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A murine model of variant late infantile ceroid lipofuscinosis recapitulates behavioral and pathological phenotypes of human disease.

Morgan JP, Magee H, Wong A, Nelson T, Koch B, Cooper JD, Weimer JM - PLoS ONE (2013)

Bottom Line: Pathologically, we demonstrate loss of neurons within specific subregions and lamina of the cortex that correlate to behavioral phenotypes.As in other NCL models, this model displays selective loss of GABAergic interneuron sub-populations in the cortex and the hippocampus with profound, early-onset glial activation.Finally, we demonstrate a novel deficit in memory and learning, including a dramatic reduction in dendritic spine density in the cerebral cortex, which suggests a reduction in synaptic strength following disruption in CLN6.

View Article: PubMed Central - PubMed

Affiliation: Division of Basic Biomedical Sciences. Sanford School of Medicine at the University of South Dakota, Vermillion, South Dakota, United States of America ; Children's Health Research Center, Sanford Research/USD, Sioux Falls, South Dakota, United States of America.

ABSTRACT
Neuronal ceroid lipofuscinoses (NCLs; also known collectively as Batten Disease) are a family of autosomal recessive lysosomal storage disorders. Mutations in as many as 13 genes give rise to ∼10 variants of NCL, all with overlapping clinical symptomatology including visual impairment, motor and cognitive dysfunction, seizures, and premature death. Mutations in CLN6 result in both a variant late infantile onset neuronal ceroid lipofuscinosis (vLINCL) as well as an adult-onset form of the disease called Type A Kufs. CLN6 is a non-glycosylated membrane protein of unknown function localized to the endoplasmic reticulum (ER). In this study, we perform a detailed characterization of a naturally occurring Cln6 mutant (Cln6(nclf)) mouse line to validate its utility for translational research. We demonstrate that this Cln6(nclf) mutation leads to deficits in motor coordination, vision, memory, and learning. Pathologically, we demonstrate loss of neurons within specific subregions and lamina of the cortex that correlate to behavioral phenotypes. As in other NCL models, this model displays selective loss of GABAergic interneuron sub-populations in the cortex and the hippocampus with profound, early-onset glial activation. Finally, we demonstrate a novel deficit in memory and learning, including a dramatic reduction in dendritic spine density in the cerebral cortex, which suggests a reduction in synaptic strength following disruption in CLN6. Together, these findings highlight the behavioral and pathological similarities between the Cln6(nclf) mouse model and human NCL patients, validating this model as a reliable format for screening potential therapeutics.

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Retinal degeneration and vision loss in the Cln6nclf mouse.Cell loss and structural degenerative changes occur in the retina of Cln6nclf mice. (A) Comparison of gross morphological changes over time in retina of Cln6nclf mice and their respective age-matched WT controls was done to determine mechanism of degeneration. (B) Micrographs (4X) show a section of one retinal hemisphere. (C) At P0 all layers are present and of equal thickness. By 3 months of age the Cln6nclf retina has begun to narrow and shows a distinct loss of the rods and cones while the overall cytoarchitecture remains intact. By 9 months the rods and cones are nearly absent and the outer plexiform layer is virtually nonexistent with the merging of a much thinned outer and inner nuclear layers. Additionally, there is a distinct narrowing of the inner plexiform layer. [RC-Rode/Cone layer; ONL-Outer nuclear layer; OPL-Outer plexiform later; INL-Inner nuclear layer; IPL-Inner plexiform layer; GCL-Ganglion cell layer]. (D) At 8 months of age, Cln6nclf mice displayed a significant reduction in visual acuity in a visual cliff assay. Mutant mice were unable to distinguish between a “safe” region of the visual cliff box versus the “unsafe” cliffed portion, spending equal time between the two regions. [Mean (in seconds) +/- SEM, n = 6–9 mice per group (**p≤0.01)].
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pone-0078694-g001: Retinal degeneration and vision loss in the Cln6nclf mouse.Cell loss and structural degenerative changes occur in the retina of Cln6nclf mice. (A) Comparison of gross morphological changes over time in retina of Cln6nclf mice and their respective age-matched WT controls was done to determine mechanism of degeneration. (B) Micrographs (4X) show a section of one retinal hemisphere. (C) At P0 all layers are present and of equal thickness. By 3 months of age the Cln6nclf retina has begun to narrow and shows a distinct loss of the rods and cones while the overall cytoarchitecture remains intact. By 9 months the rods and cones are nearly absent and the outer plexiform layer is virtually nonexistent with the merging of a much thinned outer and inner nuclear layers. Additionally, there is a distinct narrowing of the inner plexiform layer. [RC-Rode/Cone layer; ONL-Outer nuclear layer; OPL-Outer plexiform later; INL-Inner nuclear layer; IPL-Inner plexiform layer; GCL-Ganglion cell layer]. (D) At 8 months of age, Cln6nclf mice displayed a significant reduction in visual acuity in a visual cliff assay. Mutant mice were unable to distinguish between a “safe” region of the visual cliff box versus the “unsafe” cliffed portion, spending equal time between the two regions. [Mean (in seconds) +/- SEM, n = 6–9 mice per group (**p≤0.01)].

