The Cerebellum

, Volume 16, Issue 1, pp 40–54 | Cite as

Abnormalities in the Structure and Function of Cerebellar Neurons and Neuroglia in the Lc/+ Chimeric Mouse Model of Variable Developmental Purkinje Cell Loss

  • James Cairns
  • Doug Swanson
  • Joanna Yeung
  • Anna Sinova
  • Ronny Chan
  • Praneetha Potluri
  • Price Dickson
  • Guy Mittleman
  • Dan Goldowitz
Original Paper

Abstract

Autism spectrum disorders (ASDs) are a group of neurodevelopmental disorders characterized by impaired and disordered language, decreased social interactions, stereotyped and repetitive behaviors, and impaired fine and gross motor skills. It has been well established that cerebellar abnormalities are one of the most common structural changes seen in the brains of people diagnosed with autism. Common cerebellar pathology observed in autistic individuals includes variable loss of cerebellar Purkinje cells (PCs) and increased numbers of reactive neuroglia in the cerebellum and cortical brain regions. The Lc/+ mutant mouse loses 100 % of cerebellar PCs during the first few weeks of life and provided a valuable model to study the effects of developmental PC loss on underlying structural and functional changes in cerebellar neural circuits. Lurcher (Lc) chimeric mice were also generated to explore the link between variable cerebellar pathology and subsequent changes in the structure and function of cerebellar neurons and neuroglia. Chimeras with the most severe cerebellar pathology (as quantified by cerebellar PC counts) had the largest changes in cFos expression (an indirect reporter of neural activity) in cerebellar granule cells (GCs) and cerebellar nucleus (CN) neurons. In addition, Lc chimeras with the fewest PCs also had numerous reactive microglia and Bergmann glia located in the cerebellar cortex. Structural and functional abnormalities observed in the cerebella of Lc chimeras appeared to be along a continuum, with the degree of pathology related to the number of PCs in individual chimeras.

Keywords

Autism Mouse Cerebellum Pathology Neuroglia Chimeras 

Supplementary material

12311_2015_756_MOESM1_ESM.doc (106 kb)
ESM 1(DOC 105 kb)
12311_2015_756_MOESM2_ESM.ppt (8.2 mb)
Supplementary Figure S1Expression of the pro-inflammatory marker iNOS in Iba1 positive microglia in the cerebellar cortex of Lurcher chimeras with the fewest Purkinje cells. iNOS, Iba1 and DAPI immunofluorescence showing the expression of iNOS in cerebellar PCs in a Lurcher wildtype (+/+) mouse (top left panel) and a Group #4 chimera (bottom left panel) (with mild loss of cerebellar PCs) in comparison to iNOS expression in Iba1 positive microglia in the cerebellar cortex of a Lc/+ mutant mouse (top right panel) and an ataxic Group #2 chimera (bottom right panel) (with severe loss of cerebellar PCs). (PPT 8389 kb)
12311_2015_756_MOESM3_ESM.ppt (2.8 mb)
Supplementary Figure S2Co-localization of the pro-inflammatory marker iNOS with Iba1 positive microglia in the cerebellar cortex of Lurcher chimeras with the fewest Purkinje cells. Photomicrograph showing Iba1 positive microglia expressing the pro-inflammatory marker iNOS. Expression of iNOS is not microglia specific and iNOS also appears to be abundantly expressed throughout the molecular layer of Lc/+ mutant mice and chimeras with the most severe cerebellar pathology. Insets show higher magnification photomicrographs of the expression of iNOS and Iba1 in amoeboid-like microglia found in the molecular layer of the cerebellar cortex. (PPT 2819 kb)
12311_2015_756_MOESM4_ESM.ppt (144 kb)
Supplementary Figure S3Regression Analyses show inverse relationships between left cerebellum PC numbers and cFos expression in the IGL, CN neurons and the density of Iba1 positive microglia in the cerebellar cortex. The top left panel shows the relationship between left cerebellum PC numbers and the density of cFos positive cells in the IGL of 39 Lurcher chimeras. The top right panel shows the relationship between left cerebellum PC numbers and the density of cFos positive CN neurons in 13 Lurcher chimeras. The bottom panel shows the relationship between left cerebellum PC numbers and the density of Iba1 positive microglia in the cerebellar cortex of 19 Lurcher chimeras. r is the correlation coefficient for each of the regression analyses and is shown in each of the panels in the figure. (PPT 144 kb)

