Development Growth & Differentiation最新文献

{"title":"Universal Improvement of In Situ Hybridization Chain Reaction by Reducing Background Signals Caused by Single Probes","authors":"Yudai Kuboe,&nbsp;Kensuke Nakanishi,&nbsp;Ichiro Tazawa,&nbsp;Keisuke Nakajima","doi":"10.1111/dgd.70018","DOIUrl":"10.1111/dgd.70018","url":null,"abstract":"<div>\u0000 \u0000 <p>The in situ hybridization chain reaction (HCR) method involves designing multiple target sequences and a pair of split probes for each target. One split probe contains the complementary sequence for half of the target along with part of the initiation sequence. The other split probe contains the complementary sequence for the remaining half of the target sequence and the rest of the initiation sequence. The complete initiation sequence composed of both probes is capable of initiating a chain reaction of hairpin DNAs. This theoretical mechanism minimizes the background signal caused by a single probe; however, very low background signals have been observed in experiments. While these weak signals are not a significant problem in many cases, they can interfere with experiments if the expression of the target gene is very low, making the background signal noticeable. Reducing such background signals would benefit many scientists working with diverse species and sample types. To address this issue, we hypothesized that a single probe could bind and open the hairpin DNA through partial complementary sequences, acting as a bridge between hairpin DNA and samples through nonspecific binding. Our findings show that the addition of random oligonucleotides during the pre-hybridization and hybridization steps reduced background signals by approximately 3–90 times. This simple and easy modification of the in situ HCR technique improves the signal-to-noise ratio and facilitates the detection of mRNAs with very low expression levels.</p>\u0000 </div>","PeriodicalId":50589,"journal":{"name":"Development Growth & Differentiation","volume":"67 6","pages":"336-343"},"PeriodicalIF":1.0,"publicationDate":"2025-07-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144627639","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Meeting Report: The First Evo-Devo Young Researchers Meeting","authors":"Yui Uchida,&nbsp;Tasuku Ishida,&nbsp;Satomi Ono,&nbsp;Shunya Kuroda,&nbsp;Aimi Kobayashi,&nbsp;Naoki Konno,&nbsp;Yoshitaka Tanaka,&nbsp;Tetsuro Nishizawa,&nbsp;Junki Yoshida,&nbsp;Ken-ichiro Yoshimoto,&nbsp;Mizuho Yoneda","doi":"10.1111/dgd.70016","DOIUrl":"10.1111/dgd.70016","url":null,"abstract":"<p>The official poster for the First Evo-Devo Young Researchers Meeting. It features the meeting title, themes (“Evo-Devo so far” and “Evo-Devo in the future”), invited speakers, program highlights, and logistical details. The content is written in Japanese. \u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture><p></p>\u0000 </div>\u0000 </figure>\u0000 </p>","PeriodicalId":50589,"journal":{"name":"Development Growth & Differentiation","volume":"67 6","pages":"331-335"},"PeriodicalIF":1.0,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144555626","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"GLI3 in Archaic Humans: Possible Contributions to Human Skeletal and Brain Evolution","authors":"Tadashi Nomura,&nbsp;Ako Agata,&nbsp;Nguyen Thi My Trinh","doi":"10.1111/dgd.70015","DOIUrl":"10.1111/dgd.70015","url":null,"abstract":"<div>\u0000 \u0000 <p>Neanderthals, an extinct hominid, lived in Eurasia until about 40,000 years ago. According to fossils, Neanderthals had distinctive anatomical features compared to modern humans, including a long front-to-back cranium, low frontal bones, and strong skeletal formation. Furthermore, Neanderthals had large brains similar to those of modern humans, but their brain morphology was different from ours, suggesting that they had different cognitive abilities than modern humans. Recent archaic human genome analysis has unveiled genetic changes underlying Neanderthals' or modern human–specific anatomical and physiological traits. In this review, we focus on the role of <i>GLI3</i>, a key molecule that mediates Hedgehog signaling during vertebrate organogenesis. We discuss possible contributions of <i>GLI3</i>-mediated hedgehog signaling to human anatomical diversifications, including neocortical structures, which provide insights into the genetic and developmental bases for modern human evolution.