Oculomotor deficits, such as insufficiencies in accommodation, convergence, and saccades, are common following traumatic brain injury (TBI). Previous studies in patients with mild TBI attributed these deficits to insufficient activation of subcortical oculomotor nuclei, although the exact mechanism is unknown. A possible cause for neuronal dysfunction in these regions is biomechanically induced plasma membrane permeability. We used our established porcine model of head rotational TBI to investigate whether cell permeability changes occurred in subcortical oculomotor areas following single or repetitive TBI, with repetitive injuries separated by 15 min, 3 days, or 7 days. Swine were subjected to sham conditions or head rotational acceleration in the sagittal plane using a HYGE pneumatic actuator. Two hours prior to the final injury, the cell-impermeant dye Lucifer Yellow was injected into the ventricles to diffuse throughout the interstitial space to assess plasmalemmal permeability. Animals were sacrificed 15 min after the final injury for immunohistological analysis. Brain regions examined for cell membrane permeability included caudate, substantia nigra pars reticulata, superior colliculus, and cranial nerve oculomotor nuclei. We found that the distribution of permeabilized neurons varied depending on the number and spacing of injuries. Repetitive injuries separated by 15 min or 3 days resulted in the most permeability. Many permeabilized cells lost neuron-specific nuclear protein reactivity, although no neuronal loss occurred acutely after injury. Microglia contacted and appeared to begin phagocytosing permeabilized neurons in repetitively injured animals. These pathologies within oculomotor areas may mediate transient dysfunction and/or degeneration that may contribute to oculomotor deficits following diffuse TBI.
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References
1.
CiuffredaK.J., KapoorN., RutnerD., SuchoffI.B., HanM.E., and CraigS. (2007). Occurrence of oculomotor dysfunctions in acquired brain injury: a retrospective analysis. Optometry, 78, 155–161.
2.
GoodrichG.L., FlygH.M., KirbyJ.E., ChangC.-Y., and MartinsenG.L. (2013). Mechanisms of TBI and visual consequences in military and veteran populations. Optom. Vis. 90, 105–112.
3.
TylerC.W., LikovaL.T., MineffK.N., and NicholasS.C. (2015). Deficits in the activation of human oculomotor nuclei in chronic traumatic brain injury. Front. Neurol. 6, 1–9.
4.
CullenD.K., VernekarV.N., and LaPlacaM.C. (2011). Trauma-induced plasmalemma disruptions in three-dimensional neural cultures are dependent on strain modality and rate. J. Neurotrauma, 28, 2219–2233.
5.
GeddesD.M., LaPlacaM.C., and CargillR.S. (2003). Susceptibility of hippocampal neurons to mechanically induced injury. Exp. Neurol. 184, 420–427.
6.
GeddesD.M., CargillR.S. 2nd, and LaPlacaM.C. (2003). Mechanical stretch to neurons results in a strain rate and magnitude-dependent increase in plasma membrane permeability. J. Neurotrauma, 20, 1039–1049.
7.
LaPlacaM.C., PradoG.R., CullenD.K., and IronsH.R. (2006). High rate shear insult delivered to cortical neurons produces heterogeneous membrane permeability alterations. Conf. Proc. IEEE Eng. Med. Biol. Soc. 2006, 2384–2387
8.
LaPlacaM.C., PradoG.R., CullenD., and SimonC.M. (2009). Plasma membrane damage as a marker of neuronal injury. Conf. Proc. IEEE Eng. Med. Biol. Soc. 2009, 1113–1116.
9.
LaPlacaM.C., LessingM.C., PradoG.R., ZhouR., TateC.C., Geddes-KleinD., MeaneyD.F., and ZhangL. (2019). Mechanoporation is a potential indicator of tissue strain and subsequent degeneration following experimental traumatic brain injury. Clin. Biomech. 64, 2–13.
10.
LaPlacaM.C., LeeV.M., and ThibaultL.E. (1997). An in vitro model of traumatic neuronal injury: loading rate-dependent changes in acute cytosolic calcium and lactate dehydrogenase release. J. Neurotrauma, 14, 355–368.
11.
LaPlacaM.C. and ThibaultL.E. (1998). Dynamic mechanical deformation of neurons triggers an acute calcium response and cell injury involving the N-methyl-D-aspartate glutamate receptor. J. Neurosci. Res. 52, 220–229.
12.
WeberJ.T., RzigalinskiB.A., WilloughbyK.A., MooreS.F., and EllisE.F. (1999). Alterations in calcium-mediated signal transduction after traumatic injury of cortical neurons. Cell Calcium, 26, 289–299.
13.
McIntoshT.K., SaatmanK.E., RaghupathiR., GrahamD.I., SmithD.H., LeeV.M., and TrojanowskiJ.Q. (1998). The Dorothy Russell Memorial Lecture. The molecular and cellular sequelae of experimental traumatic brain injury: pathogenetic mechanisms. Neuropathol. Appl. Neurobiol. 24, 251–267.
