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Peripheral Nerve Transplantation Combined with Acidic Fibroblast Growth Factor and Chondroitinase Induces Regeneration and Improves Urinary Function in Complete Spinal Cord Transected Adult Mice.

DePaul MA, Lin CY, Silver J, Lee YS - PLoS ONE (2015)

Bottom Line: Cystometry analysis and external urethral sphincter electromyograms reveal that treatment with PNG+aFGF+ChABC reduced bladder weight, improved bladder and external urethral sphincter histology, and significantly enhanced LUT function, resulting in more efficient voiding.Regeneration of serotonin axons correlated with LUT recovery.These results suggest that our mouse model of LUT dysfunction recapitulates the results found in the rat model and may be used to further investigate genetic contributions to regeneration failure.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosciences, Case Western Reserve University, Cleveland, Ohio, United States of America.

ABSTRACT
The loss of lower urinary tract (LUT) control is a ubiquitous consequence of a complete spinal cord injury, attributed to a lack of regeneration of supraspinal pathways controlling the bladder. Previous work in our lab has utilized a combinatorial therapy of peripheral nerve autografts (PNG), acidic fibroblast growth factor (aFGF), and chondroitinase ABC (ChABC) to treat a complete T8 spinal cord transection in the adult rat, resulting in supraspinal control of bladder function. In the present study we extended these findings by examining the use of the combinatorial PNG+aFGF+ChABC treatment in a T8 transected mouse model, which more closely models human urinary deficits following spinal cord injury. Cystometry analysis and external urethral sphincter electromyograms reveal that treatment with PNG+aFGF+ChABC reduced bladder weight, improved bladder and external urethral sphincter histology, and significantly enhanced LUT function, resulting in more efficient voiding. Treated mice's injured spinal cord also showed a reduction in collagen scaring, and regeneration of serotonergic and tyrosine hydroxylase-positive axons across the lesion and into the distal spinal cord. Regeneration of serotonin axons correlated with LUT recovery. These results suggest that our mouse model of LUT dysfunction recapitulates the results found in the rat model and may be used to further investigate genetic contributions to regeneration failure.

No MeSH data available.


Related in: MedlinePlus

PNG+aFGF+ChABC treatment induces 5-HT and TH regeneration 18 weeks post-complete spinal cord transection.TX-Only: (A) Camera lucida tracing of 5-HT immunoreactive fibers in a TX-only animal. (B) 5-HT and GFAP immunostaining of the boxed area in (A), scale bar, 50 μm. (C) Camera lucida tracing of TH-immunoreactive fibers in a TX-only animal. (D) TH and GFAP immunostaining of the boxed area in (C). Scale bar, 50 μm. PNG+aFGF+ChABC: (E) Camera lucida tracing of 5-HT-immunoreactive fibers in PNG+aFGF+ChABC. (F) 5-HT and GFAP immunostaining of boxed areas in (E): (F’) Rostral spinal cord-PNG interface; (F”) within the PNG; (F”‘) caudal spinal cord. Scale bar, 50 μm. (G) Camera lucida tracing of TH-immunoreactive fibers in a PNG+aFGF+ChABC-treated animal. (H) TH and GFAP immunostaining of boxed areas in (G): (H’) Rostral spinal cord-PNG interface; (H”) within the PNG; (H”‘) caudal spinal cord. Scale bar, 50 μm. (I) Quantification of 5-HT-immunoreactive fibers found in caudal spinal cord, n = 7. (J) Quantification of TH-immunoreactive fibers found in caudal spinal cord, n = 7. Only animals whose spinal cord was processed and cut sagittally were included in the analysis.
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pone.0139335.g004: PNG+aFGF+ChABC treatment induces 5-HT and TH regeneration 18 weeks post-complete spinal cord transection.TX-Only: (A) Camera lucida tracing of 5-HT immunoreactive fibers in a TX-only animal. (B) 5-HT and GFAP immunostaining of the boxed area in (A), scale bar, 50 μm. (C) Camera lucida tracing of TH-immunoreactive fibers in a TX-only animal. (D) TH and GFAP immunostaining of the boxed area in (C). Scale bar, 50 μm. PNG+aFGF+ChABC: (E) Camera lucida tracing of 5-HT-immunoreactive fibers in PNG+aFGF+ChABC. (F) 5-HT and GFAP immunostaining of boxed areas in (E): (F’) Rostral spinal cord-PNG interface; (F”) within the PNG; (F”‘) caudal spinal cord. Scale bar, 50 μm. (G) Camera lucida tracing of TH-immunoreactive fibers in a PNG+aFGF+ChABC-treated animal. (H) TH and GFAP immunostaining of boxed areas in (G): (H’) Rostral spinal cord-PNG interface; (H”) within the PNG; (H”‘) caudal spinal cord. Scale bar, 50 μm. (I) Quantification of 5-HT-immunoreactive fibers found in caudal spinal cord, n = 7. (J) Quantification of TH-immunoreactive fibers found in caudal spinal cord, n = 7. Only animals whose spinal cord was processed and cut sagittally were included in the analysis.

