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Macrophage-Induced Blood Vessels Guide Schwann Cell-Mediated Regeneration of Peripheral Nerves.

Cattin AL, Burden JJ, Van Emmenis L, Mackenzie FE, Hoving JJ, Garcia Calavia N, Guo Y, McLaughlin M, Rosenberg LH, Quereda V, Jamecna D, Napoli I, Parrinello S, Enver T, Ruhrberg C, Lloyd AC - Cell (2015)

Bottom Line: Here we show that blood vessels direct the migrating cords of Schwann cells.Importantly, disrupting the organization of the newly formed blood vessels in vivo, either by inhibiting the angiogenic signal or by re-orienting them, compromises Schwann cell directionality resulting in defective nerve repair.This study provides important insights into how the choreography of multiple cell-types is required for the regeneration of an adult tissue.

View Article: PubMed Central - PubMed

Affiliation: MRC Laboratory for Molecular Cell Biology, UCL, Gower Street, London WC1E 6BT, UK.

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Redirection of the Blood Vessels Leads to the Misdirection of Migrating SCs(A) PBS- (control) and VEGF-treated rat sciatic nerve images show that placement of VEGF beads to the side of the injury site, leads to aberrant regeneration. Scale bar, 2 mm. Arrows indicate the bridge region and proximal to distal.(B) Immunofluorescence images of the regions demarcated by white boxes in (A) of a PBS- (control) and VEGF-treated animal, Day 6 following injury, longitudinal sections were immunostained to detect SCs (S100+, green) and ECs (RECA1+, red). i and ii: show that misdirected blood vessels in the VEGF-treated animals directed the SC cords toward the adjacent muscle. iii and iv: show the axons (NF+) following the SC cords, toward the muscle. Scale bar, 300 μm. White asterisks indicate the beads. For reconstruction of longitudinal sections, multiple images from the same sample were acquired using the same microscope settings.(C–F) Quantification of (B) to show the direction of blood vessels (C), SCs (D), and axons (E) relative to the proximal/distal axis and the alignment of blood vessels and SCs (F) in the rats treated with PBS or VEGF (n = 3). Graphs show the mean relative angle ± SD for each animal with the mean between animals shown by red lines. Rose plots show the distribution of cells for all animals.(G) Representative confocal images of axons (NF+) in indicated regions of regenerated nerves in Vegfafl/fl (control) or Vegfafl/flTie2-Cre mice, Day 14 after transection.(H) Quantification of (G) showing axonal growth in Vegfafl/fl (black line) and Vegfafl/fl Tie2-cre (gray line) mice (n = 5, graph shows mean value ± SEM).See also Figure S7.
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fig7: Redirection of the Blood Vessels Leads to the Misdirection of Migrating SCs(A) PBS- (control) and VEGF-treated rat sciatic nerve images show that placement of VEGF beads to the side of the injury site, leads to aberrant regeneration. Scale bar, 2 mm. Arrows indicate the bridge region and proximal to distal.(B) Immunofluorescence images of the regions demarcated by white boxes in (A) of a PBS- (control) and VEGF-treated animal, Day 6 following injury, longitudinal sections were immunostained to detect SCs (S100+, green) and ECs (RECA1+, red). i and ii: show that misdirected blood vessels in the VEGF-treated animals directed the SC cords toward the adjacent muscle. iii and iv: show the axons (NF+) following the SC cords, toward the muscle. Scale bar, 300 μm. White asterisks indicate the beads. For reconstruction of longitudinal sections, multiple images from the same sample were acquired using the same microscope settings.(C–F) Quantification of (B) to show the direction of blood vessels (C), SCs (D), and axons (E) relative to the proximal/distal axis and the alignment of blood vessels and SCs (F) in the rats treated with PBS or VEGF (n = 3). Graphs show the mean relative angle ± SD for each animal with the mean between animals shown by red lines. Rose plots show the distribution of cells for all animals.(G) Representative confocal images of axons (NF+) in indicated regions of regenerated nerves in Vegfafl/fl (control) or Vegfafl/flTie2-Cre mice, Day 14 after transection.(H) Quantification of (G) showing axonal growth in Vegfafl/fl (black line) and Vegfafl/fl Tie2-cre (gray line) mice (n = 5, graph shows mean value ± SEM).See also Figure S7.

Mentions: To address whether VEGF-A-induced blood vessels are sufficient to guide cords of SCs, we redirected the blood vessels to test whether the SCs would follow the blood vessels or continue to cross the bridge. To do this, we implanted heparin beads loaded with recombinant human VEGF165 into muscle adjacent to the proximal side of the injury site, immediately after the transection of the rat sciatic nerve. Six days later, the regenerative process was found to be abnormal in 10 out of 13 of the VEGF-treated animals compared to 1 out of 13 PBS-bead-treated controls. In five of the ten VEGF-treated animals in which abnormal regeneration was observed, a complete failure of the regenerative process was associated with misdirection of the blood vessels, SC cords and the accompanying axons, away from the bridge and into surrounding muscle towards the beads (Figures 7A, 7B, and S7A; quantified in Figures 7C–7F and S7B). Analysis of the bridges in a further five cases showed that the beads had moved into the bridge leading to the formation of disorganized blood vessels close to the beads (Figure S7C). In these cases, SCs migrated into the vascularized areas and either appeared “trapped” or deviated from the normal direction of movement, taking the axons along with them (Figure S7D). Moreover, beads implanted adjacent to the distal stump could also redirect blood vessels and SCs (Figure S7E). Together, these results demonstrate that VEGF-induced blood vessels are sufficient to guide SCs and their accompanying axons during peripheral nerve regeneration.


