Chronic cerebral hypoperfusion (CCH) has been indicated to impair cognitive and diverse brain functions. However, the neural mechanisms linking these cerebrovascular and phenotypic alterations remain unclear. Here, we investigated the effect of CCH on neuronal activity in male mice with unilateral common carotid artery occlusion using optical imaging and MRI. Our examinations revealed enhanced neuronal activity in concurrence with increased glutamate and tissue acidosis up to seven days after occlusion. At 21–28 days after occlusion, neuronal activity decreased below baseline, while the acidotic but not the hyperglutamatergic state persisted. Notably, pharmacological blockade of the N-methyl-D-aspartate-type glutamate receptor, initiated at an early stage of CCH, suppressed the onset of neuronal hyperexcitation and subsequent deficits in neuronal activity. Altogether, we provide experimental evidence that CCH induces a glutamate surge and results in neuronal hyperexcitation at an early phase, which thereafter gives rise to a non-lethal but progressive deterioration of neuronal functions.
JohnstonSCO'MearaESManolioTA, et al.
Cognitive impairment and decline are associated with carotid artery disease in patients without clinically evident cerebrovascular disease. Ann Intern Med2004;
140: 237–247.
2.
YamauchiHFukuyamaHNagahamaY, et al.
Atrophy of the corpus callosum associated with cognitive impairment and widespread cortical hypometabolism in carotid artery occlusive disease. Arch Neurol1996;
53: 1103–1109.
3.
ChengHLLinCJSoongBW, et al.
Impairments in cognitive function and brain connectivity in severe asymptomatic carotid stenosis. Stroke2012;
43: 2567–2573.
4.
YamauchiHFukuyamaHNagahamaY, et al.
Evidence of misery perfusion and risk for recurrent stroke in major cerebral arterial occlusive diseases from PET. J Neurol Neurosurg Psychiatry1996;
61: 18–25.
5.
ZuloagaKLZhangWYeiserLA, et al.
Neurobehavioral and imaging correlates of hippocampal atrophy in a mouse model of vascular cognitive impairment. Transl Stroke Res2015;
6: 390–398.
6.
ZhaoYGuJHDaiCL, et al.
Chronic cerebral hypoperfusion causes decrease of O-GlcNAcylation, hyperphosphorylation of tau and behavioral deficits in mice. Front Aging Neurosci2014;
66: 10.
7.
YoshizakiKAdachiKKataokaS, et al.
Chronic cerebral hypoperfusion induced by right unilateral common carotid artery occlusion causes delayed white matter lesions and cognitive impairment in adult mice. Exp Neurol2008;
210: 585–591.
8.
MaJBoSHLuXT, et al.
Protective effects of carnosine on white matter damage induced by chronic cerebral hypoperfusion. Neural Regen Res2016;
11: 1438–1444.
9.
NishinoATajimaYTakuwaH, et al.
Long-term effects of cerebral hypoperfusion on neural density and function using misery perfusion animal model. Sci Rep2016;
6: 25072.
10.
UrushihataTTakuwaHSekiC, et al.
Water diffusion in the brain of chronic hypoperfusion model mice: a study considering the effect of blood flow. Magn Reson Med Sci2018;
17: 318–324.
11.
TajimaYTakuwaHKokuryoD, et al.
Changes in cortical microvasculature during misery perfusion measured by two-photon laser scanning microscopy. J Cereb Blood Flow Metab2014;
34: 1363–1372.
12.
LeeJSImDSAnYS, et al.
Chronic cerebral hypoperfusion in a mouse model of Alzheimers disease: an additional contributing factor of cognitive impairment. Neurosci Lett2011;
489: 84–88.
13.
Schmidt-KastnerRAguirre-ChenCSaulI, et al.
Astrocytes react to oligemia in the forebrain induced by chronic bilateral common carotid artery occlusion in rats. Brain Res2005;
1052: 28–39.
