AAV-Mediated Anterograde Transsynaptic Tagging: Mapping Corticocollicular Input-Defined Neural Pathways for Defense Behaviors

To decipher neural circuits underlying brain functions, viral tracers are widely applied to map input and output connectivity of neuronal populations. Despite the successful application of retrograde transsynaptic viruses for identifying presynaptic neurons of transduced neurons, analogous anterograde transsynaptic tools for tagging postsynaptically targeted neurons remain under development. Here, we discovered that adeno-associated viruses (AAV1 and AAV9) exhibit anterograde transsynaptic spread properties. AAV1-Cre from transduced presynaptic neurons effectively and specifically drives Cre-dependent transgene expression in selected postsynaptic neuronal targets, thus allowing axonal tracing and functional manipulations of the latter input-defined neuronal population. Its application in superior colliculus (SC) reveals that SC neuron subpopulations receiving corticocollicular projections from auditory and visual cortex specifically drive flight and freezing, two different types of defense behavior, respectively. Together with an intersectional approach, AAV-mediated anterograde transsynaptic tagging can categorize neurons by their inputs and molecular identity, and allow forward screening of distinct functional neural pathways embedded in complex brain circuits.

[1]  K. Svoboda,et al.  The subcellular organization of neocortical excitatory connections , 2009, Nature.

[2]  Brian Zingg,et al.  Sensory Cortical Control of a Visually Induced Arrest Behavior via Corticotectal Projections , 2015, Neuron.

[3]  M. Ekstrand,et al.  The alpha-herpesviruses: molecular pathfinders in nervous system circuits. , 2008, Trends in molecular medicine.

[4]  R. Sturrock Light microscopic identification of immature glial cells in semithin sections of the developing mouse corpus callosum. , 1976, Journal of anatomy.

[5]  P. Dean,et al.  Responses resembling defensive behaviour produced by microinjection of glutamate into superior colliculus of rats , 1988, Neuroscience.

[6]  Karl Deisseroth,et al.  Midbrain circuits for defensive behaviour , 2016, Nature.

[7]  David J. Anderson,et al.  A Cre-Dependent, Anterograde Transsynaptic Viral Tracer for Mapping Output Pathways of Genetically Marked Neurons , 2011, Neuron.

[8]  E. Kremer,et al.  CAV-2--why a canine virus is a neurobiologist's best friend. , 2015, Current opinion in pharmacology.

[9]  Hong Wei Dong,et al.  Allen reference atlas : a digital color brain atlas of the C57Black/6J male mouse , 2008 .

[10]  P. Redgrave,et al.  Segregated Anatomical Input to Sub-Regions of the Rodent Superior Colliculus Associated with Approach and Defense , 2012, Front. Neuroanat..

[11]  S. Rumpel,et al.  Analysis of Transduction Efficiency, Tropism and Axonal Transport of AAV Serotypes 1, 2, 5, 6, 8 and 9 in the Mouse Brain , 2013, PloS one.

[12]  Johannes J. Letzkus,et al.  Amygdala interneuron subtypes control fear learning through disinhibition , 2014, Nature.

[13]  Karl Deisseroth,et al.  Optogenetics in Neural Systems , 2011, Neuron.

[14]  Qian Wang,et al.  A parvalbumin-positive excitatory visual pathway to trigger fear responses in mice , 2015, Science.

[15]  Hysell V. Oviedo,et al.  Long-term Cre-mediated retrograde tagging of neurons using a novel recombinant pseudorabies virus , 2014, Front. Neuroanat..

[16]  Liqun Luo,et al.  Monosynaptic Circuit Tracing with Glycoprotein-Deleted Rabies Viruses , 2015, The Journal of Neuroscience.

[17]  Xintian Hu,et al.  Processing of visually evoked innate fear by a non-canonical thalamic pathway , 2015, Nature Communications.

[18]  L. P. Morin,et al.  Retinofugal projections in the mouse , 2014, The Journal of comparative neurology.

[19]  M. S. Chapman,et al.  An essential receptor for adeno-associated virus infection , 2015, Nature.

[20]  J. Price :Allen Reference Atlas: A Digital Color Brain Atlas of the C57BL/6J Male Mouse , 2008 .

[21]  Matt Wachowiak,et al.  Transgene Expression in Target-Defined Neuron Populations Mediated by Retrograde Infection with Adeno-Associated Viral Vectors , 2013, The Journal of Neuroscience.

[22]  K. Kissa,et al.  Preferential transduction of neurons by canine adenovirus vectors and their efficient retrograde transport in vivo , 2001, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[23]  J. Silver,et al.  Glial fibrillary acidic protein is necessary for mature astrocytes to react to β‐amyloid , 1999 .

[24]  M. Castro-Alamancos,et al.  Whisker-related afferents in superior colliculus. , 2016, Journal of neurophysiology.

[25]  J. Silver,et al.  Glial fibrillary acidic protein is necessary for mature astrocytes to react to beta-amyloid. , 1999, Glia.

