Modeling Analysis of Axonal After Potential at Hippocampal Mossy Fibers

Action potentials reliably propagate along the axons, and after potential often follows the axonal action potentials. After potential lasts for several tens of millisecond and plays a crucial role in regulating excitability during repetitive firings of the axon. Several mechanisms underlying the generation of after potential have been suggested, including activation of ionotropic autoreceptors, accumulation of K+ ions in the surrounding extracellular space, the opening of slow voltage-dependent currents, and capacitive discharge of upstream action potentials passively propagated through axon cable. Among them, capacitive discharge is difficult to examine experimentally, since the quantitative evaluation of a capacitive component requires simultaneous recordings from at least two different sites on the connecting axon. In this study, a series of numerical simulation of the axonal action potential was performed using a proposed model of the hippocampal mossy fiber where morphological as well as electrophysiological data are accumulated. To evaluate the relative contribution of the capacitive discharge in axonal after potential, voltage-dependent Na+ current as well as voltage-dependent K+ current was omitted from a distal part of mossy fiber axons. Slow depolarization with a similar time course with the recorded after potential in the previous study was left after blockade of Na+ and K+ currents, suggesting that a capacitive component contributes substantially in axonal after potential following propagating action potentials. On the other hand, it has been shown that experimentally recorded after potential often showed clear voltage-dependency upon changes in the initial membrane potential, obviously deviating from voltage-independent nature of the capacitive component. The simulation revealed that activation of voltage-dependent K+ current also contributes to shape a characteristic waveform of axonal after potential and reconstitute similar voltage-dependency with that reported for the after potential recorded from mossy fiber terminals. These findings suggest that the capacitive component reflecting passive propagation of upstream action potential substantially contributes to the slow time course of axonal after potential, although voltage-dependent K+ current provided a characteristic voltage dependency of after potential waveform

神経終末内カルシウム貯蔵部位によるシナブス可塑性の調節

金沢大学医学部中枢シナブス可塑性における神経終末内カルシウム貯蔵部位の役割を検討する目的で、海馬スライス標本を用いた神経終末内カルシウム(Ca^)濃度測定法の開発を試みた。入力線維層組織内に局所的に注入した細胞膜透過型蛍光Ca^指示薬rhod2-AMが軸索内に取り込まれ神経終末まで輸送されることを利用して、CA1野シャーファー側枝シナプスおよびCA3野苔状線維シナプスにおける入力線維の選択的な蛍光標識が可能であった。特に苔状線維シナプスではCA3野透明層において直径2〜3μmと中枢シナプスとしては大型な単一苔状線維終末が観察できた。このような標本においてシナプス部位(CA1野放線層またはCA3野透明層)から単一フォトダイオードを用いて蛍光強度を測定し入力線維層に単一電気刺激を与えると一過性の蛍光強度の増加が測定された。この蛍光強度増加はグルタミン酸受容体阻害剤(CNQX,AP5およびMCPG)を用いてシナプス伝達をシナプス後性に遮断しても減弱せず、活動電位に伴うシナプス前部(神経終末)へのCa^流入を反映すると考えられた。上述した神経終末内Ca^濃度測定法を用いてこれまでに、CA1野シャーファー側枝シナプスにおける代謝型グルタミン酸受容体(mGluR)アゴニストのシナプス前抑制作用が神経終末へのCa^流入の低下によることを明らかにした。また、この問題と関連して、CA3野苔状線維シナプスではCA1野シナプスとは異なるmGluRサブタイプ(mGluR2またはmGluR3)が選択的に発現している可能性を示した。研究課題/領域番号:07780698, 研究期間(年度):1995出典：研究課題「神経終末内カルシウム貯蔵部位によるシナブス可塑性の調節」課題番号07780698（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） （https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-07780698/）を加工して作

