CAS  Google Scholar 

  • Sands H., Gorey-Feret L. J., Cocuzza A. J., Hobbs F. W., Chidester D., and Trainor G. L. (1994) Biodistribution and metabolism of internally3H-labeled oligonucleotides. I. Comparison of a phosphodiester and a phosphorothioate.Mol. Pharmacol. 45, 932–943.

    PubMed  CAS  Google Scholar 

  • Schlingensiepen K.-H., Wollnik F., Kunst M., Schlingensiepen R., Herdegen T., and Brysch W. (1994) The role of jun transcription factor expression and phosphorylation in neuronal differentiation, neuronal cell death, and plastic adaptations in vivo.Cell. Mol. Neurobiol. 14, 487–505.

    Article  PubMed  CAS  Google Scholar 

  • Simons M., Edelman E. R., DeKeyser J.-L., Langer R., and Rosenberg R. D. (1992) Antisense c-myb oligonucleotides inhibit intimal arterial smooth muscle cell accumulation in vivo.Nature 359, 67–70.

    Article  PubMed  CAS  Google Scholar 

  • Slotnick B. M. and Leonard C. M. (1975)A Stereotaxic Atlas of the Albino Mouse Forebrain. U.S. Dept. Health, Education, and Welfare, Rockville, MD, pp. 32–109.

    Google Scholar 

  • Sommer W., Bjelke B., Ganten D., and Fuxe K. (1993) Antisense oligonucleotide to c-fos induces ipsilateral rotational behaviour tod-amphetamine.Neuro-Report 5, 277–280.

    CAS  Google Scholar 

  • Standifer K. M., Chien C.-C., Wahlestedt C., Brown G. P., and Pasternak G. W. (1994) Selective loss of δ opioid analgesia and binding by antisense oligodeoxynucleotides to a δ opioid receptor.Neuron 12, 805–810.

    Article  PubMed  CAS  Google Scholar 

  • Stein C. A. and Cheng Y.-C. (1993) Antisense oligonucleotides as therapeutic agents—Is the bullet really magical.Science 261, 1004–1012.

    Article  PubMed  CAS  Google Scholar 

  • Stein C. A., Tonkinson J. L., and Yakubov L. (1991) Phosphorothioate oligodeoxynucleotides—antisense inhibitors of gene expression?Pharmacol. Ther. 52, 365–384.

    Article  PubMed  CAS  Google Scholar 

  • Thierry A. R. and Dritschilo A. (1992) Intracellular availability of unmodified, phosphorothioated and liposomally encapsulated oligodeoxynucleotides for antisense activity.Nucleic Acids Res. 20, 5691–5698.

    Article  PubMed  CAS  Google Scholar 

  • Wahlestedt C., Golanov E., Yamamoto S., Yee F., Ericson H., Yoo H., Inturrisi C. E., and Reis D. J. (1993a) Antisense oligodeoxynucleotides to NMDA-R1 receptor channel protect cortical neurons from excitotoxicity and reduce focal ischaemic infarctions.Nature 363, 260–263.

    Article  PubMed  CAS  Google Scholar 

  • Wahlestedt C., Pich E. M., Koob G. F., Yee F., and Heilig M. (1993b) Modulation of anxiety and neuropeptide Y-Y1 receptors by antisense oligodeoxynucleotides.Science 259, 528–531.

    Article  PubMed  CAS  Google Scholar 

  • Weiss B., Zhou L.-W., Zhang S.-P., and Qin Z.-H. (1993) Antisense oligodeoxynucleotide inhibits D2 dopamine receptor-mediated behavior and D2 messenger RNA.Neuroscience 55, 607–612.

    Article  PubMed  CAS  Google Scholar 

  • Whitesell L., Geselowitz D., Chavany C., Fahmy B., Walbridge S., Alger J. R., and Neckers L. M. (1993) Stability, clearance, and disposition of intraven-tricularly administered oligodeoxynucleotides: implications for therapeutic application within the central nervous system.Proc. Natl. Acad. Sci. USA 90, 4665–4669.

    Article  PubMed  CAS  Google Scholar 

  • Yakubov L. A., Deeva E. A., Zarytova V. F., Ivanova E. M., Ryte A. S., Yurchenko L. V., and Vlassov V. V. (1989) Mechanism of oligonucleotide uptake by cells: involvement of specific receptors?Proc. Natl. Acad. Sci. USA 86, 6454–6458.

    Article  PubMed  CAS  Google Scholar 

  • Yee F., Ericson H., Reis D. J., and Wahlestedt C. (1994) Cellular uptake of intracerebroventricularly administered biotin- or digoxigenin-labeled antisense oligodeoxynucleotides in the rat.Cell. Mol. Neurobiol. 14, 475–486.

    Article  PubMed  CAS  Google Scholar 

  • Zhang M. and Creese I. (1993) Antisense oligodeoxynucleotide reduces brain dopamine D2 receptors: behavioral correlates.Neurosci. Lett. 161, 223–226.

    Article  PubMed  CAS  Google Scholar 

  • Zhang S.-P., Zhou L.-W., and Weiss B. (1994) Oligodeoxynucleotide antisense to the D1 dopamine receptor mRNA inhibits D1 dopamine receptor-mediated behaviors in normal mice and in mice lesioned with 6-hydroxydopamine.J. Pharmacol. Exp. Ther. 271, 1462–1470.

    PubMed  CAS  Google Scholar 

  • Zhou L.-W., Zhang S.-P., Qin Z.-H., and Weiss B. (1994) In vivo administration of an oligodeoxy-nucleotide antisense to the D2 dopamine receptor mRNA inhibits D2 dopamine receptor-mediated behavior and the expression of D2 dopamine receptors in mouse striatum.J. Pharmacol. Exp. Ther. 268, 1015–1023.

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Pharmacology, Medical College of Pennsylvania and Hahnemann University, 3200 Henry Avenue, 19129, Philadelphia, PA

    Sui-Po Zhang, Long-Wu Zhou, Mark Morabito & Benjamin Weiss

  2. Department of Anatomy & Neurobiology, Medical College of Pennsylvania and Hahnemann University, 3200 Henry Avenue, 19129, Philadelphia, PA

    Rick C. S. Lin

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  1. Sui-Po Zhang
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Zhang, SP., Zhou, LW., Morabito, M. et al. Uptake and distribution of fluorescein-labeled D2 dopamine receptor antisense oligodeoxynucleotide in mouse brain. J Mol Neurosci 7, 13–28 (1996). https://doi.org/10.1007/BF02736845

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  • DOI: https://doi.org/10.1007/BF02736845

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