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Myo/Nog Cells: Targets For Preventing The Accumulation Of Skeletal Muscle-Like Cells In The Human Lens, Jacquelyn Gerhart, M. Greenbaum, V. Scheinfeld, P. Fitzgerald, M. Crawford, Arturo Bravo Nuevo, M. Pitts, Mindy George-Weinstein 2017 Philadelphia College of Osteopathic Medicine

Myo/Nog Cells: Targets For Preventing The Accumulation Of Skeletal Muscle-Like Cells In The Human Lens, Jacquelyn Gerhart, M. Greenbaum, V. Scheinfeld, P. Fitzgerald, M. Crawford, Arturo Bravo Nuevo, M. Pitts, Mindy George-Weinstein

Arturo Bravo-Nuevo

Posterior capsule opacification (PCO) is a vision impairing condition that arises in some patients following cataract surgery. The fibrotic form of PCO is caused by myofibroblasts that may emerge in the lens years after surgery. In the chick embryo lens, myofibroblasts are derived from Myo/Nog cells that are identified by their expression of the skeletal muscle specific transcription factor MyoD, the bone morphogenetic protein inhibitor Noggin, and the epitope recognized by the G8 monoclonal antibody. The goal of this study was to test the hypothesis that depletion of Myo/Nog cells will prevent the accumulation of myofibroblasts in human ...


Tol2 Gene Trap Integrations In The Zebrafish Amyloid Precursor Protein Genes Appa And Aplp2 Reveal Accumulation Of Secreted App At The Embryonic Veins, Hsin-Kai Liao, Ying Wang, Kristin E. Watt, Qin Wen, Justin Breitback, Chelsy K. Kemmet, Karl J. Clark, Stephen C. Ekker, Jeffrey J. Essner, Maura McGrail 2017 Iowa State University

Tol2 Gene Trap Integrations In The Zebrafish Amyloid Precursor Protein Genes Appa And Aplp2 Reveal Accumulation Of Secreted App At The Embryonic Veins, Hsin-Kai Liao, Ying Wang, Kristin E. Watt, Qin Wen, Justin Breitback, Chelsy K. Kemmet, Karl J. Clark, Stephen C. Ekker, Jeffrey J. Essner, Maura Mcgrail

Jeffrey J. Essner

Background—The single spanning transmembrane amyloid precursor protein (APP) and its proteolytic product, amyloid-beta (Aβ) peptide, have been intensely studied due to their role in the pathogenesis of Alzheimer’s disease. However, the biological role of the secreted ectodomain of APP, which is also generated by proteolytic cleavage, is less well understood. Here, we report Tol2 red fluorescent protein (RFP) transposon gene trap integrations in the zebrafish amyloid precursor protein a (appa) and amyloid precursor-like protein 2 (aplp2) genes. The transposon integrations are predicted to disrupt the appa and aplp2 genes to primarily produce secreted ectodomains of the corresponding proteins ...


Moesin1 And Ve-Cadherin Are Required In Endothelial Cells During In Vivo Tubulogenesis, Ying Wang, Mark S. Kaiser, Jon D. Larson, Aidas Nasevicius, Karl J. Clark, Shannon A. Wadman, Sharon E. Roberg-Perez, Stephen C. Ekker, Perry B. Hackett, Maura McGrail, Jeffrey J. Essner 2017 Iowa State University

Moesin1 And Ve-Cadherin Are Required In Endothelial Cells During In Vivo Tubulogenesis, Ying Wang, Mark S. Kaiser, Jon D. Larson, Aidas Nasevicius, Karl J. Clark, Shannon A. Wadman, Sharon E. Roberg-Perez, Stephen C. Ekker, Perry B. Hackett, Maura Mcgrail, Jeffrey J. Essner

Jeffrey J. Essner

Endothelial tubulogenesis is a crucial step in the formation of functional blood vessels during angiogenesis and vasculogenesis. Here, we use in vivo imaging of living zebrafish embryos expressing fluorescent fusion proteins of β-Actin, α-Catenin, and the ERM family member Moesin1 (Moesin a), to define a novel cord hollowing process that occurs during the initial stages of tubulogenesis in intersegmental vessels (ISVs) in the embryo. We show that the primary lumen elongates along cell junctions between at least two endothelial cells during embryonic angiogenesis. Moesin1-EGFP is enriched around structures that resemble intracellular vacuoles, which fuse with the luminal membrane during expansion ...