海馬興奮性および抑制性シナプスにおける伝達物質放出の薬剤感受性の相違について

取得学位 : 博士（医学）, 学位授与番号 : 医博乙第1327号, 学位授与年月日：平成6年12月7日,学位授与年：199

パッチクランプ法によるシナプス終末膜カルシウム・チャネル調節機構の研究

金沢大学医学部研究課題/領域番号:01770077, 研究期間(年度):1989出典：研究課題「パッチクランプ法によるシナプス終末膜カルシウム・チャネル調節機構の研究」課題番号01770077（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） （https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-01770077/）を加工して作

細胞内カルシウム放出機構による中枢シナプス可塑性の調節

金沢大学医学部中枢シナプス可塑性における神経終末内カルシウム貯蔵部位の役割を検討する目的で、海馬スライス標本を用いた神経終末内カルシウム(Ca^)濃度測定法の開発を試みた。入力線維層組織内に局所的に注入した細胞膜透過型蛍光Ca^指示薬rhod 2‐AM が軸索内に取り込まれ神経終末まで輸送されることを利用して、CA1野シャーファー側枝シナプスおよびCA3野苔状線維シナプスにおける入力線維の選択的な蛍光標識が可能であった。特に苔状線維シナプスではCA3野透明層において直径2〜3μmと中枢シナプスとしては大型な単一苔状線維終末が観察できた。このような標本においてシナプス部位(CA1野放線層またはCA3野透明層)から単一フォトダイオードを用いて蛍光強度を測定し入力線維層に単一電気刺激を与えると一過性の蛍光強度の増加が測定された。この蛍光強度増加はグルタミン酸受容体阻害剤(CNQX, AP5およびMCPG)を用いてシナプス伝達をシナプス後性に遮断しても減弱せず、活動電位に伴うシナプス前部(神経終末)へのCa^流入を反映すると考えられた。上述した神経終末内Ca^濃度測定法を用いてこれまでに、CA1野シャーファー側枝シナプスにおける代謝型グルタミン酸受容体(mGluR)アゴニストのシナプス前抑制作用が神経終末へのCa^流入の低下によることを明らかにした。また、この問題と関連して、CA3野苔状線維シナプスではCA1野シナプスとは異なるmGluRサブタイプ(mGluR2またはmGluR3)が選択的に発現している可能性を示した。研究課題/領域番号:07279216, 研究期間(年度):1995出典：研究課題「細胞内カルシウム放出機構による中枢シナプス可塑性の調節」課題番号07279216（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） （https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07279216/）を加工して作

長期増強におけるプロテイン・キナーゼC仮説の検討

金沢大学医学部研究課題/領域番号:63770082, 研究期間(年度):1988出典：研究課題「長期増強におけるプロテイン・キナーゼC仮説の検討」課題番号63770082（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） （https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-63770082/）を加工して作

Defective function of GABA-containing synaptic vesicles in mice lacking the AP-3B clathrin adaptor

AP-3 is a member of the adaptor protein (AP) complex family that regulates the vesicular transport of cargo proteins in the secretory and endocytic pathways. There are two isoforms of AP-3: the ubiquitously expressed AP-3A and the neuron-specific AP-3B. Although the physiological role of AP-3A has recently been elucidated, that of AP-3B remains unsolved. To address this question, we generated mice lacking μ3B, a subunit of AP-3B. μ3B−/− mice suffered from spontaneous epileptic seizures. Morphological abnormalities were observed at synapses in these mice. Biochemical studies demonstrated the impairment of γ-aminobutyric acid (GABA) release because of, at least in part, the reduction of vesicular GABA transporter in μ3B−/− mice. This facilitated the induction of long-term potentiation in the hippocampus and the abnormal propagation of neuronal excitability via the temporoammonic pathway. Thus, AP-3B plays a critical role in the normal formation and function of a subset of synaptic vesicles. This work adds a new aspect to the pathogenesis of epilepsy

Large expert-curated database for benchmarking document similarity detection in biomedical literature search

