Limits...
The temporal and spatial profiles of cell loss following experimental spinal cord injury: effect of antioxidant therapy on cell death and functional recovery.

Ling X, Bao F, Qian H, Liu D - BMC Neurosci (2013)

Bottom Line: Intraperitoneal treatment with MnTBAP + nitro-L-arginine significantly reduced motoneuron and cell loss and apoptosis in the gray and white matter compared with the vehicle-treated group.MnTBAP alone significantly reduced the number of apoptotic cells and improved functional recovery as evaluated by three behavioral tests.Our demonstration that apoptosis follows SCI and that MnTBAP alone or MnTBAP + nitro-L-arginine significantly reduces apoptosis correlates SCI-induced apoptosis with RS overproduction.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Neurology, University of Texas Medical Branch, 301 University Blvd,, Rt, 0881, Galveston, TX 77555-0881, USA. dliu@utmb.edu.

ABSTRACT

Background: Traumatic spinal cord injury (SCI)-induced overproduction of endogenous deleterious substances triggers secondary cell death to spread damage beyond the initial injury site. Substantial experimental evidence supports reactive species (RS) as important mediators of secondary cell death after SCI. This study established quantitative temporal and spatial profiles of cell loss, characterized apoptosis, and evaluated the effectiveness of a broad spectrum RS scavenger - Mn (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP) and a combination of MnTBAP plus nitro-L-arginine to prevent cell loss and neurological dysfunction following contusion SCI to the rat spinal cord.

Results: By counting the number of surviving cells in spinal cord sections removed at 1, 6, 12, 24, 48, 72 h and 1 week post-SCI and at 0 - 4 mm from the epicenter, the temporal and spatial profiles of motoneuron and glia loss were established. Motoneurons continued to disappear over a week and the losses decreased with increasing distance from the epicenter. Significant glia loss peaked at 24 to 48 h post-SCI, but only at sections 0-1.5 mm from the epicenter. Apoptosis of neurons, motoneurons and astrocytes was characterized morphologically by double immuno-staining with cell-specific markers and apoptosis indicators and confirmed by transmission electron microscopy. DNA laddering, ELISA quantitation and caspase-3 activation in the spinal cord tissue indicated more intense DNA fragments and greater caspase-3 activation in the epicenter than at 1 and 2 cm away from the epicenter or the sham-operated sections. Intraperitoneal treatment with MnTBAP + nitro-L-arginine significantly reduced motoneuron and cell loss and apoptosis in the gray and white matter compared with the vehicle-treated group. MnTBAP alone significantly reduced the number of apoptotic cells and improved functional recovery as evaluated by three behavioral tests.

Conclusions: Our temporal and spatial profiles of cell loss provide data bases for determining the time and location for pharmacological intervention. Our demonstration that apoptosis follows SCI and that MnTBAP alone or MnTBAP + nitro-L-arginine significantly reduces apoptosis correlates SCI-induced apoptosis with RS overproduction. MnTBAP significantly improved functional recovery, which strongly supports the important role of antioxidant therapy in treating SCI and the candidacy of MnTBAP for such treatment.

Show MeSH

Related in: MedlinePlus

Photomicrographs of CV-stained spinal cord sections at different times post-SCI and different distances from the epicenter. Following a laminectomy, the rat spinal cords were injured (50 g.cm), the injured cords were removed at different times post-SCI and processed for CV staining. The upper panel shows surviving cells in CV-stained sections at 1 mm caudal from the epicenter removed at 1 and 24 h post-SCI. The lower panel shows surviving cells in CV-stained sections at 0 and 2 mm caudal from the epicenter removed at 24 h post-SCI. A-C: Lower magnification, D-F: higher magnification of A-C. Scale bar: A-C, 100 μm, D-F, 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3924333&req=5

Figure 1: Photomicrographs of CV-stained spinal cord sections at different times post-SCI and different distances from the epicenter. Following a laminectomy, the rat spinal cords were injured (50 g.cm), the injured cords were removed at different times post-SCI and processed for CV staining. The upper panel shows surviving cells in CV-stained sections at 1 mm caudal from the epicenter removed at 1 and 24 h post-SCI. The lower panel shows surviving cells in CV-stained sections at 0 and 2 mm caudal from the epicenter removed at 24 h post-SCI. A-C: Lower magnification, D-F: higher magnification of A-C. Scale bar: A-C, 100 μm, D-F, 50 μm.

Mentions: Figure 1 illustrates photomicrographs of CV-stained sections at different times post-SCI (upper panel) and at different distances from the epicenter (lower panel). The sham-operated section (A, D) had many cells, including large motoneurons in the ventral gray matter which showed bright nuclei with nucleoli and enrichment in the Nissl bodies. In the upper panel, at 1 h post-SCI, many cells had been lost in the gray matter (B, E) and some neurons lost their normal morphological features. At 24 h post-SCI (C, F), almost no neurons could be observed in the ventral gray matter. In the lower panel, at the epicenter (0 mm), few cells and almost no neurons were observed (B, E). At 2 mm from the epicenter (C, F), some surviving cells including neurons were observed in the ventral gray matter.


