Abstract
MicroRNAs (miRNAs) are emerging as the most potential regulator of neuronal development. Recent studies from our lab and elsewhere have demonstrated a direct role of miRNAs in regulating neuronal differentiation and synaptogenesis. MicroRNA-145, a miRNA identified to regulate pluripotency of stem cells, downregulates the protein levels of reprogramming transcription factors (RTFs) like OCT4, SOX2, and KLF4 (cell, 137,647–658,2009). Studies have shown that miR-145 is multifunctional and crucial for fate determination of neurons. In our recently published study, we have identified a set of miRNAs including miR-145 and miR-29b families differentially expressed in SH-SY5Y cells exposed sequentially with retinoic acid + brain-derived neurotrophic factor (RA+BDNF) for differentiation into mature neurons (Mol Neurobiol (2016) doi:https://doi.org/10.1007/s12035-016-0042-9). In the present study, we have identified the role of miR-29b in upregulation of miR-145, which is upregulated after exposure of RA+BDNF in a P53-dependent manner. In differentiating SH-SY5Y cells, expression of miR-29b downregulates expression of P85α, a P53 inhibitor, which results in upregulation of miR-145 and downregulation of RTF proteins. Ectopic expression of miR-145 and miR-29b in amounts equivalent to their endogenous expression has induced G1 phase cell cycle arrest. In conclusion, our studies have identified miR-29b as an upstream regulator of miR-145 and targets its RTF genes during differentiation of SH-SY5Y cells.
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03 July 2019
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03 July 2019
The original version of this article unfortunately contained an error. In Figure 8, the image under section a) NTC, and b) NTC���+���PFT�� are copied by mistake.
Abbreviations
- BDNF:
-
brain-derived neurotrophic factor
- DTT:
-
Dithiothreitol
- HPRT:
-
hypoxanthine-guanine phosphoribosyltransferase
- IDV:
-
integrated density value
- miRNA:
-
microRNA
- mRNA:
-
messenger RNA
- PCR:
-
polymerase chain reaction
- RA:
-
retinoic acid
- RTF:
-
reprogramming transcription factor
- RQ:
-
relative quantification
- RT:
-
reverse transcription
- IR:
-
infrared
- PBS:
-
phosphate buffer saline
- PI:
-
propidium iodide
- PLL:
-
poly-l-lysine
- PVDF:
-
polyvinylidene fluoride
References
Coolen M, Bally-Cuif L (2009) MicroRNAs in brain development and physiology. Curr Opin Neurobiol 19(5):461–470
Petri R, Malmevik J, Fasching L, Åkerblom M, Jakobsson J (2014) miRNAs in brain development. Exp Cell Res 321(1):84–89
Singh T, Jauhari A, Pandey A, Singh P, B Pant A, Parmar D, Yadav S (2014) Regulatory triangle of neurodegeneration, adult neurogenesis and microRNAs. CNS Neurol Disord Drug Targets 13(1):96–103
Stiles J, Jernigan TL (2010) The basics of brain development. Neuropsychol Rev 20(4):327–348
Pandey A, Singh P, Jauhari A, Singh T, Khan F, Pant AB, Parmar D, Yadav S (2015) Critical role of the miR-200 family in regulating differentiation and proliferation of neurons. J Neurochem 133(5):640–652
Jauhari A, Singh T, Pandey A, Singh P, Singh N, Srivastava AK, Pant AB, Parmar D et al (2016) Differentiation induces dramatic changes in miRNA profile, where loss of dicer diverts differentiating SH-SY5Y cells toward senescence. Mol Neurobiol:1–10
Le MT, Xie H, Zhou B, Chia PH, Rizk P, Um M, Udolph G, Yang H et al (2009) MicroRNA-125b promotes neuronal differentiation in human cells by repressing multiple targets. Mol Cell Biol 29(19):5290–5305
