Evaluation of Disease Lesions in the Developing Canine MPS IIIA Brain

  • Leanne K. Winner
  • Neil R. Marshall
  • Robert D. Jolly
  • Paul J. Trim
  • Stephen K. Duplock
  • Marten F. Snel
  • Kim M. HemsleyEmail author
Research Report
Part of the JIMD Reports book series (JIMD, volume 43)


Mucopolysaccharidosis IIIA (MPS IIIA) is an inherited neurodegenerative disease of childhood that results in early death. Post-mortem studies have been carried out on human MPS IIIA brain, but little is known about early disease development. Here, we utilised the Huntaway dog model of MPS IIIA to evaluate disease lesion development from 2 to 24 weeks of age. A significant elevation in primarily stored heparan sulphate was observed in all brain regions assessed in MPS IIIA pups ≤9.5 weeks of age. There was a significant elevation in secondarily stored ganglioside (GM3 36:1) in ≤9.5-week-old MPS IIIA pup cerebellum, and other brain regions also exhibited accumulation of this lipid with time. The number of neural stem cells and neuronal precursor cells was essentially unchanged in MPS IIIA dog brain (c.f. unaffected) over the time course assessed, a finding corroborated by neuron cell counts. We observed early neuroinflammatory changes in young MPS IIIA pup brain, with significantly increased numbers of activated microglia recorded in all but one brain region in MPS IIIA pups ≤9.5 weeks of age (c.f. age-matched unaffected pups). In conclusion, infant-paediatric-stage MPS IIIA canine brain exhibits substantial and progressive primary and secondary substrate accumulation, coupled with early and robust microgliosis. Whilst early initiation of treatment is likely to be required to maintain optimal neurological function, the brain’s neurodevelopmental potential appears largely unaffected by the disease process; further investigations confirming this are warranted.


Brain development Dog Ganglioside Heparan sulphate Lysosome Microglia Mucopolysaccharidosis Neuroinflammation 



Cornus ammonis region 3


Coefficient of variation


Fibroblast growth factor 2

Ganglioside species

GD1, GM1, GM2, GM3, GT1








Lysosomal integral membrane protein-2


Mucopolysaccharidosis type IIIA


Neuronal nuclear antigen















Supplementary material

477606_1_En_110_MOESM1_ESM.docx (493 kb)
Supplementary Fig. 1 Quantification of GM2 38:1 and GM3 38:1 in 2–24-week-old unaffected and MPS IIIA dog brain (af) and spinal cord (g, h). Each data point reflects a single dog. Six-week-old unaffected dog hippocampus was unavailable for study. The spinal cord was prepared in a separate batch from the brain (hippocampus, cortex and cerebellum). Two QC samples (MPS IIIA dog hippocampus) were run with each batch; both batches were analysed on the same day (DOCX 492 kb)
477606_1_En_110_MOESM2_ESM.docx (84 kb)
Supplementary Fig. 2 Quantification of the ganglioside GM3 precursors: (a) glucosylceramide (GC), and (b) lactosylceramide (LC); in addition to the most abundant species of phospholipids, (c) phosphatidylethanolamine (PE), and (d) phosphatidylinositol (PI), in the 2–24-week-old MPS IIIA and unaffected dog cerebellum. Each data point reflects a single dog (DOCX 83 kb)
477606_1_En_110_MOESM3_ESM.docx (559 kb)
Supplementary Fig. 3 Quantification of lipids in cerebellar samples taken from 2- to 24-week-old unaffected and MPS IIIA dogs. Each data point reflects a single dog. (ad) Glucosylceramide (GC), (eg) lactosylceramide (LC), (h) glucosylsphingosine (GlcSph) and (ij) sphingomyelin (SM) (DOCX 559 kb)
477606_1_En_110_MOESM4_ESM.docx (619 kb)
Supplementary Fig. 4 Quantification of phospholipids in cerebellar homogenates taken from unaffected and MPS IIIA dogs aged 2–24 weeks. Each data point reflects a single dog. (ac) Phosphatidylethanolamine (PE), (dg) phosphatidylinositol (PI) and (hk) phosphatidylserine (PS). The inter-assay CV for PE species was <14.1%, for PI species <12.1% and for PS species <13% (DOCX 618 kb)
477606_1_En_110_MOESM5_ESM.docx (316 kb)
Supplementary Fig. 5 Quantification of phospholipids in 2–24-week-old unaffected and MPS IIIA dog cerebellum. Each data point reflects a single dog. (ad) Phosphatidylcholine (PC) and (ei) phosphatidylglycerol (PG) (DOCX 316 kb)
477606_1_En_110_MOESM6_ESM.docx (4.5 mb)
Supplementary Fig. 6 Quantification of LIMP-2 immunoreactivity in various brain regions with time (ae). Each data point reflects a single dog. Filled square 2-week-old pups, filled circle 4-week-old pups, filled triangle 6-week-old pups, filled diamond 9.5-week-old pups, filled inverted triangle 24-week-old pups. The photos in (f) show the location of the brain regions assessed: (a) gyrus suprasylvius anterior, (b) gyrus suprasylvius medialis, (c) polymorphic cell layer (hilus), (d) lobe 6 of the cerebellum and (e) caudate nucleus head. Photos in (g) and (h) show LIMP-2 staining in the gyrus suprasylvius anterior of 24-week-old unaffected and MPS IIIA dog brain (respectively). *p < 0.05, **p < 0.01 (DOCX 4573 kb)
477606_1_En_110_MOESM7_ESM.docx (1.5 mb)
Supplementary Fig. 7 Quantification of doublecortin immunoreactivity in various brain regions (a, b, d, e, g, h and jl). Each data point reflects a single dog. The photos show doublecortin staining in the unaffected (c) and MPS IIIA (f) gyrus suprasylvius anterior at 2 weeks of age. The photos in (i) show the location of the brain regions assessed: (a) gyrus suprasylvius anterior, (b) gyrus suprasylvius medialis, (d) CA3, (e) dentate gyrus, (g) subventricular zone, (h) fasciculus subcallosus, (jk) lobe 6 of the cerebellum and (l) caudate nucleus head (DOCX 1516 kb)
477606_1_En_110_MOESM8_ESM.docx (14 kb)
Supplementary Table 1 Ganglioside species assessed are listed in order of abundance, together with the inter-assay coefficient of variation for that species (DOCX 14 kb)
477606_1_En_110_MOESM9_ESM.docx (23 kb)
Supplementary Methods (DOCX 22 kb)


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Copyright information

© Society for the Study of Inborn Errors of Metabolism (SSIEM) 2018

Authors and Affiliations

  • Leanne K. Winner
    • 1
  • Neil R. Marshall
    • 2
  • Robert D. Jolly
    • 2
  • Paul J. Trim
    • 1
  • Stephen K. Duplock
    • 1
  • Marten F. Snel
    • 1
  • Kim M. Hemsley
    • 1
    Email author
  1. 1.Lysosomal Diseases Research UnitSouth Australian Health and Medical Research InstituteAdelaideAustralia
  2. 2.Institute of Veterinary, Animal and Biomedical Science, Massey UniversityPalmerston NorthNew Zealand

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