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Large-scale phenotyping of an accurate genetic mouse model of JNCL identifies novel early pathology outside the central nervous system.

Staropoli JF, Haliw L, Biswas S, Garrett L, Hölter SM, Becker L, Skosyrski S, Da Silva-Buttkus P, Calzada-Wack J, Neff F, Rathkolb B, Rozman J, Schrewe A, Adler T, Puk O, Sun M, Favor J, Racz I, Bekeredjian R, Busch DH, Graw J, Klingenspor M, Klopstock T, Wolf E, Wurst W, Zimmer A, Lopez E, Harati H, Hill E, Krause DS, Guide J, Dragileva E, Gale E, Wheeler VC, Boustany RM, Brown DE, Breton S, Ruether K, Gailus-Durner V, Fuchs H, de Angelis MH, Cotman SL - PLoS ONE (2012)

Bottom Line: Heart weight was slightly increased at 20 weeks of age, but no significant differences were observed in cardiac function in young adults.In a comprehensive blood analysis at 15-16 weeks of age, serum ferritin concentrations, mean corpuscular volume of red blood cells (MCV), and reticulocyte counts were reproducibly increased in homozygous Cln3(Δ) (ex7/8) mice, and male homozygotes had a relative T-cell deficiency, suggesting alterations in hematopoiesis.Finally, consistent with findings in JNCL patients, vacuolated peripheral blood lymphocytes were observed in homozygous Cln3(Δ) (ex7/8) neonates, and to a greater extent in older animals.

View Article: PubMed Central - PubMed

Affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, United States of America.

ABSTRACT
Cln3(Δex7/8) mice harbor the most common genetic defect causing juvenile neuronal ceroid lipofuscinosis (JNCL), an autosomal recessive disease involving seizures, visual, motor and cognitive decline, and premature death. Here, to more thoroughly investigate the manifestations of the common JNCL mutation, we performed a broad phenotyping study of Cln3(Δex7/8) mice. Homozygous Cln3(Δex7/8) mice, congenic on a C57BL/6N background, displayed subtle deficits in sensory and motor tasks at 10-14 weeks of age. Homozygous Cln3(Δex7/8) mice also displayed electroretinographic changes reflecting cone function deficits past 5 months of age and a progressive decline of retinal post-receptoral function. Metabolic analysis revealed increases in rectal body temperature and minimum oxygen consumption in 12-13 week old homozygous Cln3(Δex7/8) mice, which were also seen to a lesser extent in heterozygous Cln3(Δex7/8) mice. Heart weight was slightly increased at 20 weeks of age, but no significant differences were observed in cardiac function in young adults. In a comprehensive blood analysis at 15-16 weeks of age, serum ferritin concentrations, mean corpuscular volume of red blood cells (MCV), and reticulocyte counts were reproducibly increased in homozygous Cln3(Δ) (ex7/8) mice, and male homozygotes had a relative T-cell deficiency, suggesting alterations in hematopoiesis. Finally, consistent with findings in JNCL patients, vacuolated peripheral blood lymphocytes were observed in homozygous Cln3(Δ) (ex7/8) neonates, and to a greater extent in older animals. Early onset, severe vacuolation in clear cells of the epididymis of male homozygous Cln3(Δ) (ex7/8) mice was also observed. These data highlight additional organ systems in which to study CLN3 function, and early phenotypes have been established in homozygous Cln3(Δ) (ex7/8) mice that merit further study for JNCL biomarker development.

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Subtle genotypic differences in performance of young adult Cln3Δex7/8 mice in sensory and motor neurological assays. Shown are results of behavioural analyses in a vertical pole-climbing test (A), prepulse inhibition to the acoustic startle response (PPI) (B), acoustic startle response (C), and thermal nociception (D) for female (left) and male (right) littermate control (Cln3+/+), heterozygous (Cln3+/Δex7/8) and homozygous (Cln3Δex7/8/Δex7/8) mice (n = 9–10 mice per group). Data are presented as mean ± standard error of the mean (SEM). (A) Homozygous Cln3Δex7/8 female mice had an increased latency to descend the pole, compared to female wild-type or heterozygous littermates. In a Kruskal-Wallis test, the genotype effect was p<0.01 (*) for females, with or without heterozygous Cln3Δex7/8 mice included in the analysis. (B) Mean %PPI to an acoustic startle, with four prepulse intensities (67, 69, 73, 81 decibels [db]), or with all prepulse intensities averaged (‘global’) are shown. *, ANOVA, p<0.05. (C) The mean ± SEM of the acoustic startle response to 70–120 db sounds is shown for littermate control (Cln3+/+, circles), heterozygous (Cln3+/Δex7/8, squares) and homozygous (Cln3Δex7/8/Δex7/8, triangles) Cln3Δex7/8 mice. NS = no startle sound. For females, ANOVA, genotype effect was F(7,11) = 4.63, p<0.05, and post-hoc tests revealed that this was significant at 90 and 100 db (*p<0.05, ***P<0.001). No statistically significant differences were detected in the acoustic startle response of males. (D) The mean ± SEM latency to the first sign of pain (seconds = s) in a hot plate assay is shown. *, ANOVA genotype effect p<0.05.
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pone-0038310-g001: Subtle genotypic differences in performance of young adult Cln3Δex7/8 mice in sensory and motor neurological assays. Shown are results of behavioural analyses in a vertical pole-climbing test (A), prepulse inhibition to the acoustic startle response (PPI) (B), acoustic startle response (C), and thermal nociception (D) for female (left) and male (right) littermate control (Cln3+/+), heterozygous (Cln3+/Δex7/8) and homozygous (Cln3Δex7/8/Δex7/8) mice (n = 9–10 mice per group). Data are presented as mean ± standard error of the mean (SEM). (A) Homozygous Cln3Δex7/8 female mice had an increased latency to descend the pole, compared to female wild-type or heterozygous littermates. In a Kruskal-Wallis test, the genotype effect was p<0.01 (*) for females, with or without heterozygous Cln3Δex7/8 mice included in the analysis. (B) Mean %PPI to an acoustic startle, with four prepulse intensities (67, 69, 73, 81 decibels [db]), or with all prepulse intensities averaged (‘global’) are shown. *, ANOVA, p<0.05. (C) The mean ± SEM of the acoustic startle response to 70–120 db sounds is shown for littermate control (Cln3+/+, circles), heterozygous (Cln3+/Δex7/8, squares) and homozygous (Cln3Δex7/8/Δex7/8, triangles) Cln3Δex7/8 mice. NS = no startle sound. For females, ANOVA, genotype effect was F(7,11) = 4.63, p<0.05, and post-hoc tests revealed that this was significant at 90 and 100 db (*p<0.05, ***P<0.001). No statistically significant differences were detected in the acoustic startle response of males. (D) The mean ± SEM latency to the first sign of pain (seconds = s) in a hot plate assay is shown. *, ANOVA genotype effect p<0.05.

Mentions: Again in contrast to the Osorio et al study [27], in which 8-week homozygous Cln3Δex7/8 mice on a C57BL/6J background were reported to perform more poorly than wild-type mice on an accelerating rotarod, we found no genotypic differences in accelerating rotarod performance in 10- to 11-week-old Cln3Δex7/8 mice on the C57BL/6N background (Fig. S3). However, in a second test of motor coordination, the pole-climbing test, where mice were placed at the top of a round, metal bar, head upwards, and time-to-turn and time-to-descend the bar were recorded, especially female homozygous Cln3Δex7/8 mice performed significantly worse than wild-type or heterozygous female littermate mice (Fig. 1A). Notably, the methodology and apparatus used in the open field and rotarod assays in this study and in the Osorio et al. study differed [see Methods and [27]].