References

  1. 1.
    Courchesne E, Yeung-Courchesne BA, Press GA, Hesselink JR, Jernigan TL. Hypoplasia of cerebellar vermal lobules VI and VII in autism. N Engl J Med. 1988;318(21):1349–54.CrossRefPubMedGoogle Scholar
  2. 2.
    Schmahmann JD, Sherman JC. The cerebellar cognitive affective syndrome. Brain. 1998;121(4):561–79.CrossRefPubMedGoogle Scholar
  3. 3.
    Kaufmann WE, Cooper KL, Mostofsky SH, Capone GT, Kates WR, Newschaffer CJ, et al. Specificity of cerebellar vermian abnormalities in autism: a quantitative magnetic resonance imaging study. J Child Neurol. 2003;18(7):463–70.CrossRefPubMedGoogle Scholar
  4. 4.
    Martin LA, Goldowitz D, Mittleman G. The cerebellum and spatial ability: dissection of motor and cognitive components with a mouse model system. Eur J Neurosci. 2003;18:2002–10.CrossRefPubMedGoogle Scholar
  5. 5.
    Dickson PE, Rogers TD, Del Mar N, Martin LA, Heck D, Blaha CD, et al. Behavioral flexibility in a mouse model of developmental cerebellar Purkinje cell loss. Neurobiol Learn Mem. 2010;94:220–8.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Bauman ML, Kemper TL. Neuroanatomic observations of the brain in autism: a review and future directions. Int J Dev Neurosci. 2005;23(2–3):183–7.CrossRefPubMedGoogle Scholar
  7. 7.
    Bailey A, Luthert P, Dean A, Harding B, Janota I, Montgomery M, et al. A clinicopathological study of autism. Brain. 1998;121:889–905.CrossRefPubMedGoogle Scholar
  8. 8.
    Whitney ER, Kemper TL, Bauman ML, Rosene DL, Blatt GJ. Cerebellar Purkinje cells are reduced in a subpopulation of autistic brains: a stereological experiment using calbindin-D28k. Cerebellum. 2008;7(3):406–16.CrossRefPubMedGoogle Scholar
  9. 9.
    Fatemi SH et al. Consensus paper: pathological role of the cerebellum in autism. Cerebellum. 2012;11:777–807.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Vogel MW, Caston J, Yuzaki M, Mariani J. The lurcher mouse: fresh insights from an old mutant. Brain Res. 2007;1140:4–13.CrossRefPubMedGoogle Scholar
  11. 11.
    Armstrong CL, Duffin CA, McFarland R, Vogel MW. Mechanisms of compartmental Purkinje cell death and survival in the lurcher mutant mouse. Cerebellum. 2011;10(3):504–14.CrossRefPubMedGoogle Scholar
  12. 12.
    Zuo J, De Jager PL, Takahashi KA, Jiang W, Linden DJ, Heintz N. Neurodegeneration in lurcher mice caused by mutation in delta2 glutamate receptor gene. Nature. 1997;388(6644):769–73.CrossRefPubMedGoogle Scholar
  13. 13.
    Mintz B. Formation of genotypically mosaic mouse embryos. Am Zool. 1962;2:432.Google Scholar
  14. 14.
    Mintz B. Genetic mosaicism in adult mice of quadroparental lineage. Science. 1965;148:1232–3.CrossRefPubMedGoogle Scholar
  15. 15.
    McLaren A, LeDouarin N. Chimeras in developmental biology. New York: Academic; 1984.Google Scholar
  16. 16.
    Vogel MW, Sunter K, Herrup K. Numerical matching between granule and Purkinje cells in lurcher chimeric mice. J Neurosci. 1989;9(10):3454–62.PubMedGoogle Scholar
  17. 17.
    Lalonde R, Joyal CC, Guastavino JM, Botez MI. Hole poking and motor coordination in lurcher mutant mice. Physiol Behav. 1993;54:41–4.CrossRefPubMedGoogle Scholar
  18. 18.
    Ritvo ER, Freeman BJ, Scheibel AB, Duong T, Robinson H, Guthrie D, et al. Lower Purkinje cell counts in the cerebella of four autistic subjects: initial findings of the UCLA-NSAC autopsy research report. Am J Psychiatr. 1986;143:862–6.CrossRefPubMedGoogle Scholar
  19. 19.
    Amaral DG, Schumann CM, Nordahl CW. Neuroanatomy of autism. Trends Neurosci. 2008;31:137–45.CrossRefPubMedGoogle Scholar
  20. 20.
    Vargas DL, Nascimbene C, Krishnan C, Zimmerman AW, Pardo CA. Neuroglial activation and neuroinflammation in the brain of patients with autism. Ann Neurol. 2005;57(1):67–81.CrossRefPubMedGoogle Scholar
  21. 21.
    Morgan JT, Chana G, Pardo CA, Achim C, Semendeferi K, Buckwalter J, et al. Microglial activation and increased microglial density observed in the dorsolateral prefrontal cortex in autism. Biol Psychiatry. 2010;68(4):368–76.CrossRefPubMedGoogle Scholar
  22. 22.
    Dragunow M, Robertson HA, Robertson GS. Amygdala kindling and c-fos protein(s). Exp Neurol. 1988;102:261–3.CrossRefPubMedGoogle Scholar
  23. 23.
    Sagar SM, Sharp FR, Curran T. Expression of c-fos protein in brain: metabolic mapping at the cellular level. Science. 1988;240:1328–31.CrossRefPubMedGoogle Scholar
  24. 24.
    Dragunow M, Faull R. The use of c-Fos as a metabolic marker in neuronal pathway tracing. J Neurosci Methods. 1989;29(3):261–5.CrossRefPubMedGoogle Scholar
  25. 25.
    Smeyne RJ, Schilling K, Robertson L, Luk D, Oberdick J, Curran T, et al. fos-lacZ transgenic mice: mapping sites of gene induction in the central nervous system. Neuron. 1992;8(1):13–23.CrossRefPubMedGoogle Scholar
  26. 26.
    Tian JB, Bishop GA. Stimulus-dependent activation of c-Fos in neurons and glia in the rat cerebellum. J Chem Neuroanat. 2002;23:157–70.CrossRefPubMedGoogle Scholar
  27. 27.
    Abercrombie M. Estimation of nuclear population from microtome sections. Anat Rec. 1946;94:239–47.CrossRefPubMedGoogle Scholar
  28. 28.
    Liu L, Hamre K, Goldowitz D. Kainic acid-induced neuronal degeneration in Hippocampal pyramidal neurons is driven by both intrinsic and extrinsic factors: analysis of FVB/N↔C57BL/6 chimeras. J Neurosci. 2012;32(35):12093–101.CrossRefPubMedGoogle Scholar
  29. 29.
    Herrup K, Wetts R. Cerebellar Purkinje cells are descended from a small number of progenitors committed during early development: quantitative analysis of lurcher chimeric mice. J Neurosci. 1982;2(10):1494–8.PubMedGoogle Scholar
  30. 30.
    Wetts R, Herrup K. Direct correlation between Purkinje and granule cell number in the cerebella of lurcher chimeras and wild-type mice. Dev Brain Res. 1983;10(1):41–7.CrossRefGoogle Scholar
  31. 31.
    Wallace VA. Purkinje-cell-derived Sonic hedgehog regulates granule neuron precursor cell proliferation in the developing mouse cerebellum. Curr Biol. 1999;9:445–8.CrossRefPubMedGoogle Scholar
  32. 32.
    Wechsler-Reya RJ, Scott MP. Control of neuronal precursor proliferation in the cerebellum by Sonic Hedgehog. Neuron. 1999;22:103–14.CrossRefPubMedGoogle Scholar
  33. 33.
    Davis GW, Bezprozvanny I. Maintaining the stability of neural function: a homeostatic hypothesis. Annu Rev Physiol. 2001;63:847–69.CrossRefPubMedGoogle Scholar
  34. 34.
    Bushong EA, Martone ME, Jones YZ, Ellisman MH. Protoplasmic astrocytes in CA1 stratum radiatum occupy separate anatomical domains. J Neurosci. 2002;22:183–92.PubMedGoogle Scholar
  35. 35.
    Bushong EA, Martone ME, Ellisman MH. Maturation of astrocyte morphology and the establishment of astrocyte domains during postnatal hippocampal development. Int J Dev Neurosci. 2004;22:73–86.CrossRefPubMedGoogle Scholar
  36. 36.
    American Psychiatric Association Diagnostic and Statistic Manual of Mental Disorders. 4th edition (DSM-IV). Washington, D.C.: APA; 1994.Google Scholar
  37. 37.
    Ozonoff S et al. Gross motor development, movement abnormalities, and early identification of autism. J Autism Dev Disord. 2008;38(4):644–56.CrossRefPubMedGoogle Scholar
  38. 38.
    Miyake A, Friedman MP, Emerson MJ, Witzki AH, Howerter A, Wager TD. The unity and diversity of executive functions and their contributions to complex “frontal lobe” tasks: a latent variable analysis. Cogn Psychol. 2000;41:49–100.CrossRefPubMedGoogle Scholar
  39. 39.
    Arnsten AFT, Bao-Ming L. Neurobiology of executive functions: catecholamine influences on prefrontal cortical functions. Biol Psychiatry. 2005;57(11):1377–84.CrossRefPubMedGoogle Scholar
  40. 40.
    Ragozzino ME. The contribution of the medial prefrontal cortex, orbitofrontal cortex, and dorsomedial striatum to behavioral flexibility. Ann N Y Acad Sci. 2007;1121:355–75.CrossRefPubMedGoogle Scholar
  41. 41.
    Han S et al. Autistic-like behaviour in Scn1a+/− mice and rescue by enhanced GABA-mediated neurotransmission. Nature. 2012;489:385–90.CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Han S et al. Enhancement of inhibitory neurotransmission by GABAA receptors having α2,3-subunits ameliorates behavioral deficits in a mouse model of autism. Neuron. 2014;81:1282–9.CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Cobos I, Calcagnotto ME, Vilaythong AJ, Thwin MT, Noebels JL, Baraban SC, et al. Mice lacking Dlx1 show subtype-specific loss of interneurons, reduced inhibition and epilepsy. Nat Neurosci. 2005;8(8):1059–68.CrossRefPubMedGoogle Scholar
  44. 44.
    Howard MA, Rubenstein JLR, Baraban SC. Bidirectional homeostatic plasticity induced by interneuron cell death and transplantation in vivo. PNAS. 2014;111(1):492–7.CrossRefPubMedGoogle Scholar
  45. 45.
    Ibata K, Sun Q, Turrigiano GG. Rapid synaptic scaling induced by changes in postsynaptic firing. Neuron. 2008;57(6):819–26.CrossRefPubMedGoogle Scholar
  46. 46.
    Turrigiano G. Too many cooks? Intrinsic and synaptic homeostatic mechanisms in cortical circuit refinement. Annu Rev Neurosci. 2011;34:89–103.CrossRefPubMedGoogle Scholar
  47. 47.
    Caddy KWT, Martin MR, Biscoe TJ. The identification of mossy fibres and their cells of origin in the normal and lurcher mutant mouse. J Neurol Sci. 1977;34:121–9.CrossRefPubMedGoogle Scholar
  48. 48.
    Häusser M, Clark BA. Tonic synaptic inhibition modulates neuronal output pattern and spatiotemporal synaptic integration. Neuron. 1997;19(3):665–78.CrossRefPubMedGoogle Scholar
  49. 49.
    Harvey RJ, Napper RM. Quantitative studies on the mammalian cerebellum. Prog Neurobiol. 1991;36:437–63.CrossRefPubMedGoogle Scholar
  50. 50.
    Rowland NC, Jaeger D. Coding of tactile response properties in the rat deep cerebellar nuclei. J Neurophysiol. 2005;94:1236–51.CrossRefPubMedGoogle Scholar
  51. 51.
    Catz N, Dicke PW, Thier P. Cerebellar-dependent motor learning is based on pruning a Purkinje cell population response. PNAS USA. 2008;105:7309–14.CrossRefPubMedPubMedCentralGoogle Scholar
  52. 52.
    Garin N, Hornung JP, Escher G. Distribution of postsynaptic GABAA receptor aggregates in the deep cerebellar nuclei of normal and mutant mice. J Comp Neurol. 2002;447(3):210–7.CrossRefPubMedGoogle Scholar
  53. 53.
    Sultan F, König T, Möck M, Thier P. Quantitative organization of neurotransmitters in the deep cerebellar nuclei of the Lurcher mutant. J Comp Neurol. 2002;452(4):311–23.CrossRefPubMedGoogle Scholar
  54. 54.
    Linnemann C, Sultan F, Pedroarena CM, Schwarz C, Thier P. Lurcher mice exhibit potentiation of GABA(A)-receptor mediated conductance in cerebellar nuclei neurons in close temporal relationship to Purkinje cell death. J Neurophysiol. 2004;91(2):1102–7.CrossRefPubMedGoogle Scholar
  55. 55.
    Bechade C, Colasse S, Diana MA, Rouault M, Bessis A. NOS2 expression is restricted to neurons in the healthy brain but is triggered in microglia upon inflammation. Glia. 2014;62(6):956–63.CrossRefPubMedGoogle Scholar
  56. 56.
    Correa-Cerro LS, Mandell JW. Molecular mechanisms of astrogliosis: new approaches with mouse genetics. J Neuropathol Exp Neurol. 2007;66:169–76.CrossRefPubMedGoogle Scholar
  57. 57.
    Kyuhou S, Kato N, Gemba H. Emergence of endoplasmic reticulum stress and activated microglia in Purkinje cell degeneration mice. Neurosci Lett. 2006;396:91–6.CrossRefPubMedGoogle Scholar
  58. 58.
    Milligan ED, Watkins LR. Pathological and protective roles of glia in chronic pain. Nat Rev Neurosci. 2009;10:23–36.CrossRefPubMedPubMedCentralGoogle Scholar
  59. 59.
    Mullen RJ, Eicher EM, Sidman RL. Purkinje cell degeneration, a new neurological mutation in the mouse. PNAS USA. 1976;73:208–12.CrossRefPubMedPubMedCentralGoogle Scholar
  60. 60.
    Wang T, Morgan JI. The Purkinje cell degeneration (pcd) mouse: an unexpected molecular link between neuronal degeneration and regeneration. Brain Res. 2007;1140:26–40.CrossRefPubMedGoogle Scholar
  61. 61.
    Baltanas FC, Berciano MT, Valero J, Gomez C, Diaz D, Alonso JR, et al. Differential glial activation during the degeneration of Purkinje cells and mitral cells in the PCD mutant mice. Glia. 2013;61:254–72.CrossRefPubMedGoogle Scholar
  62. 62.
    Spacek J. Three-dimensional analysis of dendritic spines. III. Glial sheath. Anat Embryol. 1985;171:245–52.CrossRefPubMedGoogle Scholar
  63. 63.
    Reichenbach A, Siegel A, Rickmann M, Wolff JR, Noone D, Robinson SR. Distribution of Bergmann glial somata and processes: implications for function. J Hirnforsch. 1995;36:509–17.PubMedGoogle Scholar
  64. 64.
    Yamada K, Watanabe M. Cytodifferentiation of Bergmann glia and its relationship with Purkinje cells. Anat Sci Int. 2002;77:94–108.CrossRefPubMedGoogle Scholar
  65. 65.
    Caddy KW, Biscoe TJ. Structural and quantitative studies on the normal C3H and Lurcher mutant mouse. Philos Trans Royal Soci Lond B Biol Sci. 1979;287(1020):167–201.CrossRefGoogle Scholar
  66. 66.
    Vernet-der Garabedian B, Derer P, Bailly Y, Mariani J. Innate immunity in the Grid2Lc/+ mouse model of cerebellar neurodegeneration: glial CD95/CD95L plays a non-apoptotic role in persistent neuron loss-associated inflammatory reactions in the cerebellum. J Neuroinflammation. 2013;10(65):1–11.Google Scholar
  67. 67.
    Jankowski J, Miething A, Schilling K, Baader SL. Physiological Purkinje cell death is spatiotemporally organized in the developing mouse cerebellum. Cerebellum. 2009;8(3):277–90.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • James Cairns
    • 1
    • 2
    • 3
    • 4
  • Doug Swanson
    • 1
    • 2
    • 3
  • Joanna Yeung
    • 1
    • 2
    • 3
  • Anna Sinova
    • 1
    • 2
    • 3
    • 4
  • Ronny Chan
    • 1
    • 2
    • 3
  • Praneetha Potluri
    • 1
    • 2
    • 3
  • Price Dickson
    • 6
  • Guy Mittleman
    • 5
  • Dan Goldowitz
    • 1
    • 2
    • 3
    • 4
  1. 1.Department of Medical GeneticsUniversity of British ColumbiaVancouverCanada
  2. 2.Centre for Molecular Medicine and TherapeuticsUniversity of British ColumbiaVancouverCanada
  3. 3.Child and Family Research InstituteUniversity of British ColumbiaVancouverCanada
  4. 4.Graduate Program in Neuroscience, Djavad Mowafaghian Centre for Brain HealthUniversity of British ColumbiaVancouverCanada
  5. 5.Department of Psychological ScienceBall State UniversityMuncieUSA
  6. 6.The Jackson LaboratoryBar HarborUSA

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