FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / J Vet Intern Med / Cerebellar axonopathy in Shivers horses identified by spatia...
Journal of veterinary internal medicine2023; 37(4); 1568-1579; doi: 10.1111/jvim.16784

Cerebellar axonopathy in Shivers horses identified by spatial transcriptomic and proteomic analyses.

Abstract: Shivers in horses is characterized by abnormal hindlimb movement when walking backward and is proposed to be caused by a Purkinje cell (PC) axonopathy based on histopathology. Objective: Define region-specific differences in gene expression within the lateral cerebellar hemisphere and compare cerebellar protein expression between Shivers horses and controls. Methods: Case-control study of 5 Shivers and 4 control geldings ≥16.2 hands in height. Methods: Using spatial transcriptomics, gene expression was compared between Shivers and control horses in PC soma and lateral cerebellar hemisphere white matter, consisting primarily of axons. Tandem-mass-tag (TMT-11) proteomic analysis was performed on lateral cerebellar hemisphere homogenates. Results: Differences in gene expression between Shivers and control horses were evident in principal component analysis of axon-containing white matter but not PC soma. In white matter, there were 455/1846 differentially expressed genes (DEG; 350 ↓DEG, 105 ↑DEG) between Shivers and controls, with significant gene set enrichment of the Toll-Like Receptor 4 (TLR4) cascade, highlighting neuroinflammation. There were 50/936 differentially expressed proteins (DEP). The 27 ↓DEP highlighted loss of axonal proteins including intermediate filaments (5), myelin (3), cytoskeleton (2), neurite outgrowth (2), and Na/K ATPase (1). The 23 ↑DEP were involved in the extracellular matrix (7), cytoskeleton (7), redox balance (2), neurite outgrowth (1), signal transduction (1), and others. Conclusions: Our findings support axonal degeneration as a characteristic feature of Shivers. Combined with histopathology, these findings are consistent with the known distinctive response of PC to injury where axonal changes occur without a substantial impact on PC soma.
© 2023 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals LLC on behalf of American College of Veterinary Internal Medicine.
Get Full Text
Publication Date: 2023-06-08 PubMed ID: 37288990PubMed Central: PMC10365050DOI: 10.1111/jvim.16784Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research examines the characteristic symptoms of ‘Shivers’, an ailment in horses, and suggests that it might be caused by degeneration of Purkinje cell axons in the brain. Using advanced spatial transcriptomic and proteomic analyses, the researchers compared the gene and protein expressions in the cerebellums of shivering horses to those of healthy horses.

Objective and Methodology

  • The research aimed to discover region-specific differences in gene expression within the lateral cerebellar hemisphere and contrast cerebellar protein expression between Shivers-inflicted horses and healthy horses.
  • The study was conducted on 5 Shivers horses and 4 control horses, all geldings and all ≥16.2 hands in height.
  • Spatial transcriptomics allowed researchers to compare gene expression between sick and healthy horses in the Purkinje cell soma and the white matter of the lateral cerebellar hemisphere that primarily consists of axons.
  • Tandem-mass-tag (TMT-11) proteomic analysis was conducted on the homogenates from the lateral cerebellar hemisphere.

Findings

  • The results showed a discernable difference in gene expression between the shivering and healthy horses, especially in the principal component analysis of the axon-containing white matter.
  • Of the 1846 genes analyzed, 455 were differentially expressed genes (DEGs) including 350 downregulated and 105 upregulated genes. These findings highlighted a significant gene set enrichment of the TLR4 cascade, indicating neuroinflammation in affected horses.
  • Out of 936 proteins analyzed, 50 were differentially expressed proteins (DEPs), out of which 27 were downregulated and mostly associated with axonal proteins. This result suggests a loss of axonal proteins like intermediate filaments, myelin, cytoskeleton, neurite outgrowth, and Na/K ATPase in shivering horses.
  • Among the 23 upregulated proteins, many were involved in the extracellular matrix, signal transduction, and redox balance, indicating additional adaptive or compensatory changes.