</p>\u0000 </div>","PeriodicalId":50589,"journal":{"name":"Development Growth & Differentiation","volume":"67 6","pages":"306-313"},"PeriodicalIF":1.0,"publicationDate":"2025-06-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144477726","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Syncytin-1 Is Responsible for the Fusion Between Human Trophoblasts and Endometrial Stromal Cells","authors":"Akira Oike,&nbsp;Shun Shibata,&nbsp;Takahiro Arima,&nbsp;Hiroaki Okae","doi":"10.1111/dgd.70014","DOIUrl":"10.1111/dgd.70014","url":null,"abstract":"<div>\u0000 \u0000 <p>Syncytiotrophoblasts (STs) are multinucleated cells formed by the fusion of trophoblasts and play critical roles in placental development and function. Retrovirus-derived fusogenic proteins, known as syncytins, regulate trophoblast fusion by interacting with specific receptors. In humans, two syncytins, Syncytin-1 (Syn1) and Syncytin-2 (Syn2), have been identified. Although both are considered to be involved in ST formation, the expression patterns of Syn1, Syn2, and their receptors differ significantly, suggesting that Syn1 and Syn2 may have distinct roles. To investigate the functional differences of syncytins in human trophoblasts, we generated <i>Syn1</i> and <i>Syn2</i> knockout (KO) human trophoblast stem cells (hTSCs). ST differentiation assays revealed that Syn2 plays a predominant role in trophoblast-trophoblast fusion. We also examined the fusion between hTSCs and endometrial stromal cells (EMSCs), as trophoblasts and EMSCs interact directly during implantation, and genes encoding Syn1 and Syn2 receptors are expressed in EMSCs. This analysis revealed that hTSCs do fuse with EMSCs, and in contrast to trophoblast-trophoblast fusion, Syn1 plays a predominant role in trophoblast-EMSC fusion. Given that fusion-capable Syn1 is found only in primates whose embryos invade deep into the uterus, we hypothesize that Syn1 may be more involved in implantation rather than in trophoblast-trophoblast fusion.</p>\u0000 </div>","PeriodicalId":50589,"journal":{"name":"Development Growth & Differentiation","volume":"67 5","pages":"270-278"},"PeriodicalIF":1.7,"publicationDate":"2025-06-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144287018","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Step-by-Step Protocol for Making a Knock-In Xenopus laevis to Visualize Endogenous Gene Expression","authors":"Norie Kagawa,&nbsp;Yoshihiko Umesono,&nbsp;Ken-ichi T. Suzuki,&nbsp;Makoto Mochii","doi":"10.1111/dgd.70011","DOIUrl":"10.1111/dgd.70011","url":null,"abstract":"<div>\u0000 \u0000 <p>We established a novel knock-in technique, New and Easy <i>Xenopus</i> Targeted integration (<i>NEXTi</i>), to recapitulate endogenous gene expression by reporter expression. <i>NEXTi</i> is a CRISPR-Cas9-based method to integrate a donor DNA containing a reporter gene (<i>egfp</i>) into the target 5′ untranslated region (UTR) of the <i>Xenopus laevis</i> genome. It enables us to track eGFP expression under the regulation of endogenous promoter/enhancer activities. We obtained about 2% to 13% of knock-in vector-injected embryos showing eGFP signal in a tissue-specific manner, targeting <i>krt.12.2.L</i>, <i>myod1.S</i>, <i>sox2.L</i>, and <i>bcan</i>.<i>S</i> loci, as previously reported. In addition, F1 embryos which show stable eGFP signals were obtained by outcrossing the matured injected frogs with wild-type animals. Integrations of donor DNAs into target 5′ UTRs were confirmed by PCR amplification and sequencing. Here, we describe the step-by-step protocol for preparation of donor DNA and single guide RNA, microinjection, and genotyping of F1 animals for the <i>NEXTi</i> procedure.</p>\u0000 </div>","PeriodicalId":50589,"journal":{"name":"Development Growth & Differentiation","volume":"67 5","pages":"293-302"},"PeriodicalIF":1.7,"publicationDate":"2025-06-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144259272","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimization of Culture and Transfection Methods for Primary Snake Cells","authors":"Shoma Kuriyama,&nbsp;Keisuke Shigematsu,&nbsp;Seung June Kwon,&nbsp;Ryusei Kuwata,&nbsp;Yuji Atsuta","doi":"10.1111/dgd.70013","DOIUrl":"10.1111/dgd.70013","url":null,"abstract":"<div>\u0000 \u0000 <p>Snakes serve as important models for understanding how changes in genes and genome sequences drive vertebrate morphological evolution. However, the lack of established primary culture methods and gene delivery techniques for snake cells has