Atropa belladonna Expresses a microRNA (aba-miRNA-9497) Highly Homologous to Homo sapiens miRNA-378 (hsa-miRNA-378); both miRNAs target the 3′-Untranslated Region (3′-UTR) of the mRNA Encoding the Neurologically Relevant, Zinc-Finger Transcription Factor ZNF-691

  • Bihter Avsar
  • Yuhai Zhao
  • Wenhong Li
  • Walter J. LukiwEmail author
Brief Communication


Recent advances in ethnobotanical and neurological research indicate that ingested plants from our diet may not only be a source of nutrition but also a source of biologically relevant nucleic-acid-encoded genetic information. A major source of RNA-encoded information from plants has been shown to be derived from small non-coding RNAs (sncRNAs) such as microRNAs (miRNAs) that can transfer information horizontally between plants and humans. In human hosts, the 3′-untranslated region (3′-UTR) of messenger RNAs (mRNAs) is targeted by these miRNAs to effectively down-regulate expression of that mRNA target in the host CNS. In this paper, we provide evidence that the Atropa belladonna aba-miRNA-9497 (miRBase conserved ID: bdi-miRNA-9497) is highly homologous to the CNS-abundant Homo sapiens miRNA-378 (hsa-miRNA-378) and both target the zinc-finger transcription factor ZNF-691 mRNA 3′-UTR to down-regulate ZNF-691 mRNA abundance. We speculate that the potent neurotoxic actions of the multiple tropane alkaloids of Atropa belladonna may be supplemented by the neuroregulatory actions of aba-miRNA-9497 on ZNF-691, and this may be followed by the modulation in the expression of ZNF-691-sensitive genes. This is the first example of a human brain-enriched transcription factor, ZNF-691, targeted and down-regulated by a naturally occurring plant microRNA, with potential to modulate gene expression in the human CNS and thus contribute to the neurotoxicological-and-psychoactive properties of the Atropa belladonna species of the deadly nightshade Solanaceae family.


Atropa belladonna Atropine aba-miRNA-9497 hsa-miRNA-378 Trans-kingdom miRNA signaling Zinc-finger protein ZNF-691 



The research in this ‘Perspectives’ article was presented in part at the Vavilov Institute of General Genetics Autumn 2018 Seminar Series (Инcтитyт oбщeй гeнeтики имeни Baвилoвa Oceнь 2018 Ceминap cepии) in Moscow, Russia, October 2018, at the Society for Neuroscience (SFN) Annual Meeting, San Diego USA, November 2018. Sincere thanks are extended to Drs. L. Carver, L. Cong, F. Culicchia, C. Eicken, K. Navel, A.I. Pogue, W. Poon, and the late Drs. J.M. Hill, P.N. Alexandrov, D.R.C. McLachlan, and T.P.A. Kruck for helpful discussions in this research area, for short post-mortem interval (PMI) human brain and retinal tissues or extracts, for initial bioinformatics and data interpretation, and to A.I. Pogue and D. Guillot for expert technical assistance and medical artwork. We would like to further thank the following brain and tissue banks for access to high-quality post-mortem tissues and valuable analytical advice: the National Institute of Neurological Disorders and Stroke (NINDS), Bethesda MD USA; the Oregon Health Sciences University, Portland OR, USA; the Southern Eye Bank, Metairie LA, USA; the University of California (UCI), Irvine CA, USA; and the many neuropathologists, physicians, and researchers in the US and Canada who have provided high-quality, short PMI human brain tissue fractions for scientific analysis. Research on microRNAs, ethnobiology, botanical neurotoxins, pro-inflammatory, and pathogenic signaling in the Lukiw laboratory involving the microbiome, the innate-immune response, amyloidogenesis, synaptogenesis, and neuroinflammation in AD, prion, and in other human neurological- and plant-viroid-based diseases was supported through an unrestricted grant to the LSU Eye Center from Research to Prevent Blindness (RPB); the Louisiana Biotechnology Research Network (LBRN) and NIH grants NEI EY006311, NIA AG18031, and NIA AG038834 (WJL). The content of this manuscript is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute on Aging, the National Center for Research Resources, or the National Institutes of Health.

