Visualization of dynamics of single endogenous mRNA labeled in live mouse.
The transcription and transport of messenger RNA (mRNA) are critical steps in regulating the spatial and temporal components of gene expression, but it has not been possible to observe the dynamics of endogenous mRNA in primary mammalian tissues. We have developed a transgenic mouse in which all β-actin mRNA is fluorescently labeled. We found that β-actin mRNA in primary fibroblasts localizes predominantly by diffusion and trapping as single mRNAs. In cultured neurons and acute brain slices, we found that multiple β-actin mRNAs can assemble together, travel by active transport, and disassemble upon depolarization by potassium chloride. Imaging of brain slices revealed immediate early induction of β-actin transcription after depolarization. Studying endogenous mRNA in live mouse tissues provides insight into its dynamic regulation within the context of the cellular and tissue microenvironment.
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Hye Yoon
HY
Park
Hyungsik
H
Lim
Young J
YJ
Yoon
Antonia
A
Follenzi
Chiso
C
Nwokafor
Melissa
M
Lopez-Jones
Xiuhua
X
Meng
Robert H
RH
Singer
D000199:Actins / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D001921:Brain / Q000166:cytology / Q000378:metabolism; D002478:Cells, Cultured; D005347:Fibroblasts / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry; D051379:Mice; D008822:Mice, Transgenic; D059906:Neuroimaging / Q000379:methods*; D009474:Neurons / Q000378:metabolism; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*; D013194:Staining and Labeling
D000199:Actins; D005456:Fluorescent Dyes; D012333:RNA, Messenger