{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T21:42:40Z","timestamp":1768340560658,"version":"3.49.0"},"reference-count":51,"publisher":"Public Library of Science (PLoS)","issue":"7","license":[{"start":{"date-parts":[[2021,7,1]],"date-time":"2021-07-01T00:00:00Z","timestamp":1625097600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"JSPS KAKENHI Grant-in-Aid for Scientific Research","award":["21590204 and 15K10689"],"award-info":[{"award-number":["21590204 and 15K10689"]}]},{"name":"JSPS KAKENHI","award":["JP 16H06276 (AdAMS)"],"award-info":[{"award-number":["JP 16H06276 (AdAMS)"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Protein kinase C-delta (PKC\u03b4) has a caspase-3 recognition sequence in its structure, suggesting its involvement in apoptosis. In addition, PKC\u03b4 was recently reported to function as an anti-cancer factor. The generation of a PKC\u03b4 knockout mouse model indicated that PKC\u03b4 plays a role in B cell homeostasis. However, the <jats:italic>Pkcrd<\/jats:italic> gene, which is regulated through complex transcription, produces multiple proteins via alternative splicing. Since gene mutations can result in the loss of function of molecular species required for each tissue, in the present study, conditional PKC\u03b4 knockout mice lacking PKC\u03b4I, II, IV, V, VI, and VII were generated to enable tissue-specific deletion of PKC\u03b4 using a suitable Cre mouse. We generated PKC\u03b4-null mice that lacked whole-body expression of PKC\u03b4. PKC\u03b4+\/- parental mice gave birth to only 3.4% PKC\u03b4-\/- offsprings that deviated significantly from the expected Mendelian ratio (\u03c72(2) = 101.7, <jats:italic>P<\/jats:italic> &lt; 0.001). Examination of mice on embryonic day 11.5 (E11.5) showed the proportion of PKC\u03b4-\/- mice implanted in the uterus in accordance with Mendelian rules; however, approximately 70% of the fetuses did not survive at E11.5. PKC\u03b4-\/- mice that survived until adulthood showed enlarged spleens, with some having cardiac and pulmonary abnormalities. Our findings suggest that the lack of PKC\u03b4 may have harmful effects on fetal development, and heart and lung functions after birth. Furthermore, our study provides a reference for future studies on PKC\u03b4 deficient mice that would elucidate the effects of the multiple protein variants in mice and decipher the roles of PKC\u03b4 in various diseases.<\/jats:p>","DOI":"10.1371\/journal.pone.0253912","type":"journal-article","created":{"date-parts":[[2021,7,1]],"date-time":"2021-07-01T18:13:44Z","timestamp":1625163224000},"page":"e0253912","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":8,"title":["PKC\u03b4 deficiency inhibits fetal development and is associated with heart elastic fiber hyperplasia and lung inflammation in adult PKC\u03b4 knockout mice"],"prefix":"10.1371","volume":"16","author":[{"given":"Yuko S.","family":"Niino","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ikuo","family":"Kawashima","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yoshinobu","family":"Iguchi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hiroaki","family":"Kanda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kiyoshi","family":"Ogura","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kaoru","family":"Mita-Yoshida","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tomio","family":"Ono","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maya","family":"Yamazaki","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kenji","family":"Sakimura","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Satomi","family":"Yogosawa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kiyotsugu","family":"Yoshida","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seiji","family":"Shioda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5270-8658","authenticated-orcid":true,"given":"Takaya","family":"Gotoh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2021,7,1]]},"reference":[{"issue":"21","key":"pone.0253912.ref001","doi-asserted-by":"crossref","first-page":"7603","DOI":"10.1016\/S0021-9258(17)41010-6","article-title":"Studies on a cyclic nucleotide-independent protein kinase and its proenzyme in mammalian tissues. I. Purification and characterization of an active enzyme from bovine cerebellum","volume":"252","author":"M Inoue","year":"1977","journal-title":"J Biol Chem"},{"issue":"5082","key":"pone.0253912.ref002","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1126\/science.1411571","article-title":"Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C","volume":"258","author":"Y