Mentions: Retinal atrophy and progressive vision loss is a pathological hallmark of vLINCL patients (reviewed in [6], [53], [54]). Cln6nclf mice show a similar progressive loss of cells within the retina. At postnatal day zero (P0), all layers are present and of equal thickness (Fig. 1 A). By 3 months the Cln6nclf retina has begun to narrow and shows a distinct loss in the photoreceptor layer with both inner and outer nuclear layers showing disorganization (Fig. 1 B). By 9 months there is massive degeneration, with the rods and cones nearly absent, the outer plexiform layer (OPL) virtually nonexistent, and a merging of the outer (ONL) and inner nuclear layers (INL). There is also a distinct narrowing of the inner plexiform layer (IPL) demonstrating that by 8 months of life, the retina of the Cln6nclf is very severely deteriorated (Fig. 1 C). To assay visual performance and acuity, mice were analyzed in a visual cliff paradigm [38], [39]. 8 month old Cln6nclf mice were unable to discriminate between the safe zone and crossing over the cliff zone, spending equal amounts of time within each compartment (Fig 1 D), demonstrating a significant loss in vision at this time point.


A murine model of variant late infantile ceroid lipofuscinosis recapitulates behavioral and pathological phenotypes of human disease.

Morgan JP, Magee H, Wong A, Nelson T, Koch B, Cooper JD, Weimer JM - PLoS ONE (2013)

Retinal degeneration and vision loss in the Cln6nclf mouse.Cell loss and structural degenerative changes occur in the retina of Cln6nclf mice. (A) Comparison of gross morphological changes over time in retina of Cln6nclf mice and their respective age-matched WT controls was done to determine mechanism of degeneration. (B) Micrographs (4X) show a section of one retinal hemisphere. (C) At P0 all layers are present and of equal thickness. By 3 months of age the Cln6nclf retina has begun to narrow and shows a distinct loss of the rods and cones while the overall cytoarchitecture remains intact. By 9 months the rods and cones are nearly absent and the outer plexiform layer is virtually nonexistent with the merging of a much thinned outer and inner nuclear layers. Additionally, there is a distinct narrowing of the inner plexiform layer. [RC-Rode/Cone layer; ONL-Outer nuclear layer; OPL-Outer plexiform later; INL-Inner nuclear layer; IPL-Inner plexiform layer; GCL-Ganglion cell layer]. (D) At 8 months of age, Cln6nclf mice displayed a significant reduction in visual acuity in a visual cliff assay. Mutant mice were unable to distinguish between a “safe” region of the visual cliff box versus the “unsafe” cliffed portion, spending equal time between the two regions. [Mean (in seconds) +/- SEM, n = 6–9 mice per group (**p≤0.01)].
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3815212&req=5