Makeyev EV, Zhang J, Carrasco MA, Maniatis T (2007) The MicroRNA miR-124 promotes neuronal differentiation by triggering brain-specific alternative pre-mRNA splicing. Mol Cell 27(3):435–448
Purvis JE, Karhohs KW, Mock C, Batchelor E, Loewer A, Lahav G (2012) p53 dynamics control cell fate. Science 336(6087):1440–1444
Quadrato G, Di Giovanni S (2012) Gatekeeper between quiescence and differentiation: p53 in axonal outgrowth and neurogenesis. Int Rev Neurobiol 105:71–89
Xu N, Papagiannakopoulos T, Pan G, Thomson JA, Kosik KS (2009) MicroRNA-145 regulates OCT4, SOX2, and KLF4 and represses pluripotency in human embryonic stem cells. Cell 137(4):647–658
Roshan R, Shridhar S, Sarangdhar MA, Banik A, Chawla M, Garg M, Singh VP, Pillai B (2014) Brain-specific knockdown of miR-29 results in neuronal cell death and ataxia in mice. RNA 20(8):1287–1297
Yang G, Song Y, Zhou X, Deng Y, Liu T, Weng G, Yu D, Pan S (2015) MicroRNA-29c targets β-site amyloid precursor protein-cleaving enzyme 1 and has a neuroprotective role in vitro and in vivo. Mol Med Rep 12(2):3081–3088
Ripa R, Dolfi L, Terrigno M, Pandolfini L, Savino A, Arcucci V, Groth M, Tozzini ET et al (2017) MicroRNA miR-29 controls a compensatory response to limit neuronal iron accumulation during adult life and aging. BMC Biol 15(1):9
Ouyang YB, Xu L, Lu Y, Sun X, Yue S, Xiong XX, Giffard RG (2013) Astrocyte-enriched miR-29a targets PUMA and reduces neuronal vulnerability to forebrain ischemia. Glia 61(11):1784–1794
Hébert SS, Horré K, Nicolaï L, Papadopoulou AS, Mandemakers W, Silahtaroglu AN, Kauppinen S, Delacourte A et al (2008) Loss of microRNA cluster miR-29a/b-1 in sporadic Alzheimer’s disease correlates with increased BACE1/β-secretase expression. Proc Natl Acad Sci 105(17):6415–6420
Yadav S,Pandey A,Shukla A, Talwelkar SS,Kumar A, Pant AB,Parmar D, (2011) miR-497 and miR-302b regulate ethanol-induced neuronal cell death through BCL2 protein and cyclin D2. J Biol Chem 286(43):37347–37357
Jin HY, Gonzalez-Martin A, Miletic AV, Lai M, Knight S, Sabouri-Ghomi M, Head SR, Macauley MS et al (2015) Transfection of microRNA mimics should be used with caution. Front Genet 6:340
Jauhari A,Singh T, Singh P, Parmar D, Yadav S, (2018) Regulation of miR-34 family in neuronal development. Mol Neurobiol 55(2):936–945
Pandey A, Jauhari A, Singh T, Singh P, Singh N, Srivastava AK, Khan F, Pant AB, Parmar D, Yadav S (2015) Transactivation of P53 by cypermethrin induced miR-200 and apoptosis in neuronal cells. Toxicol Res 4(6):1578–1586
Anderson P, Kedersha N (2006) RNA granules. J Cell Biol 172(6):803–808
Cherkasov V, Hofmann S, Druffel-Augustin S, Mogk A, Tyedmers J, Stoecklin G, Bukau B (2013) Coordination of translational control and protein homeostasis during severe heat stress. Curr Biol 23(24):2452–2462
Farny NG, Kedersha NL, Silver PA (2009) Metazoan stress granule assembly is mediated by P-eIF2α-dependent and independent mechanisms. RNA 15(10):1814–1821
Riback JA, Katanski CD, Kear-Scott JL, Pilipenko EV, Rojek AE, Sosnick TR, Drummond DA (2017) Stress-triggered phase separation is an adaptive, evolutionarily tuned response. Cell 168(6):1028–1040. e1019
Fenn AM, Smith KM, Lovett-Racke AE, Guerau-de-Arellano M, Whitacre CC, Godbout JP (2013) Increased micro-RNA 29b in the aged brain correlates with the reduction of insulin-like growth factor-1 and fractalkine ligand. Neurobiol Aging 34(12):2748–2758
Kole AJ, Swahari V, Hammond SM, Deshmukh M (2011) miR-29b is activated during neuronal maturation and targets BH3-only genes to restrict apoptosis. Genes Dev 25(2):125–130
