Downregulation of CREB expression in Alzheimer's brain and in Aβ-treated rat hippocampal neurons

Molecular Neurodegeneration

Christopher B Eckman⁵  Chun-I Sze⁴  Serena Pham²  Maorong Wang³  Subbiah Pugazhenthi¹ 

[1] Department of Medicine, University of Colorado Denver, Aurora, CO, USA;Section of Endocrinology, Veterans Affairs Medical Center, Denver, CO, USA;Nanjing Bayi Hospital, Nanjing University of Traditional Chinese Medicine, Nanjing, China;Department of Cell Biology and Anatomy, National Cheng Kung University Medical College, Tainan, Taiwan;MidAtlantic Neonatal Research Institute, Atlantic Health Systems, Morristown, NJ, USA

关键词: Laser capture microdissection;    Tg2576 mice;    Apoptosis;    Oxidative stress;    CREB;    Alzheimer's disease;   

Others  :  865663
DOI  :  10.1186/1750-1326-6-60

 received in 2011-05-31, accepted in 2011-08-19,  发布年份 2011

【 摘 要 】

Background

Oxidative stress plays an important role in neuronal dysfunction and neuron loss in Alzheimer's brain. Previous studies have reported downregulation of CREB-mediated transcription by oxidative stress and Aβ. The promoter for CREB itself contains cyclic AMP response elements. Therefore, we examined the expression of CREB in the hippocampal neurons of Tg2576 mice, AD post-mortem brain and in cultured rat hippocampal neurons exposed to Aβ aggregates.

Results

Laser Capture Microdissection of hippocampal neurons from Tg2576 mouse brain revealed decreases in the mRNA levels of CREB and its target, BDNF. Immunohistochemical analysis of Tg2576 mouse brain showed decreases in CREB levels in hippocampus and cortex. Markers of oxidative stress were detected in transgenic mouse brain and decreased CREB staining was observed in regions showing abundance of astrocytes. There was also an inverse correlation between SDS-extracted Aβ and CREB protein levels in Alzheimer's post-mortem hippocampal samples. The levels of CREB-regulated BDNF and BIRC3, a caspase inhibitor, decreased and the active cleaved form of caspase-9, a marker for the intrinsic pathway of apoptosis, was elevated in these samples. Exposure of rat primary hippocampal neurons to Aβ fibrils decreased CREB promoter activity. Decrease in CREB mRNA levels in Aβ-treated neurons was reversed by the antioxidant, N-acetyl cysteine. Overexpression of CREB by adenoviral transduction led to significant protection against Aβ-induced neuronal apoptosis.

Conclusions

Our findings suggest that chronic downregulation of CREB-mediated transcription results in decrease of CREB content in the hippocampal neurons of AD brain which may contribute to exacerbation of disease progression.

