Differential Levels and Phosphorylation of Type 1 Inositol 1,4,5-Trisphosphate Receptor in Four Different Murine Models of Huntington Disease

Article type: Research Article

Authors: Post, Joakim Iver^{a; b} | Leergaard, Trygve B.^b | Ratz, Veronika^c | Walaas, S. Ivar^b | von Hörsten, Stephan^c | Nissen-Meyer, Lise Sofie H.^{a; b; d; *}

Affiliations: [a] The Biotechnology Centre of Oslo, University of Oslo, Oslo, Norway | [b] Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway | [c] Department for Experimental Therapy, Preclinical Experimental Centre, Friedrich-Alexander-University Erlangen-Nürnberg, Germany | [d] Department of Immunology and Transfusion, Oslo University Hospital, Oslo, Norway

Correspondence: [*] Correspondence to: Lise Sofie H. Nissen-Meyer, MD, PhD, Department of Immunology and Transfusion, Oslo University Hospital Ullevål, POB 4950 Nydalen, 0424 Oslo, Norway. Tel.: +47 48 28 45 25; E-mail: [email protected].

Abstract: Background:The intracellular ion channel type 1 inositol 1,4,5-trisphosphate receptor (IP3R1) releases Ca2+ from the endoplasmic reticulum upon stimulation with IP3. Perturbation of IP3R1 has been implicated in the development of several neurodegenerative disorders, including Huntington disease (HD). Objective:To elucidate the putative role of IP3R1 phosphorylation in HD, we investigated IP3R1 levels and protein phosphorylation state in the striatum, hippocampus and cerebellum of four murine HD models. Methods:Quantitative immunoblotting with antibodies to IP3R1 protein and its phosphorylated serines 1589 and 1755 was applied to brain homogenates from R6/1 mice to study early-onset aggressive HD. To determine if IP3R1 changes precede overt pathology, we immunostained tissues from the regions of interest and several control regions for IP3R1 in tgHDCAG51n rats and BACHD and zQ175DNKI mice, all recognized models for late-onset HD. Results:R6/1 mice had reduced total IP3R1 immunoreactivity, variably reduced serine1755-phosphorylation in all regions investigated, and reduced serine1589-phosphorylation in cerebellum. IP3R1 levels were decreased relative to cell-specific marker proteins. In tgHDCAG51n rats we found reduced IP3R1 levels in the cerebellum, but otherwise unchanged IP3R1 phosphorylation and protein levels. In BACHD and zQ175DNKI mice only age-dependent decline of IP3R1 was observed. Conclusion:The level and phosphorylation of IP3R1 is reduced to a variable degree in the different HD models relative to control, indicating that earlier findings in more aggressive exon 1-truncated HD models may not be replicated in models with higher construct validity. Further analysis of possible coupling of reduced IP3R1 levels with development of neuropathological responses and cell-specific degeneration is warranted.

Keywords: Cerebellum, Huntington disease, IP₃ receptor, neurodegeneration, phosphorylation, transgenic model

DOI: 10.3233/JHD-180301

Journal: Journal of Huntington's Disease, vol. 8, no. 3, pp. 271-289, 2019