The temporal and spatial profiles of cell loss following experimental spinal cord injury: effect of antioxidant therapy on cell death and functional recovery.

Ling X, Bao F, Qian H, Liu D - BMC Neurosci (2013)

Photomicrographs of CV-stained spinal cord sections at different times post-SCI and different distances from the epicenter. Following a laminectomy, the rat spinal cords were injured (50 g.cm), the injured cords were removed at different times post-SCI and processed for CV staining. The upper panel shows surviving cells in CV-stained sections at 1 mm caudal from the epicenter removed at 1 and 24 h post-SCI. The lower panel shows surviving cells in CV-stained sections at 0 and 2 mm caudal from the epicenter removed at 24 h post-SCI. A-C: Lower magnification, D-F: higher magnification of A-C. Scale bar: A-C, 100 μm, D-F, 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3924333&req=5

Figure 1: Photomicrographs of CV-stained spinal cord sections at different times post-SCI and different distances from the epicenter. Following a laminectomy, the rat spinal cords were injured (50 g.cm), the injured cords were removed at different times post-SCI and processed for CV staining. The upper panel shows surviving cells in CV-stained sections at 1 mm caudal from the epicenter removed at 1 and 24 h post-SCI. The lower panel shows surviving cells in CV-stained sections at 0 and 2 mm caudal from the epicenter removed at 24 h post-SCI. A-C: Lower magnification, D-F: higher magnification of A-C. Scale bar: A-C, 100 μm, D-F, 50 μm.
Mentions: Figure 1 illustrates photomicrographs of CV-stained sections at different times post-SCI (upper panel) and at different distances from the epicenter (lower panel). The sham-operated section (A, D) had many cells, including large motoneurons in the ventral gray matter which showed bright nuclei with nucleoli and enrichment in the Nissl bodies. In the upper panel, at 1 h post-SCI, many cells had been lost in the gray matter (B, E) and some neurons lost their normal morphological features. At 24 h post-SCI (C, F), almost no neurons could be observed in the ventral gray matter. In the lower panel, at the epicenter (0 mm), few cells and almost no neurons were observed (B, E). At 2 mm from the epicenter (C, F), some surviving cells including neurons were observed in the ventral gray matter.

Bottom Line: Intraperitoneal treatment with MnTBAP + nitro-L-arginine significantly reduced motoneuron and cell loss and apoptosis in the gray and white matter compared with the vehicle-treated group.MnTBAP alone significantly reduced the number of apoptotic cells and improved functional recovery as evaluated by three behavioral tests.Our demonstration that apoptosis follows SCI and that MnTBAP alone or MnTBAP + nitro-L-arginine significantly reduces apoptosis correlates SCI-induced apoptosis with RS overproduction.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Neurology, University of Texas Medical Branch, 301 University Blvd,, Rt, 0881, Galveston, TX 77555-0881, USA. dliu@utmb.edu.

ABSTRACT

Background: Traumatic spinal cord injury (SCI)-induced overproduction of endogenous deleterious substances triggers secondary cell death to spread damage beyond the initial injury site. Substantial experimental evidence supports reactive species (RS) as important mediators of secondary cell death after SCI. This study established quantitative temporal and spatial profiles of cell loss, characterized apoptosis, and evaluated the effectiveness of a broad spectrum RS scavenger - Mn (III) tetrakis (4-benzoic acid) porphyrin (MnTBAP) and a combination of MnTBAP plus nitro-L-arginine to prevent cell loss and neurological dysfunction following contusion SCI to the rat spinal cord.

Results: By counting the number of surviving cells in spinal cord sections removed at 1, 6, 12, 24, 48, 72 h and 1 week post-SCI and at 0 - 4 mm from the epicenter, the temporal and spatial profiles of motoneuron and glia loss were established. Motoneurons continued to disappear over a week and the losses decreased with increasing distance from the epicenter. Significant glia loss peaked at 24 to 48 h post-SCI, but only at sections 0-1.5 mm from the epicenter. Apoptosis of neurons, motoneurons and astrocytes was characterized morphologically by double immuno-staining with cell-specific markers and apoptosis indicators and confirmed by transmission electron microscopy. DNA laddering, ELISA quantitation and caspase-3 activation in the spinal cord tissue indicated more intense DNA fragments and greater caspase-3 activation in the epicenter than at 1 and 2 cm away from the epicenter or the sham-operated sections. Intraperitoneal treatment with MnTBAP + nitro-L-arginine significantly reduced motoneuron and cell loss and apoptosis in the gray and white matter compared with the vehicle-treated group. MnTBAP alone significantly reduced the number of apoptotic cells and improved functional recovery as evaluated by three behavioral tests.

Conclusions: Our temporal and spatial profiles of cell loss provide data bases for determining the time and location for pharmacological intervention. Our demonstration that apoptosis follows SCI and that MnTBAP alone or MnTBAP + nitro-L-arginine significantly reduces apoptosis correlates SCI-induced apoptosis with RS overproduction. MnTBAP significantly improved functional recovery, which strongly supports the important role of antioxidant therapy in treating SCI and the candidacy of MnTBAP for such treatment.

Show MeSH
Related in: MedlinePlus