Conclusions

  • The findings substantiate that axonal degeneration is a characteristic feature of the ‘Shivers’ ailment in horses – a condition in which horses display abnormal hindlimb movement when walking backward.
  • In combination with histopathological studies, the findings complement the known fact that injury to the Purkinje cell can lead to axonal changes without impacting the Purkinje cell soma substantially, which aligns with the observed condition in Shivers horses.

Cite This Article

APA
Valberg SJ, Williams ZJ, Henry ML, Finno CJ. (2023). Cerebellar axonopathy in Shivers horses identified by spatial transcriptomic and proteomic analyses. J Vet Intern Med, 37(4), 1568-1579. https://doi.org/10.1111/jvim.16784

Publication

Journal of veterinary internal medicine
ISSN: 1939-1676
NlmUniqueID: 8708660
Country: United States
Language: English
Volume: 37
Issue: 4
Pages: 1568-1579

Researcher Affiliations

Valberg, Stephanie J
  • Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, Michigan, USA.
Williams, Zoë J
  • C. Wayne McIlwraith Translational Medicine, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado, USA.
Henry, Marisa L
  • Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, Michigan, USA.
Finno, Carrie J
  • Department of Population Health and Reproduction, School of Veterinary Medicine, University of California-Davis, Davis, California, USA.

Grant Funding

  • Mary Anne McPhail Endowment Michigan State University
  • United States Equestrian Foundation

Conflict of Interest Statement

Dr. Valberg runs a commercial muscle biopsy service and receives royalties for genetic tests, but these have no overlap with the neurologic disease presented in this article. No other authors declare a conflict of interest.