Mentions: Given the voiding improvements in mice receiving PNG+aFGF+ChABC treatment and a reduction in collagen scarring, we next examined whether axonal regrowth extended beyond the caudal PNG-spinal cord interface. Regeneration into the bridge and well beyond the transection site in the rat model has been more extensively studied and has been found to be critical in the recovery of urinary function. In the mouse, immunostained 5-HT and TH axons were traced via camera lucida projections of consecutive serial parasagittal sections for the TX-only (Fig 4A and 4C) and PNG+aFGF+ChABC (Fig 4E and 4G) groups. 5-HT- and TH-positive fibers were identified only in the rostral penumbra of the lesion in the TX-only group. They did not penetrate deeply within the lesion nor did they ever regenerate beyond the lesion (Fig 4A–4D). In the PNG+aFGF+ChABC group, both 5-HT- (Fig 4E and 4F) and TH-positive fibers (Fig 4G and 4H) were visualized crossing the rostral spinal cord-PNG interface (Fig 4F’ and 4H’) to enter the PNG (Fig 4F”and 4H”). Importantly, some fibers continued into the distal caudal spinal cord (Fig 4F”and 4H”‘) beyond the caudal PNG-spinal cord interface. Quantitative analyses of fibers in the distal spinal cord confirmed the presence of regenerated 5-HT (Fig 4I) and TH (Fig 4J) fibers beyond the PNG.


Peripheral Nerve Transplantation Combined with Acidic Fibroblast Growth Factor and Chondroitinase Induces Regeneration and Improves Urinary Function in Complete Spinal Cord Transected Adult Mice.

DePaul MA, Lin CY, Silver J, Lee YS - PLoS ONE (2015)

PNG+aFGF+ChABC treatment induces 5-HT and TH regeneration 18 weeks post-complete spinal cord transection.TX-Only: (A) Camera lucida tracing of 5-HT immunoreactive fibers in a TX-only animal. (B) 5-HT and GFAP immunostaining of the boxed area in (A), scale bar, 50 μm. (C) Camera lucida tracing of TH-immunoreactive fibers in a TX-only animal. (D) TH and GFAP immunostaining of the boxed area in (C). Scale bar, 50 μm. PNG+aFGF+ChABC: (E) Camera lucida tracing of 5-HT-immunoreactive fibers in PNG+aFGF+ChABC. (F) 5-HT and GFAP immunostaining of boxed areas in (E): (F’) Rostral spinal cord-PNG interface; (F”) within the PNG; (F”‘) caudal spinal cord. Scale bar, 50 μm. (G) Camera lucida tracing of TH-immunoreactive fibers in a PNG+aFGF+ChABC-treated animal. (H) TH and GFAP immunostaining of boxed areas in (G): (H’) Rostral spinal cord-PNG interface; (H”) within the PNG; (H”‘) caudal spinal cord. Scale bar, 50 μm. (I) Quantification of 5-HT-immunoreactive fibers found in caudal spinal cord, n = 7. (J) Quantification of TH-immunoreactive fibers found in caudal spinal cord, n = 7. Only animals whose spinal cord was processed and cut sagittally were included in the analysis.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4591338&req=5

