Optogenetic Modulation of Neural Progenitor Cells Improves Neuroregenerative Potential
Abstract
:1. Introduction
2. Results
2.1. Optogenetic Stimulation Increases ChR2-NPC Proliferation and Neurite Outgrowth under Proliferative Conditions
2.2. Optogenetic Stimulation Enhances ChR2-NPC Differentiation into Neurons and Oligodendrocytes
2.3. Optogenetic Stimulation of ChR2-NPCs Influences Astrocyte Maturation into Distinct Morphological and Functional Subtypes
2.4. Optogenetic Stimulation Enhances Axon Growth and Branching in ChR2-NPC-Derived Neurons
3. Discussion
4. Materials and Methods
4.1. NPC Isolation, Culture, ChR2 Transduction, and Photostimulation
4.2. Drug Administrations
4.3. Immunocytochemical Staining, Image Acquisition, and Quantification
4.4. Statistical Analyses
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
AAV | Adeno-associated virus |
AAV9 | Adeno-associated virus serotype 9 |
AKT | Protein kinase B |
APC | Adenomatous-polyposis-coli |
BL | Blue light |
bFGF | Basic fibroblast growth factor |
CAG | CMV early enhancer/chicken β actin |
CaMKII | Ca2+/calmodulin-dependent protein kinase II |
cAMP | Cyclic adenosine monophosphate |
ChR2 | Channelrhodopsin-2 |
CREB | cAMP-responsive element-binding protein |
CSPG | Chondroitin sulphate proteoglycan |
DAPI | 4′,6-diamidino-2-phenylindole |
EGF | Epidermal growth factor |
EPAC2 | Exchange protein directly activated by cAMP |
ERK | Extracellular-signal-related kinase |
GFAP | Glial fibrillary acidic protein |
GSK3β | Glycogen synthase kinase 3β |
HBSS | Hanks Balanced Salt Solution |
hChR2 | Humanized channelrhodopsin-2 |
mTOR | Mammalian target of rapamycin |
NGS | Normal goat serum |
NF-κB | Nuclear Factor κB |
NPC | Neural progenitor cell |
LIF | Leukaemia inhibitory factor |
LPA | Lysophosphatidic acid |
Olig2 | Oligodendrocyte transcription factor 2 |
PI3K | Phosphoinositide 3-kinase |
SCI | Spinal cord injury |
Stat3 | Signal transducer and activator of transcription 3 |
VGCC | Voltage-gated calcium channel |
References
- Fischer, I.; Dulin, J.N.; Lane, M.A. Transplanting neural progenitor cells to restore connectivity after spinal cord injury. Nat. Rev. Neurosci. 2020, 21, 366–383. [Google Scholar] [CrossRef] [PubMed]
- De Feo, D.; Merlini, A.; Laterza, C.; Martino, G. Neural stem cell transplantation in central nervous system disorders: From cell replacement to neuroprotection. Curr. Opin. Neurol. 2012, 25, 322–333. [Google Scholar] [CrossRef] [PubMed]
- Giusto, E.; Donegà, M.; Cossetti, C.; Pluchino, S. Neuro-immune interactions of neural stem cell transplants: From animal disease models to human trials. Exp. Neurol. 2014, 260, 19–32. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ceto, S.; Sekiguchi, K.J.; Takashima, Y.; Nimmerjahn, A.; Tuszynski, M.H. Neural Stem Cell Grafts Form Extensive Synaptic Networks that Integrate with Host Circuits after Spinal Cord Injury. Cell Stem Cell 2020, 27, 430–440.e5. [Google Scholar] [CrossRef]
- Yang, B.; Zhang, F.; Cheng, F.; Ying, L.; Wang, C.; Shi, K.; Wang, J.; Xia, K.; Gong, Z.; Huang, X.; et al. Strategies and prospects of effective neural circuits reconstruction after spinal cord injury. Cell Death Dis. 2020, 11, 1–14. [Google Scholar] [CrossRef]