14.
BeppuKSasakiTTanakaKF, et al.
Optogenetic countering of glial acidosis suppresses glial glutamate release and ischemic brain damage. Neuron2014;
81: 314–320.
15.
RakersCPetzoldGC.Astrocytic calcium release mediates peri-infarct depolarizations in a rodent stroke model. J Clin Invest2017;
127: 511–516.
16.
SzalayGMartineczBLénártN, et al.
Microglia protect against brain injury and their selective elimination dysregulates neuronal network activity after stroke. Nat Commun2016;
7: 11499.
17.
WaatajaJJKimHJRoloffAM, et al.
Excitotoxic loss of post-synaptic sites is distinct temporally and mechanistically from neuronal death. J Neurochem2008;
104: 364–375.
18.
ShibukiKHishidaRMurakamiH, et al.
Dynamic imaging of somatosensory cortical activity in the rat visualized by flavoprotein autofluorescence. J Physiol2003;
549: 919–927.
19.
ChenTWWardillTJSunY, et al.
Ultrasensitive fluorescent proteins for imaging neuronal activity. Nature2013;
499: 295–300.
20.
ZhuHBarkerPB.MR spectroscopy and spectroscopic imaging of the brain. Methods Mol Biol2010;
711: 203–226.
21.
ZhouJPayenJFWilsonDA, et al.
Using the amide proton signals of intracellular proteins and peptides to detect pH effects in MRI. Nat Med2003;
9: 1085–1090.
22.
HoltmaatABonhoefferTChowDK, et al.
Long-term, high-resolution imaging in the mouse neocortex through a chronic cranial window. Nat Protoc2009;
4: 1128–1144.
23.
TomitaYKubisNCalandoY, et al.
Long-term in vivo investigation of mouse cerebral microcirculation by fluorescence confocal microscopy in the area of focal ischemia. J Cereb Blood Flow Metab2005;
25: 858–867.
24.
MinkevicieneRBanerjeePTanilaH.Memantine improves spatial learning in a transgenic mouse model of Alzheimer’s disease. J Pharmacol Exp Ther2004;
311: 677–682.
25.
TakuwaHAutioJNakayamaH, et al.
Reproducibility and variance of a stimulation-induced hemodynamic response in barrel cortex of awake behaving mice. Brain Res2011;
1369: 103–111.
26.
TakuwaHMatsuuraTNishinoA, et al.
Development of new optical imaging systems of oxygen metabolism and simultaneous measurement in hemodynamic changes using awake mice. J Neurosci Methods2014;
237: 9–15.
27.
TakahashiMUrushihataTTakuwaH, et al.
Imaging of neuronal activity in awake mice by measurements of flavoprotein autofluorescence corrected for cerebral blood flow. Front Neurosci2017;
11: 723.
28.
PnevmatikakisEAGiovannucciA.NoRMCorre: an online algorithm for piecewise rigid motion correction of calcium imaging data. J Neurosci Methods2017;
291: 83–94.
29.
JiaHRochefortNLChenX, et al.
In vivo two-photon imaging of sensory-evoked dendritic calcium signals in cortical neurons. Nat Protoc2011;
6: 28–35.
30.
TakanashiJNittaNIwasakiN, et al.
Neurochemistry in shiverer mouse depicted on MR spectroscopy. J Magn Reson Imaging2014;
39: 1550–1557.
31.
AtwoodTRobbinsMEZhuJM.Quantitative in vivo proton MR spectroscopic evaluation of the irradiated rat brain. J Magn Reson Imaging2007;
26: 1590–1595.
ShahTLuLDellKM, et al.
CEST-FISP: a novel technique for rapid chemical exchange saturation transfer MRI at 7 T. Magn Reson Med2011;
65: 432–437.
34.
KhlebnikovVvan der KempWJMHoogduinH, et al.
