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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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Abnormal hematology in peripheral blood from homozygous Cln3Δex7/8 mice. (A) Mean corpuscular volume (MCV, fL) of peripheral red blood cells from ∼12-week-old mice was measured on an automated analyzer. *, p<0.05, WT and heterozygous mutant mice vs. homozygous mutant mice, unpaired, two-tailed t test. Data shown as mean ± SEM. Percentage of reticulocytes (B) and absolute reticulocyte counts (C) on the specimens analyzed in (A) were determined manually by new methylene blue staining. *, p<0.05, WT and heterozygous mutant mice vs. homozygous mutant mice, unpaired, two-tailed t test. Data shown as mean ± SEM. (D) Linear regression analysis of data from (A) and (C). r2 = 0.32, p = 0.02. Datapoints represent individual mice.
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pone-0038310-g005: Abnormal hematology in peripheral blood from homozygous Cln3Δex7/8 mice. (A) Mean corpuscular volume (MCV, fL) of peripheral red blood cells from ∼12-week-old mice was measured on an automated analyzer. *, p<0.05, WT and heterozygous mutant mice vs. homozygous mutant mice, unpaired, two-tailed t test. Data shown as mean ± SEM. Percentage of reticulocytes (B) and absolute reticulocyte counts (C) on the specimens analyzed in (A) were determined manually by new methylene blue staining. *, p<0.05, WT and heterozygous mutant mice vs. homozygous mutant mice, unpaired, two-tailed t test. Data shown as mean ± SEM. (D) Linear regression analysis of data from (A) and (C). r2 = 0.32, p = 0.02. Datapoints represent individual mice.

Mentions: To survey Cln3Δex7/8 mice in standard clinical chemistry and hematological parameters, we collected blood samples from 12- to 19-week-old wild-type, heterozygous and homozygous littermate Cln3Δex7/8 mice. Samples were analyzed for 21 different analytes including plasma electrolytes, liver enzymes, ferritin and transferrin, and for basic hematological and immunological parameters. No differences in plasma electrolytes or liver enzyme activities were observed. In contrast, serum ferritin concentrations were consistently elevated in homozygous Cln3Δex7/8 mice compared to wild-type and heterozygous littermates (Tables 3 and S4). We also observed a consistently increased mean corpuscular volume (MCV) in the complete blood count (CBC) analysis from homozygous Cln3Δex7/8 mice, as compared to wild-type and heterozygous Cln3Δex7/8 littermate mice (Tables 3 and S4, Fig. 5A).


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)

Abnormal hematology in peripheral blood from homozygous Cln3Δex7/8 mice. (A) Mean corpuscular volume (MCV, fL) of peripheral red blood cells from ∼12-week-old mice was measured on an automated analyzer. *, p<0.05, WT and heterozygous mutant mice vs. homozygous mutant mice, unpaired, two-tailed t test. Data shown as mean ± SEM. Percentage of reticulocytes (B) and absolute reticulocyte counts (C) on the specimens analyzed in (A) were determined manually by new methylene blue staining. *, p<0.05, WT and heterozygous mutant mice vs. homozygous mutant mice, unpaired, two-tailed t test. Data shown as mean ± SEM. (D) Linear regression analysis of data from (A) and (C). r2 = 0.32, p = 0.02. Datapoints represent individual mice.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3368842&req=5

pone-0038310-g005: Abnormal hematology in peripheral blood from homozygous Cln3Δex7/8 mice. (A) Mean corpuscular volume (MCV, fL) of peripheral red blood cells from ∼12-week-old mice was measured on an automated analyzer. *, p<0.05, WT and heterozygous mutant mice vs. homozygous mutant mice, unpaired, two-tailed t test. Data shown as mean ± SEM. Percentage of reticulocytes (B) and absolute reticulocyte counts (C) on the specimens analyzed in (A) were determined manually by new methylene blue staining. *, p<0.05, WT and heterozygous mutant mice vs. homozygous mutant mice, unpaired, two-tailed t test. Data shown as mean ± SEM. (D) Linear regression analysis of data from (A) and (C). r2 = 0.32, p = 0.02. Datapoints represent individual mice.
Mentions: To survey Cln3Δex7/8 mice in standard clinical chemistry and hematological parameters, we collected blood samples from 12- to 19-week-old wild-type, heterozygous and homozygous littermate Cln3Δex7/8 mice. Samples were analyzed for 21 different analytes including plasma electrolytes, liver enzymes, ferritin and transferrin, and for basic hematological and immunological parameters. No differences in plasma electrolytes or liver enzyme activities were observed. In contrast, serum ferritin concentrations were consistently elevated in homozygous Cln3Δex7/8 mice compared to wild-type and heterozygous littermates (Tables 3 and S4). We also observed a consistently increased mean corpuscular volume (MCV) in the complete blood count (CBC) analysis from homozygous Cln3Δex7/8 mice, as compared to wild-type and heterozygous Cln3Δex7/8 littermate mice (Tables 3 and S4, Fig. 5A).

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