- Lu, P.; Graham, L.; Wang, Y.; Wu, D.; Tuszynski, M. Promotion of Survival and Differentiation of Neural Stem Cells with Fibrin and Growth Factor Cocktails after Severe Spinal Cord Injury. J. Vis. Exp. 2014, e50641. [Google Scholar] [CrossRef] [Green Version]
- Martínez-Ramos, C.; Doblado, L.R.; Mocholi, E.L.; Alastrue-Agudo, A.; Petidier, M.S.; Giraldo, E.; Pradas, M.M.; Moreno-Manzano, V. Biohybrids for spinal cord injury repair. J. Tissue Eng. Regen. Med. 2019, 13, 509–521. [Google Scholar] [CrossRef]
- Alastrue-Agudo, A.; Rodriguez-Jimenez, F.J.; López-Mocholí, E.; de Giorgio, F.; Erceg, S.; Moreno-Manzano, V. FM19G11 and Ependymal Progenitor/Stem Cell Combinatory Treatment Enhances Neuronal Preservation and Oligodendrogenesis after Severe Spinal Cord Injury. Int. J. Mol. Sci. 2018, 19, 200. [Google Scholar] [CrossRef] [Green Version]
- Requejo, R.; Alastrue-Agudo, A.; Cases-Villar, M.; Lopez-Mocholi, E.; England, R.; Vicent, M.J.; Moreno-Manzano, V. Combined polymer-curcumin conjugate and ependymal progenitor/stem cell treatment enhances spinal cord injury functional recovery. Biomaterials 2017, 113, 18–30. [Google Scholar] [CrossRef]
- Li, X.; Peng, Z.; Long, L.; Tuo, Y.; Wang, L.; Zhao, X.; Le, W.; Wan, Y. Wnt4-modified NSC transplantation promotes functional recovery after spinal cord injury. FASEB J. 2019, 34, 82–94. [Google Scholar] [CrossRef] [Green Version]
- Yu, S.P.; Tung, J.K.; Wei, Z.Z.; Chen, D.; Berglund, K.; Zhong, W.; Zhang, J.Y.; Gu, X.; Song, M.; Gross, R.E.; et al. Optochemogenetic Stimulation of Transplanted iPS-NPCs Enhances Neuronal Repair and Functional Recovery after Ischemic Stroke. J. Neurosci. 2019, 39, 6571–6594. [Google Scholar] [CrossRef] [Green Version]
- Nectow, A.R.; Nestler, E.J. Viral tools for neuroscience. Nat. Rev. Neurosci. 2020, 21, 669–681. [Google Scholar] [CrossRef] [PubMed]
- Thompson, A.C.; Wise, A.K.; Hart, W.L.; Needham, K.; Fallon, J.B.; Gunewardene, N.; Stoddart, P.R.; Richardson, R.T. Hybrid optogenetic and electrical stimulation for greater spatial resolution and temporal fidelity of cochlear activation. J. Neural Eng. 2020, 17, 056046. [Google Scholar] [CrossRef]
- Zhang, K.; Cui, B. Optogenetic control of intracellular signaling pathways. Trends Biotechnol. 2015, 33, 92–100. [Google Scholar] [CrossRef] [Green Version]
- Toettcher, J.E.; Voigt, C.A.; Weiner, O.D.; Lim, W.A. The promise of optogenetics in cell biology: Interrogating molecular circuits in space and time. Nat. Methods 2010, 8, 35–38. [Google Scholar] [CrossRef] [Green Version]
- Deisseroth, K. Optogenetics. Nat. Methods 2011, 8, 26–29. [Google Scholar] [CrossRef]
- Guru, A.; Post, R.J.; Ho, Y.-Y.; Warden, M.R. Making Sense of Optogenetics. Int. J. Neuropsychopharmacol. 2015, 18, pyv079. [Google Scholar] [CrossRef]
- Hongdusit, A.; Liechty, E.T.; Fox, J.M. Optogenetic interrogation and control of cell signaling. Curr. Opin. Biotechnol. 2020, 66, 195–206. [Google Scholar] [CrossRef]
- Weick, J.P.; Johnson, M.A.; Skroch, S.P.; Williams, J.C.; Deisseroth, K.; Zhang, S.-C. Functional Control of Transplantable Human ESC-Derived Neurons Via Optogenetic Targeting. Stem Cells 2010, 28, 2008–2016. [Google Scholar] [CrossRef] [Green Version]
