PMID- 27346077 OWN - NLM STAT- MEDLINE DCOM- 20180112 LR - 20220408 IS - 1095-9327 (Electronic) IS - 1044-7431 (Print) IS - 1044-7431 (Linking) VI - 75 DP - 2016 Sep TI - MicroRNA-1-associated effects of neuron-specific brain-derived neurotrophic factor gene deletion in dorsal root ganglia. PG - 36-43 LID - S1044-7431(16)30052-5 [pii] LID - 10.1016/j.mcn.2016.06.003 [doi] AB - BACKGROUND: MicroRNAs (miRNAs) regulate gene expression in physiological as well as in pathological processes, including chronic pain. Whether deletion of a gene can affect expression of the miRNAs that associate with the deleted gene mRNA remains elusive. We investigated the effects of brain-derived neurotrophic factor (Bdnf) gene deletion on the expression of miR-1 in dorsal root ganglion (DRG) neurons and its pain-associated downstream targets heat shock protein 60 (Hsp60) and connexin 43 (Cx43) in tamoxifen-inducible conditional knockout mice, Bdnf(fl/fl); Advillin-CreER(T2) (Bdnf cKO). RESULTS: Efficient Bdnf gene deletion was confirmed in DRG of Bdnf cKO mice by Real-Time qRT-PCR and ELISA 10days after completed tamoxifen treatment. In DRG, miR-1 expression was reduced 0.44-fold (p<0.05; Real-time qRT-PCR) in Bdnf cKO compared to floxed wildtype littermate control Bdnf(fl/fl) mice (WT). While Hsp60 protein expression was increased 1.85-fold (p<0.05; Western blot analysis), expression levels of Cx43 and the miR-1-associated transcription factors MEF2a and SRF remained unchanged. When analyzing Bdnf cKO mice 32days after complete tamoxifen treatment to investigate whether observed expression alterations remain permanently, we found no significant differences between Bdnf cKO and WT mice. However, miRNA microarray analysis revealed that 167 miRNAs altered (p<0.05) in DRG of these mice following Bdnf gene deletion. CONCLUSIONS: Our results indicate that deletion of Bdnf in DRG neurons leads to a temporary dysregulation of miR-1, suggesting an impairment of a presumable feedback loop between BDNF protein and its targeting miR-1. This appears to affect its downstream protein Hsp60 and as a consequence might influence the phenotype after inducible Bdnf gene deletion. While this appears to be a MEF2a-/SRF-independent and transient effect, expression levels of various other miRNAs may remain permanently altered. CI - Copyright (c) 2016 Elsevier Inc. All rights reserved. FAU - Neumann, Elena AU - Neumann E AD - Department of Anesthesiology, Medical Faculty, Heinrich-Heine-University Dusseldorf, Moorenstr. 5, Dusseldorf 40225, Germany. Electronic address: elena.neumann@med.uni-duesseldorf.de. FAU - Brandenburger, Timo AU - Brandenburger T AD - Department of Anesthesiology, Medical Faculty, Heinrich-Heine-University Dusseldorf, Moorenstr. 5, Dusseldorf 40225, Germany. Electronic address: timo.brandenburger@med.uni-duesseldorf.de. FAU - Santana-Varela, Sonia AU - Santana-Varela S AD - Molecular Nociception Group, Wolfson Institute for Biomedical Research (WIBR), Cruciform Building, University College London (UCL), London WC1E 6BT, UK. Electronic address: s.santana@ucl.ac.uk. FAU - Deenen, Rene AU - Deenen R AD - Biological and Medical Research Center (BMFZ), Medical Faculty, Heinrich-Heine-University Dusseldorf, Universitatsstrasse 1, Dusseldorf 40225, Germany. Electronic address: rene.deenen@hhu.de. FAU - Kohrer, Karl AU - Kohrer K AD - Biological and Medical Research Center (BMFZ), Medical Faculty, Heinrich-Heine-University Dusseldorf, Universitatsstrasse 1, Dusseldorf 40225, Germany. Electronic address: koehrer@uni-duesseldorf.de. FAU - Bauer, Inge AU - Bauer I AD - Department of Anesthesiology, Medical Faculty, Heinrich-Heine-University Dusseldorf, Moorenstr. 5, Dusseldorf 40225, Germany. Electronic address: inge.bauer@med.uni-duesseldorf.de. FAU - Hermanns, Henning AU - Hermanns H AD - Department of Anesthesiology, Academic Medical Center, Meibergdreef 9, Amsterdam 1100 DD, The Netherlands. Electronic address: h.hermanns@amc.uva.nl. FAU - Wood, John N AU - Wood JN AD - Molecular Nociception Group, Wolfson Institute for Biomedical Research (WIBR), Cruciform Building, University College London (UCL), London WC1E 6BT, UK. Electronic address: j.wood@ucl.ac.uk. FAU - Zhao, Jing AU - Zhao J AD - Molecular Nociception Group, Wolfson Institute for Biomedical Research (WIBR), Cruciform Building, University College London (UCL), London WC1E 6BT, UK. Electronic address: jing02.zhao@ucl.ac.uk. FAU - Werdehausen, Robert AU - Werdehausen R AD - Department of Anesthesiology, Medical Faculty, Heinrich-Heine-University Dusseldorf, Moorenstr. 5, Dusseldorf 40225, Germany. Electronic address: robert.werdehausen@uni-duesseldorf.de. LA - eng GR - 200183/Z/15/Z/WT_/Wellcome Trust/United Kingdom GR - 20200/ARC_/Arthritis Research UK/United Kingdom GR - 20200/VAC_/Versus Arthritis/United Kingdom GR - G0901905/MRC_/Medical Research Council/United Kingdom PT - Journal Article DEP - 20160621 PL - United States TA - Mol Cell Neurosci JT - Molecular and cellular neurosciences JID - 9100095 RN - 0 (Brain-Derived Neurotrophic Factor) RN - 0 (Chaperonin 60) RN - 0 (Connexin 43) RN - 0 (Hspd1 protein, mouse) RN - 0 (MicroRNAs) RN - 0 (Mirn1 microRNA, mouse) RN - 0 (Mitochondrial Proteins) SB - IM MH - Animals MH - Brain-Derived Neurotrophic Factor/*genetics/metabolism MH - Chaperonin 60/genetics/metabolism MH - Connexin 43/genetics/metabolism MH - Female MH - Ganglia, Spinal/*metabolism MH - Gene Deletion MH - Male MH - Mice MH - Mice, Inbred C57BL MH - MicroRNAs/*genetics/metabolism MH - Mitochondrial Proteins/genetics/metabolism PMC - PMC6675923 OTO - NOTNLM OT - BDNF-Advillin-Cre-ERT2 OT - Bdnf OT - Dorsal root ganglion OT - Gene deletion OT - Neuropathic pain OT - miR-1 OT - microRNA EDAT- 2016/06/28 06:00 MHDA- 2018/01/13 06:00 PMCR- 2016/09/01 CRDT- 2016/06/28 06:00 PHST- 2015/08/11 00:00 [received] PHST- 2016/06/08 00:00 [revised] PHST- 2016/06/20 00:00 [accepted] PHST- 2016/06/28 06:00 [entrez] PHST- 2016/06/28 06:00 [pubmed] PHST- 2018/01/13 06:00 [medline] PHST- 2016/09/01 00:00 [pmc-release] AID - S1044-7431(16)30052-5 [pii] AID - 10.1016/j.mcn.2016.06.003 [doi] PST - ppublish SO - Mol Cell Neurosci. 2016 Sep;75:36-43. doi: 10.1016/j.mcn.2016.06.003. Epub 2016 Jun 21.