Nishizuka","year":"1992","journal-title":"Science"},{"issue":"4","key":"pone.0253912.ref003","doi-asserted-by":"crossref","first-page":"1341","DOI":"10.1152\/physrev.00034.2007","article-title":"Structural basis of protein kinase c isoform function","volume":"88","author":"SF Steinberg","year":"2008","journal-title":"Physiol Rev"},{"issue":"6","key":"pone.0253912.ref004","doi-asserted-by":"crossref","first-page":"831","DOI":"10.1093\/oxfordjournals.jbchem.a003294","article-title":"Protein kinase C delta (PKC delta): activation mechanisms and functions","volume":"132","author":"U Kikkawa","year":"2002","journal-title":"J Biochem"},{"key":"pone.0253912.ref005","doi-asserted-by":"crossref","first-page":"2272","DOI":"10.1100\/tsw.2010.214","article-title":"Two faces of protein kinase C\u03b4: the contrasting roles of PKC\u03b4 in cell survival and cell death","volume":"10","author":"A Basu","year":"2010","journal-title":"ScientificWorldJournal"},{"issue":"7","key":"pone.0253912.ref006","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1007\/s10875-016-0323-0","article-title":"Protein Kinase C \u03b4: A gatekeeper of immune homeostasis","volume":"36","author":"E Salzer","year":"2016","journal-title":"J Clin Immunol"},{"key":"pone.0253912.ref007","doi-asserted-by":"crossref","first-page":"10159","DOI":"10.1073\/pnas.89.21.10159","article-title":"Cell division arrest induced by phorbol ester in CHO cells overexpressing protein kinase C-\u03b4 subspecies","volume":"89","author":"T Watanabe","year":"1992","journal-title":"Proc Natl Acad Sci U S A"},{"key":"pone.0253912.ref008","doi-asserted-by":"crossref","first-page":"6148","DOI":"10.1002\/j.1460-2075.1995.tb00305.x","article-title":"Proteolytic activation of protein kinase C \u03b4 by an ICE-like protease in apoptotic cells","volume":"14","author":"Y Emoto","year":"1995","journal-title":"EMBO J"},{"issue":"6","key":"pone.0253912.ref009","doi-asserted-by":"crossref","first-page":"2399","DOI":"10.1084\/jem.184.6.2399","article-title":"Proteolytic activation of protein kinase c \u03b4 by an ice\/ced 3-like protease induces characteristics of apoptosis","volume":"184","author":"BT Ghayur","year":"1996","journal-title":"J Exp Med"},{"key":"pone.0253912.ref010","doi-asserted-by":"crossref","first-page":"1251","DOI":"10.1038\/sj.onc.1202408","article-title":"PKC\u03b4 acts as a growth and tumor suppressor in rat colonic epithelial cells","volume":"18","author":"GP Perletti","year":"1999","journal-title":"Oncogene"},{"key":"pone.0253912.ref011","doi-asserted-by":"crossref","first-page":"3067","DOI":"10.1038\/sj.onc.1208465","article-title":"Suppression of cell migration by protein kinase C\u03b4","volume":"24","author":"D Jackson","year":"2005","journal-title":"Oncogene"},{"key":"pone.0253912.ref012","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1007\/s12094-014-1241-3","article-title":"The protein kinase C (PKC) inhibitors combined with chemotherapy in the treatment of advanced non-small cell lung cancer: meta-analysis of randomized controlled trials","volume":"17","author":"LL Zhang","year":"2015","journal-title":"Clin Transl Oncol"},{"key":"pone.0253912.ref013","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.semcancer.2017.04.017","article-title":"Protein Kinase C as a tumor suppressor","volume":"48","author":"AC Newton","year":"2018","journal-title":"Semin Cancer 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cells","volume":"7","author":"ME Reyland","year":"2000","journal-title":"Cell Death Differ"},{"key":"pone.0253912.ref017","doi-asserted-by":"crossref","first-page":"1505","DOI":"10.1172\/JCI200112902","article-title":"Exacerbated vein graft arteriosclerosis in protein kinase C\u03b4-null mice","volume":"108","author":"M Leitges","year":"2001","journal-title":"J Clin Invest"},{"key":"pone.0253912.ref018","doi-asserted-by":"crossref","first-page":"865","DOI":"10.1038\/416865a","article-title":"Increased proliferation of B cells and auto-immunity in mice lacking protein kinase C\u03b4","volume":"416","author":"A Miyamoto","year":"2002","journal-title":"Nature"},{"issue":"6883","key":"pone.0253912.ref019","doi-asserted-by":"crossref","first-page":"860","DOI":"10.1038\/416860a","article-title":"Protein kinase Cdelta controls self-antigen-induced B-cell tolerance","volume":"416","author":"I Mecklenbr\u00e4uker","year":"2002","journal-title":"Nature"},{"key":"pone.0253912.ref020","doi-asserted-by":"crossref","first-page":"973","DOI":"10.1248\/bpb.24.973","article-title":"Novel protein kinase C \u03b4 isoform insensitive to caspase-3","volume":"24","author":"Y Sakurai","year":"2001","journal-title":"Biol