hindered functional analyses of evolutionarily modified genes and genomic elements. Here, we optimized primary culture conditions and screened for efficient transfection methods using corn snake embryonic fibroblasts. Our culture optimization experiments revealed that TeSR medium, designed for stem cells, with fetal bovine serum supplementation and incubation at 28°C provided a suitable condition for primary snake fibroblasts. Transcriptome analysis further demonstrated that under this optimized condition, genes associated with cytoskeletal organization, extracellular matrix components, and sterol biosynthetic process were upregulated, likely promoting snake cell proliferation. Additionally, screening of various gene transfection methods identified two chemical transfection reagents and an electroporation technique that yielded high plasmid introduction efficiency in cultured snake fibroblasts. These findings enhance the utility of snake cells and pave the way for functional analyses of genes and genomic elements using snake cell-based systems.</p>\u0000 </div>","PeriodicalId":50589,"journal":{"name":"Development Growth & Differentiation","volume":"67 5","pages":"279-292"},"PeriodicalIF":1.7,"publicationDate":"2025-06-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144250660","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transcriptome Analysis Suggested Striking Transition Around the End of Epiboly in the Gene Regulatory Network Downstream of the Oct4-Type POU Gene in Zebrafish Embryos","authors":"Masaaki Ikeda,&nbsp;Kana Kobayashi,&nbsp;Yukiko Nakayama-Sadakiyo,&nbsp;Yuto Sato,&nbsp;Ayano Tobita,&nbsp;Mika Saito,&nbsp;Kyo Yamasu","doi":"10.1111/dgd.70012","DOIUrl":"10.1111/dgd.70012","url":null,"abstract":"<p>Zebrafish <i>pou5f3</i> encodes a Class V POU transcription factor, Pou5f3, which regulates various developmental processes, including neurogenesis and brain formation. In the current study, we attempted to comprehensively identify the Pou5f3 downstream genes around the end of epiboly, when the competence of the mid-hindbrain region to Pou5f3 suppression changes drastically, by the microarray method and a heat-inducible dominant-interference <i>pou5f3</i> gene (<i>en-pou5f3</i>) that functionally suppresses <i>pou5f3</i>. At late epiboly and early somitogenesis stages, we identified genes whose expression was altered in <i>en-pou5f3</i>-induced embryos, revealing numerous genes regulated differently by Pou5f3 at the two stages. The validity of the microarray data was confirmed by whole mount in situ hybridization and quantitative RT-PCR. Many of the downstream genes were implicated by the Gene ontology (GO) analyses in transcriptional regulation and neural development and were enriched with <i>sox</i> genes and bHLH genes such as <i>her</i> genes. Interestingly, we noticed a tendency that Notch-dependent <i>her</i> genes were activated, whereas Notch-independent <i>her</i> genes were downregulated by Pou5f3 suppression. Among the Notch-independent <i>her</i> genes, <i>her3</i>, which is orthologous to mammalian <i>Hes3</i>, was suggested to be strongly activated endogenously by Pou5f3. In the upstream DNA of this gene, we found two noncoding conserved sequences (NCRs), which harbored consensus binding sites for Pou5f3, Sox, and Nanog. We further showed in reporter assays that the transcriptional regulatory activity of the <i>her3</i> upstream DNA was strongly enhanced by SoxB1, and this SoxB1-mediated activation was weakened by Pou5f3. Deletion experiments showed that both upstream NCRs were involved in transcriptional repression.</p>","PeriodicalId":50589,"journal":{"name":"Development Growth & Differentiation","volume":"67 5","pages":"245-269"},"PeriodicalIF":1.7,"publicationDate":"2025-06-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/dgd.70012","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144259273","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Measuring intraventricular pressure in developing mouse embryos: Uncovering a repetitive mechanical cue for brain development","authors":"Mami Akaike,&nbsp;Jun Hatakeyama,&nbsp;Yuta Nakashima,&nbsp;Kenji Shimamura","doi":"10.1111/dgd.70010","DOIUrl":"10.1111/dgd.70010","url":null,"abstract":"<p>In living organisms, including humans, the developmental processes that construct their morphology from a single fertilized egg are influenced not only by genetic regulation but also by various external factors. One such