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Retinal ganglion cell degeneration is topological but not cell type specific in DBA/2J mice.

Jakobs TC, Libby RT, Ben Y, John SW, Masland RH - J. Cell Biol. (2005)

Bottom Line: Regions of cell death or survival radiated from the optic nerve head in fan-shaped sectors.Collectively, the data suggest axon damage at the optic nerve head as an early lesion, and damage to axon bundles would cause this pattern of degeneration.However, the architecture of the mouse eye seems to preclude a commonly postulated source of mechanical damage within the nerve head.

View Article: PubMed Central - PubMed

Affiliation: Howard Hughes Medical Institute, Harvard Medical School, Boston, MA 02114.

ABSTRACT
Using a variety of double and triple labeling techniques, we have reevaluated the death of retinal neurons in a mouse model of hereditary glaucoma. Cell-specific markers and total neuron counts revealed no cell loss in any retinal neurons other than the ganglion cells. Within the limits of our ability to define cell types, no group of ganglion cells was especially vulnerable or resistant to degeneration. Retrograde labeling and neurofilament staining showed that axonal atrophy, dendritic remodeling, and somal shrinkage (at least of the largest cell types) precedes ganglion cell death in this glaucoma model. Regions of cell death or survival radiated from the optic nerve head in fan-shaped sectors. Collectively, the data suggest axon damage at the optic nerve head as an early lesion, and damage to axon bundles would cause this pattern of degeneration. However, the architecture of the mouse eye seems to preclude a commonly postulated source of mechanical damage within the nerve head.

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Related in: MedlinePlus

Amacrine cells in glaucomatous DBA/2J mice. (A) Single confocal vertical section through the retina of a C57BL/6J mouse, stained with the nuclear dye TOPRO. For the sake of comparison, images in A–C have all been taken in the central part of the retina. (B) Vertical section through a DBA/2J retina (nerve grade severe) under the same conditions. No change in thickness of either the INL or the photoreceptors are observed, but note the sparseness of cell bodies in the GCL in B. (C) Immunostaining for ChAT (green) in a vertical section of a DBA/2J mouse (nerve grade severe) reveals the typical two bands in the IPL and starburst cell bodies in the INL and GCL. (D) Maximum-intensity projection of a stack of 10 confocal sections at 1-μm step size through a starburst amacrine cell labeled biolistically with rhodamine-dextran. The cell shows the characteristic radially symmetric morphology. The arrow indicates a nearby Mueller cell. (E and F) Percentage of ChAT+ amacrine cells of the total cells in the GCL in the central (E) and peripheral (F) retina. Open circles, C57BL/6J; closed circles, 3-mo-old DBA/2J; blue rectangles, DBA/2J nerve grade unaffected; red triangles, DBA/2J nerve grade severe. (G and H) Whole-mounted retina (focus on the GCL) from a 3-mo-old control DBA/2J prediseased animal stained for ChAT (G) and cell nuclei (H). (I and J) Whole-mounted retina from a glaucomatous DBA/2J (nerve grade severe) animal. Though the numbers of ChAT+ starburst amacrine cells in the control and the diseased animals (135 vs. 138 in these two fields) are similar, note the obvious reduction of cells in the GCL in J compared with H. Images in G–J are position matched. (K) Single confocal section of a DBA/2J retina (nerve grade severe) stained for calbindin 28D (green) and cell nuclei (blue). The arrows indicate horizontal cell bodies. Note the sparseness of cell bodies in the GCL in all panels. (L) Staining for GABA (green). (M) Staining for b-NOS (green). The arrowhead indicates a b-NOS+ cell body. (N) Staining for synaptophysin (green). Note the intense synaptic stain in the outer plexiform layer (arrowhead) and throughout the IPL. (O) Staining for PKCα (green) and CD15 (red). The sections in K–O are from the same eye. Bars: (A–C) 50 μm; (D) 50 μm; (G–J) 100 μm; (K–O) 50 μm.
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fig2: Amacrine cells in glaucomatous DBA/2J mice. (A) Single confocal vertical section through the retina of a C57BL/6J mouse, stained with the nuclear dye TOPRO. For the sake of comparison, images in A–C have all been taken in the central part of the retina. (B) Vertical section through a DBA/2J retina (nerve grade severe) under the same conditions. No change in thickness of either the INL or the photoreceptors are observed, but note the sparseness of cell bodies in the GCL in B. (C) Immunostaining for ChAT (green) in a vertical section of a DBA/2J mouse (nerve grade severe) reveals the typical two bands in the IPL and starburst cell bodies in the INL and GCL. (D) Maximum-intensity projection of a stack of 10 confocal sections at 1-μm step size through a starburst amacrine cell labeled biolistically with rhodamine-dextran. The cell shows the characteristic radially symmetric morphology. The arrow indicates a nearby Mueller cell. (E and F) Percentage of ChAT+ amacrine cells of the total cells in the GCL in the central (E) and peripheral (F) retina. Open circles, C57BL/6J; closed circles, 3-mo-old DBA/2J; blue rectangles, DBA/2J nerve grade unaffected; red triangles, DBA/2J nerve grade severe. (G and H) Whole-mounted retina (focus on the GCL) from a 3-mo-old control DBA/2J prediseased animal stained for ChAT (G) and cell nuclei (H). (I and J) Whole-mounted retina from a glaucomatous DBA/2J (nerve grade severe) animal. Though the numbers of ChAT+ starburst amacrine cells in the control and the diseased animals (135 vs. 138 in these two fields) are similar, note the obvious reduction of cells in the GCL in J compared with H. Images in G–J are position matched. (K) Single confocal section of a DBA/2J retina (nerve grade severe) stained for calbindin 28D (green) and cell nuclei (blue). The arrows indicate horizontal cell bodies. Note the sparseness of cell bodies in the GCL in all panels. (L) Staining for GABA (green). (M) Staining for b-NOS (green). The arrowhead indicates a b-NOS+ cell body. (N) Staining for synaptophysin (green). Note the intense synaptic stain in the outer plexiform layer (arrowhead) and throughout the IPL. (O) Staining for PKCα (green) and CD15 (red). The sections in K–O are from the same eye. Bars: (A–C) 50 μm; (D) 50 μm; (G–J) 100 μm; (K–O) 50 μm.

