{"title":"Mild cryoinjury in zebrafish fin induces regenerative response without blastema formation","authors":"Takafumi Yoshida, Atsushi Kawakami","doi":"10.1111/dgd.12962","DOIUrl":null,"url":null,"abstract":"<p>Previous studies have shown that tissue regeneration induces expression of genes that play important roles in regeneration. Recently, several studies have identified regeneration-response enhancers (RREs) that activate gene expression by tissue injury. Particularly, we showed that RREs contain two transcription factor-binding motifs: a bHLH transcription factor-binding motif, an E-box, and an AP-1/bZIP transcription factor-binding motif, a 12-O-Tetradecanoylphorbol 13-acetate response element (TRE). However, the triggers and subsequent signals generated by injury are still unclear. In this study, we analyzed RRE activation using various injury models. Although inter-ray incisions and skin exfoliation injuries did not activate RREs or regeneration genes, the fin puncture injury activated RREs and several regeneration-response genes. After fin puncture injury, <i>msxc</i> was activated only on the proximal side of the hole where blastema-like tissue was formed, whereas RREs, <i>junbb</i>, and <i>fibronectin 1b</i> (<i>fn1b</i>) were activated on both the proximal and distal sides, implying that activation of RREs, <i>junbb</i>, and <i>fn1b</i> is independent of blastema formation. Here, we also established a mild cryoinjury method. After this injury, transient vascular destruction, an increase in cell death, and an accumulation of myeloid cells were observed; however, no major morphological damage was observed. Importantly, <i>msxc</i> was not induced by cryoinjury, whereas <i>fn1b</i>, <i>junbb</i>, and <i>1.8 k</i> RRE (−<i>1.8 kb promoter</i> of <i>fn1b</i>) were activated, suggesting that cryoinjury induces the responses of <i>fn1b</i>, <i>junbb</i>, and <i>1.8 k</i> RRE without forming the blastema. Thus, our study shows that the cryoinjury model and the RRE transgenic (Tg) zebrafish may provide a useful platform for exploring injury signals.</p>","PeriodicalId":50589,"journal":{"name":"Development Growth & Differentiation","volume":"67 3","pages":"174-181"},"PeriodicalIF":1.7000,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/dgd.12962","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Development Growth & Differentiation","FirstCategoryId":"99","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/dgd.12962","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"CELL BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Previous studies have shown that tissue regeneration induces expression of genes that play important roles in regeneration. Recently, several studies have identified regeneration-response enhancers (RREs) that activate gene expression by tissue injury. Particularly, we showed that RREs contain two transcription factor-binding motifs: a bHLH transcription factor-binding motif, an E-box, and an AP-1/bZIP transcription factor-binding motif, a 12-O-Tetradecanoylphorbol 13-acetate response element (TRE). However, the triggers and subsequent signals generated by injury are still unclear. In this study, we analyzed RRE activation using various injury models. Although inter-ray incisions and skin exfoliation injuries did not activate RREs or regeneration genes, the fin puncture injury activated RREs and several regeneration-response genes. After fin puncture injury, msxc was activated only on the proximal side of the hole where blastema-like tissue was formed, whereas RREs, junbb, and fibronectin 1b (fn1b) were activated on both the proximal and distal sides, implying that activation of RREs, junbb, and fn1b is independent of blastema formation. Here, we also established a mild cryoinjury method. After this injury, transient vascular destruction, an increase in cell death, and an accumulation of myeloid cells were observed; however, no major morphological damage was observed. Importantly, msxc was not induced by cryoinjury, whereas fn1b, junbb, and 1.8 k RRE (−1.8 kb promoter of fn1b) were activated, suggesting that cryoinjury induces the responses of fn1b, junbb, and 1.8 k RRE without forming the blastema. Thus, our study shows that the cryoinjury model and the RRE transgenic (Tg) zebrafish may provide a useful platform for exploring injury signals.
先前的研究表明,组织再生诱导了在再生中起重要作用的基因的表达。最近,一些研究发现再生反应增强因子(RREs)可以通过组织损伤激活基因表达。特别地,我们发现RREs包含两个转录因子结合基序:bHLH转录因子结合基序,一个E-box,和AP-1/bZIP转录因子结合基序,一个12- o -十四烷基phorbol 13-醋酸酯响应元件(TRE)。然而,损伤的触发因素和随后产生的信号仍不清楚。在本研究中,我们使用不同的损伤模型分析RRE激活。虽然射线间切口和皮肤剥离损伤没有激活RREs或再生基因,但鳍穿刺损伤激活了RREs和几个再生反应基因。在鱼鳍穿刺损伤后,msxc仅在形成囊胚样组织的孔的近端被激活,而RREs、junbb和纤维连接蛋白1b (fn1b)在近端和远端都被激活,这表明RREs、junbb和fn1b的激活与囊胚形成无关。在这里,我们也建立了一种轻度冷冻损伤方法。损伤后,观察到短暂的血管破坏、细胞死亡增加和骨髓细胞积聚;然而,没有观察到重大的形态学损伤。重要的是,msxc没有被低温损伤诱导,而fn1b、junbb和1.8 k RRE (fn1b的-1.8 kb启动子)被激活,这表明低温损伤诱导了fn1b、junbb和1.8 k RRE的反应,而没有形成胚基。因此,我们的研究表明,低温损伤模型和RRE转基因(Tg)斑马鱼可能为探索损伤信号提供了一个有用的平台。
期刊介绍:
Development Growth & Differentiation (DGD) publishes three types of articles: original, resource, and review papers.
Original papers are on any subjects having a context in development, growth, and differentiation processes in animals, plants, and microorganisms, dealing with molecular, genetic, cellular and organismal phenomena including metamorphosis and regeneration, while using experimental, theoretical, and bioinformatic approaches. Papers on other related fields are also welcome, such as stem cell biology, genomics, neuroscience, Evodevo, Ecodevo, and medical science as well as related methodology (new or revised techniques) and bioresources.
Resource papers describe a dataset, such as whole genome sequences and expressed sequence tags (ESTs), with some biological insights, which should be valuable for studying the subjects as mentioned above.
Submission of review papers is also encouraged, especially those providing a new scope based on the authors’ own study, or a summarization of their study series.