NGF Modulates Cholesterol Metabolism and Stimulates ApoE Secretion in Glial Cells Conferring Neuroprotection against Oxidative Stress
Abstract
:1. Introduction
2. Results
2.1. U373 Express Both the NGF Receptors TrkA and p75NTR
2.2. NGF Modulates the Protein Network Involved in Cholesterol Homeostasis in Glial Cells
2.3. NGF Increases Cholesterol Secretion by U373 into the Culture Medium
2.4. p75NTR Is Partially Involved in the NGF-Mediated Effects on Cholesterol Protein Network
2.5. Conditioned Medium Derived from NGF-Treated U373 Cells Does Not Influence Neuronal Differentiation
2.6. NGF-Mediated Secretion of ApoE by U373 Protects Neuronal Cells from Oxidative Insult
3. Discussion
4. Material and Methods
4.1. Cell Culture
4.2. Lysate Preparation and Western Blot Analysis
4.3. Oil Red O Staining
4.4. Filipin Staining
4.5. Cholesterol Quantification
4.6. ELISA
4.7. Immunofluorescence
4.8. ApoE Silencing
4.9. RNA Extraction and Real-Time PCR
4.10. Quantitative Assessment of Neuronal Morphology
4.11. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Björkhem, I.; Meaney, S.; Fogelman, A.M. Brain Cholesterol: Long Secret Life behind a Barrier. Arterioscler. Thromb. Vasc. Biol. 2004, 24, 806–815. [Google Scholar] [CrossRef] [PubMed]
- Colardo, M.; Martella, N.; Pensabene, D.; Siteni, S.; Di Bartolomeo, S.; Pallottini, V.; Segatto, M. Neurotrophins as key regulators of cell metabolism: Implications for cholesterol homeostasis. Int. J. Mol. Sci. 2021, 22, 5692. [Google Scholar] [CrossRef] [PubMed]
- Goritz, C.; Mauch, D.H.; Pfrieger, F.W. Multiple mechanisms mediate cholesterol-induced synaptogenesis in a CNS neuron. Mol. Cell. Neurosci. 2005, 29, 190–201. [Google Scholar] [CrossRef] [PubMed]
- Pfenninger, K.H. Plasma membrane expansion: A neuron’s Herculean task. Nat. Rev. Neurosci. 2009, 10, 251–261. [Google Scholar] [CrossRef]
- Takamori, S.; Holt, M.; Stenius, K.; Lemke, E.A.; Grønborg, M.; Riedel, D.; Urlaub, H.; Schenck, S.; Brügger, B.; Ringler, P.; et al. Molecular Anatomy of a Trafficking Organelle. Cell 2006, 127, 831–846. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Suzuki, S.; Kiyosue, K.; Hazama, S.; Ogura, A.; Kashihara, M.; Hara, T.; Koshimizu, H.; Kojima, M. Brain-derived neurotrophic factor regulates cholesterol metabolism for synapse development. J. Neurosci. 2007, 27, 6417–6427. [Google Scholar] [CrossRef]
- Segatto, M.; Leboffe, L.; Trapani, L.; Pallottini, V. Cholesterol Homeostasis Failure in the Brain: Implications for Synaptic Dysfunction and Cognitive Decline. Curr. Med. Chem. 2014, 21, 2788–2802. [Google Scholar] [CrossRef] [PubMed]
- Jeske, D.J.; Dietschy, J.M. Regulation of rates of cholesterol synthesis in vivo in the liver and carcass of the rat measured using [3H]water. J. Lipid Res. 1980, 21, 364–376. [Google Scholar] [CrossRef]
- Pfrieger, F.W.; Ungerer, N. Cholesterol metabolism in neurons and astrocytes. Prog. Lipid Res. 2011, 50, 357–371. [Google Scholar] [CrossRef]