Author Contributions

BA performed the original microRNA sequence analysis on Atropa belladonna; BA, WL, and WJL collected, analyzed, and performed bioinformatics analysis and summarized the data and reviewed the current neurologically relevant miRNA literature; YZ, WL, and WJL performed the experiments and were involved in additional data extraction and bioinformatics; WJL wrote the article.

Compliance with Ethical Standards

Conflict of interest

All authors declare that they have no conflict of interest in any of the experiments performed.

Ethical Approval

This article does not contain any studies with live human participants or animals performed by any of the authors of this manuscript. Human neuronal-glial (HNG) primary cells (Fig. 2e) for all transfection experiments were obtained from commercial sources (Lonza Biosciences, Houston, TX USA); brain gene expression data were derived from archived sources and the references listed in this manuscript; all human cell acquisition and handling procedures involving HNG cells were carried out in strict accordance with the ethics review board policies at donor institutions. The experimental work in this investigative study was reviewed and approved by the Institutional Biosafety Committee/Institutional Review Board (IBC/IRB) in strict accordance with the Institutional Biosafety Committee and the Institutional Review Board Committee (IBC/IRBC) ethical guidelines IBC#18059 and IRBC#6774 at the Louisiana State University Health Sciences Center, New Orleans LA 70112 USA.

Supplementary material

10571_2019_729_MOESM1_ESM.xlsx (14 kb)
Supplementary material 1 (XLSX 14 kb)


