틸팅, 점핑 및 가속 마우스 비교: 와이드 디스플레이를 위한 새로운 마우스 인터랙션

Atherosclerosis is an immuno-inflammatory disease of arteries, involving complex intercellular interactions. Direct communication through hemichannels and gap junction channels formed by connexins is important in atherosclerosis. This thesis first reviews the current knowledge on vascular connexins (Cx37, Cx40 and Cx43) in atherosclerosis. It then describes the association between connexin gene polymorphisms and coronary artery disease. Atherosclerosis localization being determined by flow patterns, this work then examines the interrelation between Cx37 and shear stress. High laminar shear stress upregulates endothelial Cx37 expression. Conversely, absence of Cx37 in mouse models of flow-dependent atherosclerosis alters plaque composition. Besides their role as (hemi)channel-forming proteins, connexins can also interact with other proteins. We unveil two interactions of Cx37: In endothelial cells, Cx37 interacts with eNOS, which is crucial for endothelial quiescence. In pDCs, Cx37 could interact with CIITA, which plays a central role in antigen presentation. Finally, these topics are integrated, and clinical implications of this work are discussed.

Behavioral Genetics The Role of Molecular Genetics in a Post Genomics World Recent Developments in Quantitative Trait Loci Analysis Practical Barriers to Identifying Complex Trait Loci Assessing Genotype X Environment Interactions and Correlations in a Postgenomic World UK Population Biomedical Collections Genetic Studies of Learning and Memory in Mouse Models An Integrative Neuroscience Program Linking Genes to Cognition and Disease Genetic Contributions to Anatomical, Behavioural and Neurophysiological Indices of Cognition Genetic Covariance Between Processing Speed and IQ General Cognitive Ability Isolation of the Genetic Factors Underlying Speech and Language Disorders Genetic Etiology of Comorbid Reading Difficulties and ADHD Epistasis and the Genetics of Complex Traits The Genetics of Autistic Disorder Finding Genes for Complex Behaviors: Progress in Mouse Models of the Addictions Genetic and Genotype X Environment Interaction Effects on Risk of Dependence on Alcohol, Tobacco, and Other Drugs: New Research Challenges Pharmacogenetics in a Postgenomic World Personality Neuroticism and Serotonin: A Developmental Genetic Perspective Animal Models of Anxiety Attention Deficit Hyperactivity Disorder: New Genetic Findings, New Directions Schizophrenia The Genetics of Affective Disorders: Current and Future Dementia Behavioral Genomics

Members of the Runx family of transcription factors play essential roles in the differentiation and development of several organ systems. Here we address the contribution of the osteoblast-related Runx gene, Runx2 , to the osteogenic and chondrogenic differentiation of mesenchymal stem cells. Using a transgenic mouse model, we observe Runx2 transcription through one of its two known promoters (designated P1 in pre-cartilaginous mesenchymal condensations as early as E9.5. Runx2 gene activity is later repressed at the onset of cartilage formation, both in vivo and in vitro , necessitating examination of the regulation and function of Runx2 in mesenchymal stem cells. We demonstrate that Runx2 gene activity is repressed by the direct interaction of the homeodomain transcription factor Nkx3.2 with the proximal Runx2 P1 promoter. This repression was found to be required for the progression of BMP-induced chondrogenesis, thereby identifying Runx2 as a modulator of BMP activity in the chondrogenic as well as osteogenic differentiation program. To further understand the regulation of the Runx2 P1 promoter and to determine the contribution of P1-derived gene product, Runx2 Type II, to the formation of mineralized tissue, we have generated a Runx2 Type II- LacZ gene replacement mouse model in which the initial coding sequences and splice donor sites of the Type II isoform are replaced with the LacZ reporter gene. Activity of the endogenous P1 promoter can therefore be monitored by β-galactosidase production. Analysis of Runx2 Type II-LacZ mice demonstrates that the P1 promoter is transcriptionally most active in mature osteoblasts, but its product, Runx2 Type II is dispensable for embryonic skeletal formation. Lastly, we examine the link between growth control and osteogenic differentiation by tissue-specific deletion of the Mdm2 proto-oncogene in developing skeletal tissues of the mouse embryo. Loss of Mdm2 results in impaired bone formation, with skeletal elements exhibiting lower bone mineral content and higher porosity. Ex vivo cultures of calvarial osteoprogenitor cells exhibit severely decreased osteoblastogenesis and bone nodule formation accompanied by a failure to activate Runx2 gene activity. These findings suggest that Mdm2 is required for inhibition of p53 activity that ultimately allows for post-confluent proliferation and induction of Runx2 during maturation of the osteogenic phenotype. Taken together, our findings suggest that Runx2 modulates the commitment of progenitor cells to the osteogenic and chondrogenic lineages, and that Runx2 activity is inextricably linked to mechanisms that control cellular proliferation.

