Fluorescent reporter transgenic mice for in vivo live imaging of angiogenesis and lymphangiogenesis.
Fluorescent reporter transgenic mice for in vivo live imaging of angiogenesis and lymphangiogenesis.
复制标题
用于血管生成和淋巴管生成的体内活体成像的荧光记者转基因小鼠。
DOI：
10.1007/s10456-018-9629-2
复制
发表时间：
2018-11
期刊：
影响因子：
9.8
通讯作者：
Azar DT
中科院分区：
文献类型：
--
作者：
Doh SJ;Yamakawa M;Santosa SM;Montana M;Guo K;Sauer JR;Curran N;Han KY;Yu C;Ema M;Rosenblatt MI;Chang JH;Azar DT
淋巴管生成的研究是一门新兴的科学，它揭示了淋巴系统在许多病理条件下的核心作用，包括癌症转移、淋巴水肿和器官移植排斥。彻底了解淋巴生长的机制将在针对这些疾病的治疗策略的发展中发挥关键作用。尽管已知该领域的潜力，淋巴学的研究历来落后于血管生成的研究。直到最近，由于缺乏淋巴特异性标记物和合适的实验模型，与那些更直接可见的血管系统相比，淋巴研究的重大进展受到阻碍。淋巴管生成也被证明是发育生物学过程中的一个关键现象，如细胞增殖、引导迁移、分化和细胞间通讯，这使得淋巴特异性可视化技术非常可取和迫切需要。成像方式包括免疫组织化学和原位杂交是有限的，需要牺牲动物模型的组织收获在每个实验时间点。此外，采集组织的安装和染色过程可能会引入干扰结果的伪影。这些研究淋巴和血管系统的传统方法存在一些问题，包括在复制淋巴生长环境时的动物变异性（例如，小鼠之间）以及长时间、劳动密集型研究的成本问题，所有这些都使动态淋巴过程的研究复杂化。随着淋巴特异性标记物的发现，研究人员已经能够开发出几种淋巴和血管特异性的、启动子驱动的、荧光报告的转基因小鼠，用于体内和体外淋巴的可视化。例如，GFP、mOrange、tdTomato和其他荧光蛋白可以在淋巴特异性标记物（如Prospero-related homeobox 1 (Prox1)）的控制下表达，Prox1是一种高度保守的转录因子，在脊椎动物中决定胚胎器官发生，与淋巴管发生以及几种人类癌症有关。重要的是，没有prox1的小鼠胚胎发育时没有淋巴管。在成人中，Prox1维持淋巴内皮细胞，上调淋巴管生成相关蛋白（如VEGFR-3），下调血管生成相关基因表达（如STAT6）。为了在血管生成的背景下观察淋巴的发育，研究人员培育了双荧光转基因报告者，如Prox1-GFP/Flt1-DsRed小鼠，以在体内同时表征淋巴和血管。在本文中，我们讨论了淋巴管可视化的发展趋势以及转基因品种在血管生成和淋巴管生成研究中的潜在应用，以了解血管发育与病理进展之间的时空相关性。
The study of lymphangiogenesis is an emerging science that has revealed the lymphatic system as a central player in many pathological conditions including cancer metastasis, lymphedema, and organ graft rejection. A thorough understanding of the mechanisms of lymphatic growth will play a key role in the development of therapeutic strategies against these conditions. Despite the known potential of this field, the study of lymphatics has historically lagged behind that of hemangiogenesis. Until recently, significant strides in lymphatic studies were impeded by a lack of lymphatic-specific markers and suitable experimental models compared to those of the more immediately visible blood vasculature. Lymphangiogenesis has also been shown to be a key phenomenon in developmental biological processes, such as cell proliferation, guided migration, differentiation, and cell-to-cell communication, making lymphatic-specific visualization techniques highly desirable and desperately needed. Imaging modalities including immunohistochemistry and in-situ hybridization are limited by the need to sacrifice animal models for tissue harvesting at every experimental time point. Moreover, the processes of mounting and staining harvested tissues may introduce artifacts that can confound results. These traditional methods for investigating lymphatic and blood vasculature are associated with several problems including animal variability (e.g., between mice) when replicating lymphatic growth environments and the cost concerns of prolonged, labor-intensive studies, all of which complicate the study of dynamic lymphatic processes. With the discovery of lymphatic-specific markers, researchers have been able to develop several lymphatic and blood vessel-specific, promoter-driven, fluorescent-reporter transgenic mice for visualization of lymphatics in vivo and in vitro. For instance, GFP, mOrange, tdTomato, and other fluorescent proteins can be expressed under control of a lymphatic-specific marker like Prospero-related homeobox 1 (Prox1), which is a highly conserved transcription factor for determining embryonic organogenesis in vertebrates that is implicated in lymphangiogenesis as well as several human cancers. Importantly, Prox1-null mouse embryos develop without lymphatic vessels. In human adults, Prox1 maintains lymphatic endothelial cells and upregulates proteins associated with lymphangiogenesis (e.g., VEGFR-3) and downregulates angiogenesis-associated gene expression (e.g., STAT6). To visualize lymphatic development in the context of angiogenesis, dual fluorescent-transgenic reporters, like Prox1-GFP/Flt1-DsRed mice, have been bred to characterize lymphatic and blood vessels simultaneously in vivo. In this review, we discuss the trends in lymphatic visualization and the potential usage of transgenic breeds in hemangiogenesis and lymphangiogenesis research to understand spatial and temporal correlations between vascular development and pathological progression.
登录
查看更多内容
改善绿色和红色荧光蛋白的亮度和光稳定性，用于活细胞成像和FRET报告。
影响因子：
4.6
作者：
Bajar BT;Wang ES;Lam AJ;Kim BB;Jacobs CL;Howe ES;Davidson MW;Lin MZ;Chu J
通讯作者：
Chu J
DOI：
10.1007/s00428-016-1945-6
发表时间：
2016-07
期刊：
Virchows Archiv : an international journal of pathology
影响因子：
--
作者：
Adamczyk LA;Gordon K;Kholová I;Meijer-Jorna LB;Telinius N;Gallagher PJ;van der Wal AC;Baandrup U
通讯作者：
Baandrup U
影响因子：
6
作者：
Hirakawa, S;Hong, YK;Detmar, M
通讯作者：
Detmar, M
影响因子：
4.8
作者：
Iljin, K;Petrova, TV;Alitalo, K
通讯作者：
Alitalo, K
影响因子：
2.7
作者：
Dash, Amit K.;Yende, Ashutosh S.;Tyagi, Rakesh K.
通讯作者：
Tyagi, Rakesh K.