- Steinbeck, J.A.; Choi, S.J.; Mrejeru, A.; Ganat, Y.; Deisseroth, K.; Sulzer, D.; Mosharov, E.V.; Studer, L. Optogenetics enables functional analysis of human embryonic stem cell-derived grafts in a Parkinson’s disease model. Nat. Biotechnol. 2015, 33, 204–209. [Google Scholar] [CrossRef]
- Daadi, M.M.; Klausner, J.Q.; Bajar, B.; Goshen, I.; Lee-Messer, C.; Lee, S.Y.; Winge, M.C.G.; Ramakrishnan, C.; Lo, M.; Sun, G.; et al. Optogenetic Stimulation of Neural Grafts Enhances Neurotransmission and Downregulates the Inflammatory Response in Experimental Stroke Model. Cell Transplant. 2016, 25, 1371–1380. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Bott, C.J.; Johnson, C.G.; Yap, C.C.; Dwyer, N.D.; Litwa, K.A.; Winckler, B. Nestin in immature embryonic neurons affects axon growth cone morphology and Semaphorin3a sensitivity. Mol. Biol. Cell 2019, 30, 1214–1229. [Google Scholar] [CrossRef] [PubMed]
- Tabata, H. Diverse subtypes of astrocytes and their development during corticogenesis. Front. Neurosci. 2015, 9, 114. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Liddelow, S.A.; Barres, B.A. Reactive Astrocytes: Production, Function, and Therapeutic Potential. Immunity 2017, 46, 957–967. [Google Scholar] [CrossRef] [Green Version]
- Chang, K.A.; Kim, J.W.; Kim, J.A.; Lee, S.E.; Kim, S.; Suh, W.H.; Kim, H.S.; Kwon, S.; Kim, S.J.; Suh, Y.H. Biphasic electrical currents stimulation promotes both proliferation and differentiation of fetal neural stem cells. PLoS ONE 2011, 6, e18738. [Google Scholar] [CrossRef] [Green Version]
- Yu, P.; Jin, J.; Tilve, S.; Huang, Z.; Zhou, L.; Geller, H.M. Effect of chondroitin sulfate proteoglycans on neuronal cell adhesion, spreading and neurite growth in culture. Neural Regen. Res. 2018, 13, 289–297. [Google Scholar] [CrossRef]
- Toettcher, J.E.; Weiner, O.D.; Lim, W.A. Using Optogenetics to Interrogate the Dynamic Control of Signal Transmission by the Ras/Erk Module. Cell 2013, 155, 1422–1434. [Google Scholar] [CrossRef] [Green Version]
- Nagel, G.; Szellas, T.; Huhn, W.; Kateriya, S.; Adeishvili, N.; Berthold, P.; Ollig, D.; Hegemann, P.; Bamberg, E. Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. Proc. Natl. Acad. Sci. USA 2003, 100, 13940–13945. [Google Scholar] [CrossRef] [Green Version]
- Boyden, E.S.; Zhang, F.; Bamberg, E.; Nagel, G.; Deisseroth, K. Millisecond-timescale, genetically targeted optical control of neural activity. Nat. Neurosci. 2005, 8, 1263–1268. [Google Scholar] [CrossRef]
- Lonze, B.E.; Ginty, D.D. Function and Regulation of CREB Family Transcription Factors in the Nervous System. Neuron 2002, 35, 605–623. [Google Scholar] [CrossRef] [Green Version]
- Kahl, C.R.; Means, A.R. Regulation of Cell Cycle Progression by Calcium/Calmodulin-Dependent Pathways. Endocr. Rev. 2003, 24, 719–736. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kajikawa, K.; Imaizumi, K.; Shinozaki, M.; Shibata, S.; Shindo, T.; Kitagawa, T.; Shibata, R.; Kamata, Y.; Kojima, K.; Nagoshi, N.; et al. Cell therapy for spinal cord injury by using human iPSC-derived region-specific neural progenitor cells. Mol. Brain 2020, 13, 120. [Google Scholar] [CrossRef] [PubMed]
