Evolution of Axial Patterning - Functional Approaches

奖学金培训和职业目标我的论文研究对一个生物学问题采用了描述性的方法，克隆了关键的发育调控基因，并检查了它们的表达模式，以推断发育功能。我和马丁代尔博士提出的这个项目与我的研究生培训非常不同，因为它将从功能的角度来回答有关后生动物基础发展项目演变的问题。我将提出可验证的假设，并使用新的实验方法来操纵和研究体内的基因功能。这将需要更复杂的智力方法来解决这个问题，并将利用现代分子技术，包括RNAi、吗啉、异位表达和显微注射。NRSA博士后奖学金将在马丁代尔博士的指导下为我提供一个宝贵的研究和职业发展机会。马丁代尔博士是进化发育生物学领域的顶尖专家之一。这将极大地促进我作为一名全面研究科学家的发展，朝着我的最终职业目标--在一所研究能力很强的大学成为一名富有成效的教员--迈进。I 19.姓名(S)马丁代尔，学士，博士20.职位/职级副教授21.研究兴趣/领域图案形成的细胞、分子和进化方面]-[，，_，-f，i]r=.l=ms]=r*L？r*‘[._*_22.描述(不超过所提供的篇幅)本研究的目的是调查多细胞动物发育过程中关键调控基因在轴向区域化中的功能。这一目标属于解决后生动物分子构型机制基本问题的长期综合目标。在这项研究中，我们将使用功能遗传学分析来确定Hox/ParaHox基因在蛇类动物轴型中的作用。HOX/ParaHox基因在三倍体动物前后构型中的保守功能决定了双侧动物的“动物型”。这些基因是否在蛇形目动物中发挥类似的作用仍有待确定。这项拟议的研究有几个具体的目标。首先，从矢状线虫(线虫纲：珊瑚虫纲)中分离出一个后ParaHox基因，并对其表达特征进行了分析。接下来，我们将使用两种互补的功能分析方法，即异位基因表达和基因沉默，来研究蚊虫Hox/ParaHox基因在发育过程中的功能。最后，我们将研究人类HOX/ParaHOX基因在再生过程中的功能，以阐明个体发育和再生机制之间的潜在同源性。确定线虫HoxParaHox基因的功能将使我们能够在线虫和双线动物之间建立轴向同源性。这些结果将揭示Hoxand ParaHox基因在后生动物早期历史中的祖先作用，并将为复杂身体计划的进化提供有价值的见解。PHS416-1(12/98版)表第2页BB抄送名称(姓氏、首字母、中间字母)个人非专利申请目录========================================Section End===========================================

GOALS FOR FELLOWSHIP TRAINING AND CAREER My dissertation research employed a descriptive approach to a biological question, cloning key developmental regulatory genes and examining their expression patterns to infer developmental functions. The project I am proposing with Dr. Martindale is very different from my graduate training as it will take a functional approach to questions about the evolution of basic metazoan developmental programs. I will propose testable hypotheses and use new experimental approaches to manipulate and investigate gene function in vivo. This will require more sophisticated intellectual approaches to the question, and will utilize modern molecular techniques, including RNAi, morpholinos, ectopic expression, and microinjection. An NRSA Postdoctoral Fellowship will provide me with an invaluable research- and career- development opportunity under the mentorship of Dr. Martindale, one of the foremost experts in evolutionary developmental biology. It will greatly advance my development as a well-rounded research scientist toward my ultimate career objective of becoming a productive faculty member at a university with a strong research [)ro_ram. / / li']iF1"]i 19. NAMEANDDEGREE(S) Mark Q. Martindale, B.A., Ph.D. 20. POSITION/RANK Associate Professor 21. RESEARCH INTERESTS/AREAS Cellular, molecular, and evolutionary aspects of pattern formation ] -[,,_ ,-f,i .]r=.l = ms .] =r*l ¿r*'[._*_ 22. DESCRIPTION (Do not exceed space provided) The objective of this research is to investigate functions of key regulatory genes in axial regionalization during development of multicellular animals. This objective falls within the long-range comprehensive goal of addressing fundamental questions of molecular patterning mechanisms in metazoans. In this study, we will use functional genetic assays to determine the roles of the Hox/ParaHox genes in axial patterning in cnidarians. The conserved function of Hox/ParaHox genes in anterioposterior patterning of triploblastic animals defines the bilaterian 'zootype'. Whether these genes perform comparable roles in cnidarians remains to be determined. There are several specific aims in this proposed study. First, a posterior ParaHox gene will be isolated from Nematostella vectensis (Cnidaria: Anthozoa) and its expression 3attern characterized. Next, the function of cnidarian Hox/ParaHox genes during development will be examined in N. vectensis using two complementary functional assays, ectopic gene expression and gene silencing. Finally, the function of cnidarian Hox/ParaHox genes during the process of regeneration will be examined to elucidate potential homology between the mechanistic events in ontogeny and regeneration. Determining HoxParaHox gene function in cnidarians will allow us to establish axial homologies between cnidarians and bilaterians. These results will reveal the ancestral roles of Hoxand ParaHox genes in early metazoan history and will provide valuable insight to the evolution of complex body plans. PHS 416-1 (Rev. 12/98) Form Page 2 BB CC NAME (Last, first, middle initial) Individual NRSA Application Table of Contents ========================================Section End===========================================