Mutations In Tubb8 And Human Oocyte Meiotic Arrest, Ruizhi Feng, Qing Sang, Yanping Kuang, Xiaoxi Sun, Zheng Yan, Shaozhen Zhang, Juanzi Shi, Guoling Tian, Anna Luchniak, Yusuke Fukuda, Bin Li, Min Yu, Junling Chen, Yao Xu, Luo Guo, Ronggui Qu, Xueqian Wang, Zhaogui Sun, Miao Liu, Huijuan Shi, Hongyan Wang, Yi Feng, Ruijin Shao, Renjie Chai, Qiaoli Li, Qinghe Xing, Rui Zhang, Eva Nogales, Li Jin, Lin He, Mohan L. Gupta Jr., Nicholas J. Cowan, Lei Wang 2017 Fudan University

Mutations In Tubb8 And Human Oocyte Meiotic Arrest, Ruizhi Feng, Qing Sang, Yanping Kuang, Xiaoxi Sun, Zheng Yan, Shaozhen Zhang, Juanzi Shi, Guoling Tian, Anna Luchniak, Yusuke Fukuda, Bin Li, Min Yu, Junling Chen, Yao Xu, Luo Guo, Ronggui Qu, Xueqian Wang, Zhaogui Sun, Miao Liu, Huijuan Shi, Hongyan Wang, Yi Feng, Ruijin Shao, Renjie Chai, Qiaoli Li, Qinghe Xing, Rui Zhang, Eva Nogales, Li Jin, Lin He, Mohan L. Gupta Jr., Nicholas J. Cowan, Lei Wang

Mohan Gupta, Jr.

BACKGROUND Human reproduction depends on the fusion of a mature oocyte with a sperm cell to form a fertilized egg. The genetic events that lead to the arrest of human oocyte maturation are unknown.

METHODS We sequenced the exomes of five members of a four-generation family, three of whom had infertility due to oocyte meiosis I arrest. We performed Sanger sequencing of a candidate gene, TUBB8, in DNA samples from these members, additional family members, and members of 23 other affected families. The expression of TUBB8 and all other β-tubulin isotypes was assessed in human oocytes, early embryos, sperm cells ...


Platelet Derived Growth Factor Receptor B (Pdgfrb) Expressing Cells During Zebrafish Coronary Vessel Development, Juancarlos Fierros 2017 California State University, San Bernardino

Platelet Derived Growth Factor Receptor B (Pdgfrb) Expressing Cells During Zebrafish Coronary Vessel Development, Juancarlos Fierros

Electronic Theses, Projects, and Dissertations

Coronary heart disease is a prevalent issue in developed countries throughout the world. It can have crippling effects on the quality of life and even lead to mortality, in the case of myocardial infarction. Part of the problem is the lack of a robust regenerative response in mammals after injury. Zebrafish have an amazing ability to regenerate after injury, and studies have demonstrated that the regenerative response recapitulates embryonic development. Our lab previously reported the first analysis of coronary vessel development in zebrafish and demonstrated that coronary endothelial cells undergo angiogenesis to form a vascular network. The roles of perivascular ...


Epitaxially Grown Collagen Fibrils Reveal Diversity In Contact Guidance Behavior Among Cancer Cells, Juan Wang, Joseph W. Petefish, Andrew C. Hillier, Ian C. Schneider 2017 Iowa State University

Epitaxially Grown Collagen Fibrils Reveal Diversity In Contact Guidance Behavior Among Cancer Cells, Juan Wang, Joseph W. Petefish, Andrew C. Hillier, Ian C. Schneider

Andrew C. Hillier

Invasion of cancer cells into the surrounding tissue is an important step during cancer progression and is driven by cell migration. Cell migration can be random, but often it is directed by various cues such as aligned fibers composed of extracellular matrix (ECM), a process called contact guidance. During contact guidance, aligned fibers bias migration along the long axis of the fibers. These aligned fibers of ECM are commonly composed of type I collagen, an abundant structural protein around tumors. In this paper, we epitaxially grew several different patterns of organized type I collagen on mica and compared the morphology ...