Pharm Bull"},{"key":"pone.0253912.ref021","doi-asserted-by":"crossref","first-page":"2458","DOI":"10.1016\/j.febslet.2006.03.084","article-title":"New PKC\u03b4 family members, PKC\u03b4IV, \u03b4V, \u03b4VI, and \u03b4VII are specifically expressed in mouse testis","volume":"580","author":"T Kawaguchi","year":"2006","journal-title":"FEBS Lett"},{"key":"pone.0253912.ref022","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1016\/j.bbrc.2011.04.096","article-title":"A novel mouse PKC\u03b4 splice variant, PKC\u03b4IX, inhibits etoposide-induced apoptosis","volume":"410","author":"JD Kim","year":"2011","journal-title":"Biochem Biophys Res 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mouse embryonic stem cells","volume":"13","author":"NA Patel","year":"2005","journal-title":"Gene Expr"},{"key":"pone.0253912.ref026","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1021\/bi7019782","article-title":"Identification of a novel antiapoptotic human protein kinase C delta isoform, PKCdeltaVIII in NT2 cells","volume":"47","author":"K Jiang","year":"2008","journal-title":"Biochemistry"},{"key":"pone.0253912.ref027","doi-asserted-by":"crossref","first-page":"9299","DOI":"10.1074\/jbc.M111.313080","article-title":"Insulin promotes neuronal survival via the alternatively spliced protein kinase C\u03b4II isoform","volume":"287","author":"A Apostolatos","year":"2012","journal-title":"J Biol Chem"},{"key":"pone.0253912.ref028","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/S0888-7543(03)00072-7","article-title":"Genomic structure and promoter analysis of PKC-delta","volume":"82","author":"S Suh Kwang","year":"2003","journal-title":"Genomics"},{"key":"pone.0253912.ref029","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/j.neures.2007.01.004","article-title":"Conditional gene targeting on the pure C57BL\/6 genetic background","volume":"58","author":"M Mishina","year":"2007","journal-title":"Neurosci Res"},{"key":"pone.0253912.ref030","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1111\/j.1440-169x.2004.00775.x","article-title":"The testicular fatty acid binding protein PERF15 regulates the fate of germ cells in PERF15 transgenic mice","volume":"47","author":"T Kido","year":"2005","journal-title":"Dev Growth Differ"},{"issue":"3","key":"pone.0253912.ref031","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1111\/j.1474-9726.2009.00478.x","article-title":"Aging in inbred strains of mice: study design and interim report on median lifespans and circulating IGF1 levels","volume":"8","author":"R Yuan","year":"2009","journal-title":"Aging 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Lett"},{"key":"pone.0253912.ref035","doi-asserted-by":"crossref","first-page":"6050","DOI":"10.1093\/emboj\/cdf606","article-title":"Nuclear import of PKC\u03b4 is required for apoptosis: Identification of a novel nuclear import sequence","volume":"21","author":"TA DeVries","year":"2002","journal-title":"EMBO J"},{"issue":"3","key":"pone.0253912.ref036","first-page":"1","article-title":"PLOD3 suppression exerts an anti-tumor effect on human lung cancer cells by modulating the PKC-delta signaling pathway","volume":"10","author":"JH Baek","year":"2019","journal-title":"Cell Death Dis"},{"key":"pone.0253912.ref037","doi-asserted-by":"crossref","first-page":"9728","DOI":"10.1074\/jbc.M507851200","article-title":"Suppression of apoptosis in the protein kinase C\u03b4 null mouse in vivo","volume":"281","author":"MJ Humphries","year":"2006","journal-title":"J Biol 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kinase C-delta (PKC\u03b4) disrupts fertilization and embryonic development","volume":"82","author":"W Ma","year":"2015","journal-title":"Mol Reprod Dev"},{"key":"pone.0253912.ref042","doi-asserted-by":"crossref","first-page":"2062","DOI":"10.1152\/ajpheart.2001.281.5.H2062","article-title":"Protein kinase C isoform expression and activity in the mouse heart","volume":"281","author":"KL Schreiber","year":"2001","journal-title":"Am J Physiol\u2014Hear Circ Physiol"},{"key":"pone.0253912.ref043","doi-asserted-by":"crossref","DOI":"10.1126\/scisignal.aaa1855","article-title":"Combined cardiomyocyte PKC\u03b4 and PKC\u03b5 gene deletion uncovers their central role in restraining developmental and reactive heart growth","volume":"8","author":"M Song","year":"2015","journal-title":"Sci Signal"},{"key":"pone.0253912.ref044","doi-asserted-by":"crossref","first-page":"2161","DOI":"10.1002\/art.38008","article-title":"protein kinase c\u03b4 deficiency causes mendelian sle with b cell-defective 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