{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,7]],"date-time":"2025-10-07T14:35:04Z","timestamp":1759847704248,"version":"3.41.2"},"reference-count":24,"publisher":"The Company of Biologists","issue":"9","license":[{"start":{"date-parts":[[2023,8,21]],"date-time":"2023-08-21T00:00:00Z","timestamp":1692576000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100000071","name":"National Institute of Child Health and Human Development","doi-asserted-by":"crossref","award":["R15HD094324","R15CA227573","R15HD111986"],"award-info":[{"award-number":["R15HD094324","R15CA227573","R15HD111986"]}],"id":[{"id":"10.13039\/100000071","id-type":"DOI","asserted-by":"crossref"}]},{"name":"National Cancer Institute"},{"DOI":"10.13039\/100013185","name":"Florida Institute of Technology","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100013185","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2023,9,15]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n               <jats:p>Fertilization is a critical step in development, yet internal fertilization events are notoriously difficult to visualize. Taking advantage of the calcium response that is a hallmark of sperm-egg fusion, we adapted the genetically encoded calcium indicator jGCaMP7s to visualize the moment of fertilization in Caenorhabditis elegans using fluorescence. We termed this tool the \u2018CaFE\u2019 reporter, for \u2018calcium during fertilization in C. elegans\u2019. The CaFE reporter produced a robust signal that recapitulated the previously reported, biphasic nature of the calcium wave and had no significant deleterious effects on worm physiology or fecundity. Calcium waves were not observed at the restrictive temperature in the spe-9(hc88) strain, in which sperm can still trigger meiotic maturation but can no longer fuse with the oocyte. Demonstrating the utility of the CaFE reporter, we analyzed polyspermy induced by inhibition of egg-3 via RNAi and observed late calcium waves in the uterus. This finding provides support to the idea that calcium release is not restricted to the first sperm fusion event during polyspermy. Establishment of the CaFE reporter in the genetically tractable and optically transparent worm provides a powerful tool to dissect the oocyte-to-embryo transition inside a living animal.<\/jats:p>","DOI":"10.1242\/bio.059832","type":"journal-article","created":{"date-parts":[[2023,8,10]],"date-time":"2023-08-10T12:33:43Z","timestamp":1691670823000},"source":"Crossref","is-referenced-by-count":3,"title":["Visualization of the biphasic calcium wave during fertilization in <i>Caenorhabditis elegans<\/i> using a genetically encoded calcium indicator"],"prefix":"10.1242","volume":"12","author":[{"given":"Katie M.","family":"Toperzer","sequence":"first","affiliation":[{"name":"Florida Institute of Technology, Biomedical Sciences Program 1 , 150 W. University Blvd, Melbourne, FL 32901 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Savannah J.","family":"Brennan","sequence":"additional","affiliation":[{"name":"Florida Institute of Technology, Biomedical Sciences Program 1 , 150 W. University Blvd, Melbourne, FL 32901 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1334-3518","authenticated-orcid":false,"given":"David J.","family":"Carroll","sequence":"additional","affiliation":[{"name":"Midwestern University 2 , Department of Biochemistry and Molecular Genetics, 19555 N 59th Ave, Glendale, AZ 85308 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1901-5598","authenticated-orcid":false,"given":"Eric A.","family":"Guisbert","sequence":"additional","affiliation":[{"name":"Florida Institute of Technology, Biomedical Sciences Program 1 , 150 W. University Blvd, Melbourne, FL 32901 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9907-9502","authenticated-orcid":false,"given":"Karen S.","family":"Kim Guisbert","sequence":"additional","affiliation":[{"name":"Florida Institute of Technology, Biomedical Sciences Program 1 , 150 W. University Blvd, Melbourne, FL 32901 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"237","published-online":{"date-parts":[[2023,8,21]]},"reference":[{"key":"2023082110532848200_BIO059832C1","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1007\/978-3-319-44820-6_4","article-title":"Signaling-mediated regulation of meiotic prophase I and transition during oogenesis","volume":"59","author":"Arur","year":"2017","journal-title":"Results Probl. Cell Differ."