Method And Apparatus For Solid State Molecular Analysis, Eric Henderson, Curtis Mosher, Michael P. Lynch 2017 Iowa State University

Method And Apparatus For Solid State Molecular Analysis, Eric Henderson, Curtis Mosher, Michael P. Lynch

Eric Henderson

The invention is a method for the formation and analysis of novel miniature deposition domains. These deposition domains are placed on a surface to form a molecular array. The molecular array is scanned with an AFM to analyze molecular recognition events and the effect of introduced agents on defined molecular interactions. this approach can be carried out in a high throughput format, allowing rapid screening of thousands of molecular species in a sold state array. The procedures described here have the added benefit of allowing the measurement of changes in the analysis environment or introduction of additional effector molecules to ...


Decontamination Device And Method Thereof, Eric Henderson, James Vesenka 2017 Iowa State University

Decontamination Device And Method Thereof, Eric Henderson, James Vesenka

Eric Henderson

One embodiment of a decontamination device for a surface, such as a probe, and method thereof in accordance with the present invention includes a first electrode, a second electrode, and a transformer. The first electrode comprises a housing with an opening and one end of the second electrode is positioned in the opening. The second electrode has an elongated-shape which tapers to a point at the one end adjacent to the opening. The transformer is mounted in the housing and has first and second pairs of leads. One of the second pair of leads is coupled to the first electrode ...


Device And Method Of Use For Detection And Characterization Of Pathogens And Biological Materials, Eric R. Henderson, Saju R. Nettikadan, Curtis L. Mosher 2017 Iowa State University

Device And Method Of Use For Detection And Characterization Of Pathogens And Biological Materials, Eric R. Henderson, Saju R. Nettikadan, Curtis L. Mosher

Eric Henderson

The present invention includes a method and apparatus for the detection of a target material. The method and apparatus includes providing a substrate with a surface and forming a domains of deposited materials thereon. The deposited material can be placed on the surface and bound directly and non-specifically to the surface, or it may be specifically or non-specifically bound to the surface. The deposited material has an affinity for a specific target material. The domains thus created are termed affinity domains or deposition domains. Multiple affinity domains of deposited materials can be deposited on a single surface, creating a plurality ...


Macrophages Are Necessary For Epimorphic Regeneration In African Spiny Mice, Jennifer Simkin, Thomas R. Gawriluk, John C. Gensel, Ashley W. Seifert 2017 University of Kentucky

Macrophages Are Necessary For Epimorphic Regeneration In African Spiny Mice, Jennifer Simkin, Thomas R. Gawriluk, John C. Gensel, Ashley W. Seifert

Biology Faculty Publications

How the immune system affects tissue regeneration is not well understood. In this study, we used an emerging mammalian model of epimorphic regeneration, the African spiny mouse, to examine cell-based inflammation and tested the hypothesis that macrophages are necessary for regeneration. By directly comparing inflammatory cell activation in a 4 mm ear injury during regeneration (Acomys cahirinus) and scarring (Mus musculus), we found that both species exhibited an acute inflammatory response, with scarring characterized by stronger myeloperoxidase activity. In contrast, ROS production was stronger and more persistent during regeneration. By depleting macrophages during injury, we demonstrate a functional requirement for ...


Excitatory Neurons Sculpt Gabaergic Neuronal Connectivity In The C. Elegans Motor Circuit, Belinda Barbagallo, Alison Philbrook, Denis Touroutine, Navonil Banerjee, Devyn Oliver, Christopher M. Lambert, Michael M. Francis 2017 University of Massachusetts Medical School

Excitatory Neurons Sculpt Gabaergic Neuronal Connectivity In The C. Elegans Motor Circuit, Belinda Barbagallo, Alison Philbrook, Denis Touroutine, Navonil Banerjee, Devyn Oliver, Christopher M. Lambert, Michael M. Francis

Neurobiology Publications and Presentations

Establishing and maintaining the appropriate number of GABA synapses is key for balancing excitation and inhibition in the nervous system, though we have only a limited understanding of the mechanisms controlling GABA circuit connectivity. Here, we show that disrupting cholinergic innervation of GABAergic neurons in the C. elegans motor circuit alters GABAergic neuron synaptic connectivity. These changes are accompanied by a reduced frequency and increased amplitude of GABAergic synaptic events. Acute genetic disruption in early development-during the integration of post-embryonic born GABAergic neurons into the circuit-produces irreversible effects on GABAergic synaptic connectivity that mimic those produced by chronic manipulations. In ...


