Intravital Imaging of Candida albicans Identifies Differential In Vitro and In Vivo Filamentation Phenotypes for Transcription Factor Deletion Mutants.
Intravital Imaging of Candida albicans Identifies Differential In Vitro and In Vivo Filamentation Phenotypes for Transcription Factor Deletion Mutants.
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DOI：
10.1128/msphere.00436-21
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发表时间：
2021-06-30
期刊：
影响因子：
4.8
通讯作者：
Krysan DJ
中科院分区：
文献类型：
--
作者：
Wakade RS;Huang M;Mitchell AP;Wellington M;Krysan DJ
白色念珠菌是引起人类真菌感染的重要原因。广泛研究的C.白色念珠菌的一个重要特征是它能够进行菌丝和假菌丝的转化。虽然酵母菌，假菌丝，和菌丝存在于感染的哺乳动物组织的病理样品，它一直具有挑战性的特点，在体内表达过程中的调控网络和特定基因的作用。此外，C.白念珠菌临床分离株正日益被认识，而将这种异质性与发病机制相关联仍然是一个重要的目标。在这里，我们描述了使用活体成像方法来表征C。在哺乳动物感染模型中，通过利用小鼠耳廓（耳朵）的透光性，利用该模型，我们发现不同的C.白色念珠菌分离株可以显著变化，特别是当将体内表达与基于固体琼脂的测定进行比较时。我们还表明，良好的特点转录调节Efg1和Brg1似乎在体内和体外发挥重要作用。相比之下，Ume6在体外比在体内重要得多。最后，依赖于Bcr 1体外表达的菌株能够在缺乏Bcr 1的情况下在体内形成细丝。这种活体成像方法提供了一种新的方法，在哺乳动物感染的这一重要的毒力性状的系统表征。我们的初步研究为C.白念珠菌体内诱导不同于体外诱导。重要性白色念珠菌是人类真菌感染的最常见原因之一。C.白色念珠菌在感染期间经历从圆形酵母形式到丝状形式的转变，这对于其引起疾病的能力是至关重要的。虽然这种转变已经在实验室中研究了多年，但在动物感染模型中这样做的方法有限。我们已经开发了一种显微镜方法来可视化荧光标记的C。白色念珠菌在小鼠皮下组织中经历这种转变。我们的研究表明，C。在感染过程中白色念珠菌的出现与在实验室条件下观察到的不同。
Candida albicans is an important cause of human fungal infections. A widely studied virulence trait of C. albicans is its ability to undergo filamentation to hyphae and pseudohyphae. Although yeast, pseudohyphae, and hyphae are present in pathological samples of infected mammalian tissue, it has been challenging to characterize the role of regulatory networks and specific genes during in vivo filamentation. In addition, the phenotypic heterogeneity of C. albicans clinical isolates is becoming increasingly recognized, while correlating this heterogeneity with pathogenesis remains an important goal. Here, we describe the use of an intravital imaging approach to characterize C. albicans filamentation in a mammalian model of infection by taking advantage of the translucence of mouse pinna (ears). Using this model, we have found that the in vitro and in vivo filamentation phenotypes of different C. albicans isolates can vary significantly, particularly when in vivo filamentation is compared to solid agar-based assays. We also show that the well-characterized transcriptional regulators Efg1 and Brg1 appear to play important roles both in vivo and in vitro. In contrast, Ume6 is much more important in vitro than in vivo. Finally, strains that are dependent on Bcr1 for in vitro filamentation are able to form filaments in vivo in its absence. This intravital imaging approach provides a new approach to the systematic characterization of this important virulence trait during mammalian infection. Our initial studies provide support for the notion that the regulation and initiation of C. albicans filamentation in vivo is distinct from in vitro induction. IMPORTANCE Candida albicans is one of the most common causes of fungal infections in humans. C. albicans undergoes a transition from a round yeast form to a filamentous form during infection, which is critical for its ability to cause disease. Although this transition has been studied in the laboratory for years, methods to do so in an animal model of infection have been limited. We have developed a microscopy method to visualize fluorescently labeled C. albicans undergoing this transition in the subcutaneous tissue of mice. Our studies indicate that the regulation of C. albicans filamentation during infection is distinct from that observed in laboratory conditions.