- Nieweg, K.; Schaller, H.; Pfrieger, F.W. Marked differences in cholesterol synthesis between neurons and glial cells from postnatal rats. J. Neurochem. 2009, 109, 125–134. [Google Scholar] [CrossRef]
- Oram, J.F.; Heinecke, J.W. ATP-binding cassette transporter A1: A cell cholesterol exporter that protects against cardiovascular disease. Physiol. Rev. 2005, 85, 1343–1372. [Google Scholar] [CrossRef]
- Pfrieger, F.W. Outsourcing in the brain: Do neurons depend on cholesterol delivery by astrocytes? BioEssays 2003, 25, 72–78. [Google Scholar] [CrossRef]
- Christopherson, K.S.; Ullian, E.M.; Stokes, C.C.A.; Mullowney, C.E.; Hell, J.W.; Agah, A.; Lawler, J.; Mosher, D.F.; Bornstein, P.; Barres, B.A. Thrombospondins are astrocyte-secreted proteins that promote CNS synaptogenesis. Cell 2005, 120, 421–433. [Google Scholar] [CrossRef] [Green Version]
- Mauch, D.H.; Nägier, K.; Schumacher, S.; Göritz, C.; Müller, E.C.; Otto, A.; Pfrieger, F.W. CNS synaptogenesis promoted by glia-derived cholesterol. Science 2001, 294, 1354–1357. [Google Scholar] [CrossRef]
- Valenza, M.; Marullo, M.; Di Paolo, E.; Cesana, E.; Zuccato, C.; Biella, G.; Cattaneo, E. Disruption of astrocyte-neuron cholesterol cross talk affects neuronal function in Huntington’s disease. Cell Death Differ. 2015, 22, 690–702. [Google Scholar] [CrossRef] [Green Version]
- Tensaouti, Y.; Stephanz, E.P.; Yu, T.S.; Kernie, S. ApoE Regulates the Development of Adult Newborn Hippocampal Neurons. eNeuro 2018, 5, ENEURO.0155-18.2018. [Google Scholar] [CrossRef] [Green Version]
- Gao, H.; Zheng, W.; Li, C.; Xu, H. Isoform-Specific Effects of Apolipoprotein E on Hydrogen Peroxide-Induced Apoptosis in Human Induced Pluripotent Stem Cell (iPSC)-Derived Cortical Neurons. Int. J. Mol. Sci. 2021, 22, 11582. [Google Scholar] [CrossRef]
- Lee, Y.; Aono, M.; Laskowitz, D.; Warner, D.S.; Pearlstein, R.D. Apolipoprotein E protects against oxidative stress in mixed neuronal-glial cell cultures by reducing glutamate toxicity. Neurochem. Int. 2004, 44, 107–118. [Google Scholar] [CrossRef]
- Chaldakov, G.N. The metabotrophic NGF and BDNF: An emerging concept. Arch. Ital. Biol. 2011, 149, 257–263. [Google Scholar] [CrossRef]
- Chaldakov, G.N.; Fiore, M.; Stankulov, I.S.; Manni, L.; Hristova, M.G.; Antonelli, A.; Ghenev, P.I.; Aloe, L. Neurotrophin presence in human coronary atherosclerosis and metabolic syndrome: A role for NGF and BDNF in cardiovascular disease? Prog. Brain Res. 2004, 146, 279–289. [Google Scholar] [CrossRef]
- Chaldakov, G.N.; Fiore, M.; Hristova, M.G.; Aloe, L. Metabotrophic potential of neurotrophins: Implication in obesity and related diseases? Med. Sci. Monit. 2003, 9, 19–22. [Google Scholar]
- Pallottini, V.; Colardo, M.; Tonini, C.; Martella, N.; Strimpakos, G.; Colella, B.; Tirassa, P.; Di Bartolomeo, S.; Segatto, M. ProNGF/p75NTR Axis Drives Fiber Type Specification by Inducing the Fast-Glycolytic Phenotype in Mouse Skeletal Muscle Cells. Cells 2020, 9, 2232. [Google Scholar] [CrossRef] [PubMed]
