Two-photon imaging of cellular dynamics in the mouse spinal cord.
Two-photon (2P) microscopy is utilized to reveal cellular dynamics and interactions deep within living, intact tissues. Here, we present a method for live-cell imaging in the murine spinal cord. This technique is uniquely suited to analyze neural precursor cell (NPC) dynamics following transplantation into spinal cords undergoing neuroinflammatory demyelinating disorders. NPCs migrate to sites of axonal damage, proliferate, differentiate into oligodendrocytes, and participate in direct remyelination. NPCs are thereby a promising therapeutic treatment to ameliorate chronic demyelinating diseases. Because transplanted NPCs migrate to the damaged areas on the ventral side of the spinal cord, traditional intravital 2P imaging is impossible, and only information on static interactions was previously available using histochemical staining approaches. Although this method was generated to image transplanted NPCs in the ventral spinal cord, it can be applied to numerous studies of transplanted and endogenous cells throughout the entire spinal cord. In this article, we demonstrate the preparation and imaging of a spinal cord with enhanced yellow fluorescent protein-expressing axons and enhanced green fluorescent protein-expressing transplanted NPCs.
Molecular Biology and Biochemistry, University of California, Irvine.
Jason G
JG
Weinger
Physiology and Biophysics, University of California, Irvine.
Milton L
ML
Greenberg
University of California San Francisco Diabetes Center, University of California, San Francisco.
Melanie P
MP
Matheu
Neurobiology and Behavior, University of California, Irvine.
Ian
I
Parker
Molecular Biology and Biochemistry, University of California, Irvine.
Craig M
CM
Walsh
Pathology, University of Utah.
Thomas E
TE
Lane
Physiology and Biophysics, University of California, Irvine; mcahalan@uci.edu.
Michael D
MD
Cahalan
D000818:Animals; D001369:Axons / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D003711:Demyelinating Diseases; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D058953:Neural Stem Cells / Q000166:cytology*; D009474:Neurons / Q000166:cytology; D009836:Oligodendroglia / Q000166:cytology; D013116:Spinal Cord / Q000166:cytology*
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins