Skip to main content
SpringerLink
Log in

Significantly differential diffusion of neuropathological aggregates in the brain of transgenic mice carrying N-terminal mutant huntingtin fused with green fluorescent protein

  • Original Article
  • Published:
Brain Structure and Function Aims and scope Submit manuscript

Abstract

Huntington’s disease (HD) is a genetically neurodegenerative disease, affecting the central nervous system and leading to mental and motor dysfunctions. To date, there is no cure for HD; as a result, HD patients gradually suffer devastating symptoms, such as chorea, weight loss, depression and mood swings, until death. According to previous studies, the exon 1 region of the huntingtin (HTT) gene with expanded CAG trinucleotide repeats plays a critical role in causing HD. In vitro studies using exon 1 of HTT fused with green fluorescent protein (GFP) gene have facilitated discovering several mechanisms of HD. However, whether this chimera construct exerts similar functions in vivo is still not clear. Here, we report the generation of transgenic mice carrying GFP fused with mutant HTT exon 1 containing 84 CAG trinucleotide repeats, and the evaluation of phenotypes via molecular, neuropathological and behavioral analyses. Results show that these transgenic mice not only displayed neuropathological characteristics, observed either by green fluorescent signals or by immunohistochemical staining, but also progressively developed pathological and behavioral symptoms of HD. Most interestingly, these transgenic mice showed significantly differential expression levels of nuclear aggregates between cortex and striatum regions, highly mimicking selective expression of mutant HTT in HD patients. To the best of our knowledge, this is the first report showing different nuclear diffusion profiling in mouse models with transgenic mice carrying the exon 1 region of mutant HTT. Our model will be beneficial for tracing the expression of mutant HTT and accelerating the understanding of selective pathological progression in HD.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Carter RJ, Lione LA, Humby T et al (1999) Characterization of progressive motor deficits in mice transgenic for the human Huntington’s disease mutation. J Neurosci 19:3248–3257

    PubMed  CAS  Google Scholar 

  • Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC (1994) Green fluorescent protein as a marker for gene expression. Science 263:802–805

    Article  PubMed  CAS  Google Scholar 

  • Chan AW, Yang SH (2009) Generation of transgenic monkeys with human inherited genetic disease. Methods 49:78–84

    Article  PubMed  CAS  Google Scholar 

  • Chiang MC, Chern Y, Juo CG (2011) The dysfunction of hepatic transcriptional factors in mice with Huntington’s Disease. Biochim Biophys Acta 1812:1111–1120

    Article  PubMed  CAS  Google Scholar 

  • Ciammola A, Sassone J, Sciacco M et al (2011) Low anaerobic threshold and increased skeletal muscle lactate production in subjects with Huntington’s disease. Mov Disord 26:130–137

    Article  PubMed  Google Scholar 

  • Cornett J, Cao F, Wang CE et al (2005) Polyglutamine expansion of huntingtin impairs its nuclear export. Nat Genet 37:198–204

    Article  PubMed  CAS  Google Scholar 

  • Davies SW, Turmaine M, Cozens BA et al (1997) Formation of neuronal intranuclear inclusions underlies the neurological dysfunction in mice transgenic for the HD mutation. Cell 90:537–548

    Article  PubMed  CAS  Google Scholar 

  • DiFiglia M, Sapp E, Chase K et al (1995) Huntingtin is a cytoplasmic protein associated with vesicles in human and rat brain neurons. Neuron 14:1075–1081

    Article  PubMed  CAS  Google Scholar 

  • Hodgson JG, Agopyan N, Gutekunst CA et al (1999) A YAC mouse model for Huntington’s disease with full-length mutant huntingtin, cytoplasmic toxicity, and selective striatal neurodegeneration. Neuron 23:181–192

    Article  PubMed  CAS  Google Scholar 

  • Hogan B (1994) Manipulating the mouse embryo: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

  • Li S, Li XJ (2006) Multiple pathways contribute to the pathogenesis of Huntington disease. Mol Neurodegener 1:19

    Article  PubMed  Google Scholar 

  • Lin CH, Tallaksen-Greene S, Chien WM et al (2001) Neurological abnormalities in a knock-in mouse model of Huntington’s disease. Hum Mol Genet 10:137–144

