Large-Scale Phenotyping of an Accurate Genetic Mouse Model of JNCL Identifies Novel Early Pathology Outside the Central Nervous System John F. Staropoli 1,2 , Larissa Haliw 1 , Sunita Biswas 1 , Lillian Garrett 3 , Sabine M. Ho ¨ lter 3 , Lore Becker 4,5 , Sergej Skosyrski 6 , Patricia Da Silva-Buttkus 7 , Julia Calzada-Wack 7 , Frauke Neff 7 , Birgit Rathkolb 5,8 , Jan Rozman 5,9 , Anja Schrewe 5 , Thure Adler 5,10 , Oliver Puk 3 , Minxuan Sun 3 , Jack Favor 11 , Ildiko ´ Racz 12 , Raffi Bekeredjian 13 , Dirk H. Busch 10 , Jochen Graw 3 , Martin Klingenspor 9 , Thomas Klopstock 4 , Eckhard Wolf 8 , Wolfgang Wurst 3,14,15,16 , Andreas Zimmer 12 , Edith Lopez 1 , Hayat Harati 1,17 , Eric Hill 18 , Daniela S. Krause 2 , Jolene Guide 1 , Ella Dragileva 1 , Evan Gale 1 , Vanessa C. Wheeler 1 , Rose- Mary Boustany 17 , Diane E. Brown 2,19 , Sylvie Breton 18 , Klaus Ruether 20 , Vale ´ rie Gailus-Durner 5 , Helmut Fuchs 5 , Martin Hrabe ˇ de Angelis 5,21 , Susan L. Cotman 1 * 1 Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, United States of America, 2 Department of Pathology, Massachusetts General Hospital, Boston, Massachusetts, United States of America, 3 Institute of Developmental Genetics, Helmholtz Zentrum Mu ¨ nchen, Neuherberg/Munich, Germany, 4 Department of Neurology, Friedrich-Baur-Institut, Ludwig-Maximilians-Universita ¨t Mu ¨nchen, Munich, Germany, 5 German Mouse Clinic, Institute of Experimental Genetics, Helmholtz Zentrum Mu ¨nchen, Neuherberg/Munich, Germany, 6 Charite ´-Eye Hospital, Campus Virchow-Klinikum, Berlin, Germany, 7 Institute of Pathology, Helmholtz Zentrum Mu ¨nchen, Neuherberg/Munich, Germany, 8 Chair for Molecular Animal Breeding and Biotechnology, Gene Center, Ludwig-Maximilians- Universita ¨t Mu ¨nchen, Munich, Germany, 9 Molecular Nutritional Medicine, Else Kro ¨ner-Fresenius Center, TUM, Freising-Weihenstephan, Germany, 10 Institute of Medical Microbiology, Immunology, and Hygiene, TUM, Mu ¨ nchen, Germany, 11 Institute of Human Genetics, Helmholtz Zentrum Mu ¨nchen, Neuherberg/Munich, Germany, 12 Institute of Molecular Psychiatry, University of Bonn, Bonn, Germany, 13 Department of Medicine III, Division of Cardiology, University of Heidelberg, Otto-Meyerhof-Zentrum, Heidelberg, Germany, 14 Lehrstuhl fu ¨r Entwicklungsgenetik, TUM, Freising-Weihenstephan, Germany, 15 Max-Planck-Institute of Psychiatry, Munich, Germany, 16 Deutsches Zentrum fu ¨r Neurodegenerative Erkrankungen e. V. Site Munich, Munich, Germany, 17 Neurogenetics Program and Division of Pediatric Neurology, Departments of Pediatrics and Biochemistry, American University of Beirut, Beirut, Lebanon, 18 Center for Systems Biology, Program in Membrane Biology/Nephrology Division, Massachusetts General Hospital, Boston, Massachusetts, United States of America, 19 Center for Comparative Medicine, Massachusetts General Hospital, Boston, Massachusetts, United States of America, 20 Augenabteilung Sankt Gertrauden Krankenhaus, Berlin, Germany, 21 Lehrstuhl fu ¨r Experimentelle Genetik, TUM, Freising-Weihenstephan, Germany Abstract Cln3 Dex7/8 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 Dex7/8 mice. Homozygous Cln3 Dex7/8 mice, congenic on a C57BL/6N background, displayed subtle deficits in sensory and motor tasks at 10–14 weeks of age. Homozygous Cln3 Dex7/8 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 Dex7/8 mice, which were also seen to a lesser extent in heterozygous Cln3 Dex7/8 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 Dex7/8 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 Dex7/8 neonates, and to a greater extent in older animals. Early onset, severe vacuolation in clear cells of the epididymis of male homozygous Cln3 Dex7/8 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 Dex7/8 mice that merit further study for JNCL biomarker development. Citation: Staropoli JF, Haliw L, Biswas S, Garrett L, Ho ¨ lter SM, et al. (2012) Large-Scale Phenotyping of an Accurate Genetic Mouse Model of JNCL Identifies Novel Early Pathology Outside the Central Nervous System. PLoS ONE 7(6): e38310. doi:10.1371/journal.pone.0038310 Editor: Thomas Langmann, Center of Ophtalmology, Germany Received February 23, 2012; Accepted May 8, 2012; Published June 6, 2012 Copyright: ß 2012 Staropoli et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: This work was supported by the Dubai-Harvard Foundation for Medical Research [to SLC and R-MB]; NCL Stiftung [to KR]; the Batten Disease Support and Research Association [to JFS]; the National Institutes of Health: National1380 Institute of Neurological Disorders & Stroke [NS073813 to SLC and NS049206 to VCW]; the German Federal Ministry of Education and Research [DZD E.V. to the German Center for Diabetes Research and NGFN-Plus grants 01GS0850, 01GS0851, 01GS0852, 01GS0853, 01GS0854, GS0868, 01GS0869 to the German Mouse Clinic]; EU grant [EUMODIC, LSHG-2006-037188 to the German Mouse Clinic]; and by the Initiative and Networking Fund of the Helmholtz Association in the framework of the Helmholtz Alliance for Mental Research in an Ageing Society [T(HA-215) to Systems Biology/Program in Membrane Biology is partially supported National Institute of Diabetes and Digestive and Kidney Diseases in the form of an Inflammatory Bowel Disease Grant [DK43351] and a Boston Area Diabetes and Endocrinology Research Center (BADERC) Award [DK57521]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. PLoS ONE | www.plosone.org 1 June 2012 | Volume 7 | Issue 6 | e38310