14.
FarkasO., LifshitzJ., and PovlishockJ.T. (2006). Mechanoporation Induced by Diffuse Traumatic Brain Injury: An Irreversible or Reversible Response to Injury?. J. Neurosci. 26, 3130–3140.
15.
PatelT.P., VentreS.C., and MeaneyD.F. (2012). Dynamic changes in neural circuit topology following mild mechanical injury in vitro. Ann. Biomed. Eng. 40, 23–36.
16.
WhalenM.J., DalkaraT., YouZ., QiuJ., BermpohlD., MehtaN., SuterB., BhideP.G., LoE.H., EricssonM., and MoskowitzM.A. (2008). Acute plasmalemma permeability and protracted clearance of injured cells after controlled cortical impact in mice. J. Cereb. Blood Flow Metab. 28, 490–505.
17.
BailesJ.E., DashnawM.L., PetragliaA.L., and TurnerR.C. (2014). Cumulative effects of repetitive mild traumatic brain injury. Prog. Neurol. Surg. 28, 50–62.
18.
GardnerR.C. and YaffeK. (2015). Epidemiology of mild traumatic brain injury and neurodegenerative disease. Mol. Cell. Neurosci. 66, 75–80.
19.
GardnerR.C., BurkeJ.F., NettiksimmonsJ., GoldmanS., TannerC.M., and YaffeK. (2015). Traumatic brain injury in later life increases risk for Parkinson disease. Ann. Neurol. 77, 987–995.
20.
McKeeA.C., CantuR.C., NowinskiC.J., Hedley-WhyteE.T., GavettB.E., BudsonA.E., SantiniV.E., LeeH.S., KubilusC.A., and SternR.A. (2009). Chronic traumatic encephalopathy in athletes: progressive tauopathy after repetitive head injury. J. Neuropathol. Exp. Neurol. 68, 709–735.
21.
McKeeA.C. and RobinsonM.E. (2014). Military-related traumatic brain injury and neurodegeneration. Alzheimers. Dement. 10, S242–S53.
22.
LoBueC., WilmothK., CullumC.M., RossettiH.C., LacritzL.H., HynanL.S., HartJ.J., and WomackK.B. (2016). Traumatic brain injury history is associated with earlier age of onset of frontotemporal dementia. J. Neurol. Neurosurg. Psychiatry, 87, 817–820.
23.
LoBueC., WadsworthH., WilmothK., ClemM., HartJ.J., WomackK.B., DidehbaniN., LacritzL.H., RossettiH.C., and CullumC.M. (2017). Traumatic brain injury history is associated with earlier age of onset of Alzheimer disease. Clin. Neuropsychol. 31, 85–98.
24.
HikosakaO., TakikawaY., and KawagoeR. (2000). Role of the basal ganglia in the control of purposive saccadic eye movements. Physiol. Rev. 80, 953–978.
25.
SegerC.A. (2013). The visual corticostriatal loop through the tail of the caudate: circuitry and function. Front. Syst. Neurosci. 7, 1–15.
26.
RossD.T., MeaneyD.F., SabolM.K., SmithD.H., and GennarelliT.A. (1994). Distribution of forebrain diffuse axonal injury following inertial closed head injury in miniature swine. Exp. Neurol. 126, 291–298.
27.
CullenD.K., HarrisJ.P., BrowneK.D., WolfJ.A., DudaJ.E., MeaneyD.F., MarguliesS.S., and SmithD.H. (2016). A porcine model of traumatic brain injury via head rotational acceleration. Methods Mol. Biol. 1462, 289–324
28.
BaileyE.L., McCullochJ., SudlowC., and WardlawJ.M. (2009). Potential animal models of lacunar stroke: a systematic review. Stroke, 40, e451–e458.
29.
HowellsD.W., PorrittM.J., RewellS.S.J., O'CollinsV., SenaE.S., van der WorpH.B., TraystmanR.J., and MacleodM.R. (2010). Different strokes for different folks: the rich diversity of animal models of focal cerebral ischemia. J. Cereb. Blood Flow Metab. 30, 1412–1431.
30.
ZhangK. and SejnowskiT.J. (2000). A universal scaling law between gray matter and white matter of cerebral cortex. Proc. Natl. Acad. Sci. U. S. A. 97, 5621–5626.
31.
SmithD.H., ChenX.H., XuB.N., McIntoshT.K., GennarelliT.A., and MeaneyD.F. (1997). Characterization of diffuse axonal pathology and selective hippocampal damage following inertial brain trauma in the pig. J. Neuropathol. Exp. Neurol. 56, 822–834.
32.