- Segatto, M.; Fico, E.; Gharbiya, M.; Rosso, P.; Carito, V.; Tirassa, P.; Plateroti, R.; Lambiase, A. VEGF inhibition alters neurotrophin signalling pathways and induces caspase-3 activation and autophagy in rabbit retina. J. Cell. Physiol. 2019, 234, 18297–18307. [Google Scholar] [CrossRef] [PubMed]
- Haklai, R.; Lerner, S.; Kloog, Y. Nerve growth factor induces a succession of increases in isoprenylated methylated small GTP-binding proteins of PC-12 phyochromocytoma cells. Neuropeptides 1993, 24, 11–25. [Google Scholar] [CrossRef]
- Pham, D.D.; Bruelle, C.; Thi Do, H.; Pajanoja, C.; Jin, C.; Srinivasan, V.; Olkkonen, V.M.; Eriksson, O.; Jauhiainen, M.; Lalowski, M.; et al. Caspase-2 and p75 neurotrophin receptor (p75NTR) are involved in the regulation of SREBP and lipid genes in hepatocyte cells. Cell Death Dis. 2019, 10, 537. [Google Scholar] [CrossRef]
- Do, H.T.; Bruelle, C.; Pham, D.D.; Jauhiainen, M.; Eriksson, O.; Korhonen, L.T.; Lindholm, D. Nerve growth factor (NGF) and pro-NGF increase low-density lipoprotein (LDL) receptors in neuronal cells partly by different mechanisms: Role of LDL in neurite outgrowth. J. Neurochem. 2016, 136, 306–315. [Google Scholar] [CrossRef] [Green Version]
- Volknandt, W.; Küster, F.; Wilhelm, A.; Obermüller, E.; Steinmann, A.; Zhang, L.; Zimmermann, H. Expression and allocation of proteins of the exo-endocytotic machinery in U373 glioma cells: Similarities to long-term cultured astrocytes. Cell. Mol. Neurobiol. 2002, 22, 153–169. [Google Scholar] [CrossRef]
- Eddleston, M.; De La Torre, J.C.; Oldstone, M.B.A.; Loskutoff, D.J.; Edgington, T.S.; Mackman, N. Astrocytes are the primary source of tissue factor in the murine central nervous system: A role for astrocytes in cerebral hemostasis. J. Clin. Investig. 1993, 92, 349–358. [Google Scholar] [CrossRef] [Green Version]
- Janda, E.; Lascala, A.; Carresi, C.; Parafati, M.; Aprigliano, S.; Russo, V.; Savoia, C.; Ziviani, E.; Musolino, V.; Morani, F.; et al. Parkinsonian toxin-induced oxidative stress inhibits basal autophagy in astrocytes via NQO2/quinone oxidoreductase 2: Implications for neuroprotection. Autophagy 2015, 11, 1063–1080. [Google Scholar] [CrossRef]
- Singer, H.S.; Hansen, B.; Martinie, D.; Karp, C.L. Mitogenesis in glioblastoma multiforme cell lines: A role for NGF and its TrkA receptors. J. Neurooncol. 1999, 45, 1–8. [Google Scholar] [CrossRef]
- Rosso, P.; De Nicolò, S.; Carito, V.; Fiore, M.; Iannitelli, A.; Moreno, S.; Tirassa, P. Ocular Nerve Growth Factor Administration Modulates Brain-derived Neurotrophic Factor Signaling in Prefrontal Cortex of Healthy and Diabetic Rats. CNS Neurosci. Ther. 2017, 23, 198–208. [Google Scholar] [CrossRef]
- Sycheva, M. Pro-Nerve Growth Factor Induces Activation of RhoA. Brain Sci. 2019, 9, 204. [Google Scholar] [CrossRef] [Green Version]
- Liu, Z.; Cao, Y.; Liu, G.; Yin, S.; Ma, J.; Liu, J.; Zhang, M.; Wang, Y. p75 neurotrophin receptor regulates NGF-induced myofibroblast differentiation and collagen synthesis through MRTF-A. Exp. Cell Res. 2019, 383, 111504. [Google Scholar] [CrossRef]
