Abstract
Lentiviral vectors are used for a wide range of research applications in the field of neuroscience. Notably, lentiviral tropism can be manipulated by genetically engineering envelope glycoproteins that are essential for vector transduction. The structural and functional analyses of specific neural pathways are crucial to understanding the neural mechanisms underlying brain functions controlled through complex neural circuits. Viral vectors that produce gene transfer via retrograde axonal transport offer a powerful tool for the analysis of neural pathways. We have succeeded in developing novel types of lentiviral vectors for retrograde gene transfer, named “highly efficient retrograde gene transfer” (HiRet) and “neuron-specific retrograde gene transfer” (NeuRet) vectors, by pseudotyping human immunodeficiency virus type 1 with fusion envelope glycoproteins composed of rabies virus glycoprotein segments and vesicular stomatitis virus glycoprotein segments. HiRet/NeuRet vectors show highly efficient retrograde gene transfer in diverse neural pathways in animal models. These vectors have been harnessed for the analysis of specific neural pathways in combination with various genetic approaches for neuromodulation, including optogenetics and chemogenetics. In this chapter, we describe experimental procedures for producing HiRet/NeuRet vectors and injecting them into brain regions, as well as summarize points important to conduct the experiments smoothly and effectively.
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References
Naldini L, Blömer U, Gage FH et al (1996) Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector. Proc Natl Acad Sci U S A 93:11382–11388
Mitrophanous K, Yoon S, Rohll J et al (1999) Stable gene transfer to the nervous system using a non-primate lentiviral vector. Gene Ther 6:1808–1818
Butler SL, Johnson EP, Bushman FD (2002) Human immunodeficiency virus cDNA metabolism: notable stability of two-long terminal repeat circles. J Virol 76:3739–3747
Thomas CE, Ehrhardt A, Kay MA (2003) Progress and problems with the use of viral vectors for gene therapy. Nat Rev Genet 4:346–358
Kafri T (2004) Gene delivery by lentivirus vectors an overview. Methods Mol Biol 246:367–390
Wong LF, Goodhead L, Prat C et al (2006) Lentivirus-mediated gene transfer to the central nervous system: therapeutic and research applications. Hum Gene Ther 17:1–9
Cockrell AS, Kafri T (2007) Gene delivery by lentivirus vectors. Mol Biotechnol 36:184–204
Nectow AR, Nestler EJ (2020) Viral tools for neuroscience. Nat Rev Neurosci 21:669–681
Cronin J, Zhang XY, Reiser J (2005) Altering the tropism of lentiviral vectors through pseudotyping. Curr Gene Ther 5:387–398
Kobayashi K, Inoue KI, Tanabe S et al (2017) Pseudotyped lentiviral vectors for retrograde gene delivery into target brain regions. Front Neuroanat 11:65
Kobayashi K, Kato S, Kobayashi K (2018) Genetic manipulation of specific neural circuits by use of a viral vector system. J Neural Transm (Vienna) 125:67–75
Kato S, Kobayashi K (2020) Pseudotyped lentiviral vectors for tract-targeting and application for the functional control of selective neural circuits. J Neurosci Methods 344:108854
Joshi S, Joshi RL (1996) Molecular biology of human immunodeficiency virus type-1. Transfus Sci 17:351–378
Nielsen MH, Pedersen FS, Kjems J (2005) Molecular strategies to inhibit HIV-1 replication. Retrovirology 2:10
Pluta K, Kacprzak MM (2009) Use of HIV as a gene transfer vector. Acta Biochim Pol 56:531–595
Haery L, Deverman BE, Matho KS et al (2019) Adeno-associated virus technologies and methods for targeted neuronal manipulation. Front Neuroanat 13:93
Callaway EM, Luo L (2015) Monosynaptic circuit tracing with glycoprotein-deleted rabies viruses. J Neurosci 35:8979–8985
Junyent F, Kremer EJ (2015) CAV-2--why a canine virus is a neurobiologist's best friend. Curr Opin Pharmacol 24:86–93
Pomeranz LE, Reynolds AE, Hengartner CJ (2005) Molecular biology of pseudorabies virus: impact on neurovirology and veterinary medicine. Microbiol Mol Biol Rev 69:462–500
Fenno LE, Mattis J, Ramakrishnan C et al (2014) Targeting cells with single vectors using multiple-feature Boolean logic. Nat Methods 11:763–772
Kato S, Kobayashi K, Inoue K et al (2011) A lentiviral strategy for highly efficient retrograde gene transfer by pseudotyping with fusion envelope glycoprotein. Hum Gene Ther 22:197–206
Kato S, Kuramochi M, Takasumi K et al (2011) Neuron-specific gene transfer through retrograde transport of lentiviral vector pseudotyped with a novel type of fusion envelope glycoprotein. Hum Gene Ther 22:1511–1523