},{"key":"2023082110532848200_BIO059832C2","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1093\/genetics\/77.1.71","article-title":"The genetics of Caenorhabditis elegans","volume":"77","author":"Brenner","year":"1974","journal-title":"Genetics"},{"key":"2023082110532848200_BIO059832C3","doi-asserted-by":"publisher","first-page":"9192","DOI":"10.1038\/s41598-019-45517-0","article-title":"In vivo measurements reveal a single 5\u2019-intron is sufficient to increase protein expression level in Caenorhabditis elegans","volume":"9","author":"Crane","year":"2019","journal-title":"Sci. Rep."},{"key":"2023082110532848200_BIO059832C4","doi-asserted-by":"publisher","first-page":"649","DOI":"10.1038\/s41592-019-0435-6","article-title":"High-performance calcium sensors for imaging activity in neuronal populations and microcompartments","volume":"16","author":"Dana","year":"2019","journal-title":"Nat. Methods"},{"key":"2023082110532848200_BIO059832C5","doi-asserted-by":"publisher","first-page":"e61030","DOI":"10.3791\/61030","article-title":"Standardized methods for measuring induction of the heat shock response in Caenorhabditis elegans","volume":"161","author":"Golden","year":"2020","journal-title":"J. Vis. Exp."},{"key":"2023082110532848200_BIO059832C6","doi-asserted-by":"publisher","first-page":"1265","DOI":"10.1002\/dvdy.22202","article-title":"Sperm and oocyte communication mechanisms controlling C. elegans fertility","volume":"239","author":"Han","year":"2010","journal-title":"Dev. Dyn."},{"key":"2023082110532848200_BIO059832C7","doi-asserted-by":"publisher","first-page":"2","DOI":"10.1002\/(SICI)1097-0177(200005)218:1&lt;2::AID-DVDY2&gt;3.0.CO;2-W","article-title":"The Caenorhabditis elegans gonad: a test tube for cell and developmental biology","volume":"218","author":"Hubbard","year":"2000","journal-title":"Dev. Dyn."},{"key":"2023082110532848200_BIO059832C8","doi-asserted-by":"publisher","first-page":"1932","DOI":"10.1016\/j.cub.2010.09.059","article-title":"Eggshell chitin and chitin-interacting proteins prevent polyspermy in C","volume":"20","author":"Johnston","year":"2010","journal-title":"Curr. Biol."},{"key":"2023082110532848200_BIO059832C9","doi-asserted-by":"publisher","first-page":"313","DOI":"10.1016\/S1046-2023(03)00050-1","article-title":"Genome-wide RNAi screening in Caenorhabditis elegans","volume":"30","author":"Kamath","year":"2003","journal-title":"Methods"},{"key":"2023082110532848200_BIO059832C10","doi-asserted-by":"publisher","first-page":"227","DOI":"10.1093\/genetics\/146.1.227","article-title":"Distinct requirements for somatic and germline expression of a generally expressed Caernorhabditis elegans gene","volume":"146","author":"Kelly","year":"1997","journal-title":"Genetics"},{"key":"2023082110532848200_BIO059832C11","doi-asserted-by":"publisher","first-page":"215","DOI":"10.1016\/S0968-0004(03)00052-5","article-title":"How introns influence and enhance eukaryotic gene expression","volume":"28","author":"Le Hir","year":"2003","journal-title":"Trends Biochem. Sci."},{"key":"2023082110532848200_BIO059832C12","doi-asserted-by":"publisher","first-page":"435","DOI":"10.1093\/genetics\/120.2.435","article-title":"Developmental genetics of chromosome I spermatogenesis-defective mutants in the nematode Caenorhabditis elegans","volume":"120","author":"L'Hernault","year":"1988","journal-title":"Genetics"},{"key":"2023082110532848200_BIO059832C13","doi-asserted-by":"publisher","first-page":"1555","DOI":"10.1016\/j.cub.2007.08.011","article-title":"EGG-3 regulates cell-surface and cortex rearrangements during egg activation in Caenorhabditis elegans","volume":"17","author":"Maruyama","year":"2007","journal-title":"Curr. Biol."},{"key":"2023082110532848200_BIO059832C14","doi-asserted-by":"publisher","first-page":"121","DOI":"10.1006\/dbio.1996.8429","article-title":"Soma-germ cell interactions in Caenorhabditis elegans: multiple events of hermaphrodite germline development require the somatic sheath and spermathecal lineages","volume":"181","author":"Mccarter","year":"1997","journal-title":"Dev. Biol."