References

This article includes 50 references
  1. Baird JDF, Firschman A, Valberg SJ. Shivers (Shivering) in the Horse. San Antonio, TX: American Association of Equine Practitioners; 2006:359‐364.
  2. Draper AC, Trumble TN, Firshman AM, Baird JD, Reed S, Mayhew IG, MacKay R, Valberg SJ. Posture and movement characteristics of forward and backward walking in horses with shivering and acquired bilateral stringhalt.. Equine Vet J 2015 Mar;47(2):175-81.
    pubmed: 24612176doi: 10.1111/evj.12259google scholar: lookup
  3. Draper AC, Bender JB, Firshman AM, Baird JD, Reed S, Mayhew IG, Valberg SJ. Epidemiology of shivering (shivers) in horses.. Equine Vet J 2015 Mar;47(2):182-7.
    pubmed: 24802303doi: 10.1111/evj.12296google scholar: lookup
  4. Seino KK, Secord T, Vig M, Kyllonen S, DeClue AJ. Three-Dimensional Kinematic Motion Analysis of Shivers in Horses: A Pilot Study.. J Equine Vet Sci 2019 Aug;79:13-22.
    pubmed: 31405492doi: 10.1016/j.jevs.2019.03.006google scholar: lookup
  5. Valberg SJ, Lewis SS, Shivers JL, Barnes NE, Konczak J, Draper AC, Armién AG. The Equine Movement Disorder "Shivers" Is Associated With Selective Cerebellar Purkinje Cell Axonal Degeneration.. Vet Pathol 2015 Nov;52(6):1087-98.
    pubmed: 25714471doi: 10.1177/0300985815571668google scholar: lookup
  6. Shima A, Miyake T, Tanaka K, Ogawa A, Omae E, Nagamori Y, Miyata Y, Ohata K, Maki T, Ono Y, Mima T, Takahashi R, Koganemaru S. Case report: A novel approach of closed-loop brain stimulation combined with robot gait training in post-stroke gait disturbance.. Front Hum Neurosci 2023;17:1082556.
    pmc: PMC9911819pubmed: 36778037doi: 10.3389/fnhum.2023.1082556google scholar: lookup
  7. Myers PS, McNeely ME, Pickett KA, Duncan RP, Earhart GM. Effects of exercise on gait and motor imagery in people with Parkinson disease and freezing of gait.. Parkinsonism Relat Disord 2018 Aug;53:89-95.
    pmc: PMC6120800pubmed: 29754837doi: 10.1016/j.parkreldis.2018.05.006google scholar: lookup
  8. Aman JE, Valberg SJ, Elangovan N, Nicholson A, Lewis SS, Konczak J. Abnormal locomotor muscle recruitment activity is present in horses with shivering and Purkinje cell distal axonopathy.. Equine Vet J 2018 Sep;50(5):636-643.
    pubmed: 29356055doi: 10.1111/evj.12813google scholar: lookup
  9. Caligiore D, Pezzulo G, Baldassarre G, Bostan AC, Strick PL, Doya K, Helmich RC, Dirkx M, Houk J, Jörntell H, Lago-Rodriguez A, Galea JM, Miall RC, Popa T, Kishore A, Verschure PF, Zucca R, Herreros I. Consensus Paper: Towards a Systems-Level View of Cerebellar Function: the Interplay Between Cerebellum, Basal Ganglia, and Cortex.. Cerebellum 2017 Feb;16(1):203-229.
    pmc: PMC5243918pubmed: 26873754doi: 10.1007/s12311-016-0763-3google scholar: lookup
  10. Mayhew IG. Disorders of posture and movement. In: Mayhew IG, ed. Large Animal Neurology. 2nd ed. Chichester, West Sussex: Willey‐Blackwell; 2009:133‐142.
  11. Zimmerman SM, Fropf R, Kulasekara BR, Griswold M, Appelbe O, Bahrami A, Boykin R, Buhr DL, Fuhrman K, Hoang ML, Huynh Q, Isgur L, Klock A, Kutchma A, Lasley AE, Liang Y, McKay-Fleisch J, Nelson JS, Nguyen K, Piazza E, Rininger A, Zollinger DR, Rhodes M, Beechem JM. Spatially resolved whole transcriptome profiling in human and mouse tissue using Digital Spatial Profiling.. Genome Res 2022 Oct;32(10):1892-1905.
    pmc: PMC9712633pubmed: 36100434doi: 10.1101/gr.276206.121google scholar: lookup
