Acid/base sensing and regulation of multiple physiological processes in fish

鱼类多种生理过程的酸/碱传感和调节

• 批准号: 1754994
• 负责人: Martin Tresguerres
• 金额: $ 61.2万
• 依托单位: University of California-San Diego Scripps Inst of Oceanography
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1754994&HistoricalAwards=false
• 关键词: Acid; base; sensing; regulation; multiple

All organisms must be able to sense and regulate the pH level in their fluids to avoid cell malfunction that can lead to sickness and, in extreme cases, death. Prior work identified the a soluble enzyme in gills from sharks and rays as a novel molecular sensor and regulator of high blood pH. The current project will explore the putative role of this soluble enzyme in sensing and regulating low blood pH in shark gills, and will expand those studies to gills of other aquatic vertebrates, bony fishes. In addition, it will explore novel functions in red blood cells and heart muscle cells from both shark and bony fishes. Because those two cell types are greatly affected by pH, this soluble enzyme may play important roles in essential physiological processes such as oxygen uptake and delivery by red blood cells, and in contraction and heart beat rate by heart muscle cells. This project will develop biochemical, cellular, and physiological tools, many of which could be subsequently adapted to test similar hypotheses in other organisms. This information will help understand and predict how organisms respond to environmental and metabolic stress, as well as informing the management of wild and farmed populations in changing ocean conditions. This project will provide research training opportunities for undergraduate and graduate students and postdoctoral researchers and develop onsite and online educational outreach activities for K-12 students in collaboration with SeaCamp San Diego, a local marine science camp for kids and teenagers.The enzyme soluble adenylyl cyclase (sAC) produces the ubiquitous messenger molecule cyclic AMP in response to changes in CO2, pH and HCO3- levels. Since its discovery in 1999, sAC has been found to regulate diverse physiological processes in multiple animal phyla ranging from coral to mammals, and is therefore deemed an evolutionarily conserved acid/base sensor. The proposed research will: (1) continue research on elasmobranch gill cells to elucidate if sAC regulates acid secretion; (2) perform the initial molecular and biochemical characterization of sAC in a teleost fish (the rainbow trout); (3) explore the presence of multiple sAC splice variants in trout, and their putative differential subcellular localization; (4) characterize the putative role of sAC in trout gill cells in sensing and regulating blood acid/base status; (5) study a potential role of sAC in regulating oxygen binding in erythrocytes from shark and trout; (6) explore the role of sAC in cardiomyocytes, where it is hypothesized to regulate contractibility. To achieve those aims, a variety of experimental methods will be used including primary cell cultures, expression of GFP-tagged proteins in cultured trout fibroblasts, intracellular pH and Ca2+ measurements, erythrocyte oxygen-binding curves, sarcomere shortening, and immunocytochemistry. Given the established importance of acid/base status and cAMP on the physiology of gill cells, erythrocytes and cardiomyocytes, this research has the potential to find novel sAC-dependent mechanisms that regulate physiological functions essential to fish, and which potentially extend to other animals. The simultaneous study of elasmobranch and teleost adds a comparative and evolutionary component.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

