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Mechanisms of pH regulation in rat trigeminal ganglion neurons
DC Field | Value | Language |
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dc.contributor.advisor | 박경표 | - |
dc.contributor.author | 황성민 | - |
dc.date.accessioned | 2017-07-14T05:47:39Z | - |
dc.date.available | 2017-07-14T05:47:39Z | - |
dc.date.issued | 2014-02 | - |
dc.identifier.other | 000000018751 | - |
dc.identifier.uri | https://hdl.handle.net/10371/125165 | - |
dc.description | 학위논문 (박사)-- 서울대학교 대학원 : 치의학과, 2014. 2. 박경표. | - |
dc.description.abstract | The effect of intracellular acidification and subsequent pH recovery in sensory neurons has not been well characterized. I have studied the mechanisms underlying Ca2+-induced acidification and subsequent recovery of intracellular pH (pHi) in rat trigeminal ganglion (TG) neurons and report their effects on neuronal excitability. Glutamate (500 µM) and capsaicin (1 µM) increased intracellular Ca2+ concentration ([Ca2+]i) with a following decrease in pHi. The recovery of [Ca2+]i to the pre-stimulus level was inhibited by LaCl3 (1 mM) and O-vanadate (10 mM), a plasma membrane Ca2+/ATPase (PMCA) inhibitor. Removal of extracellular Ca2+ also completely inhibited the acidification induced by capsaicin. TRPV1 was expressed only in small and medium sized TG neurons. mRNA for Na+/H+ exchanger type 1 (NHE1), pancreatic Na+-HCO3- cotransporter type 1 (pNBC1), NBC3, NBC4 and PMCA types 1-3 were detected by RT-PCR. pHi recovery was significantly inhibited by pretreatment with NHE1 or pNBC1 siRNA. I found that the frequency of action potentials (AP) was dependent on pHi. Application of the NHE1 inhibitor 5-(N-ethyl-N-isopropyl) amiloride (5 μM | - |
dc.description.abstract | EIPA) or the pNBC1 inhibitor 4,4-di-isothiocyano-stilbene-2,2-sulfonic acid (500 µM | - |
dc.description.abstract | DIDS) delayed pHi recovery and decreased AP frequency. Simultaneous application of EIPA and DIDS almost completely inhibited APs.
In summary, These results demonstrate that the rise in [Ca2+]i in sensory neurons by glutamate and capsaicin causes intracellular acidification by activation of PMCA type 3, that the pHi recovery from acidification is mediated by membrane transporters NHE1 and pNBC1 specifically, and that the activity of these transporters has direct consequences for neuronal excitability. | - |
dc.description.tableofcontents | Contents
Abstract……………………………………………………….………………ⅰ Contents ……………………………………………………………………... ⅲ List of Figures……………………………………………………………..…..ⅳ Abbreviations………………………………………………………………….ⅴ 1. Introduction………………………………………………………………….1 2. Materials and Methods……………………………………………………....4 3. Results……………………………………………………………………...11 4. Discussion…………………………………………………………………. 42 5. References………………………………………………………………… 47 Abstract in Korean…………………………………………………………… 54 List of Figures Figure 1. Identification of membrane transporters and TRPV1 in TG neurons……………………………………………………………………….. 12 Figure 2. The effect of glutamate and capsaicin on [Ca2+]i and pHi in single TG neurons ………………………………………………………………………..16 Figure 3. The effects of LaCl3 on [Ca2+]i and pHi evoked by glutamate or capsaicin stimulation in TG neurons…………………………………………..19 Figure 4. Inhibitory effects of PMCA inhibitors on the capsaicin-induced changes in [Ca2+]i and pHi………………………………………………….….22 Figure 5. Inhibitory effects of PMCA3 siRNA on the capsaicin-induced changes in [Ca2+]i and pHi…………………………………………….…….…25 Figure 6. The pHi recovery of TG neurons from acidification induced by NH4Cl pulse…………………………………………………………….……….……..29 Figure 7. pHi recovery in TG neurons transfected with control siRNA, NHE1 siRNA and pNBC1 siRNA………………………………………………….…32 Figure 8. The effect of pHi on the frequency of action potentials (AP) in TG neurons in BBS……………………………………………….…………….…35 Figure 9. The effect of EIPA and/or DIDS on AP frequency in TG neurons in BBS……………………………………………………………...…………….38 Figure 10. Schematic diagram of pH regulation mechanism in trigeminal ganglion……………………………………………….……………………….40 | - |
dc.format | application/pdf | - |
dc.format.extent | 2381388 bytes | - |
dc.format.medium | application/pdf | - |
dc.language.iso | en | - |
dc.publisher | 서울대학교 대학원 | - |
dc.subject | Trigeminal ganglion | - |
dc.subject | Intracellular pH | - |
dc.subject | Action potentials | - |
dc.subject | Transient receptor potential vanilloid 1 | - |
dc.subject | Plasma membrane Ca2+/ATPase | - |
dc.subject | Na+/H+ exchangers | - |
dc.subject | Na+/HCO3- cotransporters | - |
dc.subject.ddc | 617 | - |
dc.title | Mechanisms of pH regulation in rat trigeminal ganglion neurons | - |
dc.type | Thesis | - |
dc.description.degree | Doctor | - |
dc.citation.pages | vi, 56 | - |
dc.contributor.affiliation | 치과대학 치의학과 | - |
dc.date.awarded | 2014-02 | - |
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