This document will detail the nature of performance / object interaction, and the roles of documentation and language in this interaction. In addition, I will explore the body / object as a site of multiplicities, the quality of simultaneity in the appearance and apprehension of such multiplicities, and traverse the triumphs and travails of the body / object’s journey from one state of being to the next. Within this text there are windows Within the window of how text performs there are windows Within the window of windows there are windows Wonder of wonders: a wellspring of windows most mast moist master measure muster miss move massage mauve masseuse muff mousse miffed mouse move muse must This document is performance. Not performance document. You are performing the act of looking and reading, however your experience tells you, and with what equipment your culture provides. We are not in the same place at the same time, but we are performing together.

Neuronal nicotinic acetylcholine receptors (nAChRs) are involved in a plethora of fundamental biological processes ranging from muscle contraction to the formation of memories. The studies described in this work focus on the transcriptional regulation of the CHRNB4 gene, which encodes the !4 subunit of neuronal nAChRs. We previously identified a regulatory sequence (5 – CCACCCCT –3 ), or “CA box”, critical for CHRNB4 promoter activity in vitro. Here I report transcription factor interaction at the CA box along with an in vivo analysis of CA box transcriptional activity. My data indicate that Sp1, Sp3, Sox10 and c-Jun interact with the CHRNB4 CA box in the context of native chromatin. Using an in vivo transgenic approach in mice, I demonstrated that a 2.3-kb fragment of the CHRNB4 promoter region, containing the CA box, is capable of directing cell-type specific expression of a reporter gene to many of the brain regions that endogenously express the CHRNB4 gene. Site-directed mutagenesis was used to test the hypothesis that the CA box is critical for CHRNB4 promoter activity in vivo. Transgenic animals were generated in which LacZ expression is driven by a mutant form of the CA box. Reporter gene expression was not detected in any tissue or cell type at ED18.5. Similarly, I observed dramatically reduced reporter gene expression at PD30 when compared to wild type transgenic animals, indicating that the CA box is an important regulatory feature of the CHRNB4 promoter. ChIP analysis of brain tissue from mutant transgenic animals demonstrated that CA box mutation results in decreased interaction of the transcription factor Sp1 with the CHRNB4 promoter.

Metastasis is a step during cancer progression that is clinically challenging to determine and treat. The association of tumor cells with leukocytes influences successful tumor cell seeding and their survival in the metastatic niche. Selectins are vascular adhesion receptors that facilitate the recruitment of leukocytes to metastatic sites, and mediate interactions between tumor and stromal cells. Particularly the absence of L-selectin on leukocytes decreases the infiltration of immune cells in the lungs and reduces pulmonary metastasis. Our aim was to elucidate the contribution of L-selectin during metastatic development in lungs and liver. While pulmonary metastasis was attenuated, liver metastasis was increased in L-selectin deficient mice. We determined that the selectin-dependent recruitment of inflammatory monocytes induced vascular permeability, promoted the extravasation of tumor cells and supported tumor cell survival in the lungs. The adoptive transfer of wild type bone marrow-derived monocytes into L-selectin deficient mice ameliorated metastatic development in the lungs. In the liver, we observed increased amounts of Kupffer and γδ T cells in the absence of L-selectin. Furthermore, L-selectin facilitated the recruitment of neutrophils into the metastatic liver. We showed that liver neutrophils produce reactive oxygen species in vivo, and increased tumor cell apoptosis in vitro. Depleting neutrophils at later metastatic stages increased liver metastasis, confirming the cytotoxic function of neutrophils. In summary, we identified an opposing role of L-selectin on metastatic development in the lungs and the liver. In the lungs, L-selectin mediates the recruitment of monocytes that promote tumor cell extravasation and support metastatic development. Our study demonstrated the contribution of myeloid cell populations to tissue-specific metastasis and defined a novel role of L-selectin during metastatic progression.