Macrophage-Induced Blood Vessels Guide Schwann Cell-Mediated Regeneration of Peripheral Nerves.

Cattin AL, Burden JJ, Van Emmenis L, Mackenzie FE, Hoving JJ, Garcia Calavia N, Guo Y, McLaughlin M, Rosenberg LH, Quereda V, Jamecna D, Napoli I, Parrinello S, Enver T, Ruhrberg C, Lloyd AC - Cell (2015)

Redirection of the Blood Vessels Leads to the Misdirection of Migrating SCs(A) PBS- (control) and VEGF-treated rat sciatic nerve images show that placement of VEGF beads to the side of the injury site, leads to aberrant regeneration. Scale bar, 2 mm. Arrows indicate the bridge region and proximal to distal.(B) Immunofluorescence images of the regions demarcated by white boxes in (A) of a PBS- (control) and VEGF-treated animal, Day 6 following injury, longitudinal sections were immunostained to detect SCs (S100+, green) and ECs (RECA1+, red). i and ii: show that misdirected blood vessels in the VEGF-treated animals directed the SC cords toward the adjacent muscle. iii and iv: show the axons (NF+) following the SC cords, toward the muscle. Scale bar, 300 μm. White asterisks indicate the beads. For reconstruction of longitudinal sections, multiple images from the same sample were acquired using the same microscope settings.(C–F) Quantification of (B) to show the direction of blood vessels (C), SCs (D), and axons (E) relative to the proximal/distal axis and the alignment of blood vessels and SCs (F) in the rats treated with PBS or VEGF (n = 3). Graphs show the mean relative angle ± SD for each animal with the mean between animals shown by red lines. Rose plots show the distribution of cells for all animals.(G) Representative confocal images of axons (NF+) in indicated regions of regenerated nerves in Vegfafl/fl (control) or Vegfafl/flTie2-Cre mice, Day 14 after transection.(H) Quantification of (G) showing axonal growth in Vegfafl/fl (black line) and Vegfafl/fl Tie2-cre (gray line) mice (n = 5, graph shows mean value ± SEM).See also Figure S7.
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fig7: Redirection of the Blood Vessels Leads to the Misdirection of Migrating SCs(A) PBS- (control) and VEGF-treated rat sciatic nerve images show that placement of VEGF beads to the side of the injury site, leads to aberrant regeneration. Scale bar, 2 mm. Arrows indicate the bridge region and proximal to distal.(B) Immunofluorescence images of the regions demarcated by white boxes in (A) of a PBS- (control) and VEGF-treated animal, Day 6 following injury, longitudinal sections were immunostained to detect SCs (S100+, green) and ECs (RECA1+, red). i and ii: show that misdirected blood vessels in the VEGF-treated animals directed the SC cords toward the adjacent muscle. iii and iv: show the axons (NF+) following the SC cords, toward the muscle. Scale bar, 300 μm. White asterisks indicate the beads. For reconstruction of longitudinal sections, multiple images from the same sample were acquired using the same microscope settings.(C–F) Quantification of (B) to show the direction of blood vessels (C), SCs (D), and axons (E) relative to the proximal/distal axis and the alignment of blood vessels and SCs (F) in the rats treated with PBS or VEGF (n = 3). Graphs show the mean relative angle ± SD for each animal with the mean between animals shown by red lines. Rose plots show the distribution of cells for all animals.(G) Representative confocal images of axons (NF+) in indicated regions of regenerated nerves in Vegfafl/fl (control) or Vegfafl/flTie2-Cre mice, Day 14 after transection.(H) Quantification of (G) showing axonal growth in Vegfafl/fl (black line) and Vegfafl/fl Tie2-cre (gray line) mice (n = 5, graph shows mean value ± SEM).See also Figure S7.
Mentions: To address whether VEGF-A-induced blood vessels are sufficient to guide cords of SCs, we redirected the blood vessels to test whether the SCs would follow the blood vessels or continue to cross the bridge. To do this, we implanted heparin beads loaded with recombinant human VEGF165 into muscle adjacent to the proximal side of the injury site, immediately after the transection of the rat sciatic nerve. Six days later, the regenerative process was found to be abnormal in 10 out of 13 of the VEGF-treated animals compared to 1 out of 13 PBS-bead-treated controls. In five of the ten VEGF-treated animals in which abnormal regeneration was observed, a complete failure of the regenerative process was associated with misdirection of the blood vessels, SC cords and the accompanying axons, away from the bridge and into surrounding muscle towards the beads (Figures 7A, 7B, and S7A; quantified in Figures 7C–7F and S7B). Analysis of the bridges in a further five cases showed that the beads had moved into the bridge leading to the formation of disorganized blood vessels close to the beads (Figure S7C). In these cases, SCs migrated into the vascularized areas and either appeared “trapped” or deviated from the normal direction of movement, taking the axons along with them (Figure S7D). Moreover, beads implanted adjacent to the distal stump could also redirect blood vessels and SCs (Figure S7E). Together, these results demonstrate that VEGF-induced blood vessels are sufficient to guide SCs and their accompanying axons during peripheral nerve regeneration.

Bottom Line: Here we show that blood vessels direct the migrating cords of Schwann cells.Importantly, disrupting the organization of the newly formed blood vessels in vivo, either by inhibiting the angiogenic signal or by re-orienting them, compromises Schwann cell directionality resulting in defective nerve repair.This study provides important insights into how the choreography of multiple cell-types is required for the regeneration of an adult tissue.

View Article: PubMed Central - PubMed

Affiliation: MRC Laboratory for Molecular Cell Biology, UCL, Gower Street, London WC1E 6BT, UK.

Show MeSH
Related in: MedlinePlus