Plasticity and Regulation in Xenopus Anterior-Posterior Patterning

非洲爪蟾前后图案的可塑性和调节

• 批准号: 8580604
• 负责人: MARGARET S SAHA
• 金额: $ 36.75万
• 依托单位: COLLEGE OF WILLIAM AND MARY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8580604
• 关键词: Address; Amphibia; Anterior; Back; Behavior; Candidate Disease Gene; Characteristics; Data; Development; Developmental Biology; Disease; Educational process of instructing; Embryo; Embryonic Development; Ensure; Environment; Event; FGF8 gene; Gastrula; Geminin; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic; Goals; Knowledge; Lead; Light; Mediating; Microarray Analysis; Molecular; Molecular Profiling; Nervous system structure; Neuronal Plasticity; Neurons; Organism; Pathway interactions; Pattern; Process; Regenerative Medicine; Regulation; Risk; Rotation; Signal Transduction; Signaling Molecule; Sorting - Cell Movement; Specificity; Staging; Stem cells; Students; System; Techniques; Time; Tissue Donors; Tissue Engineering; Tissues; Transplantation; Work; Xenopus; Xenopus laevis; body system; cell type; cellular engineering; knock-down; neural plate; novel; public health relevance; relating to nervous system; research study; restoration; transcriptome sequencing; undergraduate student

DESCRIPTION (provided by applicant): The development of a functional organ system not only entails the determinative events that lead to cell type specificity and appropriate patterning of those cell types, but also the ability to maintain this identity and pattern in the face of geneic and environmental perturbations that occur throughout embryogenesis. This ability to compensate for potentially disruptive alterations in development, which is often referred to regulation, is a near-universal characteristic of embryos, one that is required to ensure normal development. However, there has been relatively little focus on elucidating the mechanisms governing the process of regulation following a developmental perturbation, despite its clear importance for a complete understanding of development as well as its implications for regenerative medicine. Here we employ the classic amphibian embryological system of Xenopus laevis to examine regulative ability during early embryogenesis. Previous experiments from our lab have demonstrated that when the presumptive neural plate is removed from a gastrula-stage embryo, rotated 180o, and transplanted back into a host embryo from which the equivalent region was removed, there is a near total regulation, with the resulting embryo giving rise to a nervous system with appropriate regional gene expression and functional capabilities. The overall goal of this proposal is to examine the mechanisms governing this profound regulative ability. The first specific aim will assess the temporal and spatial expression profile f candidate genes important for the process of regulation, with the goal of obtaining baseline information to determine precisely when and how this regulative process occurs. These genes include: regional markers (XCG-1, Otx-2, En-2, Krox-20, HoxB9); later differentiation genes (GAD, xvGlut1); genes expressed earlier in the neural determination pathway (FoxD5, Geminin, Sox2); and genes encoding key anterior-posterior signaling molecules (Frzb-1, xWnt8, and FGF8). The second aim will determine if specific signaling cascades mediate the process of regulation by perturbing their function using a targeted morpholino knockdown approach. Finally, unbiased global gene expression approaches, specifically microarray analysis and an RNA-seq approach, will identify additional molecular components of the regulation process. Taken together, the experiments proposed in these three aims will engage an eager cadre of undergraduate students at all levels in an effort to address a fundamental and poorly studied problem in developmental biology that has broader implications for regenerative medicine.

描述(申请人提供)：功能器官系统的发展不仅包括导致细胞类型特异性和适当模式的决定性事件 这些细胞类型，而且有能力在面对整个胚胎发育过程中发生的遗传和环境扰动时保持这种身份和模式。这种补偿发育过程中潜在破坏性变化的能力，通常被称为监管，这是胚胎的一个几乎普遍的特征，是确保正常发育所必需的。然而，人们对阐明发育扰动后调控过程的机制的关注相对较少，尽管它对于全面了解发育及其对再生医学的影响具有明显的重要性。在这里，我们利用非洲爪哇经典的两栖动物胚胎学系统来检测早期胚胎发生的调控能力。我们实验室以前的实验已经证明，当假设的神经板从原肠胚期胚胎中移除，旋转180度，并移植回宿主胚胎中，移除了相应区域，就有了几乎全部的调节，所得到的胚胎产生了一个具有适当区域基因表达和功能的神经系统。这项提案的总体目标是研究管理这一深刻调控能力的机制。第一个具体目标将评估对调控过程重要的候选基因的时间和空间表达谱，目的是获得基线信息，以准确确定这一调控过程发生的时间和方式。这些基因包括：区域标记基因(XCG-1、OTX-2、EN-2、KROX-20、HOXB9)；晚期分化基因(GAD、xvGlut1)；在神经决定通路早期表达的基因(FoxD5、Ginin、Sox2)；以及编码关键前后信号分子的基因(Frzb-1、xWnt8和FGF8)。第二个目标将确定特定的信号级联是否通过使用靶向的吗啉基因敲除方法干扰它们的功能来调节调节过程。最后，无偏见的全球基因表达方法，特别是微阵列分析和RNA-SEQ方法，将识别调控过程的其他分子成分。总而言之，这三个目标中提出的实验将吸引各级本科生的热切干部，努力解决发育生物学中一个基本且研究较少的问题，该问题对再生医学具有更广泛的影响。

项目成果

Calcium Signaling in Vertebrate Development and Its Role in Disease.

• DOI: 10.3390/ijms19113390
• 发表时间: 2018-10-30
• 期刊: International journal of molecular sciences
• 影响因子: 5.6
• 作者: Paudel S;Sindelar R;Saha M
• 通讯作者: Saha M

A Markovian Entropy Measure for the Analysis of Calcium Activity Time Series.

• DOI: 10.1371/journal.pone.0168342
• 发表时间: 2016
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Marken JP;Halleran AD;Rahman A;Odorizzi L;LeFew MC;Golino CA;Kemper P;Saha MS
• 通讯作者: Saha MS

Transcriptome of Xenopus andrei, an octoploid frog, during embryonic development.

• DOI: 10.1016/j.dib.2018.05.017
• 发表时间: 2018-08
• 期刊: Data in brief
• 影响因子: 1.2
• 作者: Pownall ME;Cutler RR;Saha MS
• 通讯作者: Saha MS

Genome Sequences of 19 Rhodococcus erythropolis Cluster CA Phages.

19 种红平红球菌簇 CA 噬菌体的基因组序列。

• DOI: 10.1128/genomea.01201-17
• 发表时间: 2017
• 期刊: Genome announcements
• 作者: Bonilla,JAlfred;Isern,Sharon;Findley,AnnM;Klyczek,KarenK;Michael,ScottF;Saha,MargaretS;Buchser,WilliamJ;Forsyth,MarkH;Paudel,Sudip;Gissendanner,ChristopherR;Wiedemeier,AllisonMD;Alonzo,FernandaL;UniversityofWisconsin–R
• 通讯作者: UniversityofWisconsin–R

Embryonic transplantation experiments: Past, present, and future.

胚胎移植实验：过去、现在和未来。

• 发表时间: 2017
• 期刊: Trends in developmental biology
• 作者: Solini,GraceE;Dong,Chen;Saha,Margaret
• 通讯作者: Saha,Margaret