The ERK Cascade As a Prototype of MAPK Signaling Pathways Hadara Rubinfeld and Rony Seger Determination of ERK Activity: Antiphospho-ERK Antibodies, In Vitro Phosphorylation, and In-Gel Kinase Assay Sarah Kraus and Rony Seger Detection of ERK1/2 Activities Using Affinity Reagents Rafael Pulido, Angel Zuniga, and Axel Ullrich Activation of SAPKs/JNKs and p38s In Vitro John M. Kyriakis, Hong Liu, and Deborah N. Chadee Investigating the Cellular BMK1/ERK5 Signaling Pathway Richard I. Tapping, Kato Yutaka, Ta-Hsiang Chao, Masaaki Hayashi, Jeng-Fan Lo, Sung-Woo Kim, and Jiing-Dwan Lee Pull-Down Assays for Guanosine 5'-Triphosphate-Bound Ras-Like Guanosine 5'-Triphosphatases Miranda van Triest and Johannes L. Bos Regulation of MAPK Cascades by Protein Tyrosine Phosphatases Josema Torres, Carmen Blanco-Aparicio, and Rafael Pulido Use of Inhibitors in the Study of MAPK Signaling Yoav D. Shaul and Rony Seger Structure of MAPKs Elizabeth J. Goldsmith, Melanie H. Cobb, and Chung-I Chang Subcellular Localization of MAPKs Makoto Adachi and Eisuke Nishida Study of MAPK Signaling Using Knockout Mice Gilles Pages and Jacques Pouyssegur Computer Simulation of MAPK Signal Transduction Baltazar D. Aguda and Herbert M. Sauro Signaling by Growth Factor Receptors Bose S. Kochupurakkal and Yosef Yarden Activation of MAPK by G Protein-Coupled Receptors Piero Crespo and J. Silvio Gutkind Identification of MAPK Substrates by Expression Screening With Solid-Phase Phosphorylation Rikiro Fukunaga and Tony Hunter Study of Substrate Specificity of MAPKs Using Oriented Peptide Libraries Michael B. Yaffe Ras Signaling Pathway for Analysis of Protein-Protein Interactions in Yeast and Mammalian Cells Ami Aronheim Methods in Functional Proteomics: Two-Dimensional Polyacrylamide Gel Electrophoresis With Immobilized pH Gradients, In-Gel Digestion, and Identification of Proteins by Mass Spectrometry Karine R. Bernard, Karen R. Jonscher, Katheryn A. Resing, and Natalie G. Ahn Practical Methods for Deuterium Exchange/Mass Spectrometry Andrew N. Hoofnagle, Katheryn A. Resing, and Natalie G. Ahn Use of Xenopus Oocytes and Early Embryos to Study MAPK Signaling Eusebio Perdiguero and Angel R. Nebreda MAPK Cascades in the Brain: Lessons From Learning Diego E. Berman and Yadin Dudai Index

STUDY OF GAIN OF FUNCTION MUTATION IN P53-NULL ASTROCYTES By Sang H. Choi A thesis submitted in partial fulfillment of the requirement for the degree of Master of Science at Virginia Commonwealth University. Virginia Commonwealth University, 2000. Major Director: Oliver Bogler, Ph.D., Assistant Professor, Department of Anatomy and Division of Neurosurgery. A number of recent studies suggest that expression of mutant p53 with mutations in certain codons show a gain of function and some of the characteristics of an oncoprotein. In order to study gain of function mutation and eliminate the potential of a dominant negative interaction with endogenous wild type p53 protein, p53 knockout mouse astrocytes were used. A retrovirus system was used to introduce mutant p53 genes into these p53-null astrocytes. Immunohistochemical staining and western blot experiments showed the expression of mutant p53 protein in these cells after infection with the mutant p53 retroviruses. Cell growth experiment did not suggest growth advantages for mutant p53 expressing astrocytes over vector control cells. Data from clonogenic survival assays following exposure to etoposide or cisplatin suggested that mutant p53 expressing cells with a point mutation at codon 273 may be resistant to