factor is mechanical stimulation. Although mechanical forces are suggested to contribute to brain formation during development, quantitative information on intraventricular pressure during neurogenesis remains limited. We developed a high time-resolution system efficiently using a piezoresistive sensor to measure brain intraventricular pressure in mouse embryos from E12.5 to E16.5 (embryonic stages in days). Ex utero measurements revealed intraventricular pressure increasing from 53.76 ± 4.16 Pa at E12.5 to 158.10 ± 19.94 Pa by E16.5. In utero analyses uncovered striking periodicity in sync with uterine contractions, reaching up to 1430 ± 195.2 Pa at E12.5, indicating dynamic mechanical stimuli beyond ex utero observations. Additionally, perforation experiments at E9.0–E15.5 showed rapid neuroepithelial thickening and apical surface contraction upon pressure release, indicative of a tensile effect by the positive intraventricular pressure. This effect diminished after E15.5, implying that tension wanes or the neuroepithelium becomes more robust. These results highlight the dynamic nature of embryonic intraventricular pressure, governed by internal fluid production and uterine forces, and emphasize the importance of mechanical cues in neuroepithelial architecture. Our findings provide a steppingstone to clarify how mechanical forces integrate with genetic and molecular processes to shape normal brain development and may render new perspectives on brain evolution.</p>","PeriodicalId":50589,"journal":{"name":"Development Growth & Differentiation","volume":"67 5","pages":"230-244"},"PeriodicalIF":1.7,"publicationDate":"2025-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/dgd.70010","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144052031","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The long-term survival of enteroendocrine cells depends on their subtype and is linked to peripheral sensory innervation","authors":"Salsabila Luthfi Sesotyosari,&nbsp;Masato Kinoshita,&nbsp;Mukhamad Sunardi,&nbsp;Mo Lihan,&nbsp;Akimasa Orii,&nbsp;Takaya Abe,&nbsp;Hiroshi Kiyonari,&nbsp;Tatsuya Nakai,&nbsp;Toshihiro Uesaka,&nbsp;Yuzo Kodama,&nbsp;Hideki Enomoto","doi":"10.1111/dgd.70009","DOIUrl":"10.1111/dgd.70009","url":null,"abstract":"<p>Enteroendocrine cells (EECs) are sensory epithelial cells that sense the gut luminal environment and convey sensory information to the brain via the visceral afferent pathway. Although EECs are a part of gut epithelial cells, which generally undergo rapid turnover, some EECs have been reported to be long-lived. EECs consist of multiple subtypes, each of which displays distinct hormone production and distribution patterns. It remains unknown whether a long lifespan is a characteristic shared by all EEC subtypes. To address this issue, we conducted genetic pulse labeling of three EEC subtypes expressing serotonin (5-HT), peptide YY (PYY), and gastric inhibitory polypeptide (GIP) in mice and tracked their survival. In the proximal small intestine, all labeled GIP⁺ EECs disappeared completely within 5 days, whereas some PYY⁺ EECs survived for more than 7 days. In the proximal colon, some labeled 5-HT⁺ EECs lived for more than 28 days, whereas no PYY⁺ cells survived beyond 14 days. These long-lived 5-HT⁺ EECs were almost exclusively found in the upper half of the crypt in the mucosal fold, where visceral sensory fibers were enriched. This study reveals subtype- and region-dependent survival of EECs and suggests that EEC–nerve communication may underlie the long lifespan of certain EECs.</p>","PeriodicalId":50589,"journal":{"name":"Development Growth & Differentiation","volume":"67 4","pages":"205-214"},"PeriodicalIF":1.7,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144059600","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Waves of innovation and collaboration: Zebrafish research converges in Kyoto","authors":"Takuya Kaneko","doi":"10.1111/dgd.70008","DOIUrl":"10.1111/dgd.70008","url":null,"abstract":"<p>The 18th International Zebrafish Conference “IZFC2024” was held in Kyoto from August 17 to 21, 2024.\u0000 <figure>\u0000 <div><picture>\u0000 <source></source></picture><p></p>\u0000 </div>\u0000 </figure></p>","PeriodicalId":50589,"journal":{"name":"Development Growth & Differentiation","volume":"67 4","pages":"192-194"},"PeriodicalIF":1.7,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144063195","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}