Simon J. Rhodes - US grants

9729669 RHODES The pituitary gland is a small organ located at the base of the brain in mammals. In response to signals from the brain and the bloodstream, the pituitary secretes hormones that regulate growth, reproductive development and status, milk production, the response to stress and control of functions such as body temperature and metabolism. During development, genes are turned on and off in a regulated program that allows the specialized hormone-releasing cells to be derived from simple, common precursor cells. Molecules known as transcription factors act as switches to coordinate this program of gene activation. Using state-of-the-art molecular technology, Dr. Rhodes will determine the specific mechanism of a pituitary transcription factor called P-Lim. P-Lim is essential for pituitary development since individuals with mutated P-Lim do not develop pituitary glands and thus, die shortly after birth. The results from these fundamental studies will significantly further our understanding of pituitary development and function. Moreover, the knowledge gained from work on the pituitary can impact on related processes, such as the development of the nervous system, the blood and the limbs.

9877094

Abstract

This project involves the acquisition of a phosphoimager system for molecular biology research and training. The instrument will be housed in the Department of Biology at Indiana University-Purdue University at Indianapolis and will serve 5 major and two minor users in the department of Biology and two major users in the Department of Chemistry. All major and minor users employ conventional autoradiography, fluorography and/or chemiluminescent detection with film on a routine basis. However, these techniques are limited by the narrow linear response range of film and by the lengthy exposure times required for the fluorographic detection of weak energy isotopes such as 3H and 14C. Phosphoimager analysis has several key advantages over autoradiography and fluorography. First, phosphoimager screens have a linear response range of five orders of magnitude as opposed to only two for autoradiography. Second, phosphoimager exposure times are generally only one tenth those required for autoradiography and fluorography. This is especially advantageous for the detection of weak energy isotopes where fluorography exposure times of weeks or months are often required. Third, the phosphoimager data can be quantified and imported into computer drawing programs for accurate quantitative analysis. This system will complement existing molecular biology equipment in the laboratories of the major and minor users. Moreover, the high degree of accuracy and efficiency afforded by phosphoimager analysis will improve the quality and quantity of molecular biology research and teaching performed by the investigators.

The explosive growth of biological information sources, available over the Internet, has given rise to both opportunities and challenges for biological and medical researchers. The opportunities they provide are both scientific (e.g., understanding the information encoded in elementary biological structures) as well as technological (e.g., new drug discovery). The challenges, on the other hand, lie in how to efficiently discover, among the vast volume of information, the items that are relevant or interesting to a given researcher. The objective of the proposed research is to investigate related basic research problems and develop a biological information delivery system in a collaborative project between computer scientists, information scientists, and biological researchers. The specific plans include developing methods to make the proposed system pro-active (surveying evolving on-line sources for relevant information), personalized (cognizant of a particular researcher's interests), adaptive (able to react to changes in the information sources as well as user interests or objectives), and capable of integrating multi-format data. The impact of this research is a significant enhancement in the ability of students and researchers in biological sciences to efficiently utilize on-line resources, while generating methods for computerized analysis of biological data and providing computerized support for new scientific discovery.

0093092
Chernoff

This award is to Indiana University Purdue University-Indianapolis to support the activity described below for 36 months. The proposal was submitted in response to the Partnerships for Innovation Program Solicitation (NSF 0082).

Partners
The partners include Indiana University Purdue University - Indianapolis; Indiana University; Terre Haute Center for Medical Partner Organizations; Indiana University School of Medicine; Eli Lilly & Company; Indiana 21st Century Fund for Research and Technology; Indiana Business Modernization and Technology Corporation.

Proposed Activities
The activities for this award include innovative research in regenerative biology; technology transfer leading to development of therapies; training the workforce for regenerative biology.

Proposed Innovation
The goals of this innovation are research on regenerative biology to understand the regeneration process and identify proteins of therapeutic value for healing, and technology transfer to private companies for healthcare delivery.

Potential Economic Impact
The potential economic impacts are creation of an estimated 7000 new jobs over the next 10 years from the partner companies alone (10% of these will be in Indiana), and increased participation of minorities in the health care industry.

Potential Societal Impact
The development of therapies for injured and degenerating tissues will have medical and health benefits to society beyond anything known today. The plan also includes strong involvement of minorities in the workforce training programs with subsequent participation in the anticipated expanded healthcare job market.