    Article  PubMed  CAS  Google Scholar 

  • Maat-Schieman M, Roos R, Losekoot M et al (2007) Neuronal intranuclear and neuropil inclusions for pathological assessment of Huntington’s disease. Brain Pathol 17:31–37

    Article  PubMed  CAS  Google Scholar 

  • Mangiarini L, Sathasivam K, Seller M et al (1996) Exon 1 of the HD gene with an expanded CAG repeat is sufficient to cause a progressive neurological phenotype in transgenic mice. Cell 87:493–506

    Article  PubMed  CAS  Google Scholar 

  • McGuire JR, Rong J, Li SH, Li XJ (2006) Interaction of Huntingtin-associated protein-1 with kinesin light chain: implications in intracellular trafficking in neurons. J Biol Chem 281:3552–3559

    Article  PubMed  CAS  Google Scholar 

  • Moffitt H, McPhail GD, Woodman B, Hobbs C, Bates GP (2009) Formation of polyglutamine inclusions in a wide range of non-CNS tissues in the HdhQ150 knock-in mouse model of Huntington’s disease. PLoS ONE 4:e8025

    Article  PubMed  Google Scholar 

  • Schilling G, Becher MW, Sharp AH et al (1999) Intranuclear inclusions and neuritic aggregates in transgenic mice expressing a mutant N-terminal fragment of huntingtin. Hum Mol Genet 8:397–407

    Article  PubMed  CAS  Google Scholar 

  • The Huntington’s Disease Collaborative Research Group (1993) A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington’s disease chromosomes. Cell 72:971–983

    Google Scholar 

  • von Horsten S, Schmitt I, Nguyen HP et al (2003) Transgenic rat model of Huntington’s disease. Hum Mol Genet 12:617–624

    Article  Google Scholar 

  • Walker FO (2007) Huntington’s disease. Lancet 369:218–228

    Article  PubMed  CAS  Google Scholar 

  • Wang CE, Tydlacka S, Orr AL et al (2008) Accumulation of N-terminal mutant huntingtin in mouse and monkey models implicated as a pathogenic mechanism in Huntington’s disease. Hum Mol Genet 17:2738–2751

    Article  PubMed  CAS  Google Scholar 

  • Williams A, Sarkar S, Cuddon P et al (2008) Novel targets for Huntington’s disease in an mTOR-independent autophagy pathway. Nat Chem Biol 4:295–305

    Article  PubMed  CAS  Google Scholar 

  • Yang SH, Chan AW (2011) Transgenic animal models of Huntington’s Disease. Curr Top Behav Neurosci 7:61–85

    Article  PubMed  Google Scholar 

  • Yang SH, Cheng PH, Banta H et al (2008) Towards a transgenic model of Huntington’s disease in a non-human primate. Nature 453:921–924

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Jonathan Courtenay for critical reading of the manuscript, Dr. Xiao-Jiang Li for providing mEM48 antibodies, Dr. Chauying Jen and Pi-Hsueh Shirley Li for providing equipment and Dr. Shaw-Jeng Tsai and Dr. H. Sunny Sun for support and suggestions. This work was supported by National Science Council grants (NSC 99-2320-B-006-026-MY3 and NSC 100-2627-B-006-023) and in part by grant of the Ministry of Education, Taiwan, Republic of China, under the ATU plan.

Conflict of interest

None.

Author information

Authors and Affiliations

  1. Department of Physiology, College of Medicine, National Cheng Kung University, Tainan, 70101, Taiwan

    Pei-Hsun Cheng, Yu-Fan Chang & Shang-Hsun Yang

  2. Institute of Basic Medical Sciences, National Cheng Kung University, Tainan, 70101, Taiwan

    Chia-Ling Li & Shang-Hsun Yang

  3. Department of Life Sciences, College of Bioscience and Biotechnology, National Cheng Kung University, Tainan, 70101, Taiwan

    Lu-Shiun Her

  4. Department of Human Genetics, Emory University School of Medicine, Atlanta, GA, 30329, USA

    Anthony W. S. Chan

  5. Department of Life Sciences, Agricultural Biotechnology Center, National Chung Hsing University, Taichung, 40227, Taiwan

    Chuan-Mu Chen

Authors
  1. Pei-Hsun Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Chia-Ling Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lu-Shiun Her
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yu-Fan Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Anthony W. S. Chan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Chuan-Mu Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Shang-Hsun Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shang-Hsun Yang.