pone-0078694-g001: Retinal degeneration and vision loss in the Cln6nclf mouse.Cell loss and structural degenerative changes occur in the retina of Cln6nclf mice. (A) Comparison of gross morphological changes over time in retina of Cln6nclf mice and their respective age-matched WT controls was done to determine mechanism of degeneration. (B) Micrographs (4X) show a section of one retinal hemisphere. (C) At P0 all layers are present and of equal thickness. By 3 months of age the Cln6nclf retina has begun to narrow and shows a distinct loss of the rods and cones while the overall cytoarchitecture remains intact. By 9 months the rods and cones are nearly absent and the outer plexiform layer is virtually nonexistent with the merging of a much thinned outer and inner nuclear layers. Additionally, there is a distinct narrowing of the inner plexiform layer. [RC-Rode/Cone layer; ONL-Outer nuclear layer; OPL-Outer plexiform later; INL-Inner nuclear layer; IPL-Inner plexiform layer; GCL-Ganglion cell layer]. (D) At 8 months of age, Cln6nclf mice displayed a significant reduction in visual acuity in a visual cliff assay. Mutant mice were unable to distinguish between a “safe” region of the visual cliff box versus the “unsafe” cliffed portion, spending equal time between the two regions. [Mean (in seconds) +/- SEM, n = 6–9 mice per group (**p≤0.01)].
Mentions: Retinal atrophy and progressive vision loss is a pathological hallmark of vLINCL patients (reviewed in [6], [53], [54]). Cln6nclf mice show a similar progressive loss of cells within the retina. At postnatal day zero (P0), all layers are present and of equal thickness (Fig. 1 A). By 3 months the Cln6nclf retina has begun to narrow and shows a distinct loss in the photoreceptor layer with both inner and outer nuclear layers showing disorganization (Fig. 1 B). By 9 months there is massive degeneration, with the rods and cones nearly absent, the outer plexiform layer (OPL) virtually nonexistent, and a merging of the outer (ONL) and inner nuclear layers (INL). There is also a distinct narrowing of the inner plexiform layer (IPL) demonstrating that by 8 months of life, the retina of the Cln6nclf is very severely deteriorated (Fig. 1 C). To assay visual performance and acuity, mice were analyzed in a visual cliff paradigm [38], [39]. 8 month old Cln6nclf mice were unable to discriminate between the safe zone and crossing over the cliff zone, spending equal amounts of time within each compartment (Fig 1 D), demonstrating a significant loss in vision at this time point.

Bottom Line: Pathologically, we demonstrate loss of neurons within specific subregions and lamina of the cortex that correlate to behavioral phenotypes.As in other NCL models, this model displays selective loss of GABAergic interneuron sub-populations in the cortex and the hippocampus with profound, early-onset glial activation.Finally, we demonstrate a novel deficit in memory and learning, including a dramatic reduction in dendritic spine density in the cerebral cortex, which suggests a reduction in synaptic strength following disruption in CLN6.

View Article: PubMed Central - PubMed

Affiliation: Division of Basic Biomedical Sciences. Sanford School of Medicine at the University of South Dakota, Vermillion, South Dakota, United States of America ; Children's Health Research Center, Sanford Research/USD, Sioux Falls, South Dakota, United States of America.

ABSTRACT
Neuronal ceroid lipofuscinoses (NCLs; also known collectively as Batten Disease) are a family of autosomal recessive lysosomal storage disorders. Mutations in as many as 13 genes give rise to ∼10 variants of NCL, all with overlapping clinical symptomatology including visual impairment, motor and cognitive dysfunction, seizures, and premature death. Mutations in CLN6 result in both a variant late infantile onset neuronal ceroid lipofuscinosis (vLINCL) as well as an adult-onset form of the disease called Type A Kufs. CLN6 is a non-glycosylated membrane protein of unknown function localized to the endoplasmic reticulum (ER). In this study, we perform a detailed characterization of a naturally occurring Cln6 mutant (Cln6(nclf)) mouse line to validate its utility for translational research. We demonstrate that this Cln6(nclf) mutation leads to deficits in motor coordination, vision, memory, and learning. Pathologically, we demonstrate loss of neurons within specific subregions and lamina of the cortex that correlate to behavioral phenotypes. As in other NCL models, this model displays selective loss of GABAergic interneuron sub-populations in the cortex and the hippocampus with profound, early-onset glial activation. Finally, we demonstrate a novel deficit in memory and learning, including a dramatic reduction in dendritic spine density in the cerebral cortex, which suggests a reduction in synaptic strength following disruption in CLN6. Together, these findings highlight the behavioral and pathological similarities between the Cln6(nclf) mouse model and human NCL patients, validating this model as a reliable format for screening potential therapeutics.

Show MeSH
Related in: MedlinePlus