Mitochondrial Damage Accumulation In Oocytes __ A Potential Link Between Maternal Obesity And Increased Cardiometabolic Disease Risk In Offspring., Anna Louise Boudoures 2017 Washington University in St. Louis

Mitochondrial Damage Accumulation In Oocytes __ A Potential Link Between Maternal Obesity And Increased Cardiometabolic Disease Risk In Offspring., Anna Louise Boudoures

Arts & Sciences Electronic Theses and Dissertations

The developmental origins of health and disease (DoHAD) hypothesis suggests that negative maternal lifestyle choices, such as obesity, affect the health of her offspring. Clinical and laboratory studies support this hypothesis __ offspring born to obese mothers are at increased risk for health conditions including cardiometabolic syndrome and congenital abnormalities. Maternal obesity damages the oocytes, contributing to the increased disease risk by transmitting damaged organelles and epigenetic modifications to the offspring. Mitochondria, the most abundant organelle in the oocyte, are damaged in oocytes from obese females. However, we do not understand if mitochondrial damage in oocytes is reversible nor why ...


Two Tetrahymena G-Dna-Binding Proteins, Tgp1 And Tgp3, Share Novel Motifs And May Play A Role In Micronuclear Division, Quan Lu, Eric Henderson 2017 Iowa State University

Two Tetrahymena G-Dna-Binding Proteins, Tgp1 And Tgp3, Share Novel Motifs And May Play A Role In Micronuclear Division, Quan Lu, Eric Henderson

Eric Henderson

G-DNA is a four-stranded DNA structure with diverse putative biological roles. We have previously purified and cloned a novel G-DNA-binding protein TGP1 from the ciliate Tetrahymena thermophila. Here we report the molecular cloning of TGP3, an additional G-DNA-binding protein from the same organism. The TGP3 cDNA encodes a 365 amino acid protein that is homologous to TGP1 (34% identity and 44% similarity). The proteins share a sequence pattern that contains two novel repetitive and homologous motifs flanking an extensively hydrophilic and basic region. A nuclear fractionation experiment showed that TGP1 and TGP3 activities are localized predominantly in the nuclear fraction ...


Localization Of Individual Calcium Channels At The Release Face Of A Presynaptic Nerve Terminal, Philip G. Haydon, Eric Henderson, Elis F. Stanley 2017 Iowa State University

Localization Of Individual Calcium Channels At The Release Face Of A Presynaptic Nerve Terminal, Philip G. Haydon, Eric Henderson, Elis F. Stanley

Eric Henderson

Studies using biophysical techniques suggest a highly structured organization of calcium channels at the presynaptic transmitter release face (Llinás et al., 1981; Stanley, 1993), but it has not as yet proved possible to localize identified channels at the required nanometer level of resolution. We have used atomic force microscopy on the calyx-type nerve terminal of the chick ciliary ganglion to localize single calcium channels tagged via biotinylated ω-conotoxin GVIA to avidin-coated 30 nm gold particles. Calcium channels were in low (modal value approximately ⩽ 1 per μm2) and high (modal value55 per μm2) density areas and exhibited a prominent interchannel spacing ...


Ultrastructural Characterization Of Colloidal Metal Films For Bioanalytical Applications By Scanning Force Microscopy, Wolfgang Fritzsche, Konstantin Sokolov, George D. Chumanov, Therese M. Cotton, Eric Henderson 2017 Iowa State University

Ultrastructural Characterization Of Colloidal Metal Films For Bioanalytical Applications By Scanning Force Microscopy, Wolfgang Fritzsche, Konstantin Sokolov, George D. Chumanov, Therese M. Cotton, Eric Henderson

Eric Henderson

Colloidal metal films (CMFs) are prepared by the attachment of silver, gold, or platinum (or other metal) particles to a glass slide modified by silanization with 3‐mercaptopropyl silane. The covalent attachment of the metal particles occurs through the metal–sulfur bond. In these samples the local electromagnetic field is enhanced near the surface of the CMF due to excitation of plasmon resonances. This phenomenon can be used for a variety of analytical applications. Because the optical properties are strongly dependent on the morphology of the film, its structural characterization becomes of great importance. To further characterize CMFs we have ...