  12. Bergholtz H, Carter JM, Cesano A, Cheang MCU, Church SE, Divakar P, Fuhrman CA, Goel S, Gong J, Guerriero JL, Hoang ML, Hwang ES, Kuasne H, Lee J, Liang Y, Mittendorf EA, Perez J, Prat A, Pusztai L, Reeves JW, Riazalhosseini Y, Richer JK, Sahin Ö, Sato H, Schlam I, Sørlie T, Stover DG, Swain SM, Swarbrick A, Thompson EA, Tolaney SM, Warren SE, On Behalf Of The GeoMx Breast Cancer Consortium. Best Practices for Spatial Profiling for Breast Cancer Research with the GeoMx(®) Digital Spatial Profiler.. Cancers (Basel) 2021 Sep 4;13(17).
    pmc: PMC8431590pubmed: 34503266doi: 10.3390/cancers13174456google scholar: lookup
  13. Hänzelmann S, Castelo R, Guinney J. GSVA: gene set variation analysis for microarray and RNA-seq data.. BMC Bioinformatics 2013 Jan 16;14:7.
    pmc: PMC3618321pubmed: 23323831doi: 10.1186/1471-2105-14-7google scholar: lookup
  14. Nesvizhskii AI, Keller A, Kolker E, Aebersold R. A statistical model for identifying proteins by tandem mass spectrometry.. Anal Chem 2003 Sep 1;75(17):4646-58.
    pubmed: 14632076doi: 10.1021/ac0341261google scholar: lookup
  15. Shadforth IP, Dunkley TP, Lilley KS, Bessant C. i-Tracker: for quantitative proteomics using iTRAQ.. BMC Genomics 2005 Oct 20;6:145.
    pmc: PMC1276793pubmed: 16242023doi: 10.1186/1471-2164-6-145google scholar: lookup
  16. Oberg AL, Mahoney DW, Eckel-Passow JE, Malone CJ, Wolfinger RD, Hill EG, Cooper LT, Onuma OK, Spiro C, Therneau TM, Bergen HR 3rd. Statistical analysis of relative labeled mass spectrometry data from complex samples using ANOVA.. J Proteome Res 2008 Jan;7(1):225-33.
    pmc: PMC2528956pubmed: 18173221doi: 10.1021/pr700734fgoogle scholar: lookup
  17. Yuan A, Rao MV, Veeranna, Nixon RA. Neurofilaments and Neurofilament Proteins in Health and Disease.. Cold Spring Harb Perspect Biol 2017 Apr 3;9(4).
    pmc: PMC5378049pubmed: 28373358doi: 10.1101/cshperspect.a018309google scholar: lookup
  18. Chen X, Lu W, Wu D. Sirtuin 2 (SIRT2): Confusing Roles in the Pathophysiology of Neurological Disorders.. Front Neurosci 2021;15:614107.
    pmc: PMC8180884pubmed: 34108853doi: 10.3389/fnins.2021.614107google scholar: lookup
  19. Utsumi S, Sakamoto K, Yamashita T, Tomita H, Sugano E, Ishida K, Ishiyama E, Ozaki T. Presence of ES1 homolog in the mitochondrial intermembrane space of porcine retinal cells.. Biochem Biophys Res Commun 2020 Apr 9;524(3):542-548.
    pubmed: 32014251doi: 10.1016/j.bbrc.2020.01.127google scholar: lookup
  20. Zhu JW, Li YF, Wang ZT, Jia WQ, Xu RX. Toll-Like Receptor 4 Deficiency Impairs Motor Coordination.. Front Neurosci 2016;10:33.
    pmc: PMC4754460pubmed: 26909014doi: 10.3389/fnins.2016.00033google scholar: lookup
  21. Rossi F, Gianola S, Corvetti L. The strange case of Purkinje axon regeneration and plasticity.. Cerebellum 2006;5(2):174-82.
    pubmed: 16818392doi: 10.1080/14734220600786444google scholar: lookup
  22. Bravin M, Savio T, Strata P, Rossi F. Olivocerebellar axon regeneration and target reinnervation following dissociated Schwann cell grafts in surgically injured cerebella of adult rats.. Eur J Neurosci 1997 Dec;9(12):2634-49.
    pubmed: 9517469doi: 10.1111/j.1460-9568.1997.tb01693.xgoogle scholar: lookup
  23. Lieberman AR. The axon reaction: a review of the principal features of perikaryal responses to axon injury.. Int Rev Neurobiol 1971;14:49-124.
    pubmed: 4948651doi: 10.1016/s0074-7742(08)60183-xgoogle scholar: lookup
  24. Chan‐Palay V. The Cerebellar Dentate Nucleus. Berlin, Heidelberg: Springer; 1977.