Additional information

P.-H. Cheng and C.-L. Li contributed equally to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

429_2012_401_MOESM1_ESM.tif

Online resource 1 Copy numbers of transgene in Ubi-HTT84Q transgenic founders. Copy numbers of transgene in different transgenic founders were determined by using Southern blotting. Top panel shows the original image of Southern blotting, and numbers listed above represent the tag number of different transgenic founders. The bottom table shows the estimated copy numbers of each transgenic founder based on the Southern blotting result. (TIFF 2975 kb)

429_2012_401_MOESM2_ESM.tif

Online resource 2 Neuropathological phenotypes of HD in the brain of Ubi-G-HTT84Q transgenic mice at 2 and 6 months of age. Brain sections were sampled from Ubi-G-HTT84Q transgenic mice at 2 and 6 months of age, and stained with the mEM48 antibody via DAB immunohistochemistry staining. (a-c) Representative brain sections from Ubi-G-HTT84Q transgenic mice at 2 months of age. (d-f) Representative brain sections from Ubi-G-HTT84Q transgenic mice at 6 months of age. There are no aggregates in striatum (STR), cortex (CTX) and white matter (WM) regions at 2 months of age; however, weaker signals of neuropil aggregates are observed at 6 months of age as indicated in arrows. (TIFF 3268 kb)

429_2012_401_MOESM3_ESM.tif

Online resource 3 No neuropathological phenotypes of HD in the brains of non-transgenic littermates at 12 months of age. Brain sections were sampled from non-transgenic littermates at 12 months of age, and stained with the mEM48 antibody via DAB immunohistochemistry staining. (a) Lower magnification shows no aggregates in striatum (STR), cortex (CTX) and white matter (WM) regions. (b) A higher power image of STR displays no aggregates. (c) A higher power image shows no aggregates in CTX. (TIFF 1598 kb)

429_2012_401_MOESM4_ESM.tif

Online resource 4 Neuropathological phenotypes of HD in the brain of Ubi-G-HTT84Q transgenic mice Brain sections were sampled from one representative transgenic mouse at 12 months of age, and stained with the mEM48 antibody via DAB immunohistochemistry staining. Nucleus staining was performed using hematoxylin. (a) STR displays nuclear aggregates, intranuclear aggregates and neuropil aggregates. (b) Neuropil aggregates are dominant in CTX. Arrow heads indicate nuclear aggregates, arrows indicate intranuclear aggregates and stars indicate neuropil aggregates. (TIFF 2424 kb)

429_2012_401_MOESM5_ESM.tif

Online resource 5 Behavioral phenotypes of Ubi-G-HTT84Q transgenic mice before the onset of HD. (a) Footprinting (top panel) and rota rod (bottom panel) tests at 4 months of age. (b) Footprinting (top panel) and rota rod (bottom panel) tests at 6 months of age. There are no statistical difference (p > 0.05) between Ubi-G-HTT84Q transgenic mice (GHD) and non-transgenic mice (WT). Data represented mean ± SD. (TIFF 3064 kb)

429_2012_401_MOESM6_ESM.tif

Online resource 6 Genotyping of Ubi-G-HTT19Q and Ubi-G-HTT84Q transgenic mice. PCR with specific primers was used to confirm the transgenic status. 8-1 – 5 are Ubi-G-HTT19Q transgenic mice showing shorter PCR amplicons, whereas 2-1 – 5 are Ubi-G-HTT84Q transgenic mice showing longer amplicons due to longer CAG repeats. (TIFF 1158 kb)

429_2012_401_MOESM7_ESM.tif

Online resource 7 Neuropathological phenotypes of HD in the brain of Ubi-HTT84Q transgenic mice. Brain sections were sampled from one representative transgenic mouse at 6 months of age, and stained with the mEM48 antibody via DAB immunohistochemistry staining. Images show more nuclear aggregates (small spots) in striatum (STR) and cortex (CTX) region than in white matter region (WM). (TIFF 3597 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cheng, PH., Li, CL., Her, LS. et al. Significantly differential diffusion of neuropathological aggregates in the brain of transgenic mice carrying N-terminal mutant huntingtin fused with green fluorescent protein. Brain Struct Funct 218, 283–294 (2013). https://doi.org/10.1007/s00429-012-0401-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00429-012-0401-x

Keywords

Search

Navigation