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Spinal nociceptive circuit analysis with recombinant adeno‐associated viruses: the impact of serotypes and promoters
Author(s) -
Haenraets Karen,
Foster Edmund,
Johannssen Helge,
Kandra Vinnie,
Frezel Noémie,
Steffen Timothy,
Jaramillo Valeria,
Paterna JeanCharles,
Zeilhofer Hanns Ulrich,
Wildner Hendrik
Publication year - 2017
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/jnc.14124
Subject(s) - transduction (biophysics) , biology , adeno associated virus , neuroscience , spinal cord , rostral ventromedial medulla , microbiology and biotechnology , recombinant dna , gene , nociception , genetics , vector (molecular biology) , hyperalgesia , biochemistry , receptor
Recombinant adeno‐associated virus ( rAAV ) vector‐mediated gene transfer into genetically defined neuron subtypes has become a powerful tool to study the neuroanatomy of neuronal circuits in the brain and to unravel their functions. More recently, this methodology has also become popular for the analysis of spinal cord circuits. To date, a variety of naturally occurring AAV serotypes and genetically modified capsid variants are available but transduction efficiency in spinal neurons, target selectivity, and the ability for retrograde tracing are only incompletely characterized. Here, we have compared the transduction efficiency of seven commonly used AAV serotypes after intraspinal injection. We specifically analyzed local transduction of different types of dorsal horn neurons, and retrograde transduction of dorsal root ganglia ( DRG ) neurons and of neurons in the rostral ventromedial medulla (RVM) and the somatosensory cortex (S1). Our results show that most of the tested rAAV vectors have similar transduction efficiency in spinal neurons. All serotypes analyzed were also able to transduce DRG neurons and descending RVM and S1 neurons via their spinal axon terminals. When comparing the commonly used rAAV serotypes to the recently developed serotype 2 capsid variant rAAV 2retro, a > 20‐fold increase in transduction efficiency of descending supraspinal neurons was observed. Conversely, transgene expression in retrogradely transduced neurons was strongly reduced when the human synapsin 1 ( hS yn1) promoter was used instead of the strong ubiquitous hybrid cytomegalovirus enhancer/chicken β‐actin promoter (CAG) or cytomegalovirus (CMV) promoter fragments. We conclude that the use of AAV 2retro greatly increases transduction of neurons connected to the spinal cord via their axon terminals, while the hS yn1 promoter can be used to minimize transgene expression in retrogradely connected neurons of the DRG or brainstem. Cover Image for this issue: doi. 10.1111/jnc.13813 .