Large-scale phenotyping of an accurate genetic mouse model of JNCL identifies novel early pathology outside the central nervous system.

Staropoli JF, Haliw L, Biswas S, Garrett L, Hölter SM, Becker L, Skosyrski S, Da Silva-Buttkus P, Calzada-Wack J, Neff F, Rathkolb B, Rozman J, Schrewe A, Adler T, Puk O, Sun M, Favor J, Racz I, Bekeredjian R, Busch DH, Graw J, Klingenspor M, Klopstock T, Wolf E, Wurst W, Zimmer A, Lopez E, Harati H, Hill E, Krause DS, Guide J, Dragileva E, Gale E, Wheeler VC, Boustany RM, Brown DE, Breton S, Ruether K, Gailus-Durner V, Fuchs H, de Angelis MH, Cotman SL - PLoS ONE (2012)

Subtle genotypic differences in performance of young adult Cln3Δex7/8 mice in sensory and motor neurological assays. Shown are results of behavioural analyses in a vertical pole-climbing test (A), prepulse inhibition to the acoustic startle response (PPI) (B), acoustic startle response (C), and thermal nociception (D) for female (left) and male (right) littermate control (Cln3+/+), heterozygous (Cln3+/Δex7/8) and homozygous (Cln3Δex7/8/Δex7/8) mice (n = 9–10 mice per group). Data are presented as mean ± standard error of the mean (SEM). (A) Homozygous Cln3Δex7/8 female mice had an increased latency to descend the pole, compared to female wild-type or heterozygous littermates. In a Kruskal-Wallis test, the genotype effect was p<0.01 (*) for females, with or without heterozygous Cln3Δex7/8 mice included in the analysis. (B) Mean %PPI to an acoustic startle, with four prepulse intensities (67, 69, 73, 81 decibels [db]), or with all prepulse intensities averaged (‘global’) are shown. *, ANOVA, p<0.05. (C) The mean ± SEM of the acoustic startle response to 70–120 db sounds is shown for littermate control (Cln3+/+, circles), heterozygous (Cln3+/Δex7/8, squares) and homozygous (Cln3Δex7/8/Δex7/8, triangles) Cln3Δex7/8 mice. NS = no startle sound. For females, ANOVA, genotype effect was F(7,11) = 4.63, p<0.05, and post-hoc tests revealed that this was significant at 90 and 100 db (*p<0.05, ***P<0.001). No statistically significant differences were detected in the acoustic startle response of males. (D) The mean ± SEM latency to the first sign of pain (seconds = s) in a hot plate assay is shown. *, ANOVA genotype effect p<0.05.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3368842&req=5

pone-0038310-g001: Subtle genotypic differences in performance of young adult Cln3Δex7/8 mice in sensory and motor neurological assays. Shown are results of behavioural analyses in a vertical pole-climbing test (A), prepulse inhibition to the acoustic startle response (PPI) (B), acoustic startle response (C), and thermal nociception (D) for female (left) and male (right) littermate control (Cln3+/+), heterozygous (Cln3+/Δex7/8) and homozygous (Cln3Δex7/8/Δex7/8) mice (n = 9–10 mice per group). Data are presented as mean ± standard error of the mean (SEM). (A) Homozygous Cln3Δex7/8 female mice had an increased latency to descend the pole, compared to female wild-type or heterozygous littermates. In a Kruskal-Wallis test, the genotype effect was p<0.01 (*) for females, with or without heterozygous Cln3Δex7/8 mice included in the analysis. (B) Mean %PPI to an acoustic startle, with four prepulse intensities (67, 69, 73, 81 decibels [db]), or with all prepulse intensities averaged (‘global’) are shown. *, ANOVA, p<0.05. (C) The mean ± SEM of the acoustic startle response to 70–120 db sounds is shown for littermate control (Cln3+/+, circles), heterozygous (Cln3+/Δex7/8, squares) and homozygous (Cln3Δex7/8/Δex7/8, triangles) Cln3Δex7/8 mice. NS = no startle sound. For females, ANOVA, genotype effect was F(7,11) = 4.63, p<0.05, and post-hoc tests revealed that this was significant at 90 and 100 db (*p<0.05, ***P<0.001). No statistically significant differences were detected in the acoustic startle response of males. (D) The mean ± SEM latency to the first sign of pain (seconds = s) in a hot plate assay is shown. *, ANOVA genotype effect p<0.05.
Mentions: Again in contrast to the Osorio et al study [27], in which 8-week homozygous Cln3Δex7/8 mice on a C57BL/6J background were reported to perform more poorly than wild-type mice on an accelerating rotarod, we found no genotypic differences in accelerating rotarod performance in 10- to 11-week-old Cln3Δex7/8 mice on the C57BL/6N background (Fig. S3). However, in a second test of motor coordination, the pole-climbing test, where mice were placed at the top of a round, metal bar, head upwards, and time-to-turn and time-to-descend the bar were recorded, especially female homozygous Cln3Δex7/8 mice performed significantly worse than wild-type or heterozygous female littermate mice (Fig. 1A). Notably, the methodology and apparatus used in the open field and rotarod assays in this study and in the Osorio et al. study differed [see Methods and [27]].

