Figure 1.
Timelines for infusion experiments.
(A) For sequential delivery of growth factors during cuprizone challenge, mini-osmotic pumps were implanted after 4-weeks of cuprizone challenge to deliver either vehicle or BMP4 into the lateral ventricle for 7 days. Next, the mini-osmotic pump delivering vehicle or BMP4 was replaced with a new pump delivering vehicle, Noggin or IGF-1 for the final 7 days of a six-week cuprizone challenge. BrdU was added to the drinking water for the final 3 days of the first infusion. (B) The infusion paradigm was used as described above in A, however, the mice were allowed to recover for 1-week by removing cuprizone from their diet.
Figure 2.
BMP4 infusion increases numbers of oligodendroglia and astrocytes during cuprizone challenge but does not influence the level of demyelination.
(A–B) Immunostaining of BrdU-Olig2 (A) and GFAP (B) of the midline corpus callosum of the infused mice after 6-weeks cuprizone challenge. Arrows indicate double positive cells in A. Nuclei were counterstained with Hoechst in B. (C–D) Quantification of Olig2+ (C) and GFAP+ (D) in the midline corpus callosum of the infused mice after 6-weeks cuprizone challenge. (E–F) Quantification of the density (E) and average g ratio (F) of myelinated axons. (G) G ratios of individual axons as a function of axonal diameter in the corpus callosum of the infused mice after 6-weeks cuprizone challenge. n = 4–6 animals per group. *p<0.05,** p<0.01; Dunnett's multiple comparison post test in (C). Scale bar in B, 50 µm.
Figure 3.
Sequential delivery of BMP4 and Noggin during cuprizone challenge does not alter numbers of glial cells or myelination.
(A–B) Quantification of BrdU+, Olig2+, BrdU+Olig2+, IBA1+ and GFAP+ cells in the midline corpus callosum of the sequentially infused mice after 6-weeks cuprizone challenge. (C) Quantification average g ratio of myelinated axons. (D) G ratios of individual axons as a function of axonal diameter in the corpus callosum of the infused mice after 6-weeks cuprizone challenge. n = 4 animals per group.
Figure 4.
Sequential delivery of BMP4 and Noggin during cuprizone challenge does not alter numbers of oligodendrocytes and astrocytes following recovery.
(A–C) Immunostaining of Olig2 (A), Olig2 and CC1 (B), and GFAP (C) in the midline corpus callosum of the sequentially infused mice following 1-week recovery. Nuclei were counterstained with Hoechst. (D–F) Quantification of Olig2+, Olig2+CC1+ and GFAP+ in the midline corpus callosum of the sequentially infused mice following 1-week recovery. n = 4–6 animals per group. *p<0.05,** p<0.01. Scale bars in C, 50 µm.
Figure 5.
Sequential delivery of BMP4 and Noggin during cuprizone challenge does not enhance remyelination following recovery.
(A) Representative electron micrographs in the caudal corpus callosum of unchallenged and sequentially infused mice after 1-week recovery. (B–C) Quantification of the density (B) and average g ratio (C) of myelinated axons. (D) G ratios of individual axons as a function of axonal diameter in the corpus callosum of the infused mice after 1-week recovery. n = 3–4 animals per group. *p<0.05. **p<0.01 Scale bar, 1 µm.
Figure 6.
Sequential delivery of BMP4 and IGF-1 during cuprizone challenge increases numbers of oligodendroglia in the corpus callosum.
(A) Immunostaining of Olig2 in the corpus callosum of the sequentially infused mice after 6-weeks of cuprizone challenge. (B) Quantification of the density of Olig2+cells in the corpus callosum of the sequentially infused mice after 6-weeks of cuprizone challenge. n = 3 animals per group. *p<0.05. Scale bar, 50 µm.
Figure 7.
Sequential delivery of BMP4 and IGF-1 during cuprizone challenge increases numbers of mature oligodendrocytes following recovery.
(A–B) Immunostaining of BrdU-Olig2 (A) and Olig2-CC1 (B) in the corpus callosum of the sequentially infused mice after 1-week recovery. (C,D) Quantification of the density of BrdU+Olig2+ (C) and Olig2+CC1+ (D). n = 3–4 animals per group. *p<0.05. Scale bar, 50 µm.
Figure 8.
Sequential delivery of BMP4 and IGF-1 during cuprizone challenge does not enhance remyelination.
(A) Representative electron micrographs in the caudal corpus callosum of unchallenged and sequentially infused mice after 1-week recovery. (B–C) Quantification of the density (B) and average g ratio (C) of myelinated axons. (D) G ratios of individual axons as a function of axonal diameter in the corpus callosum of the infused mice after 1-week recovery. n = 3 animals per group Scale bar, 1 µm.