- Smolič, T.; Tavčar, P.; Horvat, A.; Černe, U.; Halužan Vasle, A.; Tratnjek, L.; Kreft, M.E.; Scholz, N.; Matis, M.; Petan, T.; et al. Astrocytes in stress accumulate lipid droplets. Glia 2021, 69, 1540–1562. [Google Scholar] [CrossRef]
- Velebit, J.; Horvat, A.; Smolič, T.; Prpar Mihevc, S.; Rogelj, B.; Zorec, R.; Vardjan, N. Astrocytes with TDP-43 inclusions exhibit reduced noradrenergic cAMP and Ca2+ signaling and dysregulated cell metabolism. Sci. Rep. 2020, 10, 6003. [Google Scholar] [CrossRef] [Green Version]
- Simmons, D.A.; Belichenko, N.P.; Ford, E.C.; Semaan, S.; Monbureau, M.; Aiyaswamy, S.; Holman, C.M.; Condon, C.; Shamloo, M.; Massa, S.M.; et al. A small molecule p75NTR ligand normalizes signalling and reduces Huntington’s disease phenotypes in R6/2 and BACHD mice. Hum. Mol. Genet. 2016, 25, 4920–4938. [Google Scholar] [CrossRef] [Green Version]
- Fagan AM, H.D. Astrocyte lipoproteins, effects of apoE on neuronal function, and role of apoE in amyloid-beta deposition in vivo. Microsc. Res Tech. 2000, 50, 297–304. [Google Scholar] [CrossRef]
- Spagnuolo, M.S.; Donizetti, A.; Iannotta, L.; Aliperti, V.; Cupidi, C.; Bruni, A.C.; Cigliano, L. Brain-derived neurotrophic factor modulates cholesterol homeostasis and Apolipoprotein E synthesis in human cell models of astrocytes and neurons. J. Cell. Physiol. 2018, 233, 6925–6943. [Google Scholar] [CrossRef]
- Vashi, N.; Justice, M.J. Treating Rett syndrome: From mouse models to human therapies. Mamm. Genome 2019, 30, 90–110. [Google Scholar] [CrossRef] [Green Version]
- Strachan-Whaley, M.R.; Reilly, K.; Dobson, J.; Kalisch, B.E. Map kinase and PKC signaling pathways modulate NGF-mediated apoE transcription. Neurosci. Lett. 2015, 595, 54–59. [Google Scholar] [CrossRef]
- Korade, Z.; Kenchappa, R.S.; Mirnics, K.; Carter, B.D. NRIF is a regulator of neuronal cholesterol biosynthesis genes. J. Mol. Neurosci. 2009, 38, 152–158. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Chang, Y.C.; Sheu, W.H.H.; Chien, Y.S.; Tseng, P.C.; Lee, W.J.; Chiang, A.N. Hyperglycemia accelerates ATP-binding cassette transporter A1 degradation via an ERK-dependent pathway in macrophages. J. Cell. Biochem. 2013, 114, 1364–1373. [Google Scholar] [CrossRef] [PubMed]
- Mulay, V.; Wood, P.; Manetsch, M.; Darabi, M.; Cairns, R.; Hoque, M.; Chan, K.C.; Reverter, M.; Álvarez-Guaita, A.; Rye, K.A.; et al. Inhibition of Mitogen-Activated Protein Kinase Erk1/2 Promotes Protein Degradation of ATP Binding Cassette Transporters A1 and G1 in CHO and HuH7 Cells. PLoS ONE 2013, 8, e62667. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Siddiqui, M.A.; Ahmad, J.; Farshori, N.N.; Saquib, Q.; Jahan, S.; Kashyap, M.P.; Ahamed, M.; Musarrat, J.; Al-Khedhairy, A.A. Rotenone-induced oxidative stress and apoptosis in human liver HepG2 cells. Mol. Cell. Biochem. 2013, 384, 59–69. [Google Scholar] [CrossRef]