Mentions: Here, we initially compared vertical sections of a severely affected DBA/2J with a control (C57BL/6J) retina. The reduced cell density in the GCL was obvious, but otherwise the retinas were indistinguishable (Fig. 2, A and B). In accordance with this, our previous comparison of vertical sections of several severely affected DBA/2J retinas with those of young DBA/2J mice does not support the loss of retinal neurons other than ganglion cells in the majority of eyes (Libby et al., 2005a). These finding argue against heavy loss of neurons other than ganglion cells, but the overall thickness of layers is a coarse method of evaluation. We therefore stained for a variety of amacrine and bipolar cell markers in both whole-mounted retinas and vertical sections, taking the cholinergic starburst type of amacrine cell (Masland et al., 1984) as a well-characterized prototype.


Retinal ganglion cell degeneration is topological but not cell type specific in DBA/2J mice.

Jakobs TC, Libby RT, Ben Y, John SW, Masland RH - J. Cell Biol. (2005)

Amacrine cells in glaucomatous DBA/2J mice. (A) Single confocal vertical section through the retina of a C57BL/6J mouse, stained with the nuclear dye TOPRO. For the sake of comparison, images in A–C have all been taken in the central part of the retina. (B) Vertical section through a DBA/2J retina (nerve grade severe) under the same conditions. No change in thickness of either the INL or the photoreceptors are observed, but note the sparseness of cell bodies in the GCL in B. (C) Immunostaining for ChAT (green) in a vertical section of a DBA/2J mouse (nerve grade severe) reveals the typical two bands in the IPL and starburst cell bodies in the INL and GCL. (D) Maximum-intensity projection of a stack of 10 confocal sections at 1-μm step size through a starburst amacrine cell labeled biolistically with rhodamine-dextran. The cell shows the characteristic radially symmetric morphology. The arrow indicates a nearby Mueller cell. (E and F) Percentage of ChAT+ amacrine cells of the total cells in the GCL in the central (E) and peripheral (F) retina. Open circles, C57BL/6J; closed circles, 3-mo-old DBA/2J; blue rectangles, DBA/2J nerve grade unaffected; red triangles, DBA/2J nerve grade severe. (G and H) Whole-mounted retina (focus on the GCL) from a 3-mo-old control DBA/2J prediseased animal stained for ChAT (G) and cell nuclei (H). (I and J) Whole-mounted retina from a glaucomatous DBA/2J (nerve grade severe) animal. Though the numbers of ChAT+ starburst amacrine cells in the control and the diseased animals (135 vs. 138 in these two fields) are similar, note the obvious reduction of cells in the GCL in J compared with H. Images in G–J are position matched. (K) Single confocal section of a DBA/2J retina (nerve grade severe) stained for calbindin 28D (green) and cell nuclei (blue). The arrows indicate horizontal cell bodies. Note the sparseness of cell bodies in the GCL in all panels. (L) Staining for GABA (green). (M) Staining for b-NOS (green). The arrowhead indicates a b-NOS+ cell body. (N) Staining for synaptophysin (green). Note the intense synaptic stain in the outer plexiform layer (arrowhead) and throughout the IPL. (O) Staining for PKCα (green) and CD15 (red). The sections in K–O are from the same eye. Bars: (A–C) 50 μm; (D) 50 μm; (G–J) 100 μm; (K–O) 50 μm.
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Related In: Results  -  Collection

