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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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Upregulation of microglial marker CD68 in the thalamus and cerebral cortex of Cln6nclf mice.Quantitative thresholding analysis in Cln6nclf mice was compared to age matched WT, revealing a significant increase in the expression of the microglia marker CD68 in the VPM/VPL (A), M1 (C), S1BF (B), and V1 (D) regions in the Cln6nclf mouse over WT. [Mean% immunoreactivity +/− SEM, n = 3 (*p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001)].
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pone-0078694-g008: Upregulation of microglial marker CD68 in the thalamus and cerebral cortex of Cln6nclf mice.Quantitative thresholding analysis in Cln6nclf mice was compared to age matched WT, revealing a significant increase in the expression of the microglia marker CD68 in the VPM/VPL (A), M1 (C), S1BF (B), and V1 (D) regions in the Cln6nclf mouse over WT. [Mean% immunoreactivity +/− SEM, n = 3 (*p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001)].

Mentions: Another pathological hallmark of the NCLs is an early onset, robust activation of glia activation. To quantify the activation of astrocytes and microglia, threshold analyses of GFAP (glial fibrillary acidic protein, astrocytes) and CD68 (microglia) immunoreactivity were performed. Levels of GFAP staining were measured in the S1BF and V1 regions of the cerebral cortex and the medial and lateral parts of the ventroposterior nucleus of the thalamus (VPM/VPL) (Fig. 7 A–C), another brain region often affected early in NCL mouse models [47], [50], [52], [57], [58], [62], [63]. Within the S1BF and V1 regions, there was a marked increase in GFAP by 6 months of age (Fig. 7 B–C). When analyzing VPM/VPL thalamic nuclei, GFAP immunoreactivity was massively upregulated at 4 months, persisting with age (Fig. 7 A). Microglial activation was apparent in all of the regions examined, appearing by 4 months of age and remaining elevated throughout the life of the animal [VPM/VPL, S1BF, M1, and V1, (Fig. 8 A–D)]. Although glial activation has been reported previously in Cln6nclf mice, these findings demonstrate earlier onset pathology, with previous studies showing astrocytosis at 21 weeks and microglial activation at 54 weeks of age [32].


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)

Upregulation of microglial marker CD68 in the thalamus and cerebral cortex of Cln6nclf mice.Quantitative thresholding analysis in Cln6nclf mice was compared to age matched WT, revealing a significant increase in the expression of the microglia marker CD68 in the VPM/VPL (A), M1 (C), S1BF (B), and V1 (D) regions in the Cln6nclf mouse over WT. [Mean% immunoreactivity +/− SEM, n = 3 (*p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001)].
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3815212&req=5

pone-0078694-g008: Upregulation of microglial marker CD68 in the thalamus and cerebral cortex of Cln6nclf mice.Quantitative thresholding analysis in Cln6nclf mice was compared to age matched WT, revealing a significant increase in the expression of the microglia marker CD68 in the VPM/VPL (A), M1 (C), S1BF (B), and V1 (D) regions in the Cln6nclf mouse over WT. [Mean% immunoreactivity +/− SEM, n = 3 (*p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001)].
Mentions: Another pathological hallmark of the NCLs is an early onset, robust activation of glia activation. To quantify the activation of astrocytes and microglia, threshold analyses of GFAP (glial fibrillary acidic protein, astrocytes) and CD68 (microglia) immunoreactivity were performed. Levels of GFAP staining were measured in the S1BF and V1 regions of the cerebral cortex and the medial and lateral parts of the ventroposterior nucleus of the thalamus (VPM/VPL) (Fig. 7 A–C), another brain region often affected early in NCL mouse models [47], [50], [52], [57], [58], [62], [63]. Within the S1BF and V1 regions, there was a marked increase in GFAP by 6 months of age (Fig. 7 B–C). When analyzing VPM/VPL thalamic nuclei, GFAP immunoreactivity was massively upregulated at 4 months, persisting with age (Fig. 7 A). Microglial activation was apparent in all of the regions examined, appearing by 4 months of age and remaining elevated throughout the life of the animal [VPM/VPL, S1BF, M1, and V1, (Fig. 8 A–D)]. Although glial activation has been reported previously in Cln6nclf mice, these findings demonstrate earlier onset pathology, with previous studies showing astrocytosis at 21 weeks and microglial activation at 54 weeks of age [32].

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