The Journal of Neuroscience, April 19, 2006, ():
Prostaglandin D2-Mediated Microglia/Astrocyte Interaction Enhances Astrogliosis and Demyelination in twitcher
J. Neurosci. Mohri et al.
26: 4383
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplemental Figure 1
A, B: Structures of the targeting vector (Top), the wild-type allele (Middle), and the null allele (Bottom) for HPGDS (A) and DP1 (B) are shown in the upper panels. Restriction sites: N, NcoI; E, Eco47III; X, XbaI; B, BstXI. TK, Herpes simplex virus thymidine kinase gene; Neo, neomycin-resistance gene. The probes used for PCR analysis are shown below the arrows. The combination of FW1/RV1primers or H-FW1/NeoR8 primers was used to detect the respective wild-type or null allele of HPGDS; and that of DP1/DP2 primer or Neo1/DP2 primer was used to detect the respective wild-type or null allele for DP1. Results of gene typing of HPGDS, DP1, and GALC are shown in the lower panels.
- supplemental material
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Supplemental Figure 2
Proposed model of HPGDS/PGD2/DP1-mediated astrogliosis in GALCtwi/twi. First, myelin debris produced by apoptosis of OLs trigger microglial activation. PGD2 produced by HPGDS (red dots) in activated microglia stimulates DP1 (shown in pink) and DP2 (shown in blue) receptors on astrocytes in a paracrine manner. PGD2 binding to DP1 and DP2 increases intracellular cAMP and Ca2+ respectively, in astrocytes and causes GFAP up-regulation. DP1 expression is up-regulated by DP1 stimulation, thus accelerating this signaling pathway of astrogliosis. PGD2 stimulates microglia/macrophages in an autocrine fashion and induces production of pro-inflammatory molecules, such as iNOS, TNFα, and IL-1β. Astrocytes produce IL-6 after PGD2 stimulation. These pro-inflammatory molecules cause activation of microglia and astrocytes, apoptosis of OLs and secondary demyelination.
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Supplemental movie legend A
A: HPGD+/+GALCtwi/twi and HPGDS-/-GALCtwi/twi are shown. Whereas both HPGDS-/-GALCtwi/twi (indicated by the yellow arrow) and HPGDS+/+GALCtwi/twi show hindlimb paralysis (dragging and shuffling), HPGDS-/-GALCtwi/twi moves faster than HPGDS+/+GALCtwi/twi. When HPGDS-/-GALCtwi/twi (left) and HPGDS+/+GALCtwi/twi (right) are suspended in the air by their tail, HPGDS-/-GALCtwi/twi shows the righting of the trunk and the head with very mild tremor of the trunk in contrast to HPGDS+/+GALCtwi/twi, which animal lacks the righting reflex and has marked tremor of the trunk and limbs.
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Supplemental movie legend B
B: Two HQL-79-treated GALCtwi/twi at PND 45 are shown. Both show staggering gait with hindlimb paralysis; however, they can move faster and more smoothly than untreated GALCtwi/twi of the same age. While severer tremor of the trunk and limbs than in HPGDS-/-GALCtwi/twi is recognized, they retain the righting reflex when suspended by the tail.