- Shea, T.B.; Rogers, E.; Ashline, D.; Ortiz, D.; Sheu, M.S. Apolipoprotein E deficiency promotes increased oxidative stress and compensatory increases in antioxidants in brain tissue. Free Radic. Biol. Med. 2002, 33, 1115–1120. [Google Scholar] [CrossRef]
- Shi, Y.; Manis, M.; Long, J.; Wang, K.; Sullivan, P.M.; Serrano, J.R.; Hoyle, R.; Holtzman, D.M. Microglia drive APOE-dependent neurodegeneration in a tauopathy mouse model. J. Exp. Med. 2019, 216, 2546–2561. [Google Scholar] [CrossRef]
- Hayashi, H.; Campenot, R.B.; Vance, D.E.; Vance, J.E. Protection of neurons from apoptosis by apolipoprotein e-containing lipoproteins does not require lipoprotein uptake and involves activation of phospholipase Cγ1 and inhibition of calcineurin. J. Biol. Chem. 2009, 284, 29605–29613. [Google Scholar] [CrossRef] [Green Version]
- Xiong, N.; Xiong, J.; Jia, M.; Liu, L.; Zhang, X.; Chen, Z.; Huang, J.; Zhang, Z.; Hou, L.; Luo, Z.; et al. The role of autophagy in Parkinson’s disease: Rotenone-based modeling. Behav. Brain Funct. 2013, 9, 13. [Google Scholar] [CrossRef] [Green Version]
- Johnson, M.E.; Bobrovskaya, L. An update on the rotenone models of Parkinson’s disease: Their ability to reproduce the features of clinical disease and model gene-environment interactions. Neurotoxicology 2015, 46, 101–116. [Google Scholar] [CrossRef]
- Li, X.; Liu, Z.; Tamashiro, K.; Shi, B.; Rudnicki, D.D.; Ross, C.A.; Moran, T.H.; Smith, W.W. Synphilin-1 exhibits trophic and protective effects against Rotenone toxicity. Neuroscience 2010, 162, 455–462. [Google Scholar] [CrossRef]
- Polazzi, E.; Mengoni, I.; Peña-Altamira, E.; Massenzio, F.; Virgili, M.; Petralla, S.; Monti, B. Neuronal Regulation of Neuroprotective Microglial Apolipoprotein E Secretion in Rat In Vitro Models of Brain Pathophysiology. J. Neuropathol. Exp. Neurol. 2015, 74, 818–834. [Google Scholar] [CrossRef] [Green Version]
- Eu, W.Z.; Chen, Y.J.; Chen, W.T.; Wu, K.Y.; Tsai, C.Y.; Cheng, S.J.; Carter, R.N.; Huang, G.J. The effect of nerve growth factor on supporting spatial memory depends upon hippocampal cholinergic innervation. Transl. Psychiatry 2021, 11, 162. [Google Scholar] [CrossRef]
- Chaturvedi, R.K.; Shukla, S.; Seth, K.; Agrawal, A.K. Nerve growth factor increases survival of dopaminergic graft, rescue nigral dopaminergic neurons and restores functional deficits in rat model of Parkinson’s disease. Neurosci. Lett. 2006, 398, 44–49. [Google Scholar] [CrossRef]
- Goss, J.R.; O’Malley, M.E.; Zou, L.; Styren, S.D.; Kochanek, P.M.; DeKosky, S.T. Astrocytes are the major source of nerve growth factor upregulation following traumatic brain injury in the rat. Exp. Neurol. 1998, 49, 301–309. [Google Scholar] [CrossRef]
- Lindsay, R.M. Adult rat brain astrocytes support survival of both NGF-dependent and NGF-insensitive neurones. Nature 1979, 282, 80–82. [Google Scholar] [CrossRef]
- Linnerbauer, M.; Rothhammer, V. Protective Functions of Reactive Astrocytes Following Central Nervous System Insult. Front. Immunol. 2020, 11, 573256. [Google Scholar] [CrossRef]