pone.0139335.g004: PNG+aFGF+ChABC treatment induces 5-HT and TH regeneration 18 weeks post-complete spinal cord transection.TX-Only: (A) Camera lucida tracing of 5-HT immunoreactive fibers in a TX-only animal. (B) 5-HT and GFAP immunostaining of the boxed area in (A), scale bar, 50 μm. (C) Camera lucida tracing of TH-immunoreactive fibers in a TX-only animal. (D) TH and GFAP immunostaining of the boxed area in (C). Scale bar, 50 μm. PNG+aFGF+ChABC: (E) Camera lucida tracing of 5-HT-immunoreactive fibers in PNG+aFGF+ChABC. (F) 5-HT and GFAP immunostaining of boxed areas in (E): (F’) Rostral spinal cord-PNG interface; (F”) within the PNG; (F”‘) caudal spinal cord. Scale bar, 50 μm. (G) Camera lucida tracing of TH-immunoreactive fibers in a PNG+aFGF+ChABC-treated animal. (H) TH and GFAP immunostaining of boxed areas in (G): (H’) Rostral spinal cord-PNG interface; (H”) within the PNG; (H”‘) caudal spinal cord. Scale bar, 50 μm. (I) Quantification of 5-HT-immunoreactive fibers found in caudal spinal cord, n = 7. (J) Quantification of TH-immunoreactive fibers found in caudal spinal cord, n = 7. Only animals whose spinal cord was processed and cut sagittally were included in the analysis.
Mentions: Given the voiding improvements in mice receiving PNG+aFGF+ChABC treatment and a reduction in collagen scarring, we next examined whether axonal regrowth extended beyond the caudal PNG-spinal cord interface. Regeneration into the bridge and well beyond the transection site in the rat model has been more extensively studied and has been found to be critical in the recovery of urinary function. In the mouse, immunostained 5-HT and TH axons were traced via camera lucida projections of consecutive serial parasagittal sections for the TX-only (Fig 4A and 4C) and PNG+aFGF+ChABC (Fig 4E and 4G) groups. 5-HT- and TH-positive fibers were identified only in the rostral penumbra of the lesion in the TX-only group. They did not penetrate deeply within the lesion nor did they ever regenerate beyond the lesion (Fig 4A–4D). In the PNG+aFGF+ChABC group, both 5-HT- (Fig 4E and 4F) and TH-positive fibers (Fig 4G and 4H) were visualized crossing the rostral spinal cord-PNG interface (Fig 4F’ and 4H’) to enter the PNG (Fig 4F”and 4H”). Importantly, some fibers continued into the distal caudal spinal cord (Fig 4F”and 4H”‘) beyond the caudal PNG-spinal cord interface. Quantitative analyses of fibers in the distal spinal cord confirmed the presence of regenerated 5-HT (Fig 4I) and TH (Fig 4J) fibers beyond the PNG.

Bottom Line: Cystometry analysis and external urethral sphincter electromyograms reveal that treatment with PNG+aFGF+ChABC reduced bladder weight, improved bladder and external urethral sphincter histology, and significantly enhanced LUT function, resulting in more efficient voiding.Regeneration of serotonin axons correlated with LUT recovery.These results suggest that our mouse model of LUT dysfunction recapitulates the results found in the rat model and may be used to further investigate genetic contributions to regeneration failure.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurosciences, Case Western Reserve University, Cleveland, Ohio, United States of America.

ABSTRACT
The loss of lower urinary tract (LUT) control is a ubiquitous consequence of a complete spinal cord injury, attributed to a lack of regeneration of supraspinal pathways controlling the bladder. Previous work in our lab has utilized a combinatorial therapy of peripheral nerve autografts (PNG), acidic fibroblast growth factor (aFGF), and chondroitinase ABC (ChABC) to treat a complete T8 spinal cord transection in the adult rat, resulting in supraspinal control of bladder function. In the present study we extended these findings by examining the use of the combinatorial PNG+aFGF+ChABC treatment in a T8 transected mouse model, which more closely models human urinary deficits following spinal cord injury. Cystometry analysis and external urethral sphincter electromyograms reveal that treatment with PNG+aFGF+ChABC reduced bladder weight, improved bladder and external urethral sphincter histology, and significantly enhanced LUT function, resulting in more efficient voiding. Treated mice's injured spinal cord also showed a reduction in collagen scaring, and regeneration of serotonergic and tyrosine hydroxylase-positive axons across the lesion and into the distal spinal cord. Regeneration of serotonin axons correlated with LUT recovery. These results suggest that our mouse model of LUT dysfunction recapitulates the results found in the rat model and may be used to further investigate genetic contributions to regeneration failure.

No MeSH data available.


Related in: MedlinePlus