Analysis of chemical exchange saturation transfer contributions from brain metabolites to the Z-spectra at various field strengths and pH. Sci Rep2019;
99: 1089.
35.
KimMGillenJLandmanBA, et al.
Water saturation shift referencing (WASSR) for chemical exchange saturation transfer (CEST) experiments. Magn Reson Med2009;
61: 1441–1450.
36.
SagiyamaKMashimoTTogaoO, et al.
In vivo chemical exchange saturation transfer imaging allows early detection of a therapeutic response in glioblastoma. Proc Natl Acad Sci U S A2014;
111: 4542–4547.
37.
RuitenbergADen HeijerTBakkerSLM, et al.
Cerebral hypoperfusion and clinical onset of dementia: the Rotterdam study. Ann Neurol2005;
57: 789–794.
38.
AustinBPNairVAMeierTB, et al.
Effects of hypoperfusion in Alzheimers disease. J Alzheimers Dis2011;
26 Suppl 3: 123–133.
39.
TangHGaoYZhangQ, et al.
Chronic cerebral hypoperfusion independently exacerbates cognitive impairment within the pathopoiesis of parkinson’s disease via microvascular pathologys. Behav Brain Res2017;
333: 286–294.
40.
CarmichaelST.Brain excitability in stroke: the yin and yang of stroke progression. Arch Neurol2012;
69: 161–167.
41.
MameniškienėRWolfP.Epilepsia partialis continua: a review. Seizure2017;
44: 74–80.
42.
BentesCFrancoACPeraltaAR, et al.
Epilepsia partialis continua after an anterior circulation ischaemic stroke. Eur J Neurol2017;
24: 929–934.
43.
CataldoAMBroadwellRD.Cytochemical identification of cerebral glycogen and glucose-6-phosphatase activity under normal and experimental conditions. II. Choroid plexus and ependymal epithelia, endothelia and pericytes. J Neurocytol1986;
15: 511–524.
44.
SchurrAPayneRSMillerJJ, et al.
Glia are the main source of lactate utilized by neurons for recovery of function posthypoxia. Brain Res1997;
774: 221–224.
45.
MutchWACHansenAJ.Extracellular pH changes during spreading depression and cerebral ischemia: Mechanisms of brain pH regulation. J Cereb Blood Flow Metab1984;
4: 17–27.
46.
TóthOMMenyhártÁFrankR, et al.
Tissue acidosis associated with ischemic stroke to guide neuroprotective drug delivery. Biology (Basel)2020;
9: 1–15.
47.
ArundineMTymianskiM.Molecular mechanisms of glutamate-dependent neurodegeneration in ischemia and traumatic brain injury. Cell Mol Life Sci2004;
61: 657–668.
48.
WangLDuYWangK, et al.
Chronic cerebral hypoperfusion induces memory deficits and facilitates Aβ generation in C57BL/6J mice. Exp Neurol2016;
283: 353–364.
49.
EhrlichIKleinMRumpelS, et al.
PSD-95 is required for activity-driven synapse stabilization. Proc Natl Acad Sci U S A2007;
104: 4176–4181.
50.
EliasGMFunkeLSteinV, et al.
Synapse-Specific and developmentally regulated targeting of AMPA receptors by a family of MAGUK scaffolding proteins. Neuron2006;
52: 307–320.
ShimojoMTakuwaHTakadoY, et al.
Selective disruption of inhibitory synapses leading to neuronal hyperexcitability at an early stage of tau pathogenesis in a mouse model. J Neurosci2020;
40: 3491–3501.
53.
YemisciMGursoy-OzdemirYVuralA, et al.
Pericyte contraction induced by oxidative-nitrative stress impairs capillary reflow despite successful opening of an occluded cerebral artery. Nat Med2009;
15: 1031–1037.
54.
Taskiran-SagAYemisciMGursoy-OzdemirY, et al.
Improving microcirculatory reperfusion reduces parenchymal oxygen radical formation and provides neuroprotection. Stroke2018;
49: 1267–1275.
