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Developmental stage of oligodendrocytes determines their response to activated microglia in vitro.

Miller BA, Crum JM, Tovar CA, Ferguson AR, Bresnahan JC, Beattie MS - J Neuroinflammation (2007)

Bottom Line: Lipopolysaccharide was used to activate microglia and microglial activation was confirmed by TNFalpha ELISA.Activated microglia reduced OPC survival, but increased survival and reduced apoptosis of mature oligodendrocytes.Activated microglia may have divergent effects on OPCs and mature oligodendrocytes, reducing OPC survival and increasing mature oligodendrocyte survival.

View Article: PubMed Central - HTML - PubMed

Affiliation: Brain and Spinal Injury Center, Department of Neurological Surgery, University of California San Francisco, 1001 Potrero Ave, Building 1, Room 101, San Francisco, CA 94143, USA. Brandon.Miller@osumc.edu

ABSTRACT

Background: Oligodendrocyte progenitor cells (OPCs) and mature oligodendrocytes are both lost in central nervous system injury and disease. Activated microglia may play a role in OPC and oligodendrocyte loss or replacement, but it is not clear how the responses of OPCs and oligodendrocytes to activated microglia differ.

Methods: OPCs and microglia were isolated from rat cortex. OPCs were induced to differentiate into oligodendrocytes with thyroid hormone in defined medium. For selected experiments, microglia were added to OPC or oligodendrocyte cultures. Lipopolysaccharide was used to activate microglia and microglial activation was confirmed by TNFalpha ELISA. Cell survival was assessed with immunocytochemistry and cell counts. OPC proliferation and oligodendrocyte apoptosis were also assessed.

Results: OPCs and oligodendrocytes displayed phenotypes representative of immature and mature oligodendrocytes, respectively. Activated microglia reduced OPC survival, but increased survival and reduced apoptosis of mature oligodendrocytes. Activated microglia also underwent cell death themselves.

Conclusion: Activated microglia may have divergent effects on OPCs and mature oligodendrocytes, reducing OPC survival and increasing mature oligodendrocyte survival. This may be of importance because activated microglia are present in several disease states where both OPCs and mature oligodendrocytes are also reacting to injury. Activated microglia may simultaneously have deleterious and helpful effects on different cells after central nervous system injury.

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Oligodendrocyte morphology. Oligodendrocytes grown in media containing thyroid hormone labeled for oligodendrocyte markers at day 3 in vitro. Oligodendrocytes did not label positively for the immature OPC markers NG2 (A) or A2B5 (B), but did label for the mature oligodendrocyte markers GalC (C) and MBP (D).
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Figure 5: Oligodendrocyte morphology. Oligodendrocytes grown in media containing thyroid hormone labeled for oligodendrocyte markers at day 3 in vitro. Oligodendrocytes did not label positively for the immature OPC markers NG2 (A) or A2B5 (B), but did label for the mature oligodendrocyte markers GalC (C) and MBP (D).

Mentions: Oligodendrocytes grown continuously with thyroid hormone matured rapidly and were seen to have multiple processes by day 1 in vitro. Oligodendrocytes were labeled with antibodies to GalC and MBP on day 3 in vitro (Fig. 5C, D). Oligodendrocytes grown in thyroid hormone did not stain positively for NG2 or A2B5 (Fig. 5A,B) indicating that these oligodendrocytes were mature.


Developmental stage of oligodendrocytes determines their response to activated microglia in vitro.

Miller BA, Crum JM, Tovar CA, Ferguson AR, Bresnahan JC, Beattie MS - J Neuroinflammation (2007)

Oligodendrocyte morphology. Oligodendrocytes grown in media containing thyroid hormone labeled for oligodendrocyte markers at day 3 in vitro. Oligodendrocytes did not label positively for the immature OPC markers NG2 (A) or A2B5 (B), but did label for the mature oligodendrocyte markers GalC (C) and MBP (D).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2214724&req=5

Figure 5: Oligodendrocyte morphology. Oligodendrocytes grown in media containing thyroid hormone labeled for oligodendrocyte markers at day 3 in vitro. Oligodendrocytes did not label positively for the immature OPC markers NG2 (A) or A2B5 (B), but did label for the mature oligodendrocyte markers GalC (C) and MBP (D).
Mentions: Oligodendrocytes grown continuously with thyroid hormone matured rapidly and were seen to have multiple processes by day 1 in vitro. Oligodendrocytes were labeled with antibodies to GalC and MBP on day 3 in vitro (Fig. 5C, D). Oligodendrocytes grown in thyroid hormone did not stain positively for NG2 or A2B5 (Fig. 5A,B) indicating that these oligodendrocytes were mature.

Bottom Line: Lipopolysaccharide was used to activate microglia and microglial activation was confirmed by TNFalpha ELISA.Activated microglia reduced OPC survival, but increased survival and reduced apoptosis of mature oligodendrocytes.Activated microglia may have divergent effects on OPCs and mature oligodendrocytes, reducing OPC survival and increasing mature oligodendrocyte survival.

View Article: PubMed Central - HTML - PubMed

Affiliation: Brain and Spinal Injury Center, Department of Neurological Surgery, University of California San Francisco, 1001 Potrero Ave, Building 1, Room 101, San Francisco, CA 94143, USA. Brandon.Miller@osumc.edu

ABSTRACT

Background: Oligodendrocyte progenitor cells (OPCs) and mature oligodendrocytes are both lost in central nervous system injury and disease. Activated microglia may play a role in OPC and oligodendrocyte loss or replacement, but it is not clear how the responses of OPCs and oligodendrocytes to activated microglia differ.

Methods: OPCs and microglia were isolated from rat cortex. OPCs were induced to differentiate into oligodendrocytes with thyroid hormone in defined medium. For selected experiments, microglia were added to OPC or oligodendrocyte cultures. Lipopolysaccharide was used to activate microglia and microglial activation was confirmed by TNFalpha ELISA. Cell survival was assessed with immunocytochemistry and cell counts. OPC proliferation and oligodendrocyte apoptosis were also assessed.

Results: OPCs and oligodendrocytes displayed phenotypes representative of immature and mature oligodendrocytes, respectively. Activated microglia reduced OPC survival, but increased survival and reduced apoptosis of mature oligodendrocytes. Activated microglia also underwent cell death themselves.

Conclusion: Activated microglia may have divergent effects on OPCs and mature oligodendrocytes, reducing OPC survival and increasing mature oligodendrocyte survival. This may be of importance because activated microglia are present in several disease states where both OPCs and mature oligodendrocytes are also reacting to injury. Activated microglia may simultaneously have deleterious and helpful effects on different cells after central nervous system injury.

Show MeSH
Related in: MedlinePlus