Regulation of Calcium Signaling by the Intrinsic Cell Death Pathway in C. elegans

钙及其浓度的调节对生命至关重要。钙浓度的变化被用来在细胞内传递各种类型的信息（细胞内钙信号）。钙信号级联的特定生理输出非常依赖于空间和时间限制。多种蛋白质有助于形成钙信号，在哺乳动物中，这种调节可能极其复杂。秀丽隐杆线虫是一种模式生物，广泛用于遗传方法，并表现出许多与哺乳动物相同的钙信号限制。该模型的使用使研究人员能够绕过哺乳动物系统的复杂性，并通过先进的基因组编辑方法和综合生理学更直接地显示因果关系。具体来说，该项目将研究钙信号传导中心的两类蛋白质之间的相互作用如何调节生理和行为输出。Nehrke实验室坚定地致力于促进社区互动和科学多样性，并培养下一代科学家。为了实现这些目标，该项目将继续通过基督教青年会的外展计划向罗切斯特市学区的三年级学生提供STEM教育，并将与门罗社区学院启动一个联合培训计划，允许学生在学年期间在罗切斯特大学医学中心的各种实验室之间轮换，然后在其中一个实验室进行为期十周的暑期研究奖学金。肌醇1,4,5 -三磷酸受体（IP3R）是一种内质网状（ER） Ca2+释放通道，其活性是Ca2+信号传导的核心。其他蛋白质对IP3R活性的微调使其信号库多样化。最近的证据表明，抗凋亡的Bcl-2家族成员与IP3R相互作用，这可能促进er到线粒体的Ca2+转移，促进振荡Ca2+信号传导，而不是凋亡的Ca2+过载。在哺乳动物中，研究IP3R/Bcl-2相互作用的作用是复杂的，因为多个IP3R亚型和Bcl-2家族成员与受体共享接触位点，因此很难确定具体的因果关系。相比之下，秀丽隐杆线虫只有一个单一的异构体的每一种蛋白质，然而，执行类似的功能，并具有几乎相同的氨基酸序列与哺乳动物的相互作用部位。复杂的遗传工具，包括CRISPR-Cas9介导的基因组编辑，以创建“相互作用丧失”模型，以及使用生物传感器测量局部钙信号的尖端方法，将用于验证这一假设并询问潜在的机制。研究结果将增加对细胞器间通讯及其与正常生理的相关性的理解。该奖项支持一项成功的长期推广计划，目标是罗切斯特市学区的三年级学生，他们是全国最贫困的学生之一。此外，一项新的倡议将与现有的当地基础设施协同作用，为未被充分代表的门罗社区学院学生继续进入四年制大学提供实习和暑期研究培训。还将为一名博士后提供研究培训，并为两名研究生提供指导。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Calcium, and regulating its concentration, are essential for life. Changes in calcium concentration are used to convey diverse types of information within cells (intracellular calcium signaling). The particular physiological outputs of calcium signaling cascades are exquisitely dependent upon spatial and temporal constraints. A variety of proteins help to shape calcium signals, and in mammals this regulation can be extremely complex. The nematode worm Caenorhabditis elegans is a model organism that is widely used for genetic approaches and exhibits many of the same constraints on calcium signaling as mammals. The use of this model allows researchers to circumvent the complexities of mammalian systems and more directly show cause and effect through advanced genomic editing approaches and integrative physiology. Specifically, this project will investigate how the interaction between two classes of proteins that are central to calcium signaling regulates physiological and behavioral outputs. The Nehrke Lab is strongly committed to promoting community interactions and diversity in science and to training the next generation of scientists. Toward these ends, the project will continue to provide STEM education to Rochester City School District third-graders through an outreach program at the YMCA and will initiate a joint training program with Monroe Community College that allows students to rotate among a variety of University of Rochester Medical Center laboratories during the academic year and then to undertake a ten-week summer research fellowship in one of those labs.The inositol 1, 4, 5-trisphosphate receptor (IP3R) is an endoplasmic reticular (ER) Ca2+ release channel whose activity is central to Ca2+ signaling. Fine-tuning of IP3R activity by other proteins diversifies its signaling repertoire. Recent evidence indicates that anti-apoptotic Bcl-2 family members interact with the IP3R and that this may facilitate ER-to-mitochondrial Ca2+ transfer to promote oscillatory Ca2+ signaling rather than apoptotic Ca2+ overload. In mammals, studying the role of IP3R/Bcl-2 interactions is complicated by multiple IP3R isoforms and Bcl-2 family members that share contact sites with the receptor, making it difficult to address specific cause and effect. In contrast, C. elegans has only a single isoform of each protein, which nevertheless perform similar functions and possess nearly identical amino acid sequences to their mammalian counterparts at the sites of interaction. Sophisticated genetic tools, including CRISPR-Cas9 mediated genome editing to create "loss-of-interaction" models and cutting-edge approaches using biosensors to measure localized calcium signaling, will be used to test this hypothesis and to interrogate the underlying mechanisms. Research results will increase understanding of inter-organelle communication and its relevance to normal physiology. The award supports a successful and long-standing outreach program aimed at Rochester City School District third-grade students, who are among the most impoverished in the country. In addition, a new initiative will synergize with existing local infrastructure to create shadowing and summer research traineeships for underrepresented Monroe Community College students continuing on to four-year universities. Research training for a postdoctoral fellow and mentoring experience for two graduate students will also be provided.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.