Vitamin A (VA) is an essential micronutrient. Numerous studies have confirmed that VA deficiency (VAD) leads to a decline in learning and memory function. Our previous studies have demonstrated that retinoic acid nuclear receptor α (RARα) in the hippocampus plays a crucial role in learning and memory, but the exact mechanism for this process is unclear. Epigenetic modifications, particularly histone acetylation, are involved in nervous system development, learning and memory function, and the pathogenesis of neurodegenerative diseases. Histone acetyltransferases (HATs), such as CREB-binding protein (CBP), E1A-binding protein p300 (p300), and p300/CBP-associated factor (PCAF), are critical for regulating memory function. The current study uses RARα and CBP as examples to study the connections between the RA signaling pathway and histone acetylation modification and to reveal the epigenetic mechanism in VAD-induced learning and memory impairment. This study examined the expression of RARα, HATs, acetylated histone H3/H4, and memory-related genes (Zif268, cFos, FosB), as well as the interaction of RARα and CBP in the hippocampus of 8-week-old rats. Additionally, the changes shown in vivo were further assessed in primary cultured neurons with the inhibition or overexpression of RARα. We found significantly lower levels of histone acetylation in the VAD rats. Furthermore, this downregulation, which impairs learning and memory, is induced by the dysregulation of CBP-dependent histone acetylation that is mediated by RARα. This work provides a solid theoretical foundation and experimental basis for the importance of ensuring sufficient nutritional VA during pregnancy and early life to prevent impairments of learning and memory in adulthood.
Histone acetylation Vitamin A RARα CBP HATs Learning and memory impairment
Vitamin A normal
Vitamin A defiency
Retinoid acid nuclear receptor α
Histone deacetylase inhibitors
Retinoic acid response element
This is a preview of subscription content, log in to check access.
This work was supported by the Specialized Research Fund for the National Natural Science Foundation of China (No. 81161120498, 81271221) and the Research Fund for the Doctoral Program of Higher Education of China (No. 20115503110003).
Conflict of Interest
The authors declare no competing financial interests.
Nali Hou, Min Gong, Yang Bi, Zhifang Dong, Jie Chen, and Tingyu Li designed the research. Nali Hou performed experiments. Lan Ren and Yan Gu helped Nali Hou in experiments. Nali Hou, Min Gong, Jie Chen, and Youxue Liu analyzed the data. Nali Hou, Min Gong, Jie Chen, and Tingyu Li wrote the paper. All authors read and approved the final manuscript.
Quantification of fluorescence intensity for the colocalization of RARα and CBP in the rat hippocampus (CA1, CA3, DG). The coexpression of RARα and CBP was weaker in the CA1, CA3 and DG region of the hippocampus in VAD than that in VAN, respectively. The reported values are shown as the mean ± SEM, and the above data were confirmed in at least six samples for each region and each sample was tested in triplicate (n = 6, ***p<0.001). (GIF 23 kb)
Identification and infection of the primary cultured neurons. a Immunofluorescence staining of neuronal marker NSE in neurons was used to identify the primary cultured cells, and the purity of greater than 90 % was achieved. TD bright light, green expression of NSE, blue DAPI, multicolor merged images of TD, NSE and DAPI. bTD neurons before adenovirus infection, growing well. Primary neurons on day 6 after infection with RFP, siRARα or Ad-RARα, respectively, were positive for RFP under the fluorescence microscope and had similar infection levels. c Primary neurons on day 6 after co-infection with Ad-RARα and siCBP or the negative control GFP lentivirus (NC) were positive for GFP and RFP, respectively, under the fluorescence microscope and showed similar infection levels. Green expression of NC or siCBP, red Ad-RARα, multicolor merged images of TD, Ad-RARRα and siCBP or NC. The original magnification is 200×; scale bars are 20 μm. (GIF 260 kb)
