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dc.contributor.advisorKARATAS KURSUN, Hulya
dc.contributor.authorDEHGHANİ MOHAMMADİ, Anisa
dc.date.accessioned2019-01-18T11:59:40Z
dc.date.issued2019-01
dc.date.submitted2019-01-08
dc.identifier.citationAnisa Dehghani, 2019, Parenchymal neuroinflammation in familial hemiplegic migraine type 1 transgenic mouse model after cortical spreading depression, PhD thesis, Hacettepe Universitytr_TR
dc.identifier.urihttp://hdl.handle.net/11655/5706
dc.description.abstractCortical spreading depression (CSD) is the likely cause of the migraine aura. CSD causes a signaling pathway between stressed neurons and trigeminal afferents via transient opening of neuronal Pannexin-1 (Panx1) mega-channels followed by high mobility group box 1 (HMGB1) release from neurons and nuclear factor kappa B (NF-κB) relocation in astrocytes. Familial hemiplegic migraine type 1 (FHM1) is a rare monogenic subtype of migraine with aura caused by mutations in the CACNA1A gene. Transgenic knock-in mice that carry human FHM1 missense mutations R192Q or S218L exhibit an increased susceptibility to CSD and other features relevant to migraine. Here, we investigated the basal and experimentally CSD-induced parenchymal neuroinflammation in female mutant mice and wild-type (WT), and studied whether CSD-induced inflammation shows a particular regional pattern, which may highlight areas that are particularly relevant to disease pathology. Our data revealed a basal parenchymal neuroinflammatory state in R192Q mutant mice as revealed by higher neuronal HMGB1 release and NF-κB activation in astrocytes. There was a different regional distribution pattern in the inflammatory signaling upon CSD in the frontal/parietal cortices and striatum/thalamus of both R192Q and WT mice. In addition, in mutant mice this inflammatory pattern was bilateral. There was a rise in HMGB1 release in subcortical areas of mutant mice compared to that in WT mice which was particularly pronounced in thalamus compared to striatum. Immunohistochemical data was supported by Western blotting in which cerebrospinal fluid (CSF) HMGB1 amount and brain nuclear translocation of NF-κB was studied. S218L mutant mice showed more HMGB1 in CSF compared to WT and brain NF-κB translocation was increased in mutant and WT CSD and mutant sham-operated groups compared to WT sham. Electrophysiological analysis showed that CSD speed is higher in mutant groups, however CSD half maximum amplitude duration and CSD frequency did not change significantly between groups. Derivative of CSD depolarisation phase demonstrated that both derivative of the first CSD and subsequent CSDs were increased in FHM1 mutant groups compared to WT mice reflecting the hyperexcitability of mutant ones. Here for the first time in the literature, we presented that there is basal parenchymal neuroinflammation in R192Q mutant mice brain both in cortical and subcortical levels. Our findings indicate that CSD triggers parenchymal inflammatory processes in both WT and mutant mouse brains, which is mostly ipsilateral to the side of CSD induction in WT mice, and bilateral in the mutant mice. Future studies may indicate if basal neuroinflammation contributes to cortical hyperexcitability seen in FHM1 mutants in addition to the increased neuronal calcium influx or increased calcium and glutamate cause basal neuroinflammation. These findings indicate that CSD-induced parenchymal neuroinflammation spreads through cortex, striatum, and thalamus of both hemispheres in genetically susceptible brains, which may explain the prolonged hemiplegia, coma, and seizure phenotype in this variant of migraine with aura.en