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fig2: Amacrine cells in glaucomatous DBA/2J mice. (A) Single confocal vertical section through the retina of a C57BL/6J mouse, stained with the nuclear dye TOPRO. For the sake of comparison, images in A–C have all been taken in the central part of the retina. (B) Vertical section through a DBA/2J retina (nerve grade severe) under the same conditions. No change in thickness of either the INL or the photoreceptors are observed, but note the sparseness of cell bodies in the GCL in B. (C) Immunostaining for ChAT (green) in a vertical section of a DBA/2J mouse (nerve grade severe) reveals the typical two bands in the IPL and starburst cell bodies in the INL and GCL. (D) Maximum-intensity projection of a stack of 10 confocal sections at 1-μm step size through a starburst amacrine cell labeled biolistically with rhodamine-dextran. The cell shows the characteristic radially symmetric morphology. The arrow indicates a nearby Mueller cell. (E and F) Percentage of ChAT+ amacrine cells of the total cells in the GCL in the central (E) and peripheral (F) retina. Open circles, C57BL/6J; closed circles, 3-mo-old DBA/2J; blue rectangles, DBA/2J nerve grade unaffected; red triangles, DBA/2J nerve grade severe. (G and H) Whole-mounted retina (focus on the GCL) from a 3-mo-old control DBA/2J prediseased animal stained for ChAT (G) and cell nuclei (H). (I and J) Whole-mounted retina from a glaucomatous DBA/2J (nerve grade severe) animal. Though the numbers of ChAT+ starburst amacrine cells in the control and the diseased animals (135 vs. 138 in these two fields) are similar, note the obvious reduction of cells in the GCL in J compared with H. Images in G–J are position matched. (K) Single confocal section of a DBA/2J retina (nerve grade severe) stained for calbindin 28D (green) and cell nuclei (blue). The arrows indicate horizontal cell bodies. Note the sparseness of cell bodies in the GCL in all panels. (L) Staining for GABA (green). (M) Staining for b-NOS (green). The arrowhead indicates a b-NOS+ cell body. (N) Staining for synaptophysin (green). Note the intense synaptic stain in the outer plexiform layer (arrowhead) and throughout the IPL. (O) Staining for PKCα (green) and CD15 (red). The sections in K–O are from the same eye. Bars: (A–C) 50 μm; (D) 50 μm; (G–J) 100 μm; (K–O) 50 μm.
Mentions: Here, we initially compared vertical sections of a severely affected DBA/2J with a control (C57BL/6J) retina. The reduced cell density in the GCL was obvious, but otherwise the retinas were indistinguishable (Fig. 2, A and B). In accordance with this, our previous comparison of vertical sections of several severely affected DBA/2J retinas with those of young DBA/2J mice does not support the loss of retinal neurons other than ganglion cells in the majority of eyes (Libby et al., 2005a). These finding argue against heavy loss of neurons other than ganglion cells, but the overall thickness of layers is a coarse method of evaluation. We therefore stained for a variety of amacrine and bipolar cell markers in both whole-mounted retinas and vertical sections, taking the cholinergic starburst type of amacrine cell (Masland et al., 1984) as a well-characterized prototype.

Bottom Line: Regions of cell death or survival radiated from the optic nerve head in fan-shaped sectors.Collectively, the data suggest axon damage at the optic nerve head as an early lesion, and damage to axon bundles would cause this pattern of degeneration.However, the architecture of the mouse eye seems to preclude a commonly postulated source of mechanical damage within the nerve head.

View Article: PubMed Central - PubMed

Affiliation: Howard Hughes Medical Institute, Harvard Medical School, Boston, MA 02114.

ABSTRACT
Using a variety of double and triple labeling techniques, we have reevaluated the death of retinal neurons in a mouse model of hereditary glaucoma. Cell-specific markers and total neuron counts revealed no cell loss in any retinal neurons other than the ganglion cells. Within the limits of our ability to define cell types, no group of ganglion cells was especially vulnerable or resistant to degeneration. Retrograde labeling and neurofilament staining showed that axonal atrophy, dendritic remodeling, and somal shrinkage (at least of the largest cell types) precedes ganglion cell death in this glaucoma model. Regions of cell death or survival radiated from the optic nerve head in fan-shaped sectors. Collectively, the data suggest axon damage at the optic nerve head as an early lesion, and damage to axon bundles would cause this pattern of degeneration. However, the architecture of the mouse eye seems to preclude a commonly postulated source of mechanical damage within the nerve head.

Show MeSH
Related in: MedlinePlus