- Cartocci, V.; Segatto, M.; Di Tunno, I.; Leone, S.; Pfrieger, F.W.; Pallottini, V. Modulation of the Isoprenoid/Cholesterol Biosynthetic Pathway During Neuronal Differentiation In Vitro. J. Cell. Biochem. 2016, 117, 2036–2044. [Google Scholar] [CrossRef]
- Harris, F.M.; Tesseur, I.; Brecht, W.J.; Xu, Q.; Mullendorff, K.; Chang, S.; Wyss-Coray, T.; Mahley, R.W.; Huang, Y. Astroglial regulation of apolipoprotein E expression in neuronal cells: Implications for Alzheimer’s disease. J. Biol. Chem. 2004, 279, 3862–3868. [Google Scholar] [CrossRef] [Green Version]
- Ioannou, M.S.; Liu, Z.; Lippincott-Schwartz, J. A Neuron-Glia Co-culture System for Studying Intercellular Lipid Transport. Curr. Protoc. Cell Biol. 2019, 84, e95. [Google Scholar] [CrossRef] [Green Version]
- Tonini, C.; Colardo, M.; Colella, B.; Di Bartolomeo, S.; Berardinelli, F.; Caretti, G.; Pallottini, V.; Segatto, M. Inhibition of bromodomain and extraterminal domain (BET) proteins by JQ1 unravels a novel epigenetic modulation to control lipid homeostasis. Int. J. Mol. Sci. 2020, 21, 1297. [Google Scholar] [CrossRef] [Green Version]
- Trapani, L.; Segatto, M.; Simeoni, V.; Balducci, V.; Dhawan, A.; Parmar, V.S.; Prasad, A.K.; Saso, L.; Incerpi, S.; Pallottini, V. Short- and long-term regulation of 3-hydroxy 3-methylglutaryl coenzyme A reductase by a 4-methylcoumarin. Biochimie 2011, 93, 1165–1171. [Google Scholar] [CrossRef]
- Pesiri, V.; Totta, P.; Segatto, M.; Bianchi, F.; Pallottini, V.; Marino, M.; Acconcia, F. Estrogen receptor α L429 and A430 regulate 17β-estradiol-induced cell proliferation via CREB1. Cell. Signal. 2015, 27, 2380–2388. [Google Scholar] [CrossRef] [PubMed]
- Segatto, M.; Szokoll, R.; Fittipaldi, R.; Bottino, C.; Nevi, L.; Mamchaoui, K.; Filippakopoulos, P.; Caretti, G. BETs inhibition attenuates oxidative stress and preserves muscle integrity in Duchenne muscular dystrophy. Nat. Commun. 2020, 11, 6108. [Google Scholar] [CrossRef]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Colardo, M.; Petraroia, M.; Lerza, L.; Pensabene, D.; Martella, N.; Pallottini, V.; Segatto, M. NGF Modulates Cholesterol Metabolism and Stimulates ApoE Secretion in Glial Cells Conferring Neuroprotection against Oxidative Stress. Int. J. Mol. Sci. 2022, 23, 4842. https://doi.org/10.3390/ijms23094842
Colardo M, Petraroia M, Lerza L, Pensabene D, Martella N, Pallottini V, Segatto M. NGF Modulates Cholesterol Metabolism and Stimulates ApoE Secretion in Glial Cells Conferring Neuroprotection against Oxidative Stress. International Journal of Molecular Sciences. 2022; 23(9):4842. https://doi.org/10.3390/ijms23094842
Chicago/Turabian StyleColardo, Mayra, Michele Petraroia, Letizia Lerza, Daniele Pensabene, Noemi Martella, Valentina Pallottini, and Marco Segatto. 2022. "NGF Modulates Cholesterol Metabolism and Stimulates ApoE Secretion in Glial Cells Conferring Neuroprotection against Oxidative Stress" International Journal of Molecular Sciences 23, no. 9: 4842. https://doi.org/10.3390/ijms23094842