【 授权许可】

   
2011 Pugazhenthi et al; licensee BioMed Central Ltd.

【 图 表 】

【 参考文献 】

• [1]Benito E, Barco A: CREB's control of intrinsic and synaptic plasticity: implications for CREB-dependent memory models. Trends Neurosci 33:230-240.
• [2]Sakamoto K, Karelina K, Obrietan K: CREB: a multifaceted regulator of neuronal plasticity and protection. J Neurochem 116:1-9.
• [3]Lonze BE, Ginty DD: Function and regulation of CREB family transcription factors in the nervous system. Neuron 2002, 35:605-623.
• [4]Impey S, Smith DM, Obrietan K, Donahue R, Wade C, Storm DR: Stimulation of cAMP response element (CRE)-mediated transcription during contextual learning. Nat Neurosci 1998, 1:595-601.
• [5]Frank DA, Greenberg ME: CREB: a mediator of long-term memory from mollusks to mammals. Cell 1994, 79:5-8.
• [6]Impey S, Mark M, Villacres EC, Poster S, Chavkin C, Storm DR: Induction of CRE-mediated gene expression by stimuli that generate long-lasting LTP in area CA1 of the hippocampus. Neuron 1996, 16:973-982.
• [7]Guzowski JF, McGaugh JL: Antisense oligodeoxynucleotide-mediated disruption of hippocampal cAMP response element binding protein levels impairs consolidation of memory for water maze training. Proc Natl Acad Sci USA 1997, 94:2693-2698.
• [8]Finkbeiner S: CREB couples neurotrophin signals to survival messages. Neuron 2000, 25:11-14.
• [9]Shaywitz AJ, Greenberg ME: CREB: a stimulus-induced transcription factor activated by a diverse array of extracellular signals. Annu Rev Biochem 1999, 68:821-861.
• [10]Pugazhenthi S, Boras T, O'Connor D, Meintzer MK, Heidenreich KA, Reusch JE: Insulin-like growth factor I-mediated activation of the transcription factor cAMP response element-binding protein in PC12 cells. Involvement of p38 mitogen-activated protein kinase-mediated pathway. J Biol Chem 1999, 274:2829-2837.
• [11]Kwok RPS, Lundblad JR, Chrivia JC, Richards JP, Bächlinger HP, Brennan RG, Roberts SGE, Green MR, Goodman RH: Nuclear protein CBP is a coactivator for the transcription factor CREB. Nature 1994, 370:223-229.
• [12]Pugazhenthi S, Miller E, Sable C, Young P, Heidenreich KA, Boxer LM, Reusch JE-B: Insulin-like growth factor-I induces bcl-2 promoter through the transcription factor cAMP-response element binding protein. J Biol Chem 1999, 274:27529-27535.
• [13]Pugazhenthi S, Nesterova A, Sable C, Heidenreich K, Boxer L, Heasley L, Reusch J: Akt/Protein kinase B up-regulates Bcl-2 expression through cAMP-response element-binding protein. J Biol Chem 2000, 275:10761-10766.
• [14]Echeverria V, Ducatenzeiler A, Dowd E, Janne J, Grant SM, Szyf M, Wandosell F, Avila J, Grimm H, Dunnett SB, et al.: Altered mitogen-activated protein kinase signaling, tau hyperphosphorylation and mild spatial learning dysfunction in transgenic rats expressing the beta-amyloid peptide intracellularly in hippocampal and cortical neurons. Neuroscience 2004, 129:583-592.
• [15]Gong B, Vitolo OV, Trinchese F, Liu S, Shelanski M, Arancio O: Persistent improvement in synaptic and cognitive functions in an Alzheimer mouse model after rolipram treatment. J Clin Invest 2004, 114:1624-1634.
• [16]Phillips HS, Hains JM, Armanini M, Laramee GR, Johnson SA, Winslow JW: BDNF mRNA is decreased in the hippocampus of individuals with Alzheimer's disease. Neuron 1991, 7:695-702.
• [17]Vitolo OV, Sant'Angelo A, Costanzo V, Battaglia F, Arancio O, Shelanski M: Amyloid beta -peptide inhibition of the PKA/CREB pathway and long-term potentiation: reversibility by drugs that enhance cAMP signaling. Proc Natl Acad Sci USA 2002, 99:13217-13221.
• [18]Tong L, Thornton PL, Balazs R, Cotman CW: Beta-amyloid-(1-42) impairs activity-dependent cAMP response element-binding protein signaling in neurons at concentrations in which cell is not compromised. J Biol Chem 2001, 276:17301-17306.
• [19]Tong L, Balazs R, Thornton PL, Cotman CW: Beta-amyloid peptide at sublethal concentrations downregulates brain-derived neurotrophic factor functions in cultured cortical neurons. J Neurosci 2004, 24:6799-6809.
• [20]McDonald D, Bamberger M, Combs C, Landreth G: b-Amyloid fibrils activate parallel mitogen-activated kinase pathways in microglia and THP1 monocytes. J Neurosci 1998, 18:4451-4460.
• [21]Sato N, Kamino K, Tateishi K, Satoh T, Nishiwaki Y, Yoshiiwa A, Miki T, Ogihara T: Elevated amyloid beta protein(1-40) level induces CREB phosphorylation at serine-133 via p44/42 MAP kinase (Erk1/2)-dependent pathway in rat pheochromocytoma PC12 cells. Biochem Biophys Res Commun 1997, 232:637-642.
• [22]Pugazhenthi S, Phansalkar K, Audesirk G, West A, Cabell L: Differential regulation of c-jun and CREB by acrolein and 4-hydroxynonenal. Free Radic Biol Med 2006, 40:21-34.
• [23]Pugazhenthi S, Nesterova A, Jambal P, Audesirk G, Kern M, Cabell L, Eves E, Rosner MR, Boxer LM, Reusch JE: Oxidative stress-mediated down-regulation of bcl-2 promoter in hippocampal neurons. J Neurochem 2003, 84:982-996.
• [24]Takuma K, Baba A, Matsuda T: Astrocyte apoptosis: implications for neuroprotection. Prog Neurobiol 2004, 72:111-127.
• [25]Gonzalez GA, Montminty MR: Cyclic AMP stimulates somatostatin gene transcription by phosphorylation of CREB at Serine 133. Cell 1989, 59:675-680.
