Acetylated Histone Expressions of the Primary Hippocampal Neurons in Rats Reduced by siCBP Lentivirus_Journal of Biomedical Engineering

Authors：

HOUNali , WUXiaofeng , RENLan , GUOMin , BIYang , LIUYouxue , CHENJie , HUANGHongmei ,  LITingyu

Children's Nutrition Research Center, Key Laboratory of Developmental Diseases in Childhood of Ministry of Education, Chongqing International Science and Technology Cooperation Center for Child Development and Disorder, Children's Hospital of Chongqing Medical University, Chongqing 400014, China;

Corresponding author：

LITingyu, Email: tyli@vip.sina.com

Keywords：

CREB binding protein; lentilvirus vector; siRNA; hippocampal neuron; histone acetylation

DOI：

10.7507/1001-5515.20150151

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

This study aims to construct the recombinant lentivirus vector containing specific small interfering RNA (siRNA) targeting rat CREB binding protein(CBP)gene and to identify its function of inhibiting the expressions of acetylated histone in primarily cultured hippocampal neurons. Firstly, we constructed four kinds of recombinant lentivirus siCBP. And then we used them to infect the primarily cultured hippocampal neurons, and performed real-time PCR, western blot respectively to detect the expressions of CBP. Afterwards, the most effective lentivirus siCBP was used to infect the primarily cultured hippocampal neurons, and then the HAT activity and protein expressions of acetylated histone Ac-H3, Ac-H4 of the neurons were examined. By using PCR, endonuclease cutting and gene sequencing, we confirmed that the target genes were correctly cloned in lentivirus vector. Besides, CBP mRNA and protein expressions in neurons were found to be with varying degrees of decreases after infections of the four kinds of lentivirus siCBP. Furthermore, the representative and most effective lentivirus GR806 could effectively inhibit the HAT activity and the protein expressions of Ac-H3, Ac-H4 in neurons. It provides the experimental basis for the subsequent application of siCBP to clarify the effects and corresponding molecular mechanism of the CBP-dependent histone acetylation on learning and memory function in hippocampus.

Citation： HOUNali, WUXiaofeng, RENLan, GUOMin, BIYang, LIUYouxue, CHENJie, HUANGHongmei, LITingyu. Acetylated Histone Expressions of the Primary Hippocampal Neurons in Rats Reduced by siCBP Lentivirus. Journal of Biomedical Engineering, 2015, 32(4): 838-846. doi: 10.7507/1001-5515.20150151 Copy

1.	SHARMA S K.Protein acetylation in synaptic plasticity and memory[J].Neurosci Biobehav Rev,2010,34(8):1234-1240.
2.	LEE S,LEE S K.Crucial roles of histone-modifying enzymes in mediating neural cell-type specification[J].Curr Opin Neurobiol,2010,20(1):29-36.
3.	BOUSIGES O,DE VASCONCELOS A P,NEIDL R A,et al.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[J].Neuropsychopharmacology,2010,35(13):2521-2537.
4.	VALOR L M,VIOSCA J,LOPEZ-ATALAYA J P,et al.Lysine acetyltransferases CBP and p300 as therapeutic targets in cognitive and neurodegenerative disorders[J].Curr Pharm Des,2013,19(28):5051-5064.
5.	BARRETT R M,MALVAEZ M,KRAMAR E,et al.Hippocampal focal knockout of CBP affects specific histone modifications,long-term potentiation,and long-term memory[J].Neuropsychopharmacology,2011,36(8):1545-1556.
6.	CHATTERJEE S,MIZAR P,CASSEL R,et al.A novel activator of CBP/p300 acetyltransferases promotes neurogenesis and extends memory duration in adult mice[J].J Neurosci,2013,33(26):10698-10712.
7.	CHEN Guiquan,ZOU Xiaoyan,WATANABE H,et al.CREB binding protein is required for both short-term and long-term memory formation[J].J Neurosci,2010,30(39):13066-13077.
8.	VALOR L M,PULOPULOS M M,JIMENEZ-MINCHAN M,et al.Ablation of CBP in forebrain principal neurons causes modest memory and transcriptional defects and a dramatic reduction of histone acetylation but does not affect cell viability[J].J Neurosci,2011,31(5):1652-1663.
9.	HENNIG A K,PENG Guanghua,CHEN Shiming.Transcription coactivators p300 and CBP are necessary for photoreceptor-specific chromatin organization and gene expression[J].PLoS One,2013,8(7):e69721.
10.	YANG Jian,JIANG Hong,CHEN Si-si,et al.Lentivirus-mediated RNAi targeting CREB binding protein attenuates neointimal formation and promotes re-endothelialization in balloon injured rat carotid artery[J].Cell Physiol Biochem,2010,26(3):441-448.
11.	CHEN Sisi,JIANG Hong,YANG Jian,et al.cAMP-response-element-binding-protein-binding protein silencing inhibits thrombin-induced endothelial progenitor cell migration via downregulation of CXCR4 expression[J].Biol Pharm Bull,2010,33(5):792-795.
12.	YANG Jian,JIANG Hong,CHEN Sisi,et al.CBP knockdown inhibits angiotensin Ⅱ-induced vascular smooth muscle cells proliferation through downregulating NF-κB transcriptional activity[J].Mol Cell Biochem,2010,340(1-2):55-62.
13.	KIRILYUK A,SHIMOJI M,CATANIA J,et al.An intrinsically disordered region of the acetyltransferase p300 with similarity to prion-like domains plays a role in aggregation[J].PLoS One,2012,7(11):e48243.
14.	LIU Rongyu,ZHANG Yili,BAXTER D A,et al.Deficit in long-term synaptic plasticity is rescued by a computationally predicted stimulus protocol[J].J Neurosci,2013,33(16):6944-6949.
15.	GAUB P,TEDESCHI A,PUTTAGUNTA R,et al.HDAC inhibition promotes neuronal outgrowth and counteracts growth cone collapse through CBP/p300 and P/CAF-dependent p53 acetylation[J].Cell Death Differ,2010,17(9):1392-1408.
16.	CACCAMO A,MALDONADO M A,BOKOV A F,et al.CBP gene transfer increases BDNF levels and ameliorates learning and memory deficits in a mouse model of Alzheimer's disease[J].Proc Natl Acad Sci U S A,2010,107(52):22687-22692.
17.	YILDIRIM F,JI Shengbo,KRONENBERG G,et al.Histone acetylation and CREB binding protein are required for neuronal resistance against ischemic injury[J].PLoS One,2014,9(4):e95465.

