Visualization of cortical projection neurons with retrograde TET-off lentiviral vector.
We are interested in identifying and characterizing various projection neurons that constitute the neocortical circuit. For this purpose, we developed a novel lentiviral vector that carries the tetracycline transactivator (tTA) and the transgene under the TET Responsive Element promoter (TRE) on a single backbone. By pseudotyping such a vector with modified rabies G-protein, we were able to express palmitoylated-GFP (palGFP) or turboFP635 (RFP) in corticothalamic, corticocortical, and corticopontine neurons of mice. The high-level expression of the transgene achieved by the TET-Off system enabled us to observe characteristic elaboration of neuronal processes for each cell type. At higher magnification, we were able to observe fine structures such as boutons and spines as well. We also injected our retrograde TET-Off vector to the marmoset cortex and proved that it can be used to label the long-distance cortical connectivity of millimeter scale. In conclusion, our novel retrograde tracer provides an attractive option to investigate the morphologies of identified cortical projection neurons of various species.
Division of Brain Biology, National Institute for Basic Biology, Okazaki, Japan. watakabe@nibb.ac.jp
Akiya
A
Watakabe
Shigeki
S
Kato
Kazuto
K
Kobayashi
Masafumi
M
Takaji
Yuki
Y
Nakagami
Osamu
O
Sadakane
Masanari
M
Ohtsuka
Hiroyuki
H
Hioki
Takeshi
T
Kaneko
Hiroyuki
H
Okuno
Takashi
T
Kawashima
Haruhiko
H
Bito
Yoshihiro
Y
Kitamura
Tetsuo
T
Yamamori
D000818:Animals; D002540:Cerebral Cortex / Q000166:cytology*; D005456:Fluorescent Dyes; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D009474:Neurons / Q000166:cytology*; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000235:genetics*; D019076:Transgenes
D005456:Fluorescent Dyes; D012097:Repressor Proteins; C034811:tetracycline resistance-encoding transposon repressor protein; D049452:Green Fluorescent Proteins