Purification and characterization of adult oligodendrocyte precursor cells from the rat optic nerve - PubMed (original) (raw)
Purification and characterization of adult oligodendrocyte precursor cells from the rat optic nerve
J Shi et al. J Neurosci. 1998.
Abstract
Oligodendrocyte precursor cells (OPCs) persist in substantial numbers in the adult brain in a quiescent state suggesting that they may provide a source of new oligodendrocytes after injury. To determine whether adult OPCs have the capacity to divide rapidly, we have developed a method to highly purify OPCs from adult optic nerve and have directly compared their properties with their perinatal counterparts. When cultured in platelet-derived growth factor (PDGF), an astrocyte-derived mitogen, perinatal OPCs divided approximately once per day, whereas adult OPCs divided only once every 3 or 4 d. The proliferation rate of adult OPCs was not increased by addition of fibroblast growth factor (FGF) or of the neuregulin glial growth factor 2 (GGF2), two mitogens that are normally produced by retinal ganglion cells. cAMP elevation has been shown previously to be essential for Schwann cells to survive and divide in response to GGF2 and other mitogens. Similarly we found that when cAMP levels were elevated, GGF2 alone was sufficient to induce perinatal OPCs to divide slowly, approximately once every 4 d, but adult OPCs still did not divide. When PDGF was combined with GGF2 and cAMP elevation, however, the adult OPCs began to divide rapidly. These findings indicate that adult OPCs are intrinsically different than perinatal OPCs. They are not senescent cells, however, because they retain the capacity to divide rapidly. Thus, after demyelinating injuries, enhanced axonal release of GGF2 or a related neuregulin might collaborate with astrocyte-derived PDGF to induce rapid division of adult OPCs.
Figures
Fig. 1.
The differentiation of adult OPC cells.A, B, Immunofluorescence micrographs of purified adult OPCs that were labeled after 4 d of culture with a monoclonal anti-GC antibody (A) or a polyclonal GFAP antiserum (B). When cultured in serum-free medium containing insulin and CNTF (A), nearly all cells differentiated into GC+ oligodendrocytes. When cultured in medium containing 10% fetal calf serum (B), nearly all cells differentiated into GFAP+ type-2 astrocytes. Scale bar, 50 μm.
Fig. 2.
Hoffman micrograph of an adult OPC clone. Purified cells were plated at clonal density and cultured for 21 d in serum-free medium containing PDGF, NT-3, CNTF, and insulin. Adult OPCs have a bipolar morphology that is indistinguishable from perinatal OPCs. Scale bar, 100 μm.
Fig. 3.
The proliferation rate of adult and perinatal OPCs in culture. Purified adult (P60) (A, C) and perinatal (P8) (B, D) OPCs were cultured at clonal density in serum-free medium containing PDGF and insulin. The number of cells per clone was determined after 4 d (A, B) and 8 d (C,D) of culture. Under these culture conditions, which lack T3, most of the clones consisted predominantly of OPCs. Note that the average size of adult OPC clones is significantly smaller than is that of the perinatal clones.
Fig. 4.
Comparison of the proliferation rate of adult and perinatal OPCs in vitro and in vivo. A, OPCs were cultured in serum-free medium containing PDGF, NT-3, CNTF, and insulin for 4 d before a 90 min incubation with BrdU (10 μ
m
). The percentage of cells that incorporated BrdU was determined by immunostaining. Values are mean ± SEM (n = 3 coverslips).B, BrdU was injected intraperitoneally into adult and P8 rats. After 90 min, OPCs were purified from the optic nerves of the injected rats and cultured for 1 hr before BrdU staining.
Fig. 5.
Comparison of the proliferation rate of adult and perinatal OPCs in cryosections. A, B, Immunofluorescence micrographs of optic nerve cryosections from P8 (A) and adult (B) optic nerves. The cryosections were obtained from optic nerves that were fixed 90 min after an intraperitoneal injection of BrdU and double labeled with NG-2 (green) and BrdU (red) antibodies. Note that in the P8 section, there are many NG-2+ cells that are also BrdU+. In contrast, in the adult section, none of the NG-2+ cells are BrdU+. Scale bar, 50 μm.
Fig. 6.
Effects of T3 on the rate of oligodendrocyte generation by OPCs. Purified P60 (A) and P8 (B) OPCs were cultured at clonal density in serum-free medium containing PDGF, NT-3, CNTF, and insulin in the presence (solid circles) and absence (open circles) of T3. The percentages of clones containing predominantly oligodendrocytes (>50% of cells) were counted after 4, 6, 8, and 12 d of culture. All values are mean ± SEM. T3 strongly enhanced the rate of oligodendrocyte generation from both P60 and P8 OPCs.
Fig. 7.
Effects of GGF2 on adult OPCs. Purified P60 and P8 OPCs were cultured for 8 d at clonal density in serum-free medium containing PDGF, GGF2,IBMX (0.1 m
m
), and forskolin (5 μ
m
), as indicated. A, The division rate of P8 and P60 was calculated from the clone size. B, The rate of oligodendrocyte generation by P60 OPCs was determined by the percentage of clones that primarily consisted of oligodendrocytes. GGF2 enhances the rate of proliferation of P60 OPCs and inhibits their differentiation into oligodendrocytes. All values are mean ± SEM.FORSK, Forskolin; IBMX, isobutylmethylxanthine.