1 Abstract Understanding the expression changes that come with age is an important step in understanding the ageing process as a whole. By combining such transcriptomic data with other sources of information, for instance protein-protein interaction (PPI) data, it is possible to make inferences about the functional changes that occur with age. To address this, we conducted a meta-analysis on 127 publicly available microarray and RNA-Seq datasets from mice, rats and humans, to identify genes that are commonly differentially expressed with age in mammals. We also conducted analyses on subsets of these datasets, to produce transcriptomic signatures for brain, heart and muscle tissues, all of which are important tissues in the pathophysiology of ageing. This approach identified the transcriptomic signatures of the ageing system, as well as brain, heart and muscle tissues. We then applied enrichment analysis and machine learning to functionally describe those signatures. This revealed a typical ageing signature including the overexpression of immune and stress response genes and the underexpression of metabolic and developmental genes. Further analysis of the ageing expression signatures revealed that genes differentially expressed with age tend to be broadly expressed across tissues, rather than be tissue-specific, and that the ageing expression signatures (particularly the overexpressed signatures) of the whole system, brain and muscle tend to include genes that are central in PPI networks. We also show that genes underexpressed in the brain are highly central in a co-expression map, suggesting that underexpression of these genes may play a part in cognitive ageing. In sum, we show numerous functional similarities between the ageing transcriptomes of these important tissues, a broad non-specific expression pattern in genes differentially expressed with age, along with altered network properties of these genes in both a PPI and co-expression network.

Establishment and maintenance of pregnancy in a number of mammalian species depends upon a tightly regulated interaction between the semiallogeneic conceptus and the maternal uterine endometrium.The term "Maternal Recognition of Pregnancy" is attributed to Roger V. Short's paper titled "Implantation and the Maternal Recognition of Pregnancy" which was published in proceedings from the 1969 Symposium on Foetal Autonomy.Professor Short's landmark paper stimulated increased interest in elucidating how the conceptus signals its presence to assure maintenance of the corpus luteum beyond the normal length of the estrous or menstrual cycle to allow pregnancy to be established and maintained.Ten years following publication of Professor Short's paper, a Ciba Foundation Symposium entitled "Maternal Recognition of Pregnancy" brought together leading scientists to discuss the multiple mechanisms and pathways by which different viviparous species establish a successful pregnancy.The present volume on "Regulation of Implantation and Establishment of Pregnancy in Mammals" brings together current reviews from leading experts to address the diversity of mechanisms by which species establish and maintain pregnancy.Implantation in mice, dogs, pigs, cattle, sheep, horses, primates, humans and species in which embryonic diapause occurs are discussed.Reviews will provide current knowledge on the role of endometrial steroid receptors, adhesion factors, cytokines, interferons, steroids, prostaglandins, growth factors and immune cells involved with regulation of conceptus development.

Host cell attachment by pathogenic bacteria can play very different roles in the course of infection. The pathogenic spirochetes Borrelia hermsii and Borrelia burgdorferi sensu lato which cause relapsing fever and Lyme disease, respectively, are transmitted by the bite of infected ticks. After transmission, these spirochetes can cause systemic infection. Relapsing fever spirochetes remain largely in the bloodstream causing febrile episodes, while Lyme disease will often colonize a variety of tissues, such as the heart, joint and nervous system, resulting in a chronic multisystemic disorder. Borrelia species have the ability to bind to various cell types, a process which plays a crucial role in pathogenesis and may influence spirochetal clearance from the bloodstream. Colonization of multiple tissues and cell types is likely promoted by the ability to bind to components found in target tissues, and many B. burgdorferi adhesins have been shown to promote attachment to a wide variety of cells and extracellular matrix components. Different Lyme disease strains have been shown to preferentially colonize certain tissues, although the basis of this tissue tropism is not well understood. In this study we found that among different Lyme disease strains, allelic variation of the adhesin DbpA contributes to variation in its in vitro binding activities raising the possibility that this variation contributes to tissue tropism in vivo. In studying B. hermsii infection, we found evidence by both histological and fluorescence in situ hybridization (FISH) analysis of tissues that indicated that red blood cells were removed by tissue resident macrophages in infected mice. Spirochetes in the spleen and liver were often visualized associated with RBCs, lending support to the hypothesis that direct interaction of B. hermsii spirochetes with RBCs leads to clearance of bacteria from the bloodstream. Our findings indicate that host cell attachment play a key role in the establishment of Lyme disease infection, and in contrast contributes to the clearance of relapsing fever infection.