Document recommendation systems for locating relevant literature have mostly relied on methods developed a decade ago. This is largely due to the lack of a large offline gold-standard benchmark of relevant documents that cover a variety of research fields such that newly developed literature search techniques can be compared, improved and translated into practice. To overcome this bottleneck, we have established the RElevant LIterature SearcH consortium consisting of more than 1500 scientists from 84 countries, who have collectively annotated the relevance of over 180 000 PubMed-listed articles with regard to their respective seed (input) article/s. The majority of annotations were contributed by highly experienced, original authors of the seed articles. The collected data cover 76% of all unique PubMed Medical Subject Headings descriptors. No systematic biases were observed across different experience levels, research fields or time spent on annotations. More importantly, annotations of the same document pairs contributed by different scientists were highly concordant. We further show that the three representative baseline methods used to generate recommended articles for evaluation (Okapi Best Matching 25, Term Frequency-Inverse Document Frequency and PubMed Related Articles) had similar overall performances. Additionally, we found that these methods each tend to produce distinct collections of recommended articles, suggesting that a hybrid method may be required to completely capture all relevant articles. The established database server located at https://relishdb.ict.griffith.edu.au is freely available for the downloading of annotation data and the blind testing of new methods. We expect that this benchmark will be useful for stimulating the development of new powerful techniques for title and title/abstract-based search engines for relevant articles in biomedical research.Peer reviewe

Photochemical Inactivation Analysis of Temporal Dynamics of Postsynaptic Native AMPA Receptors in Hippocampal Slices

Postsynaptic expression of AMPA-type glutamate receptors (AMPAR) is more mobile than previously thought. Much evidence suggests that AMPAR are delivered from intracellular reserved pools to postsynaptic sites in a constitutive, as well as activity-dependent manner by exocytosis, lateral diffusion, or diffusional trapping. These notions were supported by optical monitoring of AMPAR subunits labeled with macromolecular tags such as GFP or Immunobeads, although it remains uncertain whether the mode and rate of synaptic delivery are similar to native "unlabeled" receptors. To reveal the real-time dynamics of native AMPAR in situ, photochemical inactivation of surface receptors using 6-azido-7-nitro-1,4-dihydroquinoxaline-2,3-dione (ANQX), a photoreactive AMPAR blocker, was adopted for acute hippocampal slices of mice. Because of the irreversible block due to cross-link formation between ANQX and surface AMPAR, recovery of EPSPs after photoinactivation reflects the time course of synaptic delivery of intracellular AMPAR. BriefUV illumination with fast application of ANQX resulted in persistent suppression of EPSPs for a prolonged period of up to 3 h, suggesting minimal synaptic delivery of AMPAR by exocytosis in the resting condition. Kinetic analysis of EPSP recovery clarified that the supply of postsynaptic AMPAR from the intracellular pool is dominated in the initial, but not in the later, phase of long-term potentiation (LTP). These results suggest that constitutive synaptic delivery is minimal in the resting condition at intact hippocampal synapses in a time scale of hours, while postsynaptic AMPAR are replaced with those in intracellular pools almost exclusively in an activity-dependent manner, typically shortly after LTP induction

Assessing the roles of presynaptic ryanodine receptors and adenosine receptors in caffeine-induced enhancement of hippocampal mossy fiber transmission

Caffeine robustly enhances transmitter release from the hippocampal mossy fiber terminals, although it remains uncertain whether calcium mobilization through presynaptic ryanodine receptors mediates this enhancement. In this study, we adopted a selective adenosine A1 blocker to assess relative contribution of A1 receptors and ryanodine receptors in caffeine-induced synaptic enhancement. Application of caffeine further enhanced transmission at the hippocampal mossy fiber synapse even after full blockade of adenosine A1 receptors. This result suggests that caffeine enhances mossy fiber synaptic transmission by two distinct presynaptic mechanisms, i.e., removal of A1 receptor-mediated tonic inhibition and ryanodine receptor-mediated calcium release from intracellular stores