所有生物体都必须能够感知和调节体液中的 pH 值，以避免细胞功能障碍，从而导致疾病，在极端情况下甚至导致死亡。此前的研究发现，鲨鱼和鳐鱼的鳃中存在一种可溶性酶，可作为一种新型分子传感器和高血液 pH 值调节剂。当前的项目将探索这种可溶性酶在感知和调节鲨鱼鳃低血液 pH 值方面的假定作用，并将这些研究扩展到其他水生脊椎动物、硬骨鱼的鳃。此外，还将探索鲨鱼和硬骨鱼的红细胞和心肌细胞的新功能。由于这两种细胞类型受 pH 值影响很大，因此这种可溶性酶可能在重要的生理过程中发挥重要作用，例如红细胞的氧气吸收和输送，以及心肌细胞的收缩和心率。该项目将开发生化、细胞和生理学工具，其中许多工具随后可以用于在其他生物体中测试类似的假设。这些信息将有助于了解和预测生物体如何应对环境和代谢压力，并为不断变化的海洋条件下野生和养殖种群的管理提供信息。该项目将为本科生、研究生和博士后研究人员提供研究培训机会，并与当地儿童和青少年海洋科学营 SeaCamp San Diego 合作，为 K-12 学生开展现场和在线教育推广活动。 (sAC) 响应 CO2、pH 和 HCO3- 水平的变化，产生普遍存在的信使分子环 AMP。自 1999 年被发现以来，sAC 被发现可以调节从珊瑚到哺乳动物等多种动物门的多种生理过程，因此被认为是一种进化上保守的酸/碱传感器。拟议的研究将：（1）继续对软骨鱼鳃细胞进行研究，以阐明 sAC 是否调节酸分泌； (2) 对硬骨鱼（虹鳟鱼）中的 sAC 进行初始分子和生化表征； (3) 探索鳟鱼中多个sAC剪接变体的存在，以及它们假定的差异亚细胞定位； (4) 表征鳟鱼鳃细胞中 sAC 在感知和调节血液酸/碱状态中的假定作用； (5) 研究 sAC 在调节鲨鱼和鳟鱼红细胞氧结合方面的潜在作用； (6) 探索 sAC 在心肌细胞中的作用，假设其调节收缩性。为了实现这些目标，将使用各种实验方法，包括原代细胞培养、培养鳟鱼成纤维细胞中 GFP 标记蛋白的表达、细胞内 pH 和 Ca2+ 测量、红细胞氧结合曲线、肌节缩短和免疫细胞化学。鉴于酸/碱状态和 cAMP 对鳃细胞、红细胞和心肌细胞生理学的重要性已被证实，这项研究有可能发现新的 sAC 依赖性机制，调节鱼类必需的生理功能，并有可能扩展到其他动物。对软骨鱼和硬骨鱼的同时研究增加了比较和进化的成分。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

A multi-tasking stomach: functional coexistence of acid–peptic digestion and defensive body inflation in three distantly related vertebrate lineages

多任务胃：三个远亲脊椎动物谱系中酸消化和防御性身体膨胀的功能共存

• DOI: 10.1098/rsbl.2021.0583
• 发表时间: 2022
• 期刊: Biology Letters
• 影响因子: 3.3
• 作者: Ferreira, P.;Kwan, G. T.;Haldorson, S.;Rummer, J. L.;Tashiro, F.;Castro, L. F.;Tresguerres, M.;Wilson, J. M.
• 通讯作者: Wilson, J. M.

A novel acidification mechanism for greatly enhanced oxygen supply to the fish retina

• DOI: 10.7554/elife.58995
• 发表时间: 2020-08-25
• 期刊: ELIFE
• 影响因子: 7.7
• 作者: Damsgaard, Christian;Lauridsen, Henrik;Brauner, Colin J.
• 通讯作者: Brauner, Colin J.

Differential glycogen utilization in shark acid- and base-regulatory gill cells

鲨鱼酸碱调节鳃细胞的糖原利用差异

• DOI: 10.1242/jeb.199448
• 发表时间: 2019
• 期刊: The Journal of Experimental Biology
• 作者: Roa, Jinae N.;Tresguerres, Martin
• 通讯作者: Tresguerres, Martin

Evolutionary links between intra- and extracellular acid-base regulation in fish and other aquatic animals

• DOI: 10.1002/jez.2367
• 发表时间: 2020-05-26
• 期刊: JOURNAL OF EXPERIMENTAL ZOOLOGY PART A-ECOLOGICAL AND INTEGRATIVE PHYSIOLOGY
• 影响因子: 2.8
• 作者: Tresguerres, Martin;Clifford, Alexander M.;Brauner, Colin J.
• 通讯作者: Brauner, Colin J.

Soluble adenylyl cyclase coordinates intracellular pH homeostasis and biomineralization in calcifying cells of a marine animal

• DOI: 10.1152/ajpcell.00524.2022
• 发表时间: 2023-03-01
• 期刊: AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY
• 影响因子: 5.5
• 作者: Chang，William Weijen;Thies，Angus B.;Hu，Marian Y.
• 通讯作者: Hu，Marian Y.