- Montgomery, K.L.; Yeh, A.J.; Ho, J.S.; Tsao, V.; Iyer, S.M.; Grosenick, L.; A Ferenczi, E.; Tanabe, Y.; Deisseroth, K.; Delp, S.L.; et al. Wirelessly powered, fully internal optogenetics for brain, spinal and peripheral circuits in mice. Nat. Methods 2015, 12, 969–974. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Samineni, V.K.; Yoon, J.; Crawford, K.E.; Jeong, Y.R.; McKenzie, K.C.; Shin, G.; Xie, Z.; Sundaram, S.S.; Li, Y.; Yang, M.Y.; et al. Fully implantable, battery-free wireless optoelectronic devices for spinal optogenetics. Pain 2017, 158, 2108–2116. [Google Scholar] [CrossRef]
- Sabelström, H.; Stenudd, M.; Frisén, J. Neural stem cells in the adult spinal cord. Exp. Neurol. 2014, 260, 44–49. [Google Scholar] [CrossRef]
- Oda, Y.; Tani, K.; Isozaki, A.; Haraguchi, T.; Itamoto, K.; Nakazawa, H.; Taura, Y. Effects of Polyethylene Glycol Administration and Bone Marrow Stromal Cell Transplantation Therapy in Spinal Cord Injury Mice. J. Vet. Med Sci. 2014, 76, 415–421. [Google Scholar] [CrossRef] [Green Version]
- Stroh, A.; Tsai, H.-C.; Wang, L.-P.; Zhang, F.; Kressel, J.; Aravanis, A.; Santhanam, N.; Deisseroth, K.; Konnerth, A.; Schneider, M.B. Tracking Stem Cell Differentiation in the Setting of Automated Optogenetic Stimulation. Stem Cells 2011, 29, 78–88. [Google Scholar] [CrossRef] [Green Version]
- Kobelt, L.J.; Wilkinson, A.E.; McCormick, A.M.; Willits, R.K.; Leipzig, N.D. Short Duration Electrical Stimulation to Enhance Neurite Outgrowth and Maturation of Adult Neural Stem Progenitor Cells. Ann. Biomed. Eng. 2014, 42, 2164–2176. [Google Scholar] [CrossRef]
- Ono, K.; Suzuki, H.; Yamamoto, R.; Sahashi, H.; Takido, Y.; Sawada, M. Optogenetic control of cell differentiation in channelrhodopsin-2-expressing OS3, a bipotential glial progenitor cell line. Neurochem. Int. 2017, 104, 49–63. [Google Scholar] [CrossRef]
- Qiu, J.; Cai, D.; Dai, H.; McAtee, M.; Hoffman, P.N.; Bregman, B.S.; Filbin, M.T. Spinal Axon Regeneration Induced by Elevation of Cyclic AMP. Neuron 2002, 34, 895–903. [Google Scholar] [CrossRef] [Green Version]
- Lepski, G.; Jannes, C.E.; Nikkhah, G.; Bischofberger, J. cAMP promotes the differentiation of neural progenitor cells in vitro via modulation of voltage-gated calcium channels. Front. Cell. Neurosci. 2013, 7, 155. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Chierzi, S.; Ratto, G.M.; Verma, P.; Fawcett, J.W. The ability of axons to regenerate their growth cones depends on axonal type and age, and is regulated by calcium, cAMP and ERK. Eur. J. Neurosci. 2005, 21, 2051–2062. [Google Scholar] [CrossRef] [PubMed]
- Kamber, D.; Erez, H.; Spira, M.E. Local calcium-dependent mechanisms determine whether a cut axonal end assembles a retarded endbulb or competent growth cone. Exp. Neurol. 2009, 219, 112–125. [Google Scholar] [CrossRef] [PubMed]
- Mahar, M.; Cavalli, V. Intrinsic mechanisms of neuronal axon regeneration. Nat. Rev. Neurosci. 2018, 19, 323–337. [Google Scholar] [CrossRef] [PubMed]