Inhibition of CBP did not affect the expression levels of RARα. a CBP mRNA expression was reduced significantly after infection with siCBP lentivirus (n = 6, *p < 0.05). b RARα mRNA was not obviously affected after treatment with siCBP (n = 6, ns, nonsignificant). c The expression of RARα protein levels showed no significant changes after inhibition of CBP. The results were normalized against LaminB. (GIF 30 kb)
Zhang X, Chen K, Wei XP, Qu P, Liu YX, Chen J, Li TY (2009) Perinatal vitamin A status in relation to neurodevelopmental outcome at two years of age. International journal for vitamin and nutrition research Internationale Zeitschrift fur Vitamin- und Ernahrungsforschung. J Int de vitaminologie et de Nutr 79(4):238–249. doi:10.1024/0300-9822.214.171.124CrossRefGoogle Scholar
Zhang X, Chen K, Chen J, Liu YX, Qu P, Li TY (2011) Effect of marginal vitamin A deficiency during pregnancy on retinoic acid receptors and N-methyl-D-aspartate receptor expression in the offspring of rats. J Nutr Biochem 22(12):1112–1120. doi:10.1016/j.jnutbio.2010.09.010CrossRefPubMedGoogle Scholar
Jiang W, Yu Q, Gong M, Chen L, Wen EY, Bi Y, Zhang Y, Shi Y, Qu P, Liu YX, Wei XP, Chen J, Li TY (2012) Vitamin A deficiency impairs postnatal cognitive function via inhibition of neuronal calcium excitability in hippocampus. J Neurochem 121(6):932–943. doi:10.1111/j.1471-4159.2012.07697.xCrossRefPubMedGoogle Scholar
Nomoto M, Takeda Y, Uchida S, Mitsuda K, Enomoto H, Saito K, Choi T, Watabe AM, Kobayashi S, Masushige S, Manabe T, Kida S (2012) Dysfunction of the RAR/RXR signaling pathway in the forebrain impairs hippocampal memory and synaptic plasticity. Mol Brain 5:8. doi:10.1186/1756-6606-5-8CrossRefPubMedCentralPubMedGoogle Scholar
Wood MA, Kaplan MP, Park A, Blanchard EJ, Oliveira AM, Lombardi TL, Abel T (2005) Transgenic mice expressing a truncated form of CREB-binding protein (CBP) exhibit deficits in hippocampal synaptic plasticity and memory storage. Learn Mem 12(2):111–119. doi:10.1101/lm.86605CrossRefPubMedCentralPubMedGoogle Scholar
Kotha Subbaramaiah PAC, Dannenberg AJ (2002) Retinoids and carnosol suppress cyclooxygenase-2 transcription by CREB-binding protein/p300-dependent and -independent mechanisms. Cancer Res 62:2522–2530PubMedGoogle Scholar
Lackey DE, Hoag KA (2010) Vitamin A upregulates matrix metalloproteinase-9 activity by murine myeloid dendritic cells through a nonclassical transcriptional mechanism. J Nutr 140(8):1502–1508. doi:10.3945/jn.110.122556CrossRefPubMedGoogle Scholar
Gong M, Bi Y, Jiang W, Zhang Y, Chen L, Hou N, Liu Y, Wei X, Chen J, Li T (2011) Immortalized mesenchymal stem cells: an alternative to primary mesenchymal stem cells in neuronal differentiation and neuroregeneration associated studies. J Biomed Sci 18:87. doi:10.1186/1423-0127-18-87CrossRefPubMedCentralPubMedGoogle Scholar
Gong M, Bi Y, Jiang W, Zhang Y, Chen L, Hou N, Chen J, Li T (2013) Retinoic acid receptor beta mediates all-trans retinoic acid facilitation of mesenchymal stem cells neuronal differentiation. Int J Biochem Cell Biol 45(4):866–875. doi:10.1016/j.biocel.2013.01.002CrossRefPubMedGoogle Scholar
Bousiges O, Vasconcelos AP, Neidl R, Cosquer B, Herbeaux K, Panteleeva I, Loeffler J-P, Cassel J-C, Boutillier A-L (2010) Spatial memory consolidation is associated with induction of several lysine-acetyltransferase (histone acetyltransferase) expression levels and H2B/H4 acetylation-dependent transcriptional events in the rat hippocampus. Neuropsychopharmacology 35(13):2521–2537. doi:10.1038/npp.2010.117CrossRefPubMedCentralPubMedGoogle Scholar
Ogryzko VV (2001) Mammalian histone acetyltransferases and their complexes. CMLS Cell Mol Life Sci 58:683–692CrossRefGoogle Scholar