dc.description.tableofcontentsCONTENTS Page APPROVEMENT PAGE YAYIMLAMA VE FİKRİ MÜLKİYET HAKLARI BEYANI ETHICAL DECLARATION PAGE i ii iii ACKNOWLEDGEMENTS V ABSTRACT Vi OZET Vii CONTENTS Viii ABREVIATIONS Xii FIGURE INDEX Xiv TABLE INDEX Xvii 1. INTRODUCTION 1 2. BACKGROUND 4 2.1. Epidemiology 4 2.2. History 2.3. Economic Impact 2.4. Migraine: A Noblesse and Challenge for Social Coexistence 2.5. Migraine Classification 2.6. Migraine Without Aura 2.7. Migraine With Aura 2.8. Familial and Sporadic Hemiplegic Migraine 2.9. Familial Hemiplegic Migraine Mutations as a Model for Disease 2.10. Functional Consequences of FHM Mutations 2.11. Cav2.1 Gain of Function and CSD Susceptibility Enhancement in FHM1 Mice 2.12. Neuronal Network Dependent Synaptic Plasticity in FHM1 Mutant Mice 2.13. The Effect of FHM1 Mutations on Neuroinflammation 2.14. Cortical Spreading Depression 2.15. High Mobility Group Box 1 (HMGB1) Protein 2.16. The Role of Nuclear Factor κB in Neuroinflammation 5 7 8 11 12 13 13 14 18 19 20 21 23 24 27 3. MATERIALS AND METHODS 30 3.1. Materials Used 30 3.2. Methods 31 3.2.1. Ethics 31 3.2.2. Animals 31 3.2.3. Surgical Procedure and Electrophysiological Recordings 33 3.2.4. Experimental Design 34 3.2.5. Immunohistochemistry 35 3.2.6. Western Blotting 36 3.2.6.1. Subcellular Fractionation and NF-κB Western Blotting 3.2.6.2. CSF Collection After CSD and HMGB1 Western Blotting 36 37 3.2.7. Electrophysiological Analysis 38 3.2.8. Quantification of HMGB1 Release and NF-κB Activation 38 3.2.9. Statistical Analysis 39 4. RESULTS 40 4.1. Parenchymal Neuroinflammation in Naïve Wild-type and R192Q Mutant Mice 4.1.1 HMGB1 release in naïve Wild-type and R192Q Mutant Mice 4.1.2 NF-κB Cytoplasm–To–Nucleous Translocation In Astrocytes of Naïve Wild-type and R192Q Mutant Mice 40 40 43 4.2. Effect of a Non-invasive Confounding Stimulus, Drilling the Skull, on Parenchymal Neuroinflammation and HMGB1 Release 46 4.3. Pinprick-induced CSD Causes Parenchymal Neuroinflammation in Wild-type and R192Q Mutant Mice: Does CSD Cause a Regional Specific Distribution Pattern and Increase in HMGB1 Release? 48 4.3.1 Laterality in HMGB1 Release in WT and R192Q Mutant Mouse Brains 48 4.3.2 Laterality in HMGB1 Release in Cortical Areas of Wild-type and R192Q Mutant Mouse Brains 52 4.3.3 Laterality in HMGB1 Release In Subcortical Areas of Wild-type and R192Q Mutant Mouse Brains 56 4.3.4. Laterality in Cortical and Subcortical HMGB1 Release of Wild-type and R192Q Mutant Mouse Brains 4.4. Pinprick-induced Parenchymal Neuroinflammation in Wild-type and R192Q Mutant Mouse Brains: A Regional Distribution Pattern and Increase in NF-B Translocation in Astrocytes 4.5. Neuroinflammation After Multiple CSD Events Induced By KCl in Wild-type and S218L Mutant Mouse Brains 4.5.1. HMGB1 Release in CSF in Wild-type and S218L Mutant Mice After KCl-induced Multiple CSDs 4.5.2. Translocation of NF-B from Cytoplasm to Nucleus in Astrocytes of Wild-type and S218L Mutant Mice Assessed by Western Blotting 4.6. Electrophysiological Characterization of Cortical Spreading Depolarization Events 4.6.1. CSD amplitude was not significantly different in wild-type mice compared to that in R192Q or S218L mutant mice 4.6.2. Speed of CSD was higher in R192Q and S218L mutant mice compared to that in wild-type mice 4.6.3. Total CSD duration was not significantly changed in R192Q and S218L mutant mice compared to that in wild-type mice 4.6.4. CSD half-maximum duration was not statistically different in R192Q and S218L mutant mice compared to wild-type mice 4.6.5. Frequency of CSD events in R192Q and S218L mutant mice compared to that in wild-type mice 4.6.6. The first component of the CSD derivative was higher in the first CSD event compared to subsequent events in R192Q and S218L mutant mice compared to that in wild-type mice 60 64 66 66 67 73 77 77 79 79 80 81 5. DISCUSSION 83 6. CONCLUSION 94 7. REFERENCES 95 8. ADDITORY Add-1: Ethical Approval-1 Add-2: Ethical Approval-2 Add-3: Turnitin Ekran görüntüsü Add-4: Digital Makbuz 9. ACADEMIC CVtr_TR
dc.language.isoengtr_TR
dc.publisherNörolojik Bilimler ve Psikiyatri Enstitüsütr_TR