Kato S, Kobayashi K, Kobayashi K (2014) Improved transduction efficiency of a lentiviral vector for neuron-specific retrograde gene transfer by optimizing the junction of fusion envelope glycoprotein. J Neurosci Methods 227:151–158
Kato S, Sugawara M, Kobayashi K et al (2019) Enhancement of the transduction efficiency of a lentiviral vector for neuron-specific retrograde gene delivery through the point mutation of fusion glycoprotein type E. J Neurosci Methods 311:147–155
Tanabe S, Inoue KI, Tsuge H et al (2017) The use of an optimized chimeric envelope glycoprotein enhances the efficiency of retrograde gene transfer of a pseudotyped lentiviral vector in the primate brain. Neurosci Res 120:45–52
Tanabe S, Uezono S, Tsuge H et al (2019) A note on retrograde gene transfer efficiency and inflammatory response of lentiviral vectors pseudotyped with FuG-E vs. FuG-B2 glycoproteins. Sci Rep 9:3567
Matsuda T, Hiyama TY, Niimura F et al (2017) Distinct neural mechanisms for the control of thirst and salt appetite in the subfornical organ. Nat Neurosci 20:230–241
Morishima M, Kobayashi K, Kato S et al (2017) Segregated excitatory-inhibitory recurrent subnetworks in layer 5 of the rat frontal cortex. Cereb Cortex 27:5846–5857
Kato S, Fukabori R, Nishizawa K et al (2018) Action selection and flexible switching controlled by the intralaminar thalamic neurons. Cell Rep 22:2370–2382
Nomura K, Hiyama TY, Sakuta H et al (2019) [Na+] increases in body fluids sensed by central Nax induce sympathetically mediated blood pressure elevations via H+-dependent activation of ASIC1a. Neuron 101:60–75
Isa K, Sooksawate T, Kobayashi K et al (2020) Dissecting the tectal output channels for orienting and defense responses. eNeuro 7:ENEURO.0271-20.2020
Ishida A, Kobayashi K, Ueda Y et al (2019) Dynamic interaction between cortico-brainstem pathways during training-induced recovery in stroke model rats. J Neurosci 39:7306–7320
Hayashi T, Akikawa R, Kawasaki K et al (2020) Macaques exhibit implicit gaze bias anticipating others' false-belief-driven actions via medial prefrontal cortex. Cell Rep 30:4433–4444
Kato S, Kuramochi M, Kobayashi K et al (2011) Selective neural pathway targeting reveals key roles of thalamostriatal projection in the control of visual discrimination. J Neurosci 31:17169–17179
Kobayashi K, Sano H, Kato S et al (2016) Survival of corticostriatal neurons by Rho/Rho-kinase signaling pathway. Neurosci Lett 630:45–52
Paxinos G, Franklin KBJ (2008) The mouse brain in stereotaxic coordinates, 3rd edn. Academic Press, San Diego
Kato S, Inoue K, Kobayashi K et al (2007) Efficient gene transfer via retrograde transport in rodent and primate brains using a human immunodeficiency virus type 1-based vector pseudotyped with rabies virus glycoprotein. Hum Gene Ther 18:1141–1151
Inoue K, Koketsu D, Kato S et al (2012) Immunotoxin-mediated tract targeting in the primate brain: selective elimination of the cortico-subthalamic “hyperdirect” pathway. PLoS One 7:e39149
Kinoshita M, Matsui R, Kato S et al (2012) Genetic dissection of the circuit for hand dexterity in primates. Nature 487:235–238
Sooksawate T, Isa K, Matsui R et al (2013) Viral vector-mediated selective and reversible blockade of the pathway for visual orienting in mice. Front Neural Circuits 7:162
Ishida A, Isa K, Umeda T et al (2016) Causal link between the cortico-rubral pathway and functional recovery through forced impaired limb use in rats with stroke. J Neurosci 36:455–467
Tohyama T, Kinoshita M, Kobayashi K et al (2017) Contribution of propriospinal neurons to recovery of hand dexterity after corticospinal tract lesions in monkeys. Proc Natl Acad Sci U S A 114:604–609
Kinoshita M, Kato R, Isa K et al (2019) Dissecting the circuit for blindsight to reveal the critical role of pulvinar and superior colliculus. Nat Commun 10:135
Vancraeyenest P, Arsenault JT, Li X et al (2020) Selective mesoaccumbal pathway inactivation affects motivation but not reinforcement-based learning in macaques. Neuron 108:568–581
Acknowledgments
This work was supported by a grant-in-aid from Japan Agency for Medical Research and Development (JP17dm0207052 to Ka.K.). We thank Dr. R. Fukabori for providing photographs and Dr. Y. Iguchi for his critical reading of this manuscript.
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Kobayashi, K., Kato, S., Kobayashi, K. (2023). HiRet/NeuRet Vectors: Lentiviral System for Highly Efficient Gene Transfer Through Retrograde Axonal Transport. In: Eldridge, M.A., Galvan, A. (eds) Vectorology for Optogenetics and Chemogenetics. Neuromethods, vol 195. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2918-5_2
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DOI: https://doi.org/10.1007/978-1-0716-2918-5_2
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