title: Adeno-associated viral serotypes produce differing titers and differentially transduce neurons within the rat basal and lateral amygdala.
abstract: BACKGROUND
  In recent years, there has been an increased interest in using recombinant adeno-associated viruses (AAV) to make localized genetic manipulations within the rodent brain. Differing serotypes of AAV possess divergent capsid protein sequences and these variations greatly influence each serotype's ability to transduce particular cell types and brain regions. We therefore aimed to determine the AAV serotype that is optimal for targeting neurons within the Basal and Lateral Amygdala (BLA) since the transduction efficiency of AAV has not been previously examined within the BLA. This region is desirable to genetically manipulate due to its role in emotion, learning & memory, and numerous psychiatric disorders. We accomplished this by screening 9 different AAV serotypes (AAV2/1, AAV2/2, AAV2/5, AAV2/7, AAV2/8, AAV2/9, AAV2/rh10, AAV2/DJ and AAV2/DJ8) designed to express red fluorescent protein (RFP) under the regulation of an alpha Ca2+/calmodulin-dependent protein kinase II promoter (αCaMKII).
  
  
  RESULTS
  We determined that these serotypes produce differing amounts of virus under standard laboratory production. Notably AAV2/2 consistently produced the lowest titers compared to the other serotypes examined. These nine serotypes were bilaterally infused into the rat BLA at the highest titers achieved for each serotype and at a normalized titer of 7.8E + 11 GC/ml. Twenty one days following viral infusion the degree of transduction was quantitated throughout the amygdala. These viruses exhibited differential transduction of neurons within the BLA. AAV2/7 exhibited a trend toward having the highest efficiency of transduction and AAV2/5 exhibited significantly lower transduction efficiency as compared to the serotypes examined. AAV2/5's decreased ability to transduce BLA neurons correlates with its significantly different capsid protein sequences as compared to the other serotypes examined.
  
  
  CONCLUSIONS
  For laboratories producing their own recombinant adeno-associated viruses, the use of AAV2/2 is likely less desirable since AAV2/2 produces significantly lower titers than many other serotypes of AAV. Numerous AAV serotypes appear to efficiently transduce BLA neurons, with the exception of AAV2/5. Taking into consideration the ability of certain serotypes to achieve high titers and transduce BLA neurons well, in our hands AAV2/DJ8 and AAV2/9 appear to be ideal serotypes to use when targeting neurons within the BLA.
authors.forename: Roopashri
authors.initials: R
authors.lastname: Holehonnur
authors.forename: Jonathan A
authors.initials: JA
authors.lastname: Luong
authors.forename: Dushyant
authors.initials: D
authors.lastname: Chaturvedi
authors.forename: Anthony
authors.initials: A
authors.lastname: Ho
authors.forename: Srihari K
authors.initials: SK
authors.lastname: Lella
authors.forename: Matthew P
authors.initials: MP
authors.lastname: Hosek
authors.affiliation: School of Behavioral and Brain Sciences and the Department of Molecular & Cell Biology, University of Texas at Dallas, 800 West Campbell RD, Richardson, TX 75080, USA. Jonathan.Ploski@UTDallas.edu.
authors.forename: Jonathan E
authors.initials: JE
authors.lastname: Ploski
mesh_terms: D000256:Adenoviridae / Q000145:classification* / Q000502:physiology*; D000679:Amygdala / Q000502:physiology* / Q000821:virology*; D000818:Animals; D008164:Luminescent Proteins / Q000502:physiology*; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012703:Serotyping; D014018:Tissue Distribution; D014159:Transducers; D014161:Transduction, Genetic / Q000379:methods*; D019562:Viral Load / Q000502:physiology*; D000097573:Red Fluorescent Protein
chemical_list: D008164:Luminescent Proteins; D011994:Recombinant Proteins; D000097573:Red Fluorescent Protein