dc.rightsinfo:eu-repo/semantics/closedAccesstr_TR
dc.subjectMigrentr_TR
dc.subjectAuratr_TR
dc.subjectKortikal yayılan depresyontr_TR
dc.subjectAilevi hemiplejik migren tip 1tr_TR
dc.subjectİnflamasyontr_TR
dc.subjectHMGB1tr_TR
dc.subjectNF-κBtr_TR
dc.titleParenchymal Neuroinflammation in Familial Hemiplegic Migraine Type 1 Transgenic Mouse Model After Cortical Spreading Depressiontr_TR
dc.typedoctoralThesisen
dc.description.ozetKortikal yayılan depresyon (KYD) migren aurasının nedeni olarak kabul edilmektedir. KYD stres altındaki nöronlar ile trigeminal aferentler arasında bir sinyalizasyon yolağını başlatır, bu yolak nöronal Panneksin-1 (Panx1) mega kanallarının geçici olarak açılması ile ‘high mobility group box 1’ (HMGB1)’in nörondan salıverilmesi ve astrositlerde nükleer faktör kappa B (NF-κB)’nin nükleusa yer değiştirmesinden oluşur. Ailevi hemiplejik migren tip 1 (AHM1) CACNA1A genindeki bir mutasyonun neden olduğu migrenin nadir görülen bir monogenik alt tipidir. İnsan AHM1 mutasyonu taşıyan dişi transgenik knock-in fareler (R192Q veya S218L) KYD oluşumuna yatkınlık ve migrenin diğer özelliklerini sergiler. Bu çalışmada, AHM1 mutasyonu olan ve yabanıl farelerde bazal ve KYD’nin tetiklediği parankimal nöroinflamasyon incelenmiştir, ayrıca KYD’nin tetiklediği inflamasyonun hastalığın patofizyolojisi ile ilişkili olabilecek bölgesel farklılığı olup olmadığı çalışılmıştır. Sonuçlara göre R192Q mutasyonu olan farelerde bazalde bir parankimal nöroinflamatuvar durum olduğu nöronal HMGB1 salınımı ve astrositik NF-κB translokasyonu ile gösterilmiştir. KYD hem R192Q mutasyonu olan hem de yabanıl fare gruplarında frontal/pariyetal korteks ve striatum/talamusta farklı bölgelerde inflamatuvar sinyal yolağını tetiklemektedir. Ayrıca mutasyonu olan farelerde bu inflamatuvar paternin bilateral sergilendiği saptanmıştır. AHM1 mutasyonu olan farelerde subkortikal alanlarda HMGB1 salınımı yabanıl gruba göre artmıştır ve bu artış talamusta striatuma göre daha belirgindir. Western blotlama ile immünhistokimyasal tekniklerle saptanan bulgular desteklenmiştir. S218L mutasyonu taşıyan farelerde beyin omurilik sıvısında HMGB1 protein miktarında artış olduğu ve beyin parankiminden yapılan nükleer NF-κB translokasyonu deneylerinde KYD tetiklenen yabanıl grup ve mutant grupta NF-κB translokasyonunun yabanıl kontrole göre anlamlı olarak arttığı gösterilmiştir. Elektrofizyolojik analizlerde literatürle benzer şekilde mutant fare gruplarında KYD hızının yabanıl gruba göre artmış olduğu, yarı maksimum amplitüd süresi ve KYD frekansının iki grup arasında anlamlı olarak değişmediği görülmüştür. İlk ve diğer KYD’lerin depolarizasyon fazının türevi alındığında R192Q mutasyonu taşıyan farelerde yabanıl gruba göre anlamlı artış olduğu saptanmıştır, bu bulgu mutant gruptaki hipereksitabiliteyi desteklemektedir. Bu çalışmada literatürde ilk kez R192Q mutasyonu taşıyan farelerde bazalde bir parankimal nöroinflamatuvar durum olduğu, KYD’nin AHM1 mutant farelerde bilateral kortikal ve subkortikal, yabanıl farelerde unilateral parankimal inflamatuvar süreci tetiklediği söylenebilir. KYD’ye yatkınlığı artıran AHM1 mutasyonunda görülen kortikal hipereksitabiliteye, artmış nöronal kalsiyumun yanısıra bazal inflamatuvar durumun katkıda bulunup bulunmadığı veya artmış kalsiyum ve glutamatın bazal inflamasyona yol açıp açmadığı daha sonraki çalışmaların konusu olabilir. Bulgularımız genetik olarak yatkın beyinde KYD’nin tetiklediği parankimal nöroinflamasyonun her iki hemisferde korteks, striatum ve talamusa yayıldığını göstermiş ve auralı migrenin bu alt grubunda uzamış hemipleji, koma ve nöbet fenotipinin nasıl ortaya çıktığını anlamamıza katkıda bulunmuştur.tr_TR
dc.contributor.departmentNörolojik ve Psikiyatrik Temel Bilimlertr_TR
dc.embargo.terms6 aytr_TR
dc.embargo.lift2019-07-23T11:59:40Z


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