Rouaux C, Panteleeva I, Rene F, Gonzalez de Aguilar JL, Echaniz-Laguna A, Dupuis L, Menger Y, Boutillier AL, Loeffler JP (2007) Sodium valproate exerts neuroprotective effects in vivo through CREB-binding protein-dependent mechanisms but does not improve survival in an amyotrophic lateral sclerosis mouse model. J Neurosci: Off J Soc Neurosci 27(21):5535–5545. doi:10.1523/JNEUROSCI.1139-07.2007CrossRefGoogle Scholar
Vecsey CG, Hawk JD, Lattal KM, Stein JM, Fabian SA, Attner MA, Cabrera SM, McDonough CB, Brindle PK, Abel T, Wood MA (2007) Histone deacetylase inhibitors enhance memory and synaptic plasticity via CREB: CBP-dependent transcriptional activation. J Neurosci: Off J Soc Neurosci 27(23):6128–6140. doi:10.1523/JNEUROSCI.0296-07.2007CrossRefGoogle Scholar
Irwin DJ, Cohen TJ, Grossman M, Arnold SE, Xie SX, Lee VM, Trojanowski JQ (2012) Acetylated tau, a novel pathological signature in Alzheimer's disease and other tauopathies. Brain: J Neurol 135(Pt 3):807–818. doi:10.1093/brain/aws013CrossRefGoogle Scholar
Johnson AA, Sarthi J, Pirooznia SK, Reube W, Elefant F (2013) Increasing Tip60 HAT levels rescues axonal transport defects and associated behavioral phenotypes in a Drosophila Alzheimer's disease model. J Neurosci: Off J Soc Neurosci 33(17):7535–7547. doi:10.1523/JNEUROSCI.3739-12.2013CrossRefGoogle Scholar
Luis M, Valor JV, Lopez-Atalaya JP, Barco A (2013) Lysine acetyltransferases CBP and p300 as therapeutic targets in cognitive and neurodegenerative disorders. Curr Pharm Des 19:5051–5064CrossRefGoogle Scholar
Wegert J, Bausenwein S, Kneitz S, Roth S, Graf N, Geissinger E, Gessler M (2011) Retinoic acid pathway activity in Wilms tumors and characterization of biological responses in vitro. Mol Cancer 10(136):1–12. doi:10.1186/1476-4598-10-136Google Scholar
Andrade FO, Nagamine MK, Conti AD, Chaible LM, Fontelles CC, Jordão Junior AA, Vannucchi H, Dagli MLZ, Bassoli BK, Moreno FS, Ong TP (2012) Efficacy of the dietary histone deacetylase inhibitor butyrate alone or in combination with vitamin A against proliferation of MCF-7 human breast cancer cells. Braz J Med Biol Res 45(9):841–850. doi:10.1590/s0100-879x2012007500103CrossRefPubMedCentralPubMedGoogle Scholar
Pili R, Salumbides B, Zhao M, Altiok S, Qian D, Zwiebel J, Carducci MA, Rudek MA (2012) Phase I study of the histone deacetylase inhibitor entinostat in combination with 13-cis retinoic acid in patients with solid tumours. Br J Cancer 106(1):77–84. doi:10.1038/bjc.2011.527CrossRefPubMedCentralPubMedGoogle Scholar
Fredly H, Gjertsen BT, Bruserud Ø (2013) Histone deacetylase inhibition in the treatment of acute myeloid leukemia: the effects of valproic acid on leukemic cells, and the clinical and experimental evidence for combining valproic acid with other antileukemic agents. Clin Epigenetics 5(12):1–13Google Scholar
Brouillard F, Cremisi CE (2003) Concomitant increase of histone acetyltransferase activity and degradation of p300 during retinoic acid-induced differentiation of F9 cells. J Biol Chem 278(41):39509–39516. doi:10.1074/jbc.M307123200CrossRefPubMedGoogle Scholar
Eaves CJ, Jain AK, Allton K, Iacovino M, Mahen E, Milczarek RJ, Zwaka TP, Kyba M, Barton MC (2012) p53 regulates cell cycle and microRNAs to promote differentiation of human embryonic stem cells. PLoS Biol 10(2):e1001268. doi:10.1371/journal.pbio.1001268CrossRefGoogle Scholar
Johnson R, Chen W, Jia W, Wang K, Si X, Zhu S, Duan T, Kang J (2013) Distinct roles for CBP and p300 on the RA-mediated expression of the meiosis commitment gene Stra8 in mouse embryonic stem cells. PLoS One 8(6):e66076. doi:10.1371/journal.pone.0066076CrossRefGoogle Scholar
1.Children Nutrition Research CenterChildren’s Hospital of Chongqing Medical UniversityChongqingChina
2.Ministry of Education Key Laboratory of Child Development and DisordersChongqing Key Laboratory of Translational Medical Research in Cognitive Development and Learning and Memory DisordersChongqingChina