Bottom Line: Heart weight was slightly increased at 20 weeks of age, but no significant differences were observed in cardiac function in young adults.In a comprehensive blood analysis at 15-16 weeks of age, serum ferritin concentrations, mean corpuscular volume of red blood cells (MCV), and reticulocyte counts were reproducibly increased in homozygous Cln3(Δ) (ex7/8) mice, and male homozygotes had a relative T-cell deficiency, suggesting alterations in hematopoiesis.Finally, consistent with findings in JNCL patients, vacuolated peripheral blood lymphocytes were observed in homozygous Cln3(Δ) (ex7/8) neonates, and to a greater extent in older animals.

View Article: PubMed Central - PubMed

Affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, United States of America.

ABSTRACT
Cln3(Δex7/8) mice harbor the most common genetic defect causing juvenile neuronal ceroid lipofuscinosis (JNCL), an autosomal recessive disease involving seizures, visual, motor and cognitive decline, and premature death. Here, to more thoroughly investigate the manifestations of the common JNCL mutation, we performed a broad phenotyping study of Cln3(Δex7/8) mice. Homozygous Cln3(Δex7/8) mice, congenic on a C57BL/6N background, displayed subtle deficits in sensory and motor tasks at 10-14 weeks of age. Homozygous Cln3(Δex7/8) mice also displayed electroretinographic changes reflecting cone function deficits past 5 months of age and a progressive decline of retinal post-receptoral function. Metabolic analysis revealed increases in rectal body temperature and minimum oxygen consumption in 12-13 week old homozygous Cln3(Δex7/8) mice, which were also seen to a lesser extent in heterozygous Cln3(Δex7/8) mice. Heart weight was slightly increased at 20 weeks of age, but no significant differences were observed in cardiac function in young adults. In a comprehensive blood analysis at 15-16 weeks of age, serum ferritin concentrations, mean corpuscular volume of red blood cells (MCV), and reticulocyte counts were reproducibly increased in homozygous Cln3(Δ) (ex7/8) mice, and male homozygotes had a relative T-cell deficiency, suggesting alterations in hematopoiesis. Finally, consistent with findings in JNCL patients, vacuolated peripheral blood lymphocytes were observed in homozygous Cln3(Δ) (ex7/8) neonates, and to a greater extent in older animals. Early onset, severe vacuolation in clear cells of the epididymis of male homozygous Cln3(Δ) (ex7/8) mice was also observed. These data highlight additional organ systems in which to study CLN3 function, and early phenotypes have been established in homozygous Cln3(Δ) (ex7/8) mice that merit further study for JNCL biomarker development.

Show MeSH
Related in: MedlinePlus