Evolution of Axial Patterning - Functional Approaches

我的博士论文研究采用描述性的方法来解决一个生物学问题，克隆关键的发育调节基因并检查它们的表达模式来推断发育功能。我和Martindale博士提出的这个项目与我的研究生训练非常不同，因为它将采用一种功能性的方法来解决有关基本后生动物发育程序进化的问题。我将提出可测试的假设，并使用新的实验方法来操纵和研究基因在体内的功能。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， morpholinos，异位表达和显微注射。在进化发育生物学领域最重要的专家之一Martindale博士的指导下，NRSA博士后奖学金将为我提供宝贵的研究和职业发展机会。它将极大地促进我的发展，使我成为一名全面的研究科学家，朝着我的最终职业目标——成为一所研究能力很强的大学的一名富有成效的教员——迈进。[/ /] [1] [1]姓名与学位(S) Mark Q. Martindale， B.A, Ph.D。职位/职级副教授研究方向：细胞、分子和进化方面的模式形成[，，_,-f,i .]r=。L = ms。=r*l¿r*'[.]_ * _ 22。本研究的目的是研究多细胞动物发育过程中轴向区域化关键调控基因的功能。这一目标属于解决后生动物分子模式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基因在刺胞动物轴向模式中的作用。Hox/ParaHox基因在三倍体动物的前后模式中的保守功能定义了双边“动物型”。这些基因是否在刺胞动物中发挥类似的作用还有待确定。在这项拟议的研究中有几个具体的目的。首先，从线虫菌（刺胞纲：刺虫纲）中分离出一个后副hox基因，并对其表达模式进行表征。接下来，我们将利用异位基因表达和基因沉默这两种互补的功能分析方法，来检测刺胞细胞Hox/ParaHox基因在维氏螺旋体发育过程中的功能。最后，我们将研究刺胞细胞Hox/ParaHox基因在再生过程中的功能，以阐明个体发生和再生过程中机制事件之间的潜在同源性。确定刺胞动物的HoxParaHox基因功能将使我们能够建立刺胞动物和双边动物之间的轴向同源性。这些结果将揭示Hoxand ParaHox基因在早期后生动物历史中的祖先角色，并将为复杂身体结构的进化提供有价值的见解。小灵通416 - 1(启12/98)表单页面2 BB CC的名字(最后,首先,初始)个人NRSA应用程序目录 ======================================== 节结束 ===========================================

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===========================================