  1. Agarwal V, Bell GW, Nam Jin-Wu, Bartel DP (2015) Predicting effective microRNA target sites in mammalian mRNAs. Elife 4:e05005CrossRefPubMedCentralGoogle Scholar
  2. Alexandrov PN, Dua P, Hill JM, Bhattacharjee S, Zhao Y, Lukiw WJ (2012) microRNA (miRNA) speciation in Alzheimer’s disease (AD) cerebrospinal fluid (CSF) and extracellular fluid (ECF). Int J Biochem Mol Biol 3:365–373PubMedPubMedCentralGoogle Scholar
  3. Ambros V (2008) The evolution of our thinking about microRNAs. Nat Med 14:1036–1040. CrossRefPubMedGoogle Scholar
  4. Arteaga-Vázquez M, Caballero-Pérez J, Vielle-Calzada JP (2006) A family of microRNAs present in plants and animals. Plant Cell 18:3355–3369CrossRefPubMedPubMedCentralGoogle Scholar
  5. Avsar B, Aliabadi DE (2018) In silico identification of microRNAs in 13 medicinal plants. Turk J Biochem 42(s1):57Google Scholar
  6. Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281–297CrossRefPubMedGoogle Scholar
  7. Bartel DP (2009) MicroRNAs: target recognition and regulatory functions. Cell 136:215–233. CrossRefPubMedPubMedCentralGoogle Scholar
  8. Cai Q, He B, Kogel KH, Jin H (2018) Cross-kingdom RNA trafficking and environmental RNAi-nature’s blueprint for modern crop protection strategies. Curr Opin Microbiol 46:58–64. CrossRefPubMedPubMedCentralGoogle Scholar
  9. Caksen H, Odabaş D, Akbayram S, Cesur Y, Arslan S, Uner A, Oner AF (2003) Deadly nightshade (Atropa belladonna) intoxication: an analysis of 49 children. Hum Exp Toxicol 22:665–668. CrossRefPubMedGoogle Scholar
  10. Cassandri M, Smirnov A, Novelli F, Pitolli C, Agostini M, Malewicz M et al (2017) Zinc-finger proteins in health and disease. Cell Death Discov 3:17071. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Cong L, Zhao Y, Pogue AI, Lukiw WJ (2018) Role of microRNA (miRNA) and viroids in lethal diseases of plants and animals. Potential contribution to human neurodegenerative disorders. Biochemistry 83:1018–1029. CrossRefPubMedGoogle Scholar
  12. Diener TO (2016) Viroids: “living fossils” of primordial RNAs? Biol Direct 11:15. CrossRefPubMedPubMedCentralGoogle Scholar
  13. Djami-Tchatchou AT, Sanan-Mishra N, Ntushelo K, Dubery IA (2017) Functional roles of microRNAs in agronomically important plants-potential as targets for crop improvement and protection. Front Plant Sci 8:378. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Du J, Liang Z, Xu J, Zhao Y, Li X, Zhang Y et al (2019) Plant-derived phosphocholine facilitates cellular uptake of anti-pulmonary fibrotic HJT-sRNA-m7. Sci China Life Sci 62:309–320. CrossRefPubMedGoogle Scholar
  15. Fagerberg L, Hallström BM, Oksvold P, Kampf C, Djureinovic D, Odeberg J et al (2014) Analysis of the human tissue-specific expression by genome-wide integration of transcriptomics and antibody-based proteomics. Mol Cell Proteomics 13:397–406. CrossRefPubMedGoogle Scholar
  16. Guo H, Ingolia NT, Weissman JS, Bartel DP (2010) Mammalian microRNAs predominantly act to decrease target mRNA levels. Nature 466:835–840CrossRefPubMedPubMedCentralGoogle Scholar
  17. Hatayama M, Aruga J (2018) Role of zinc family proteins in transcriptional regulation and chromatin remodeling. Adv Exp Med Biol 1046:353–380. CrossRefPubMedGoogle Scholar
  18. Hill JM, Lukiw WJ (2014) Comparing miRNAs and viroids; highly conserved molecular mechanisms for the transmission of genetic information. Front Cell Neurosci 8:45. CrossRefPubMedPubMedCentralGoogle Scholar
  19. Hill JM, Lukiw WJ (2015) Microbial-generated amyloids and Alzheimer’s disease (AD). Front Aging Neurosci 7:9CrossRefPubMedPubMedCentralGoogle Scholar
  20. Hill JM, Lukiw WJ (2016) MicroRNA (miRNA)-mediated pathogenetic signaling in Alzheimer’s disease (AD). Neurochem Res 41:96–100. CrossRefPubMedGoogle Scholar
  21. Hill JM, Zhao Y, Bhattacharjee S, Lukiw WJ (2014) miRNAs and viroids utilize common strategies in genetic signal transfer. Front Mol Neurosci 7:10CrossRefPubMedPubMedCentralGoogle Scholar
  22. Hu H, Rashotte AM, Singh NK, Weaver DB, Goertzen LR, Singh SR, Locy RD (2015) The complexity of posttranscriptional small RNA regulatory networks revealed by in silico analysis of Gossypium arboreum L. leaf, flower and boll small regulatory RNAs. PLoS ONE 10(6):e0127468CrossRefPubMedPubMedCentralGoogle Scholar
  23. Huang F, Du J, Liang Z, Xu Z, Xu J, Zhao Y et al (2018) Large-scale analysis of small RNAs derived from traditional Chinese herbs in human tissues. Sci China Life Sci. CrossRefPubMedPubMedCentralGoogle Scholar
  24. Jaber V, Zhao Y, Lukiw WJ (2017) Alterations in micro RNA-messenger RNA (miRNA-mRNA) coupled signaling networks in sporadic Alzheimer’s disease (AD) hippocampal CA1. J Alzheimers Dis Parkinsonism. 7(2):312PubMedPubMedCentralGoogle Scholar
  25. Jones-Rhoades MW, Bartel DP (2004) Computational identification of plant microRNAs and their targets, including a stress-induced miRNA. Mol Cell 14:787–799CrossRefPubMedGoogle Scholar
  26. Kozomara A, Birgaoanu M, Griffiths-Jones S (2019) miRBase: from microRNA sequences to function. Nucleic Acids Res 47:D155–D162. CrossRefPubMedGoogle Scholar
  27. Kwakye GF, Jiménez J, Jiménez JA, Aschner M (2018) Atropa belladonna neurotoxicity: implications to neurological disorders. Food Chem Toxicol 116:346–353. CrossRefPubMedGoogle Scholar
  28. Lagos-Quintana M, Rauhut R, Lendeckel W, Tuschl TI (2001) Identification of novel genes coding for small expressed RNAs. Science 294:853–858CrossRefPubMedGoogle Scholar
  29. Lee MR (2007) Solanacea IV: atropa belladonna, deadly nightshade. J R Coll Phys Edinb. 37:77–84Google Scholar
  30. Li Z, Xu R, Li N (2018) MicroRNAs from plants to animals, do they define a new messenger for communication? Nutr Metab 15:68. CrossRefGoogle Scholar
  31. Lukiw WJ (2007) Micro-RNA speciation in fetal, adult and Alzheimer’s disease hippocampus. NeuroReport 18:297–300CrossRefPubMedGoogle Scholar
  32. Lukiw WJ (2012a) NF-кB-regulated micro RNAs (miRNAs) in primary human brain cells. Exp Neurol 235:484–490. CrossRefPubMedGoogle Scholar
  33. Lukiw WJ (2012b) NF-κB-regulated, pro-inflammatory miRNAs in Alzheimer’s disease. Alzheimers Res Ther 4:47. CrossRefPubMedPubMedCentralGoogle Scholar
  34. Luo Y, Wang P, Wang X, Wang Y, Mu Z, Li Q et al (2017) Detection of dietetically absorbed maize-derived microRNAs in pigs. Sci Rep 7:645CrossRefPubMedPubMedCentralGoogle Scholar
  35. Mackeh R, Marr AK, Fadda A, Kino T (2018) C2H2-type zinc finger proteins: evolutionarily old and new partners of the nuclear hormone receptors. Nucl Recept Signal 15:1550762918801071. CrossRefPubMedGoogle Scholar
  36. Mal C, Deb A, Aftabuddin M, Kundu S (2015) A network analysis of miRNA mediated gene regulation of rice: crosstalk among biological processes. Mol BioSyst 11:2273–2280. CrossRefPubMedGoogle Scholar
  37. Mal C, Aftabuddin M, Kundu S (2018) IIKmTA: inter and intra kingdom miRNA-target analyzer. Interdiscip Sci 10:538–543. CrossRefPubMedGoogle Scholar
  38. Mallinson T (2010) Deadly nightshade: Atropa Belladonna. Focus First Aid (15): 5. Archived from the original on 2010-05-21Google Scholar
  39. Marín-Sáez J, Romero-González R, Garrido Frenich A, Egea-González FJ (2018) Screening of drugs and homeopathic products from Atropa belladonna seed extracts: tropane alkaloids determination and untargeted analysis. Drug Test Anal 10:1579–1589. CrossRefPubMedGoogle Scholar
  40. Miller WA, Shen R, Staplin W, Kanodia P (2016) Noncoding RNAs of plant viruses and viroids: sponges of host translation and RNA interference machinery. Mol Plant Microbe Interact 29:156–164. CrossRefPubMedPubMedCentralGoogle Scholar
  41. miRBase; microRNA database; University of Manchester; (last accessed 12 May 2019)
  42. Moran Y, Agron M, Praher D, Technau U (2017) The evolutionary origin of plant and animal microRNAs. Nat Ecol Evol 1:27. CrossRefPubMedGoogle Scholar
  43. Pant BD, Musialak-Lange M, Nuc P, May P, Buhtz A, Kehr J, Walther D, Scheible WR (2009) Identification of nutrient-responsive Arabidopsis and rapeseed microRNAs by comprehensive real-time polymerase chain reaction profiling and small RNA sequencing. Plant Physiol 150(3):1541–1555. CrossRefPubMedPubMedCentralGoogle Scholar
