Collaborative Research: Identifying osmosensitive molecular targets using a unique vertebrate model

合作研究：使用独特的脊椎动物模型识别渗透敏感分子靶标

• 批准号: 1755131
• 负责人: Jason Breves
• 金额: $ 20.61万
• 依托单位: Skidmore College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1755131&HistoricalAwards=false
• 关键词: Collaborative; Research; Identifying; osmosensitive; molecular

This research addresses how specialized cells sense and respond to changes in the external environment and, in turn, control the physiological systems that maintain a stable internal environment. The regulation of salt and water balance, or osmoregulation, is fundamental to most vertebrates, including humans. Detection of a change in osmotic concentration or salinity is the necessary first step in successful osmoregulation. Nevertheless, understanding of how osmoreceptor cells work remains incomplete because of their complex structure and arrangement among other cells in the brain of many animals, making it difficult to develop effective experimental approaches. This problem is solved by study of the prolactin cells of the tilapia fish, because these cells are arranged into a nearly homogeneous mass in the pituitary gland. The hormone prolactin is involved in osmoregulation in tilapia, an estuarine fish that can thrive in salinities ranging from fresh water to greater than full-strength seawater. This project uses the tilapia prolactin cell model as a means to uniquely connect the operation of osmoreceptor cells with environmental adaptation at the organismal level. The collaborative study links the research, teaching, and outreach programs at the University of Hawai'i at Manoa and Skidmore College and will reveal the mechanisms supporting osmoreception in a manner that is applicable to all vertebrates. It will immerse graduate and undergraduate students from underrepresented groups in rigorous and interdisciplinary educational programs at the collaborating institutions. The scientific outcomes of this research will be conveyed to a broad audience through institutional outreach educational programs, workshops and symposia, posters, lay press articles, guided visits to research facilities, and a website.The overarching goal of this collaborative project is to further the understanding of the molecular mechanisms of osmoreception, a fundamental, albeit understudied, sensory modality. Tilapia prolactin cells have allowed for the identification of some of the mechanisms underlying osmoreception, such as extracellular Ca2+ entry through mechanosensitive transient-receptor-potential vanilloid channels and cAMP accumulation. Nonetheless, little is known about how the prolactin (prl) gene is activated by hyposmotic stimuli. By employing a dual luciferase reporter assay system in embryonic kidney cells (HEK293) expressing a vector containing the tilapia prl promoter, the specific promoter regions that respond to secondary messengers activated by changes in extracellular osmolality will be characterized, thereby allowing for the identification of novel transcription factors that mediate hyposmotic induction of the prl gene. Based on the discovery of specific regulators of prolactin, other vertebrate genomes can be searched for conserved gene sequences that respond to changes in extracellular osmolality. The project will advance the field by identifying promoter sequences that mediate transcriptional regulation of hyposmotically-induced genes in vertebrates. Graduate and undergraduate students will benefit from involvement in the research project and the planned educational activities. The results will be broadly disseminated, including via a website and public outreach programs, including those in association with the University of Hawai'i Institute of Marine Biology and Sea Grant.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.