SmithD., ChenX., NonakaM., TrojanowskiJ., LeeV., SaatmanK., LeoniM., XuB., WolfJ., and MeaneyD. (1999). Accumulation of amyloid beta and tau and the formation of neurofilament inclusions following diffuse brain injury in the pig. J. Neuropathol. Exp. Neurol. 58, 982–992.
33.
BrowneK.D., ChenX.H., MeaneyD.F., and SmithD.H. (2011). Mild traumatic brain injury and diffuse axonal injury in Swine. J. Neurotrauma, 28, 1747–1755.
34.
WoffordK.L., HarrisJ.P., BrowneK.D., BrownD.P., GrovolaM.R., MietusC.J., WolfJ.A., DudaJ.E., PuttM.E., SpillerK.L., and CullenD.K. (2017). Rapid neuroinflammatory response localized to injured neurons after diffuse traumatic brain injury in swine. Exp. Neurol. 290, 85–94.
35.
FolkertsM.M., BermanR.F., MuizelaarJ.P., and RafolsJ.A. (1998). Disruption of MAP-2 immunostaining in rat hippocampus after traumatic brain injury. J. Neurotrauma, 15, 349–363.
36.
Unal-CevikI., KilincM., Gursoy-OzdemirY., GurerG., and DalkaraT. (2004). Loss of NeuN immunoreactivity after cerebral ischemia does not indicate neuronal cell loss: a cautionary note. Brain Res. 1015, 169–174.
37.
CollombetJ.M., MasqueliezC., FourE., BurckhartM.F., BernabeD., BaubichonD., and LallementG. (2006). Early reduction of NeuN antigenicity induced by soman poisoning in mice can be used to predict delayed neuronal degeneration in the hippocampus. Neurosci. Lett. 398, 337–342.
38.
SommerC., StraehleC., KoetheU., and HamprechtF.A. (2011). Ilastik: Interactive learning and segmentation toolkit, in: 2011 IEEE International Symposium on Biomedical Imaging: From Nano to Macro. Chicago, IL, pps. 230–233.
39.
HikosakaO., SakamotoM., and UsuiS. (1989). Functional properties of monkey caudate neurons I . Activities related to saccadic eye movements. J. Neurophysiol. 61, 780–798.
40.
HuertaM.F., and KaasJ.H. (1990). Supplementary eye field as defined by intracortical microstimulation: connections in macaques. J. Comp. Neurol. 293, 299–330.
41.
StantonG.B., GoldbergM.E., and BruceC.J. (1988). Frontal eye field efferents in the macaque monkey: I. Subcortical pathways and topography of striatal and thalamic terminal fields. J. Comp. Neurol. 271, 473–492.
42.
UpdykeB. V. (1993). Organization of visual corticostriatal projections in the cat, with observations on visual projections to claustrum and amygdala. J. Comp. Neurol. 327, 159–193.
43.
RoyceG.J. and LaineE.J. (1984). Efferent connections of the caudate nucleus, including cortical projections of the striatum and other basal ganglia: an autoradiographic and horseradish peroxidase investigation in the cat. J. Comp. Neurol. 226, 28–49.
44.
MayP.J. (2014). Superior colliculus, in: Encyclopedia of Neuroscience, Second Ed. Springer:, Berlin, Heidelberg, 351–354.
45.
ManiR., AsperL., and KhuuS.K. (2018). Deficits in saccades and smooth-pursuit eye movements in adults with traumatic brain injury: a systematic review and meta-analysis. Brain Inj. 32, 1315–1336.
46.
LonghiL., SaatmanK.E., FujimotoS., RaghupathiR., MeaneyD.F., DavisJ., McMillanA., ConteV., LaurerH.L., SteinS., StocchettiN., and McIntoshT.K. (2005). Temporal window of vulnerability to repetitive experimental concussive brain injury. Neurosurgery, 56, 364–373.
47.
LaurerH.L., BareyreF.M., LeeV.M.Y.C., TrojanowskiJ.Q., LonghiL., HooverR., SaatmanK.E., RaghupathiR., HoshinoS., GradyM.S., and McIntoshT.K. (2001). Mild head injury increasing the brain's vulnerability to a second concussive impact. J. Neurosurg. 95, 859–870.
48.
RaghupathiR., MehrM.F., HelfaerM.A., and MarguliesS.S. (2004). Traumatic axonal injury is exacerbated following repetitive closed head injury in the neonatal pig. J. Neurotrauma, 21, 307–316.
49.
FriessS.H., IchordR.N., RalstonJ., RyallK., HelfaerM.A., SmithC., and MarguliesS.S. (2009). Repeated traumatic brain injury affects composite cognitive function in piglets. J. Neurotrauma, 26, 1111–1121.
50.
RaghupathiR., GrahamD.I., and McIntoshT.K. (2000). Apoptosis after traumatic brain injury. J. Neurotrauma. 17.