• [26]Xing J, Ginty DD, Greenberg ME: Coupling of the RAS-MAPK pathway to gene activation by RSK2, a growth factor-regulated CREB kinase. Science 1996, 273:959-963.
• [27]King DL, Arendash GW: Behavioral characterization of the Tg2576 transgenic model of Alzheimer's disease through 19 months. Physiol Behav 2002, 75:627-642.
• [28]Kawarabayashi T, Younkin LH, Saido TC, Shoji M, Ashe KH, Younkin SG: Age-dependent changes in brain, CSF, and plasma amyloid (beta) protein in the Tg2576 transgenic mouse model of Alzheimer's disease. J Neurosci 2001, 21:372-381.
• [29]Wu ZL, Ciallella JR, Flood DG, O'Kane T M, Bozyczko-Coyne D, Savage MJ: Comparative analysis of cortical gene expression in mouse models of Alzheimer's disease. Neurobiol Aging 2005.
• [30]Wyss-Coray T: Inflammation in Alzheimer disease: driving force, bystander or beneficial response? Nat Med 2006, 12:1005-1015.
• [31]Glass CK, Saijo K, Winner B, Marchetto MC, Gage FH: Mechanisms underlying inflammation in neurodegeneration. Cell 140:918-934.
• [32]Jambal P, Masterson S, Nesterova A, Bouchard R, Bergman B, Hutton JC, Boxer LM, Reusch JE, Pugazhenthi S: Cytokine-mediated downregulation of the transcription factor CREB in pancreatic beta -cells. J Biol Chem 2003, 278:23055-23065.
• [33]Nicolle MM, Gonzalez J, Sugaya K, Baskerville KA, Bryan D, Lund K, Gallagher M, McKinney M: Signatures of hippocampal oxidative stress in aged spatial learning-impaired rodents. Neuroscience 2001, 107:415-431.
• [34]Hensley K, Carney JM, Mattson MP, Aksenova M, Harris M, Wu JF, Floyd RA, Butterfield DA: A model for beta-amyloid aggregation and neurotoxicity based on free radical generation by the peptide: relevance to Alzheimer's disease. Proc Natl Acad Sci USA 1994, 91:3270-3274.
• [35]Huang X, Atwood CS, Hartshorn MA, Multhaup G, Goldstein LE, Scarpa RC, Cuajungco MP, Gray DN, Lim J, Moir RD, et al.: The A beta peptide of Alzheimer's disease directly produces hydrogen peroxide through metal ion reduction. Biochemistry 1999, 38:7609-7616.
• [36]Markesbery WR: Oxidative stress hypothesis in Alzheimer's disease. Free Radical Biology & Medicine 1997, 23:134-147.
• [37]Smith MA, Harris PL, Sayre LM, Perry G: Iron accumulation in Alzheimer disease is a source of redox-generated free radicals. Proc Natl Acad Sci USA 1997, 94:9866-9868.
• [38]Falangola MF, Lee SP, Nixon RA, Duff K, Helpern JA: Histological co-localization of iron in Abeta plaques of PS/APP transgenic mice. Neurochem Res 2005, 30:201-205.
• [39]Behl C: Amyloid beta protein toxicity and oxidative stress in Alzheimer's disease. Cell Tissue Res 1997, 290:471-480.
• [40]Smith MA, Rottkamp CA, Nunomura A, Raina AK: Oxidative stress in Alzheimer's disease. Biochim Biophys Acta 2000, 1502:139-144.
• [41]Keller JN, Schmitt FA, Scheff SW, Ding Q, Chen Q, Butterfield DA, Markesbery WR: Evidence of increased oxidative damage in subjects with mild cognitive impairment. Neurology 2005, 64:1152-1156.
• [42]Roth KA: Caspases, apoptosis, and Alzheimer disease: causation, correlation, and confusion. J Neuropathol Exp Neurol 2001, 60:829-838.
• [43]Rissman RA, Poon WW, Blurton-Jones M, Oddo S, Torp R, Vitek MP, LaFerla FM, Rohn TT, Cotman CW: Caspase-cleavage of tau is an early event in Alzheimer disease tangle pathology. J Clin Invest 2004, 114:121-130.
• [44]Mattson MP: Apoptosis in neurodegenerative disorders. Nature Rev Mol Cell Biol 2000, 1:120-129.
• [45]Xia M, Huang R, Guo V, Southall N, Cho MH, Inglese J, Austin CP, Nirenberg M: Identification of compounds that potentiate CREB signaling as possible enhancers of long-term memory. Proc Natl Acad Sci USA 2009, 106:2412-2417.
• [46]Cummings TJ, Strum JC, Yoon LW, Szymanski MH, Hulette CM: Recovery and expression of messenger RNA from postmortem human brain tissue. Mod Pathol 2001, 14:1157-1161.
• [47]Loring JF, Wen X, Lee JM, Seilhamer J, Somogyi R: A gene expression profile of Alzheimer's disease. DNA Cell Biol 2001, 20:683-695.
• [48]Browning MD, Dudek EM, Rapier JL, Leonard S, Freedman R: Significant reductions in synapsin but not synaptophysin specific activity in the brains of some schizophrenics. Biol Psychiatry 1993, 34:529-535.
• [49]Blalock EM, Geddes JW, Chen KC, Porter NM, Markesbery WR, Landfield PW: Incipient Alzheimer's disease: microarray correlation analyses reveal major transcriptional and tumor suppressor responses. Proc Natl Acad Sci USA 2004, 101:2173-2178.
• [50]Mirra SS, Heyman A, McKeel D, Sumi SM, Crain BJ, Brownlee LM, Vogel FS, Hughes JP, van Belle G, Berg L: The Consortium to Establish a Registry for Alzheimer's Disease (CERAD). Part II. Standardization of the neuropathologic assessment of Alzheimer's disease. Neurology 1991, 41:479-486.
• [51]Audesirk T, Audesirk G, Ferguson C, Shugarts D: Effects of inorganic lead on the differentiation and growth of cultured hippocampal and neuroblastoma cells. Neurotoxicology 1991, 12:529-538.
• [52]Goslin K, Banker G: Rat hippocampal neurons in low-density culture. In Culturing Nerve cells. Edited by Banker G, Goslin K. Cambridge, Massachusetts: MIT Press; 1991:251-281.