},{"key":"2023082110532848200_BIO059832C15","doi-asserted-by":"publisher","first-page":"451","DOI":"10.1016\/S0091-679X(08)61399-0","article-title":"DNA transformation","volume":"48","author":"Mello","year":"1995","journal-title":"Methods Cell Biol."},{"key":"2023082110532848200_BIO059832C16","doi-asserted-by":"publisher","first-page":"1476","DOI":"10.1016\/j.cub.2008.08.013","article-title":"3\u2019 UTRs are the primary regulators of gene expression in the C. elegans germline","volume":"18","author":"Merritt","year":"2008","journal-title":"Curr. Biol."},{"key":"2023082110532848200_BIO059832C17","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1006\/dbio.1997.8728","article-title":"The POU gene ceh-18 promotes gonadal sheath cell differentiation and function required for meiotic maturation and ovulation in Caenorhabditis elegans","volume":"192","author":"Rose","year":"1997","journal-title":"Dev. Biol."},{"key":"2023082110532848200_BIO059832C18","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1186\/1471-213X-1-8","article-title":"Calcium dynamics during fertilization in C. elegans","volume":"1","author":"Samuel","year":"2001","journal-title":"BMC Dev Biol"},{"key":"2023082110532848200_BIO059832C19","doi-asserted-by":"publisher","first-page":"201","DOI":"10.1093\/genetics\/152.1.201","article-title":"Sperm competition in the absence of fertilization in Caenorhabditis elegans","volume":"152","author":"Singson","year":"1999","journal-title":"Genetics"},{"key":"2023082110532848200_BIO059832C20","doi-asserted-by":"publisher","first-page":"200118","DOI":"10.1098\/rsob.200118","article-title":"Modulators of calcium signalling at fertilization","volume":"10","author":"Stein","year":"2020","journal-title":"Open. Biol."},{"key":"2023082110532848200_BIO059832C21","doi-asserted-by":"publisher","first-page":"3775","DOI":"10.1534\/g3.120.401400","article-title":"Rapid self-selecting and clone-free integration of transgenes into engineered CRISPR safe harbor locations in Caenorhabditis elegans","volume":"10","author":"Stevenson","year":"2020","journal-title":"G3"},{"key":"2023082110532848200_BIO059832C22","doi-asserted-by":"publisher","first-page":"625","DOI":"10.1016\/j.celrep.2016.03.040","article-title":"The sperm TRP-3 channel mediates the onset of a Ca(2+) wave in the fertilized C. elegans oocyte","volume":"15","author":"Takayama","year":"2016","journal-title":"Cell Rep."},{"key":"2023082110532848200_BIO059832C23","doi-asserted-by":"publisher","first-page":"e2205","DOI":"10.21769\/BioProtoc.2205","article-title":"In vivo live imaging of calcium waves and other cellular processes during fertilization in Caenorhabditis elegans","volume":"7","author":"Takayama","year":"2017","journal-title":"Bio. Protoc."},{"key":"2023082110532848200_BIO059832C24","doi-asserted-by":"publisher","first-page":"571","DOI":"10.1002\/dvdy.20662","article-title":"Start me up: cell signaling and the journey from oocyte to embryo in C. elegans","volume":"235","author":"Yamamoto","year":"2006","journal-title":"Dev. Dyn."}],"container-title":["Biology Open"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.logists.com\/bio\/bio\/article-pdf\/doi\/10.1242\/bio.059832\/3102652\/bio059832.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/journals.logists.com\/bio\/bio\/article-pdf\/doi\/10.1242\/bio.059832\/3106222\/bio059832.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/journals.logists.com\/bio\/bio\/article-pdf\/doi\/10.1242\/bio.059832\/3106222\/bio059832.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,8,21]],"date-time":"2023-08-21T10:53:51Z","timestamp":1692615231000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.biologists.com\/bio\/article\/12\/9\/bio059832\/325943\/Visualization-of-the-biphasic-calcium-wave-during"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,8,21]]},"references-count":24,"journal-issue":{"issue":"9","published-print":{"date-parts":[[2023,9,15]]}},"URL":"https:\/\/doi.org\/10.1242\/bio.059832","relation":{},"ISSN":["2046-6390"],"issn-type":[{"type":"electronic","value":"2046-6390"}],"subject":[],"published":{"date-parts":[[2023,8,21]]},"article-number":"bio059832"}}