Park S-Y, Lee JH, Ha M, Nam J-W, Kim VN (2009) miR-29 miRNAs activate p53 by targeting p85α and CDC42. Nat Struct Mol Biol 16(1):23–29
Gil-Perotin S, Haines JD, Kaur J, Marin-Husstege M, Spinetta MJ, Kim KH, Duran-Moreno M, Schallert T et al (2011) Roles of p53 and p27 Kip1 in the regulation of neurogenesis in the murine adult subventricular zone. Eur J Neurosci 34(7):1040–1052
Zheng H, Ying H, Yan H, Kimmelman AC, Hiller DJ, Chen A-J, Perry SR, Tonon G et al (2008) p53 and Pten control neural and glioma stem/progenitor cell renewal and differentiation. Nature 455(7216):1129–1133
Meletis K, Wirta V, Hede S-M, Nistér M, Lundeberg J, Frisén J (2006) p53 suppresses the self-renewal of adult neural stem cells. Development 133(2):363–369
Armesilla-Diaz A, Bragado P, Del Valle I, Cuevas E, Lazaro I, Martin C, Cigudosa J, Silva A (2009) p53 regulates the self-renewal and differentiation of neural precursors. Neuroscience 158(4):1378–1389
Lookeren Campagne MV, Gill R (1998) Tumor-suppressor p53 is expressed in proliferating and newly formed neurons of the embryonic and postnatal rat brain: Comparison with expression of the cell cycle regulators p21Waf1/Cip1, p27Kip1, p57Kip2, p16Ink4a, cyclin G1, and the proto-oncogene bax. J Comp Neurol 397(2):181–198
Suzuki HI, Yamagata K, Sugimoto K, Iwamoto T, Kato S, Miyazono K (2009) Modulation of microRNA processing by p53. Nature 460(7254):529–533
Barros R, Pereira D, Callé C, Camilo V, Cunha AI, David L, Almeida R, Dias-Pereira A et al (2016) Dynamics of SOX2 and CDX2 expression in Barrett’s mucosa. Dis Markers 2016:1–7
Zou G, Liu T, Guo L, Huang Y, Feng Y, Huang Q, Duan T (2016) miR-145 modulates lncRNA-ROR and Sox2 expression to maintain human amniotic epithelial stem cell pluripotency and β islet-like cell differentiation efficiency. Gene 591(1):48–57
Morgado AL, Rodrigues CM, Solá S (2016) MicroRNA-145 regulates neural stem cell differentiation through the Sox2–Lin28/let-7 signaling pathway. Stem Cells 34(5):1386–1395
Ozen M, Karatas OF, Gulluoglu S, Bayrak OF, Sevli S, Guzel E, Ekici ID, Caskurlu T et al (2015) Overexpression of miR-145–5p inhibits proliferation of prostate cancer cells and reduces SOX2 expression. Cancer Investig 33(6):251–258
Liu X, Huang J, Chen T, Wang Y, Xin S, Li J, Pei G, Kang J (2008) Yamanaka factors critically regulate the developmental signaling network in mouse embryonic stem cells. Cell Res 18(12):1177–1189
Nolan K, Mitchem MR, Jimenez-Mateos EM, Henshall DC, Concannon CG, Prehn JH (2014) Increased expression of microRNA-29a in ALS mice: functional analysis of its inhibition. J Mol Neurosci 53(2):231–241
Annis RP, Swahari V, Nakamura A, Xie AX, Hammond SM, Deshmukh M (2016) Mature neurons dynamically restrict apoptosis via redundant pre-mitochondrial brakes. FEBS J 283:4569–4582
Acknowledgments
Mr. Abhishek Jauhari is grateful to UGC, New Delhi, and Ms. Tanisha Singh is grateful to DST, New Delhi, for providing research fellowships. The technical assistance of Mr. B S Pandey and Mr. Puneet Khare is also gratefully acknowledged. The CSIR-IITR communication reference number is 3493.
Funding
Funding for the work carried out in the present study had been provided by the CSIR network project (miND).
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Jauhari, A., Singh, T. & Yadav, S. Expression of miR-145 and Its Target Proteins Are Regulated by miR-29b in Differentiated Neurons. Mol Neurobiol 55, 8978–8990 (2018). https://doi.org/10.1007/s12035-018-1009-9
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DOI: https://doi.org/10.1007/s12035-018-1009-9