Mouse NG2+ oligodendrocyte precursors express mRNA for proteolipid protein but not its DM-20 variant: a study of laser microdissection-captured NG2+ cells.
Despite recent advances in our understanding of lineage of oligodendrocytes, detailed molecular characterization of this lineage in vivo is limited, primarily because of our inability to obtain a pure population of cells in situ. To define the molecular characteristics of oligodendrocyte lineage cells during development and their response to injury, we developed a strategy that uses laser capture microdissection (LCM) to isolate cells from sections and reverse transcription-PCR to determine mRNA expression. As a first step, we examined the expression of myelin-specific protein genes in NG2+ cells in cerebral cortex. We demonstrate that NG2+ cells in both developing and adult mice express NG2 mRNA but not mRNA for proteins specific for astrocytes, neurons, or microglia, indicating that a highly pure population of antigen-specific cells of the oligodendrocyte lineage can be obtained using LCM. Furthermore, we show that NG2+ cells express mRNAs for proteolipid protein (PLP), myelin basic protein, and 2',3'-cyclic nucleotide 3'-phosphodiesterase, but they dot not express DM-20 mRNA, a PLP mRNA splicing variant. Our data demonstrate that antigen-specific cells of oligodendrocyte lineage differentially express mRNA for myelin-specific proteins and their variants in vivo, partly define the gene expression in NG2+ cells, and raise questions about the cellular sites of DM-20 expression. This work also shows that LCM is a valuable tool to define and analyze gene expression in the cells of the oligodendrocyte lineage.
Ye
Ping
P
Department of Pediatrics, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7220, USA. ping_ye@med.unc.edu
Bagnell
Robert
R
D'Ercole
A Joseph
AJ
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000235:genetics; D000818:Animals; D000941:Antigens / Q000096:biosynthesis*; D019070:Cell Lineage; D002469:Cell Separation / Q000295:instrumentation; D002540:Cerebral Cortex / Q000166:cytology; D007834:Lasers; D051379:Mice; D004676:Myelin Basic Protein / Q000235:genetics; D018991:Myelin Proteolipid Protein / Q000235:genetics*; D009419:Nerve Tissue Proteins; D009836:Oligodendroglia / Q000166:cytology*; D011509:Proteoglycans / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D000099216:Chondroitin Sulfate Proteoglycan 4
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases; D000941:Antigens; D004676:Myelin Basic Protein; D018991:Myelin Proteolipid Protein; D009419:Nerve Tissue Proteins; D011509:Proteoglycans; D012333:RNA, Messenger; D000099216:Chondroitin Sulfate Proteoglycan 4; C047516:Plp1 protein, mouse