Similar articles
- Long-term culture of purified postnatal oligodendrocyte precursor cells. Evidence for an intrinsic maturation program that plays out over months.
Tang DG, Tokumoto YM, Raff MC. Tang DG, et al. J Cell Biol. 2000 Mar 6;148(5):971-84. doi: 10.1083/jcb.148.5.971. J Cell Biol. 2000. PMID: 10704447 Free PMC article. - Oligodendrocyte-type-2 astrocyte (O-2A) progenitor cells derived from adult rat spinal cord: in vitro characteristics and response to PDGF, bFGF and NT-3.
Engel U, Wolswijk G. Engel U, et al. Glia. 1996 Jan;16(1):16-26. doi: 10.1002/(SICI)1098-1136(199601)16:1<16::AID-GLIA3>3.0.CO;2-9. Glia. 1996. PMID: 8787770 - Control of division and differentiation in oligodendrocyte-type-2 astrocyte progenitor cells.
Noble M, Barnett SC, Bögler O, Land H, Wolswijk G, Wren D. Noble M, et al. Ciba Found Symp. 1990;150:227-43; discussion 244-9. doi: 10.1002/9780470513927.ch14. Ciba Found Symp. 1990. PMID: 2373025 - Redox state as a central modulator of precursor cell function.
Noble M, Smith J, Power J, Mayer-Pröschel M. Noble M, et al. Ann N Y Acad Sci. 2003 Jun;991:251-71. doi: 10.1111/j.1749-6632.2003.tb07481.x. Ann N Y Acad Sci. 2003. PMID: 12846992 Review.
Cited by
- Erythropoietin regulates developmental myelination in the brain stimulating postnatal oligodendrocyte maturation.
Muttathukunnel P, Wälti M, Aboouf MA, Köster-Hegmann C, Haenggi T, Gassmann M, Pannzanelli P, Fritschy JM, Schneider Gasser EM. Muttathukunnel P, et al. Sci Rep. 2023 Nov 9;13(1):19522. doi: 10.1038/s41598-023-46783-9. Sci Rep. 2023. PMID: 37945644 Free PMC article. - Early Postnatal Expression of Tgfβ-1 and Fgf-2 Correlates With Regenerative Functions of Unrestricted Somatic Stem Cell Infusion After Rabbit GMH-IVH.
Finkel DA, Malfa A, Liao Y, Purohit D, Hu F, Sulaymankhil D, Abhishek Narra S, Hussein K, Subbian S, Cairo MS, Vinukonda G, La Gamma EF. Finkel DA, et al. Stem Cells Transl Med. 2023 Dec 18;12(12):811-824. doi: 10.1093/stcltm/szad064. Stem Cells Transl Med. 2023. PMID: 37774396 Free PMC article. - Molecular and functional heterogeneity in dorsal and ventral oligodendrocyte progenitor cells of the mouse forebrain in response to DNA damage.
Boda E, Lorenzati M, Parolisi R, Harding B, Pallavicini G, Bonfanti L, Moccia A, Bielas S, Di Cunto F, Buffo A. Boda E, et al. Nat Commun. 2022 Apr 28;13(1):2331. doi: 10.1038/s41467-022-30010-6. Nat Commun. 2022. PMID: 35484145 Free PMC article. - mTOR Signaling Regulates Metabolic Function in Oligodendrocyte Precursor Cells and Promotes Efficient Brain Remyelination in the Cuprizone Model.
Jeffries MA, McLane LE, Khandker L, Mather ML, Evangelou AV, Kantak D, Bourne JN, Macklin WB, Wood TL. Jeffries MA, et al. J Neurosci. 2021 Oct 6;41(40):8321-8337. doi: 10.1523/JNEUROSCI.1377-20.2021. Epub 2021 Aug 20. J Neurosci. 2021. PMID: 34417330 Free PMC article. - Essential roles of plexin-B3+ oligodendrocyte precursor cells in the pathogenesis of Alzheimer's disease.
Nihonmatsu-Kikuchi N, Yu XJ, Matsuda Y, Ozawa N, Ito T, Satou K, Kaname T, Iwasaki Y, Akagi A, Yoshida M, Toru S, Hirokawa K, Takashima A, Hasegawa M, Uchihara T, Tatebayashi Y. Nihonmatsu-Kikuchi N, et al. Commun Biol. 2021 Jul 15;4(1):870. doi: 10.1038/s42003-021-02404-7. Commun Biol. 2021. PMID: 34267322 Free PMC article.
References
- Ahlgren SC, Wallace H, Bishop J, Neophytou C, Raff MC. Effects of thyroid hormone on embryonic oligodendrocyte precursor cell development in vivo and in vitro. Mol Cell Neurosci. 1997;9:420–432. - PubMed
- Allen RE, Rankin LL. Regulation of satellite cells during skeletal muscle growth and development. Proc Soc Exp Biol Med. 1990;194:81–86. - PubMed
- Archer DR, Cuddon PA, Lipsitz D, Duncan I. Myelination of the canine central nervous system by glial cell transplantation: a model for repair of human myelin disease. Nat Med. 1997;3:54–59. - PubMed
- Barres BA, Raff MC. Proliferation of oligodendrocyte precursor cells depends on electrical activity in axons. Nature. 1993;361:258–260. - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Medical