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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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Bone marrow analysis of Cln3Δex7/8 mice. Representative images are shown of Wright-Giemsa-stained bone marrow brush cytology, H&E stained sections of formalin-fixed, paraffin embedded tibias, and iron stained brush cytology, from wild-type (Cln3+/+) and homozygous mutant (Cln3Δex7/8/Δex7/8) mice (n = 3 mice per genotype). Stained iron appears blue. Note the reduced amount of stained iron in Cln3Δex7/8/Δex7/8 marrow, compared to wild-type marrow. Arrow, erythroid element; arrowhead, myeloid element; asterisk, megakaryocyte. Scale bars, top and bottom panels = 25 µm; middle panels = 100 µm.
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pone-0038310-g006: Bone marrow analysis of Cln3Δex7/8 mice. Representative images are shown of Wright-Giemsa-stained bone marrow brush cytology, H&E stained sections of formalin-fixed, paraffin embedded tibias, and iron stained brush cytology, from wild-type (Cln3+/+) and homozygous mutant (Cln3Δex7/8/Δex7/8) mice (n = 3 mice per genotype). Stained iron appears blue. Note the reduced amount of stained iron in Cln3Δex7/8/Δex7/8 marrow, compared to wild-type marrow. Arrow, erythroid element; arrowhead, myeloid element; asterisk, megakaryocyte. Scale bars, top and bottom panels = 25 µm; middle panels = 100 µm.

Mentions: Next, in order to determine whether the increased reticulocyte number was secondary to increased erythroid precursor production in primary sites of hematopoiesis or whether it was due to delayed maturation of erythroid precursors in the periphery, we analyzed liver, spleen, and bone marrow, the major hematopoietic tissues in developing and adult mice. Liver and spleen from homozygous Cln3Δex7/8 mice were not enlarged or morphologically different from the heterozygous and wild-type littermate tissues (Fig. S9), consistent with previous data [8]. Moreover, brush cytology of bone marrow from wild-type and homozygous Cln3Δex7/8 mice revealed normal trilineage hematopoiesis and a normal myeloid:erythroid ratio (∼2∶1; Fig. 6). Age-appropriate marrow cellularity (∼80%–85%) and normal hematopoietic architecture were also observed in tibia cross-sections from 12-week-old wild-type and homozygous Cln3Δex7/8 mice (Fig. 6). Taken together, these data suggest that CLN3 dysfunction affects reticulocyte maturation in the periphery, but does not exert a global effect on primary hematopoiesis. However, we cannot exclude that the grossly normal tissue pathology did not immediately follow a regenerative erythroid response.


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)

Bone marrow analysis of Cln3Δex7/8 mice. Representative images are shown of Wright-Giemsa-stained bone marrow brush cytology, H&E stained sections of formalin-fixed, paraffin embedded tibias, and iron stained brush cytology, from wild-type (Cln3+/+) and homozygous mutant (Cln3Δex7/8/Δex7/8) mice (n = 3 mice per genotype). Stained iron appears blue. Note the reduced amount of stained iron in Cln3Δex7/8/Δex7/8 marrow, compared to wild-type marrow. Arrow, erythroid element; arrowhead, myeloid element; asterisk, megakaryocyte. Scale bars, top and bottom panels = 25 µm; middle panels = 100 µm.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3368842&req=5

pone-0038310-g006: Bone marrow analysis of Cln3Δex7/8 mice. Representative images are shown of Wright-Giemsa-stained bone marrow brush cytology, H&E stained sections of formalin-fixed, paraffin embedded tibias, and iron stained brush cytology, from wild-type (Cln3+/+) and homozygous mutant (Cln3Δex7/8/Δex7/8) mice (n = 3 mice per genotype). Stained iron appears blue. Note the reduced amount of stained iron in Cln3Δex7/8/Δex7/8 marrow, compared to wild-type marrow. Arrow, erythroid element; arrowhead, myeloid element; asterisk, megakaryocyte. Scale bars, top and bottom panels = 25 µm; middle panels = 100 µm.
Mentions: Next, in order to determine whether the increased reticulocyte number was secondary to increased erythroid precursor production in primary sites of hematopoiesis or whether it was due to delayed maturation of erythroid precursors in the periphery, we analyzed liver, spleen, and bone marrow, the major hematopoietic tissues in developing and adult mice. Liver and spleen from homozygous Cln3Δex7/8 mice were not enlarged or morphologically different from the heterozygous and wild-type littermate tissues (Fig. S9), consistent with previous data [8]. Moreover, brush cytology of bone marrow from wild-type and homozygous Cln3Δex7/8 mice revealed normal trilineage hematopoiesis and a normal myeloid:erythroid ratio (∼2∶1; Fig. 6). Age-appropriate marrow cellularity (∼80%–85%) and normal hematopoietic architecture were also observed in tibia cross-sections from 12-week-old wild-type and homozygous Cln3Δex7/8 mice (Fig. 6). Taken together, these data suggest that CLN3 dysfunction affects reticulocyte maturation in the periphery, but does not exert a global effect on primary hematopoiesis. However, we cannot exclude that the grossly normal tissue pathology did not immediately follow a regenerative erythroid response.

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