  44. Pepeu, G., Giovannini M.G. (2004). “Acetylcholine: I. Muscarinic Receptors”. In Gernot Riedel; Bettina Platt. From messengers to molecules: memories are made of these. Springer. ISBN 978-0-306-47862-8Google Scholar
  45. Pogue AI, Clement C, Hill JM, Lukiw WJ (2014a) Evolution of microRNA (miRNA) structure and function in plants and animals: relevance to aging and disease. J Aging Sci 2(2):119PubMedPubMedCentralGoogle Scholar
  46. Pogue AI, Hill JM, Lukiw WJ (2014b) MicroRNA (miRNA): sequence and stability, viroid-like properties, and disease association in the CNS. Brain Res 1584:73–79. CrossRefPubMedGoogle Scholar
  47. Qin X, Chang F, Wang Z, Jiang W (2018) Correlation of circulating pro-angiogenic miRNAs with cardiotoxicity induced by epirubicin/cyclophosphamide followed by docetaxel. Cancer Biomark 23:473–484. CrossRefPubMedGoogle Scholar
  48. Slota JA, Booth SA (2019) MicroRNAs in neuroinflammation: implications in disease pathogenesis, biomarker discovery and therapeutic applications. Noncoding RNA. CrossRefPubMedPubMedCentralGoogle Scholar
  49. Steger G, Perreault JP (2016) Structure and associated biological functions of viroids. Adv Virus Res 94:141–172. CrossRefPubMedGoogle Scholar
  50. Tosar JP, Rovira C, Naya H, Cayota A (2014) Mining of public sequencing databases supports a non-dietary origin for putative foreign miRNAs. RNA 20:754–757. CrossRefPubMedPubMedCentralGoogle Scholar
  51. Ulbricht C, Basch E, Hammerness P, Vora M, Wylie J Jr, Woods J (2004) An evidence-based systematic review of belladonna by the natural standard research collaboration. J Herb Pharmacother 4:61–90CrossRefPubMedGoogle Scholar
  52. Valadi H, Ekström K, Bossios A, Sjöstrand M, Lee JJ, Lötvall JO et al (2007) Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol 9:654–659. CrossRefGoogle Scholar
  53. Xie W, Melzig MF (2018) The stability of medicinal plant microRNAs in the herb preparation process. Molecules 23(4):919CrossRefPubMedCentralGoogle Scholar
  54. Zeng J, Gupta VK, Jiang Y, Yang B, Gong L, Zhu H (2019) Cross-kingdom small RNAs among animals, plants and microbes. Cells 8(4):371CrossRefPubMedCentralGoogle Scholar
  55. Zhang L, Hou D, Chen X, Li D, Zhu L, Zhang Y et al (2012) Exogenous plant MIR168a specifically targets mammalian LDLRAP1: evidence of cross-kingdom regulation by microRNA. Cell Res 22:107–126. CrossRefPubMedGoogle Scholar
  56. Zhao Y, Lukiw WJ (2018) Microbiome-mediated upregulation of microRNA-146a in sporadic Alzheimer’s disease. Front Neurol 9:145. CrossRefPubMedPubMedCentralGoogle Scholar
  57. Zhao Y, Cong L, Lukiw WJ (2018) Plant and animal microRNAs (miRNAs) and their potential for inter-kingdom communication. Cell Mol Neurobiol 38:133–140. CrossRefPubMedGoogle Scholar
  58. Zhao Y, Jaber VR, LeBeauf A, Sharfman NM, Lukiw WJ (2019) microRNA-34a (miRNA-34a) mediated down-regulation of the post-synaptic cytoskeletal element SHANK3 in sporadic Alzheimer’s disease (AD). Front Neurol 10:28. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Bihter Avsar
    • 1
    • 2
  • Yuhai Zhao
    • 3
    • 4
  • Wenhong Li
    • 3
    • 5
  • Walter J. Lukiw
    • 3
    • 6
    • 7
    Email author
  1. 1.Sabanci University SUNUM Nanotechnology Research CentreIstanbulTurkey
  2. 2.Faculty of Arts & Science, Department of Molecular Biology and GeneticsUludag UniversityBursaTurkey
  3. 3.LSU Neuroscience Center, Louisiana State University Health Sciences CenterNew OrleansUSA
  4. 4.Departments of Anatomy and Cell BiologyLouisiana State University Health Sciences CenterNew OrleansUSA
  5. 5.Department of Pharmacology, School of PharmacyJiangxi University of TCMNanchangChina
  6. 6.Department of NeurologyLouisiana State University Health Sciences CenterNew OrleansUSA
  7. 7.Department of OphthalmologyLouisiana State University Health Sciences CenterNew OrleansUSA

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