[26]  Michael J. Castle,et al.  Adeno-associated virus serotypes 1, 8, and 9 share conserved mechanisms for anterograde and retrograde axonal transport. , 2014, Human gene therapy.

[27]  Michael J. Castle,et al.  Long-distance axonal transport of AAV9 is driven by dynein and kinesin-2 and is trafficked in a highly motile Rab7-positive compartment. , 2014, Molecular therapy : the journal of the American Society of Gene Therapy.

[28]  V. Lemmon,et al.  Afferent and efferent connections of the striate and extrastriate visual cortex of the normal and reeler mouse , 1982, The Journal of comparative neurology.

[29]  Yoshikazu Isomura,et al.  Two distinct layer-specific dynamics of cortical ensembles during learning of a motor task , 2014, Nature Neuroscience.

[30]  J. Friedman,et al.  Virus-Assisted Mapping of Neural Inputs to a Feeding Center in the Hypothalamus , 2001, Science.

[31]  L. Schenberg,et al.  Functional specializations within the tectum defense systems of the rat , 2005, Neuroscience & Biobehavioral Reviews.

[32]  Brian Zingg,et al.  Thalamocortical Innervation Pattern in Mouse Auditory and Visual Cortex: Laminar and Cell-Type Specificity. , 2016, Cerebral cortex.

[33]  F. Aboitiz,et al.  One hundred million years of interhemispheric communication: the history of the corpus callosum. , 2003, Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas.

[34]  C. Kathe,et al.  Trans-neuronal transduction of spinal neurons following cortical injection and anterograde axonal transport of a bicistronic AAV1 vector , 2015, Gene Therapy.

[35]  Allan R. Jones,et al.  A robust and high-throughput Cre reporting and characterization system for the whole mouse brain , 2009, Nature Neuroscience.

[36]  Hongkui Zeng,et al.  Adeno‐Associated Viral Vectors for Anterograde Axonal Tracing with Fluorescent Proteins in Nontransgenic and Cre Driver Mice , 2012, Current protocols in neuroscience.

[37]  Brian Zingg,et al.  Cross-Modality Sharpening of Visual Cortical Processing through Layer-1-Mediated Inhibition and Disinhibition , 2016, Neuron.

[38]  J. K. Harting,et al.  Connectional organization of the superior colliculus , 1984, Trends in Neurosciences.

[39]  David V. Schaffer,et al.  Engineering adeno-associated viruses for clinical gene therapy , 2014, Nature Reviews Genetics.

[40]  Gabriele Mittermeyer,et al.  Axonal transport of adeno-associated viral vectors is serotype-dependent , 2012, Gene Therapy.

[41]  Ian R. Wickersham,et al.  Monosynaptic circuit tracing in vivo through Cre-dependent targeting and complementation of modified rabies virus , 2010, Proceedings of the National Academy of Sciences.

[42]  G. Ugolini,et al.  Advances in viral transneuronal tracing , 2010, Journal of Neuroscience Methods.

[43]  K. Deisseroth,et al.  Molecular and Cellular Approaches for Diversifying and Extending Optogenetics , 2010, Cell.

[44]  L. Looger,et al.  A Designer AAV Variant Permits Efficient Retrograde Access to Projection Neurons , 2016, Neuron.

[45]  Kevin T. Beier,et al.  Anterograde or retrograde transsynaptic labeling of CNS neurons with vesicular stomatitis virus vectors , 2011, Proceedings of the National Academy of Sciences.

[46]  R. Gronostajski,et al.  The FLP recombinase of the Saccharomyces cerevisiae 2 microns plasmid attaches covalently to DNA via a phosphotyrosyl linkage , 1985, Molecular and cellular biology.

[47]  Lief E. Fenno,et al.  Targeting cells with single vectors using multiple-feature Boolean logic , 2014, Nature Methods.

[48]  Brian Zingg,et al.  Auditory cortex controls sound-driven innate defense behaviour through corticofugal projections to inferior colliculus , 2015, Nature Communications.

[49]  Allan R. Jones,et al.  A mesoscale connectome of the mouse brain , 2014, Nature.

[50]  Arthur W. Toga,et al.  Neural Networks of the Mouse Neocortex , 2014, Cell.

[51]  Liqun Luo,et al.  Viral-genetic tracing of the input–output organization of a central norepinephrine circuit , 2015, Nature.

[52]  C. Petersen,et al.  Long‐range connectivity of mouse primary somatosensory barrel cortex , 2010, The European journal of neuroscience.

[53]  Kevin T. Beier,et al.  Neuroanatomy goes viral! , 2015, Front. Neuroanat..

[54]  Ian R. Wickersham,et al.  Monosynaptic Restriction of Transsynaptic Tracing from Single, Genetically Targeted Neurons , 2007, Neuron.

[55]  P Redgrave,et al.  Movements resembling orientation or avoidance elicited by electrical stimulation of the superior colliculus in rats , 1986, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[56]  G. Perea,et al.  Tripartite synapses: astrocytes process and control synaptic information , 2009, Trends in Neurosciences.