Functional motor neurons differentiating from mouse multipotent spinal cord precursor cells in culture and after transplantation into transected sciatic nerve.
OBJECTIVE
One of the current challenges in neurobiology is to ensure that neural precursor cells differentiate into specific neuron types, so that they can be used for transplantation purposes in patients with neuron loss. The goal of this study was to determine if spinal cord precursor cells could differentiate into motor neurons both in culture and following transplantation into a transected sciatic nerve.


METHODS
In cultures with trophic factors, neurons differentiate from embryonic precursor cells and express motor neuronal markers such as choline acetyltransferase (ChAT), Islet-1, and REG2. Reverse transcription-polymerase chain reaction analysis has also demonstrated the expression of Islet-1 in differentiated cultures. A coculture preparation of neurospheres and skeletal myocytes was used to show the formation of neuromuscular connections between precursor cell-derived neurons and myocytes both immunohistochemically and electrophysiologically. Following various survival intervals, precursor cells transplanted distal to a transection of the sciatic nerve differentiated into neurons expressing the motor neuron markers ChAT and the alpha1 1.2 (class C, L-type) voltage-sensitive Ca++ channel subunit. These cells extended axons into the muscle, where they formed cholinergic terminals.


CONCLUSIONS
These results demonstrate that motor neurons can differentiate from spinal cord neural precursor cells grown in culture as well as following transplantation into a transected peripheral nerve.
MacDonald
Stephen C
SC
Department of Physiology, University of Manitoba, Winnipeg, Manitoba, Canada.
Fleetwood
Ian G
IG
Hochman
Shawn
S
Dodd
Janice G
JG
Cheng
Gavin K W
GK
Jordan
Larry M
LM
Brownstone
Robert M
RM
D000818:Animals; D001369:Axons / Q000000981:diagnostic imaging; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D002470:Cell Survival / Q000235:genetics / Q000502:physiology; D002795:Choline O-Acetyltransferase / Q000235:genetics; D016332:Fetal Tissue Transplantation; D018398:Homeodomain Proteins / Q000235:genetics; D060850:LIM-Homeodomain Proteins; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D009046:Motor Neurons / Q000166:cytology / Q000637:transplantation*; D039902:Multipotent Stem Cells / Q000166:cytology*; D018482:Muscle, Skeletal / Q000294:innervation; D009416:Nerve Regeneration / Q000235:genetics / Q000502:physiology; D009419:Nerve Tissue Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012584:Sciatic Nerve / Q000601:surgery*; D013116:Spinal Cord / Q000166:cytology*; D013569:Synapses / Q000000981:diagnostic imaging; D014157:Transcription Factors; D014463:Ultrasonography
D018398:Homeodomain Proteins; D060850:LIM-Homeodomain Proteins; D009419:Nerve Tissue Proteins; D014157:Transcription Factors; C063843:insulin gene enhancer binding protein Isl-1; D002795:Choline O-Acetyltransferase