  25. Abg Abd Wahab DY, Gau CH, Zakaria R, Muthu Karuppan MK, A-Rahbi BS, Abdullah Z, Alrafiah A, Abdullah JM, Muthuraju S. Review on Cross Talk between Neurotransmitters and Neuroinflammation in Striatum and Cerebellum in the Mediation of Motor Behaviour.. Biomed Res Int 2019;2019:1767203.
    pmc: PMC6877979pubmed: 31815123doi: 10.1155/2019/1767203google scholar: lookup
  26. Kim JD, Park KE, Ishida J, Kako K, Hamada J, Kani S, Takeuchi M, Namiki K, Fukui H, Fukuhara S, Hibi M, Kobayashi M, Kanaho Y, Kasuya Y, Mochizuki N, Fukamizu A. PRMT8 as a phospholipase regulates Purkinje cell dendritic arborization and motor coordination.. Sci Adv 2015 Dec;1(11):e1500615.
    pmc: PMC4672763pubmed: 26665171doi: 10.1126/sciadv.1500615google scholar: lookup
  27. Yuan Y, Kong F, Xu H, Zhu A, Yan N, Yan C. Cryo-EM structure of human glucose transporter GLUT4.. Nat Commun 2022 May 13;13(1):2671.
    pmc: PMC9106701pubmed: 35562357doi: 10.1038/s41467-022-30235-5google scholar: lookup
  28. Friede RL, Samorajski T. Axon caliber related to neurofilaments and microtubules in sciatic nerve fibers of rats and mice.. Anat Rec 1970 Aug;167(4):379-87.
    pubmed: 5454590doi: 10.1002/ar.1091670402google scholar: lookup
  29. Yuan A, Rao MV, Veeranna, Nixon RA. Neurofilaments at a glance.. J Cell Sci 2012 Jul 15;125(Pt 14):3257-63.
    pmc: PMC3516374pubmed: 22956720doi: 10.1242/jcs.104729google scholar: lookup
  30. Yum SW. Neurofilaments are required for the maintenance of myelinated axons: analysis of the first homozygous mutation in NEFL associated with Charcot‐Marie‐Tooth disease. Ann Neurol 2009;66:S17.
  31. Gravel M, Trapp B, Peterson J. CNP in myelination: overexpression alters oligodendrocyte morphogenesis. Cell Biology and Pathology of Myelin: Evolvineg Biological Concepts and Therapeutic Approaches Boston, MA: Springer; 1997:75‐82.
  32. Fard MK, van der Meer F, Sánchez P, Cantuti-Castelvetri L, Mandad S, Jäkel S, Fornasiero EF, Schmitt S, Ehrlich M, Starost L, Kuhlmann T, Sergiou C, Schultz V, Wrzos C, Brück W, Urlaub H, Dimou L, Stadelmann C, Simons M. BCAS1 expression defines a population of early myelinating oligodendrocytes in multiple sclerosis lesions.. Sci Transl Med 2017 Dec 6;9(419).
    pmc: PMC7116798pubmed: 29212715doi: 10.1126/scitranslmed.aam7816google scholar: lookup
  33. Peng L, Martin-Vasallo P, Sweadner KJ. Isoforms of Na,K-ATPase alpha and beta subunits in the rat cerebellum and in granule cell cultures.. J Neurosci 1997 May 15;17(10):3488-502.
    pmc: PMC6573685pubmed: 9133374doi: 10.1523/jneurosci.17-10-03488.1997google scholar: lookup
  34. Forrest MD, Wall MJ, Press DA, Feng J. The sodium-potassium pump controls the intrinsic firing of the cerebellar Purkinje neuron.. PLoS One 2012;7(12):e51169.
    pmc: PMC3527461pubmed: 23284664doi: 10.1371/journal.pone.0051169google scholar: lookup
  35. Roostaei T, Sadaghiani S, Park MT, Mashhadi R, Nazeri A, Noshad S, Salehi MJ, Naghibzadeh M, Moghadasi AN, Owji M, Doosti R, Taheri AP, Rad AS, Azimi A, Chakravarty MM, Voineskos AN, Nazeri A, Sahraian MA. Channelopathy-related SCN10A gene variants predict cerebellar dysfunction in multiple sclerosis.. Neurology 2016 Feb 2;86(5):410-7.
    pmc: PMC4773947pubmed: 26740675doi: 10.1212/wnl.0000000000002326google scholar: lookup
  36. Zhao WQ, Lu B. Expression of annexin A2 in GABAergic interneurons in the normal rat brain.. J Neurochem 2007 Mar;100(5):1211-23.
    pubmed: 17316400doi: 10.1111/j.1471-4159.2006.04311.xgoogle scholar: lookup
  37. Shin JH, Kim YN, Kim IY, Choi DH, Yi SS, Seong JK. Increased Cell Proliferations and Neurogenesis in the Hippocampal Dentate Gyrus of Ahnak Deficient Mice.. Neurochem Res 2015 Jul;40(7):1457-62.
