Plasticity and Regulation in Xenopus Anterior-Posterior Patterning

非洲爪蟾前后图案的可塑性和调节

基本信息

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
项目类别：
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

项目摘要

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（由申请人提供）：功能器官系统的开发不仅需要导致细胞类型特异性和适当图案的确定性事件在这些细胞类型中，以及面对整个胚胎发生的基因和环境扰动，能够维持这种身份和模式的能力。这种补偿潜在的破坏性变化的能力通常被称为调节，是胚胎的几乎全宇宙特征，胚胎是确保正常发育所必需的。然而，尽管在发展扰动之后，阐明管理调节过程的机制相对较少，尽管它对于完全理解发展及其对再生医学的影响显而易见。在这里，我们采用了爪诺乱的经典两栖动物胚胎学系统来检查早期胚胎发生过程中的调节能力。我们实验室的先前实验表明，当将推定性神经板从胃阶段的胚胎中取出，旋转180O，并将其移植到宿主胚胎中，从中去除等效区域，将近乎整体调节，并且会产生与适当的区域基因表达和功能性的胚胎相关的胚胎。该提案的总体目标是检查管理这种强烈监管能力的机制。第一个具体目的将评估对调节过程重要的时间和空间表达谱F候选基因，其目的是获得基线信息，以确定何时以及如何发生这种调节过程。这些基因包括：区域标记（XCG-1，OTX-2，EN-2，KROX-20，HOXB9）；后来的分化基因（GAD，XVGLUT1）；基因在神经测定途径（FOXD5，GEMININ，SOX2）之前表达；以及编码钥匙前后信号分子（FRZB-1，XWNT8和FGF8）的基因。第二个目的将确定特定的信号级联是否通过使用靶向的Morpholino敲低方法来扰动其函数来介导调节过程。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，将确定调节过程的其他分子成分。综上所述，这三个目标中提出的实验将吸引各个层次的渴望的本科生干部，以解决对发展生物学的基本且研究不足的问题，该问题对再生医学具有更广泛的影响。

项目成果

期刊论文数量（17）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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
影响因子：
0
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。