The mammalian anterior pituitary gland is a major organ for neuroendocrine function because it secretes multiple, important hormones. During development, proteins known as transcription factors regulate activation of the genes that encode these hormones. Two transcription factors of the LIM homeodomain group, Lhx3 and Lhx4, are involved in the specification of the hormone-secreting cells in the anterior pituitary. Very little is known about the biochemical mechanisms by which these factors exert their specific gene regulatory functions. Lhx3 exists in different isoforms, which have distinct DNA-binding and gene regulatory properties. This project uses molecular and biochemical approaches to test the hypothesis that Lhx3 is a phosphoprotein that is modified at multiple locations, and that phosphorylation of Lhx3 modulates its gene regulatory properties. Lhx3 will be expressed, purified and analyzed by mass spectrometry techniques, and modified isoforms will be changed by site-directed mutatgenesis. The DNA binding, gene regulatory properties, involvement of intracellular signaling pathways, and localization of these modified Lhx3 molecules will be tested using reporter genes, constitutively active signaling enzymes and confocal microscopy. Experiments also will test to what extent Lhx4 has similar, but distinct, transcriptional properties to Lhx3. An important issue from genomics is to understand how a limited number of genes can mediate the development of complex organisms, and this study shows an example of how multiple proteins can be generated from individual genes, and how their functions depend on post-transcriptional regulation.
Results will have an impact beyond neuroendocrinology, because LIM proteins are required for development and function of many other systems in both animals and plants. Training and mentoring of undergraduate and graduate students also are an integral part of this project.

DESCRIPTION (provided by applicant): The anterior lobe of the pituitary releases hormones that are critical for developmental and physiological functions. During embryogenesis, the actions of transcription factors govern the establishment of the hormone-secreting pituitary cell lineages. Heritable mutations in these regulatory genes cause combined pituitary hormone deficiency diseases in children. However, the molecular defects involved in the majority of pituitary disorders remain unknown. LHX3, a LIM homeodomain transcription factor, is essential for pituitary development and function. Although mutations in the human LHX3 gene cause a severe endocrine disease that features compound hormone deficiencies, the biochemical mechanism by which the LHX3 gene exerts its critical functions is not understood. The human LHX3 gene encodes protein isoforms with distinct expression patterns and gene regulatory activities. These isoforms are LHX3a, LHX3b, and a novel protein, M2-LHX3. To test the hypothesis that the human LHX3 gene produces isoforms with unique functions in pituitary development, the protein domains required for the function of the LHX3 isoforms will be characterized and partner proteins that mediate or modulate LHX3 activity by interacting with these domains will be identified. The functions of the LHX3a, LHX3b, and M2-LHX3 proteins will be analyzed by overexpressing each isoform in specific pituitary cell types during development in transgenic mice. In addition, the transcriptional mechanisms that generate the human LHX3a and LHX3b transcripts will be determined using in vitro and transgenic animal approaches. Further, to test the in vivo functions of LHX3a and LHX3b, gene-targeting techniques will be used to selectively prevent their individual expression in mice. These experiments will provide an increased understanding of the genetic pathways that control pituitary organogenesis and will, therefore, improve our ability to treat pituitary disease. Identified proteins and gene regulatory regions will provide candidates for pituitary diseases of unknown etiology. Finally, these studies will demonstrate how single genes can produce protein isoforms with different activities, providing an example of how mammals have evolved mechanisms to increase the proteome-encoding capacity of their genomes.

Graduate Teaching Fellows in K-12 Education (GK-12)
Abstract

Proposal #: 0742475
PI: Kathleen Marrs
Institution: Indiana University-Purdue University
Title: The GK-12 Urban Educators Program at IUPUI: Teaching and Learning Science through Research
NSF-supported STEM disciplines: Biology, Chemistry, Physics, Math, Earth Sciences



This project combines the research and teaching strengths of the IUPUI School of Science (SOS) and the Indiana University School of Medicine (IUSM) with requests by local teachers for inquiry-based STEM laboratory experiences for children in grades 6-12. The selected urban partner schools have core competences in medical or environmental education. The fellows will collaborate with a secondary science or math teacher in ongoing professional development to develop grade-appropriate research activities. Innovative aspects include development of layered research projects that build in complexity through several grade levels, with a theme of relevance to student?s lives.

Expected outcomes include enhanced research, teaching, leadership and communication skills of the fellows. Benefits to science teachers include opportunities for professional development, collaboration with GK-12 fellows and university faculty, and opportunities to engage their students in scientific research. Goals for middle and high school students include exposure to mentors and role models, opportunities to experience research, and exposure to science and math as exciting and desirable career options. By designing a collaborative, diverse partnership between STEM graduate researchers, faculty and STEM teachers, this project allows us to integrate research-based activities into the teaching and learning of secondary science and math, broaden the involvement and participation of a diverse inner-city student body in research, and strengthen existing ties between IUPUI and the local community.