- Wei, D.; Hurd, C.; Galleguillos, D.; Singh, J.; Fenrich, K.K.; Webber, C.A.; Sipione, S.; Fouad, K. Inhibiting cortical protein kinase A in spinal cord injured rats enhances efficacy of rehabilitative training. Exp. Neurol. 2016, 283, 365–374. [Google Scholar] [CrossRef] [PubMed]
- Park, K.K.; Liu, K.; Hu, Y.; Smith, P.D.; Wang, C.; Cai, B.; Xu, B.; Connolly, L.; Kramvis, I.; Sahin, M.; et al. Promoting Axon Regeneration in the Adult CNS by Modulation of the PTEN/mTOR Pathway. Science 2008, 322, 963–966. [Google Scholar] [CrossRef] [Green Version]
- Schöneborn, H.; Raudzus, F.; Coppey, M.; Neumann, S.; Heumann, R. Perspectives of RAS and RHEB GTPase Signaling Pathways in Regenerating Brain Neurons. Int. J. Mol. Sci. 2018, 19, 4052. [Google Scholar] [CrossRef] [Green Version]
- Van Niekerk, E.A.; Tuszynski, M.H.; Lu, P.; Dulin, J.N. Molecular and Cellular Mechanisms of Axonal Regeneration After Spinal Cord Injury. Mol. Cell. Proteom. 2016, 15, 394–408. [Google Scholar] [CrossRef] [Green Version]
- Shapiro, D.L. Morphological and Biochemical Alterations in Foetal Rat Brain Cells cultured in the Presence of Monobutyryl Cyclic AMP. Nat. Cell Biol. 1973, 241, 203–204. [Google Scholar] [CrossRef]
- Liddelow, S.A.; Guttenplan, K.A.; Clarke, L.E.; Bennett, F.C.; Bohlen, C.J.; Schirmer, L.; Bennett, M.L.; Münch, A.E.; Chung, W.-S.; Peterson, T.C.; et al. Neurotoxic reactive astrocytes are induced by activated microglia. Nature 2017, 541, 481–487. [Google Scholar] [CrossRef]
- Schiweck, J.; Eickholt, B.J.; Murk, K. Important Shapeshifter: Mechanisms Allowing Astrocytes to Respond to the Changing Nervous System During Development, Injury and Disease. Front. Cell. Neurosci. 2018, 12, 261. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Yang, T.; Dai, Y.; Chen, G.; Cui, S. Dissecting the Dual Role of the Glial Scar and Scar-Forming Astrocytes in Spinal Cord Injury. Front Cell Neurosci. 2020, 14, 78. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Nagel, G.; Brauner, M.; Liewald, J.F.; Adeishvili, N.; Bamberg, E.; Gottschalk, A. Light Activation of Channelrhodopsin-2 in Excitable Cells of Caenorhabditis elegans Triggers Rapid Behavioral Responses. Curr. Biol. 2005, 15, 2279–2284. [Google Scholar] [CrossRef] [PubMed] [Green Version]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2020 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 (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Giraldo, E.; Palmero-Canton, D.; Martinez-Rojas, B.; Sanchez-Martin, M.d.M.; Moreno-Manzano, V. Optogenetic Modulation of Neural Progenitor Cells Improves Neuroregenerative Potential. Int. J. Mol. Sci. 2021, 22, 365. https://doi.org/10.3390/ijms22010365
Giraldo E, Palmero-Canton D, Martinez-Rojas B, Sanchez-Martin MdM, Moreno-Manzano V. Optogenetic Modulation of Neural Progenitor Cells Improves Neuroregenerative Potential. International Journal of Molecular Sciences. 2021; 22(1):365. https://doi.org/10.3390/ijms22010365
Chicago/Turabian StyleGiraldo, Esther, David Palmero-Canton, Beatriz Martinez-Rojas, Maria del Mar Sanchez-Martin, and Victoria Moreno-Manzano. 2021. "Optogenetic Modulation of Neural Progenitor Cells Improves Neuroregenerative Potential" International Journal of Molecular Sciences 22, no. 1: 365. https://doi.org/10.3390/ijms22010365