Use of a rat Y chromosome probe to determine the long-term survival of glial cells transplanted into areas of CNS demyelination.
A lack of suitable markers for cells which undergo division following transplantation has hindered studies assessing the long-term survival of glial cell grafts in the CNS. A probe specific to the rat Y chromosome was used to identify male glial cells grafted into an area of ethidium bromide-induced demyelination in syngeneic adult female rat spinal cord 4 weeks, 6 months and 12 months post-transplantation. At all time points there was extensive oligodendrocyte remyelination of transplanted lesions, and graft-derived cells were present within the lesion up to 12 months post-transplantation. In order to demonstrate graft-derived oligodendrocytes in the remyelinated region at 6 and 12 months, double-labelling studies were performed using the oligodendrocyte-specific antibodies carbonic anhydrase II or phosphatidyl ethanolamine-binding protein in combination with the Y chromosome probe. It was found that the majority of oligodendrocytes in the transplanted region were graft-derived. Graft-mediated remyelination was associated with a reduction in myelin sheath thickness and increase in nodal frequency similar to that observed in spontaneous remyelination, suggesting that, like axons remyelinated spontaneously, axons remyelinated by grafted cells will be capable of secure conduction. An alteration in the immunoreactivity of oligodendrocytes from carbonic anhydrase II-negative in the unlesioned dorsal funiculus to carbonic anhydrase II-positive in the remyelinated dorsal funiculus was considered to reflect a reduction in the amount of myelin supported by each oligodendrocyte, leading to the proposal that carbonic anhydrase II immunoreactivity may provide a means of identifying areas of remyelination in normally carbonic anhydrase II-negative white matter tracts.
O'Leary
M T
MT
MRC Cambridge Centre for Brain Repair, UK.
Blakemore
W F
WF
D000818:Animals; D001369:Axons / Q000473:pathology / Q000502:physiology; D002256:Carbonic Anhydrases / Q000032:analysis; D002470:Cell Survival; D003711:Demyelinating Diseases / Q000139:chemically induced / Q000473:pathology* / Q000628:therapy; D004996:Ethidium; D005260:Female; D006085:Graft Survival; D017403:In Situ Hybridization; D008297:Male; D009186:Myelin Sheath / Q000473:pathology; D009457:Neuroglia / Q000637:transplantation*; D009836:Oligodendroglia / Q000473:pathology / Q000502:physiology; D051381:Rats; D013116:Spinal Cord / Q000201:enzymology / Q000473:pathology; D014998:Y Chromosome
D002256:Carbonic Anhydrases; D004996:Ethidium