CENTRAL RESPIRATORY CHEMORECEPTION IN AMPHIBIANS

两栖动物的中枢呼吸化学感受器

• 批准号: 7959251
• 负责人: MATTHEW J GDOVIN
• 金额: $ 7.56万
• 依托单位: UNIVERSITY OF TEXAS SAN ANTONIO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7959251
• 关键词: Acidosis; Ammonium Chloride; Amphibia; Area; Bathing; Brain Stem; Calibration; Cells; Computer Retrieval of Information on Scientific Projects Database; Data; Environmental air flow; Exposure to; Fluorescence; Frequencies; Funding; Grant; In Vitro; Institution; Lung; Nerve; Neurons; Neurosciences Research; Optics; Peer Review; Preparation; Process; Publications; Reporting; Research; Research Personnel; Resources; Rest; Source; Tadpoles; Techniques; Ultraviolet Rays; United States National Institutes of Health; interest; research study; respiratory

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. A. Specific Aims The Specific Aims in the application have not changed and experiments have been conducted in accordance with these aims. We have progressed in our abilities to perform optical recordings of intracellular pH (pHi) from neurons in the spontaneously rhythmic in vitro tadpole brainstem preparation. B. Studies and Results Upon the completion of the BCECF calibration curve, we were then able to record emitted fluorescence rations from both chemo- and non-chemosensitive regions while simultaneously recording whole nerve respiratory activities. We report significant differences in resting pHi between neurons located in known chemosensitive areas and neurons from non-chemosensitive areas. In addition, we utilized the ammonium chloride prepulse technique to induce intracellular acidification while maintaining a control bath pH of 7.8. Finally, we were able to demonstrate a significant negative correlation between decreases in pHi in chemosensitive neurons and increases in lung burst frequency. We have successfully demonstrated a correlation between pHi and ventilation, as proposed in our Specific Aims, and are in the process of submitting these data for peer review publication. We also explored a potential new technique to induce intracellular acidosis rather than use the ammonium chloride prepulse technique, as the ammonium chloride prepulse technique induces intracellular acidification to all brainstem neurons. We hoped to use "uncaging" of H+ via exposure to ultraviolet light, thereby allowing us to focally induce decreases in pHi in only those cells of interest.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 A. 具体目标 申请中的具体目标没有改变，并且已经根据这些目标进行了实验。 我们在自发节律体外蝌蚪脑干制备中对神经元细胞内 pH (pHi) 进行光学记录的能力取得了进展。 B. 研究和结果 完成 BCECF 校准曲线后，我们就能够记录化学敏感区域和非化学敏感区域的发射荧光比率，同时记录整个神经呼吸活动。 我们报告了位于已知化学敏感区域的神经元和来自非化学敏感区域的神经元之间的静息 pHi 显着差异。 此外，我们利用氯化铵预脉冲技术诱导细胞内酸化，同时保持控制浴 pH 值为 7.8。 最后，我们能够证明化学敏感神经元 pHi 的降低与肺爆发频率的增加之间存在显着的负相关。 正如我们的具体目标中所提出的，我们已经成功证明了 pHi 和通风之间的相关性，并且正在提交这些数据以供同行评审发表。 我们还探索了一种潜在的新技术来诱导细胞内酸中毒，而不是使用氯化铵预脉冲技术，因为氯化铵预脉冲技术会诱导所有脑干神经元的细胞内酸化。 我们希望通过暴露于紫外线来“释放”H+，从而使我们能够仅在那些感兴趣的细胞中集中诱导 pHi 降低。