1. SHARMA S K.Protein acetylation in synaptic plasticity and memory[J].Neurosci Biobehav Rev,2010,34(8):1234-1240.
2. LEE S,LEE S K.Crucial roles of histone-modifying enzymes in mediating neural cell-type specification[J].Curr Opin Neurobiol,2010,20(1):29-36.
3. BOUSIGES O,DE VASCONCELOS A P,NEIDL R A,et al.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[J].Neuropsychopharmacology,2010,35(13):2521-2537.
4. VALOR L M,VIOSCA J,LOPEZ-ATALAYA J P,et al.Lysine acetyltransferases CBP and p300 as therapeutic targets in cognitive and neurodegenerative disorders[J].Curr Pharm Des,2013,19(28):5051-5064.
5. BARRETT R M,MALVAEZ M,KRAMAR E,et al.Hippocampal focal knockout of CBP affects specific histone modifications,long-term potentiation,and long-term memory[J].Neuropsychopharmacology,2011,36(8):1545-1556.
6. CHATTERJEE S,MIZAR P,CASSEL R,et al.A novel activator of CBP/p300 acetyltransferases promotes neurogenesis and extends memory duration in adult mice[J].J Neurosci,2013,33(26):10698-10712.
7. CHEN Guiquan,ZOU Xiaoyan,WATANABE H,et al.CREB binding protein is required for both short-term and long-term memory formation[J].J Neurosci,2010,30(39):13066-13077.
8. VALOR L M,PULOPULOS M M,JIMENEZ-MINCHAN M,et al.Ablation of CBP in forebrain principal neurons causes modest memory and transcriptional defects and a dramatic reduction of histone acetylation but does not affect cell viability[J].J Neurosci,2011,31(5):1652-1663.
9. HENNIG A K,PENG Guanghua,CHEN Shiming.Transcription coactivators p300 and CBP are necessary for photoreceptor-specific chromatin organization and gene expression[J].PLoS One,2013,8(7):e69721.
10. YANG Jian,JIANG Hong,CHEN Si-si,et al.Lentivirus-mediated RNAi targeting CREB binding protein attenuates neointimal formation and promotes re-endothelialization in balloon injured rat carotid artery[J].Cell Physiol Biochem,2010,26(3):441-448.
11. CHEN Sisi,JIANG Hong,YANG Jian,et al.cAMP-response-element-binding-protein-binding protein silencing inhibits thrombin-induced endothelial progenitor cell migration via downregulation of CXCR4 expression[J].Biol Pharm Bull,2010,33(5):792-795.
12. YANG Jian,JIANG Hong,CHEN Sisi,et al.CBP knockdown inhibits angiotensin Ⅱ-induced vascular smooth muscle cells proliferation through downregulating NF-κB transcriptional activity[J].Mol Cell Biochem,2010,340(1-2):55-62.
13. KIRILYUK A,SHIMOJI M,CATANIA J,et al.An intrinsically disordered region of the acetyltransferase p300 with similarity to prion-like domains plays a role in aggregation[J].PLoS One,2012,7(11):e48243.
14. LIU Rongyu,ZHANG Yili,BAXTER D A,et al.Deficit in long-term synaptic plasticity is rescued by a computationally predicted stimulus protocol[J].J Neurosci,2013,33(16):6944-6949.
15. GAUB P,TEDESCHI A,PUTTAGUNTA R,et al.HDAC inhibition promotes neuronal outgrowth and counteracts growth cone collapse through CBP/p300 and P/CAF-dependent p53 acetylation[J].Cell Death Differ,2010,17(9):1392-1408.
16. CACCAMO A,MALDONADO M A,BOKOV A F,et al.CBP gene transfer increases BDNF levels and ameliorates learning and memory deficits in a mouse model of Alzheimer's disease[J].Proc Natl Acad Sci U S A,2010,107(52):22687-22692.
17. YILDIRIM F,JI Shengbo,KRONENBERG G,et al.Histone acetylation and CREB binding protein are required for neuronal resistance against ischemic injury[J].PLoS One,2014,9(4):e95465.

Journal of Biomedical Engineering

Acetylated Histone Expressions of the Primary Hippocampal Neurons in Rats Reduced by siCBP Lentivirus

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content