这项研究涉及专门的细胞如何感知和响应外部环境的变化，进而控制维持稳定内部环境的生理系统。盐和水平衡的调节，或盐调节，是大多数脊椎动物，包括人类的基础。检测渗透浓度或盐度的变化是成功调节盐度的必要的第一步。尽管如此，由于许多动物大脑中的其他细胞的复杂结构和排列，对β受体细胞如何工作的理解仍然不完整，因此很难开发有效的实验方法。这个问题通过对罗非鱼催乳素细胞的研究得到了解决，因为这些细胞在脑垂体中排列成几乎均匀的团块。激素催乳素参与了罗非鱼的新陈代谢调节，罗非鱼是一种河口鱼类，可以在从淡水到高于全浓度海水的盐度范围内茁壮成长。该项目使用罗非鱼催乳素细胞模型作为一种手段，在生物体水平上独特地将催乳素受体细胞的运作与环境适应联系起来。这项合作研究将夏威夷大学马诺阿分校和斯基德莫尔学院的研究、教学和推广项目联系起来，并将揭示以适用于所有脊椎动物的方式支持生物接收的机制。它将使来自代表性不足群体的研究生和本科生沉浸在合作机构的严格和跨学科的教育计划中。这项研究的科学成果将通过机构外展教育计划，讲习班和研讨会，海报，新闻文章，研究设施的指导参观，和一个website.The的首要目标传达给广大观众这个合作项目是进一步了解的分子机制的接收，一个基本的，虽然研究不足，感觉模态。罗非鱼催乳素细胞已经允许识别一些潜在的细胞外钙离子受体的机制，如通过机械敏感的瞬时受体电位香草酸通道和cAMP积累的细胞外Ca 2+进入。尽管如此，人们对催乳素（prl）基因是如何被低渗刺激激活的知之甚少。通过在表达含有罗非鱼prl启动子的载体的胚胎肾细胞（HEK293）中采用双荧光素酶报告基因测定系统，将表征响应于由细胞外渗透压浓度的变化激活的第二信使的特异性启动子区域，从而允许鉴定介导prl基因的低渗诱导的新型转录因子。基于催乳素的特异性调节因子的发现，可以在其他脊椎动物基因组中搜索响应细胞外渗透压变化的保守基因序列。该项目将通过鉴定介导脊椎动物低渗诱导基因转录调控的启动子序列来推进该领域。研究生和本科生将受益于参与研究项目和计划的教育活动。结果将被广泛传播，包括通过一个网站和公众宣传计划，包括那些与夏威夷大学海洋生物学和海洋赠款研究所。这个奖项反映了NSF的法定使命，并已被认为是值得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的支持。

项目成果

Molecular targets of prolactin in mummichogs (Fundulus heteroclitus): Ion transporters/channels, aquaporins, and claudins

mummichogs（眼底异斜）催乳素的分子靶标：离子转运蛋白/通道、水通道蛋白和密蛋白

• DOI: 10.1016/j.ygcen.2022.114051
• 发表时间: 2022
• 期刊: General and Comparative Endocrinology
• 影响因子: 2.7
• 作者: Breves, Jason P.;Puterbaugh, Katie M.;Bradley, Serena E.;Hageman, Annie E.;Verspyck, Adrian J.;Shaw, Lydia H.;Danielson, Elizabeth C.;Hou, Yubo
• 通讯作者: Hou, Yubo

Salinity-dependent expression of ncc2 in opercular epithelium and gill of mummichog (Fundulus heteroclitus)

Mummichog (Fundulusheteroclitus) 鳃盖上皮和鳃中 ncc2 的盐度依赖性表达

• DOI: 10.1007/s00360-020-01260-x
• 发表时间: 2020
• 期刊: Journal of Comparative Physiology B
• 作者: Breves, Jason P.;Starling, Julie A.;Popovski, Christine M.;Doud, James M.;Tipsmark, Christian K.
• 通讯作者: Tipsmark, Christian K.

Prolactin controls branchial clcn2c but not atp1a1a.2 in zebrafish Danio rerio

催乳素控制斑马鱼斑马鱼鳃 clcn2c 但不控制 atp1a1a.2

• DOI: 10.1111/jfb.13854
• 发表时间: 2018
• 期刊: Journal of Fish Biology
• 影响因子: 2
• 作者: Breves, Jason P.

Tilapia prolactin cells are thermosensitive osmoreceptors

罗非鱼催乳素细胞是热敏渗透压感受器

• DOI: 10.1152/ajpregu.00027.2022
• 发表时间: 2022
• 期刊: Integrative and Comparative Physiology
• 作者: Woo, Daniel W.;Malintha, G. H.;Celino-Brady, Fritzie T.;Yamaguchi, Yoko;Breves, Jason P.;Seale, Andre P.
• 通讯作者: Seale, Andre P.

Na+/HCO3− cotransporter 1 (nbce1) isoform gene expression during smoltification and seawater acclimation of Atlantic salmon

• DOI: 10.1007/s00360-022-01443-8
• 发表时间: 2022-06
• 期刊: Journal of Comparative Physiology B
• 作者: J. Breves;Ian S. H. McKay;Victor Koltenyuk;Nastasia N Nelson;S. Lema;S. McCormick