51.
HernandezM.L., ChatlosT., GorseK.M., and LafrenayeA.D. (2019). Neuronal membrane disruption occurs late following diffuse brain trauma in rats and involves a subpopulation of NeuN negative cortical neurons. Front. Neurol. 10, 1–13.
52.
HenryL.C., ElbinR.J., CollinsM.W., MarchettiG., and KontosA.P. (2016). Examining recovery trajectories after sport-related concussion with a multimodal clinical assessment approach. Neurosurgery, 78, 232–240.
53.
SufrinkoA.M., MuchaA., CovassinT., MarchettiG., ElbinR.J., and CollinsM.W. (2017). Sex differences in vestibular/ocular and neurocognitive outcomes after sport-related concussion. Clin. J. Sport Med. 27, 133–138.
54.
EllisM.J., CordingleyD.M., VisS., ReimerK.M., LeiterJ., and RussellK. (2017). Clinical predictors of vestibulo-ocular dysfunction in pediatric sports-related concussion. J. Neurosurg. Pediatr. 19, 38–45.
55.
WombleM.N., McAllister-DeitrickJ., MarchettiG.F., ReynoldsE., CollinsM.W., ElbinR.J., and KontosA.P. (2019). Risk factors for vestibular and oculomotor outcomes after sport-related concussion. Clin. J. Sport Med. 00, 1.
56.
ZuckermanS.L., AppleR.P., OdomM.J., LeeY.M., SolomonG.S., and SillsA.K. (2014). Effect of sex on symptoms and return to baseline in sport-related concussion: Clinical article. J. Neurosurg. Pediatr. 13, 72–81.
57.
GupteR., BrooksW., VukasR., PierceJ., and HarrisJ. (2019). Sex differences in traumatic brain injury: what we know and what we should know. J. Neurotrauma, 29, 1–29.
58.
XuL., NguyenJ. V., LeharM., MenonA., RhaE., ArenaJ., RyuJ., MarmarouN.M.-A.C.R., and KoliatsosV.E. (2016). Repetitive mild traumatic brain injury with impact acceleration in the mouse: Multifocal axonopathy, neuroinflammation, and neurodegeneration in the visual system. Exp. Neurol. 275, 436–449.
59.
WangJ., HammR.J., and PovlishockJ.T. (2011). Traumatic axonal injury in the optic nerve: evidence for axonal swelling, disconnection, dieback, and reorganization. J. Neurotrauma, 28, 1185–1198.
60.
EvansonN.K., Guilhaume-CorreaF., HermanJ.P., and GoodmanM.D. (2018). Optic tract injury after closed head traumatic brain injury in mice: A model of indirect traumatic optic neuropathy. PLoS One, 13, 1–20.
61.
DeMarJ., SharrowK., HillM., BermanJ., OliverT., and LongJ. (2016). Effects of primary blast overpressure on retina and optic tract in rats. Front. Neurol. 7, 1–12.
62.
GuleyN.H., RogersJ.T., MarN.A., DelDeng, Y., IslamR.M., D'SurneyL., FerrellJ., DengB., Hines-BeardJ., BuW., RenH., ElbergerA.J., MarchettaJ.G., RexT.S., HonigM.G., and ReinerA. (2016). A novel closed-head model of mild traumatic brain injury using focal primary overpressure blast to the cranium in mice. J. Neurotrauma FEB, 15, 403–422.
63.
PetrasJ.M., BaumanR.A., and ElsayedN.M. (1997). Visual system degeneration induced by blast overpressure. Toxicology, 121, 41–49.
64.
ReinerA., HeldtS.A., PresleyC.S., GuleyN.H., ElbergerA.J., DengY., D'SurneyL., RogersJ.T., FerrellJ., BuW., Del MarN., HonigM.G., GurleyS.N., and MooreB.M. (2015). Motor, visual and emotional deficits in mice after closed-head mild traumatic brain injury are alleviated by the novel CB2 inverse agonist SMM-189. Int. J. Mol. Sci. 16, 758–787.
65.
PettusE.H., ChristmanC.W., GiebelM.L., and PovlishockJ.T. (1994). Traumatically induced altered membrane permeability: its relationship to traumatically induced reactive axonal change. J. Neurotrauma, 11, 507–522.
66.
SingletonR.H. and PovlishockJ.T. (2004). Identification and characterization of heterogeneous neuronal injury and death in regions of diffuse brain injury: evidence for multiple independent injury phenotypes. J. Neurosci. 24, 3543–3553.
67.
StoneJ.R., OkonkwoD.O., DialoA.O., RubinD.G., MutluL.K., PovlishockJ.T., and HelmG.A. (2004). Impaired axonal transport and altered axolemmal permeability occur in distinct populations of damaged axons following traumatic brain injury. Exp. Neurol. 190, 59–69.
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