    pubmed: 26007245doi: 10.1007/s11064-015-1615-0google scholar: lookup
  38. Yabe JT, Chan WK, Wang FS, Pimenta A, Ortiz DD, Shea TB. Regulation of the transition from vimentin to neurofilaments during neuronal differentiation.. Cell Motil Cytoskeleton 2003 Nov;56(3):193-205.
    pubmed: 14569598doi: 10.1002/cm.10137google scholar: lookup
  39. Plantman S. Proregenerative properties of ECM molecules.. Biomed Res Int 2013;2013:981695.
    pmc: PMC3782155pubmed: 24195084doi: 10.1155/2013/981695google scholar: lookup
  40. Asher RA, Morgenstern DA, Moon LD, Fawcett JW. Chondroitin sulphate proteoglycans: inhibitory components of the glial scar.. Prog Brain Res 2001;132:611-9.
    pubmed: 11545024doi: 10.1016/s0079-6123(01)32106-4google scholar: lookup
  41. Ramon y Cajal S, May RM. Degeneration and Regeneration in the Nervous System. Vol 1959. Oxford UK: Oxford University Press; 1928:750.
  42. Airaksinen MS, Eilers J, Garaschuk O, Thoenen H, Konnerth A, Meyer M. Ataxia and altered dendritic calcium signaling in mice carrying a targeted null mutation of the calbindin D28k gene.. Proc Natl Acad Sci U S A 1997 Feb 18;94(4):1488-93.
    pmc: PMC19818pubmed: 9037080doi: 10.1073/pnas.94.4.1488google scholar: lookup
  43. Schwaller B, Meyer M, Schiffmann S. 'New' functions for 'old' proteins: the role of the calcium-binding proteins calbindin D-28k, calretinin and parvalbumin, in cerebellar physiology. Studies with knockout mice.. Cerebellum 2002 Dec;1(4):241-58.
    pubmed: 12879963doi: 10.1080/147342202320883551google scholar: lookup
  44. Caillard O, Moreno H, Schwaller B, Llano I, Celio MR, Marty A. Role of the calcium-binding protein parvalbumin in short-term synaptic plasticity.. Proc Natl Acad Sci U S A 2000 Nov 21;97(24):13372-7.
    pmc: PMC27231pubmed: 11069288doi: 10.1073/pnas.230362997google scholar: lookup
  45. Celio MR. Calbindin D-28k and parvalbumin in the rat nervous system.. Neuroscience 1990;35(2):375-475.
    pubmed: 2199841doi: 10.1016/0306-4522(90)90091-hgoogle scholar: lookup
  46. Chard PS, Jordán J, Marcuccilli CJ, Miller RJ, Leiden JM, Roos RP, Ghadge GD. Regulation of excitatory transmission at hippocampal synapses by calbindin D28k.. Proc Natl Acad Sci U S A 1995 May 23;92(11):5144-8.
    pmc: PMC41865pubmed: 7761464doi: 10.1073/pnas.92.11.5144google scholar: lookup
  47. Barski JJ, Hartmann J, Rose CR, Hoebeek F, Mörl K, Noll-Hussong M, De Zeeuw CI, Konnerth A, Meyer M. Calbindin in cerebellar Purkinje cells is a critical determinant of the precision of motor coordination.. J Neurosci 2003 Apr 15;23(8):3469-77.
    pmc: PMC6742296pubmed: 12716955doi: 10.1523/jneurosci.23-08-03469.2003google scholar: lookup
  48. Abel JM, Witt DM, Rissman EF. Sex differences in the cerebellum and frontal cortex: roles of estrogen receptor alpha and sex chromosome genes.. Neuroendocrinology 2011;93(4):230-40.
    pmc: PMC3128132pubmed: 21325792doi: 10.1159/000324402google scholar: lookup
  49. Jimsheleishvili S, Dididze M. Neuroanatomy, Cerebellum.. 2023 Jan;.
    pubmed: 30844194
  50. White JJ, Arancillo M, Stay TL, George-Jones NA, Levy SL, Heck DH, Sillitoe RV. Cerebellar zonal patterning relies on Purkinje cell neurotransmission.. J Neurosci 2014 Jun 11;34(24):8231-45.
    pmc: PMC4051975pubmed: 24920627doi: 10.1523/jneurosci.0122-14.2014google scholar: lookup

Citations

This article has been cited 0 times.
Subscribe to our newsletter
Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.