ErmineCMBivardAParsonsMW, et al.
The ischemic penumbra: from concept to reality. Int J Stroke2021;
16: 497–509.
57.
MöbiusHJ.Pharmacologic rationale for memantine in chronic cerebral hypoperfusion, especially vascular dementia. Alzheimer Dis Assoc Disord1999;
13 Suppl 3: S172–178.
58.
MöbiusHJStöfflerA.Memantine in vascular dementia. Int Psychogeriatr2003;
15 Suppl 1: 207–213.
59.
OrgogozoJMRigaudASStöfflerA, et al.
Efficacy and safety of memantine in patients with mild to moderate vascular dementia: a randomized, placebo-controlled trial (MMM 300). Stroke2002;
33: 1834–1839.
60.
VosselKARanasingheKGBeagleAJ, et al.
Incidence and impact of subclinical epileptiform activity in alzheimer’s disease. Ann Neurol2016;
80: 858–870.
61.
KwanACDanY.Dissection of cortical microcircuits by single-neuron stimulation in vivo. Curr Biol2012;
22: 1459–1467.
62.
HornTKleinJ.Lactate levels in the brain are elevated upon exposure to volatile anesthetics: a microdialysis study. Neurochem Int2010;
57: 940–947.
63.
AndersonRESundtTM.Brain pH in focal cerebral ischemia and the protective effects of barbiturate anesthesia. J Cereb Blood Flow Metab1983;
3: 493–497.
64.
YanGDaiZXuanY, et al.
Early metabolic changes following ischemia onset in rats: an in vivo diffusion-weighted imaging and 1H-magnetic resonance spectroscopy study at 7.0 T. Mol Med Rep2015;
11: 4109–4114.
65.
TakadoYTakuwaHSampeiK, et al.
MRS-measured glutamate versus GABA reflects excitatory versus inhibitory neural activities in awake mice. J Cereb Blood Flow Metab2022;
42: 197–212.
66.
KoepJLBondBBarkerAR, et al.
Sex modifies the relationship between age and neurovascular coupling in healthy adults. J Cereb Blood Flow Metab2023;
43: 1254–1266.
67.
CikicSChandraPKHarmanJC, et al.
Sexual differences in mitochondrial and related proteins in rat cerebral microvessels: a proteomic approach. J Cereb Blood Flow Metab2021;
41: 397–412.
68.
RutkaiIDuttaSKatakamPV, et al.
Dynamics of enhanced mitochondrial respiration in female compared with male rat cerebral arteries. Am J Physiol Heart Circ Physiol2015;
309: H1490–1500.
69.
KremerCLorenzanoSKruuseC.Editorial: Sex differences in cerebrovascular diseases. Front Neurol2022;
13: 1128177.
70.
SeaksCEWeekmanEMSudduthTL, et al.
Apolipoprotein E ε4/4 genotype limits response to dietary induction of hyperhomocysteinemia and resulting inflammatory signaling. J Cereb Blood Flow Metab2022;
42: 771–787.
71.
JiaCLovinsCMaloneHM, et al.
Female-specific neuroprotection after ischemic stroke by vitronectin-focal adhesion kinase inhibition. J Cereb Blood Flow Metab2022;
42: 1961–1974.
72.
DinhDDWanHLidingtonD, et al.
Female mice display sex-specific differences in cerebrovascular function and subarachnoid haemorrhage-induced injury. EBioMedicine2024;
102: 105058.
73.
Raman-NairJCronGMacLeodK, et al.
Sex-Specific acute cerebrovascular responses to photothrombotic stroke in mice. eNeuro2024;
11: 0400–0422.
74.
SalineroAERobisonLSGannonOJ, et al.
Sex-Specific effects of high fat diet on cognitive impairment in a mouse model of VCID. FASEB J2020;
34: 15108–15122.
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