Three-Dimensional Imaging Of Living Neurons And Glia With The Atomic Force Microscope, Vladimir Parpura, Philip G. Haydon, Eric Henderson 2017 Iowa State University

Three-Dimensional Imaging Of Living Neurons And Glia With The Atomic Force Microscope, Vladimir Parpura, Philip G. Haydon, Eric Henderson

Eric Henderson

The atomic force microscope (AFM) was used to directly image hippocampal neurons and glia. Using chemically fixed and living cells it was possible to reconstruct three-dimensional cell structure and detect sub-cellular features such as the nucleus, mitochondria and filaments. By repeatedly scanning a single living cell we observed the movement of filaments beneath the cell membrane. Furthermore, by controlling the force applied to the scanning tip, nanosurgery was performed to selectively remove cells from the culture substratum. Thus, the atomic force microscope offers the opportunity to gain three-dimensional information about living cells and to observe the behavior of cellular components ...


Detection And Quantification Of Protein Biomarkers From Fewer Than 10 Cells, Saju Nettikadan, Korinna Radke, James Johnson, Juntao Xu, Michael Lynch, Curtis Mosher, Eric Henderson 2017 BioForce Nanosciences Inc.

Detection And Quantification Of Protein Biomarkers From Fewer Than 10 Cells, Saju Nettikadan, Korinna Radke, James Johnson, Juntao Xu, Michael Lynch, Curtis Mosher, Eric Henderson

Eric Henderson

The use of antibody microarrays continues to grow rapidly due to the recent advances in proteomics and automation and the opportunity this combination creates for high throughput multiplexed analysis of protein biomarkers. However, a primary limitation of this technology is the lack of PCR-like amplification methods for proteins. Therefore, to realize the full potential of array-based protein biomarker screening it is necessary to construct assays that can detect and quantify protein biomarkers with very high sensitivity, in the femtomolar range, and from limited sample quantities. We describe here the construction of ultramicroarrays, combining the advantages of microarraying including multiplexing capabilities ...


Atomic Force Microscopy And Manipulation Of Living Glial Cells, V. Parpura, P. G. Haydon, D. S. Sakaguchi, E. Henderson 2017 Iowa State University

Atomic Force Microscopy And Manipulation Of Living Glial Cells, V. Parpura, P. G. Haydon, D. S. Sakaguchi, E. Henderson

Eric Henderson

The atomic force microscope (AFM) is capable of imaging surfaces at very high resolution. The AFM has been used to image living glial cells in culture. Typical images reveal the three‐dimensional shape of the cell and often internal cellular structures are visible. In this report, it is shown that by increasing the imaging force, cells can be removed from the surface on which they are grown. Although the forces involved in this process are complex, it is possible to compare relative adhesion of different types of living cells to a particular substrate.


Atomic Force Microscopy Of Oriented Linear Dna Molecules Labeled With 5nm Gold Spheres, Wen-Ling Shaiu, Drena D. Larson, James Vesenka, Eric Henderson 2017 Iowa State University

Atomic Force Microscopy Of Oriented Linear Dna Molecules Labeled With 5nm Gold Spheres, Wen-Ling Shaiu, Drena D. Larson, James Vesenka, Eric Henderson

Eric Henderson

The atomic force microscope (AFM;1) can image DNA and RNA in air and under solutions at resolution comparable to that obtained by electron microscopy (EM) (2–7). We have developed a method for depositing and imaging linear DNA molecules to which 5nm gold spheres have been attached. The gold spheres facilitate orientation of the DNA molecules on the mica surface to which they are adsorbed and are potentially useful as internal height standards and as high resolution gene or sequence specific tags. We show that by modulating their adhesion to the mica surface, the gold spheres can be moved ...


Identification And Characterization Of A Putative Telomere End-Binding Protein From Tetrahymena Thermophila, Hong Sheng, Zhen Hou, Ted Schierer, Drena L. Dobbs, Eric Henderson 2017 Iowa State University

Identification And Characterization Of A Putative Telomere End-Binding Protein From Tetrahymena Thermophila, Hong Sheng, Zhen Hou, Ted Schierer, Drena L. Dobbs, Eric Henderson

Eric Henderson

Telomeric DNA of Tetrahymena thermophila consists of a long stretch of (TTGGGG)n double-stranded repeats with a single-stranded (TTGGGG)2 3' overhang at the end of the chromosome. We have identified and characterized a protein that specifically binds to a synthetic telomeric substrate consisting of duplex DNA and the 3' telomeric repeat overhang. This protein is called TEP (telomere end-binding protein). A change from G to A in the third position of the TTGGGG overhang repeat converts the substrate to a human telomere analog and reduces the binding affinity approximately threefold. Changing two G's to C's in the ...


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