Developmental Biology Commons^™

Salinomycin: A Notch Signaling Antagonist - A Novel Way Of Targeting Cancer Stem Cells, Carlos U. Muzlera

The Meducator

Dr. Hassell’s research team aims to investigate the roles of therapeutically-relevant genes or gene signatures in the development of “tumour-initiating cells” or breast cancer stem cells. His research team also explores the effects of antagonistic compounds on certain regulatory receptor pathways using in vitro breast cancer cultures and transgenic mouse models. The following research focuses on validating the inhibitory effects of an anti-breast cancer stem cell agent, salinomycin, on downstream Notch signaling. It suggests the possibility of targeting cancer stem cells, the primary culprit in tumour initiation, chemoresistance, and metastasis, by inhibiting key regulatory pathways - such as Notch signaling ...

Activation Of Fgfr1 In Adult Cardiomyocytes Leads To Development Of A Hypertrophic Cardiomyopathy, Joy I. Wang

Undergraduate Research Symposium

In both humans and mice, Fibroblast Growth Factor (FGF) is upregulated following injury to the heart, and published studies have shown that FGF2 serves as a mediator in cardioprotection following cardiac stress or injury. Although FGF2 plays an important role following injury, mice that lack or overexpress FGF2 develop normally and do not have a phenotype under homeostatic conditions. It is currently unknown how FGF signaling is regulated in the adult heart and why the effects are only observed following injury. We hypothesized that FGF signaling may be repressed in the adult heart under homeostatic conditions and becomes reactivated following ...

Flash4 Dark Reference Images, George Mcnamara

George McNamara

Hamamatsu FLASH4.0 dark reference images, acquired with 10 second exposure times, no light to camera. Camera offset (set by Hamamatsu( is ~100 (the average intensity of the first image is always ~1 intensity level higher - an odd feature, but trivial in practice for a 16-bit camera).

George McNamara, Ph.D.

Single Cells Analyst at L.J.N. Cooper Lab

University of Texas M.D. Anderson Cancer Center

Β-Catenin Overexpression Within The Metanephric Mesenchyme Causes Renal Dysplasia Via Upregulation Of The Gdnf Signalling Axis, Sanjay Sarin

Open Access Dissertations and Theses

Renal dysplasia, a developmental disorder characterized by defective nephrogenesis and branching morphogenesis, ranks as one of the major causes of renal failure among the pediatric population. The molecular mechanisms underlying the pathogenesis of renal dysplasia are not well understood; however, changes in gene expression are a major contributing factor. In this study, we demonstrate that the levels of activated β-catenin, a transcriptional co-regulator, are elevated in the nuclei of ureteric, stromal, and mesenchymal cells within dysplastic human kidney tissue. To determine the mechanisms by which mesenchymal β-catenin over-expression leads to renal dysplasia, we generated a conditional mouse model in which ...

Sex Determination In Beetles: Production Of All Male Progeny By Parental Rnai Knockdown Of Transformer, Jayendra Nath Shukla, Subba Reddy Palli

Entomology Faculty Publications

Sex in insects is determined by a cascade of regulators ultimately controlling sex-specific splicing of a transcription factor, Doublesex (Dsx). We recently identified homolog of dsx in the red flour beetle, Tribolium castaneum (Tcdsx). Here, we report on the identification and characterization of a regulator of Tcdsx splicing in T. castaneum. Two male-specific and one female-specific isoforms of T. castaneum transformer (Tctra) were identified. RNA interference-aided knockdown of Tctra in pupa or adults caused a change in sex from females to males by diverting the splicing of Tcdsx pre-mRNA to male-specific isoform. All the pupa and adults developed from Tctra ...

Doublesex Target Genes In The Red Flour Beetle, Tribolium Castaneum, Jayendra Nath Shukla, Subba Reddy Palli

Entomology Faculty Publications

Sex determination cascade in insects terminates with the production of sex-specific protein, Doublesex (Dsx). We identified the dsx homolog (Tcdsx) in Tribolium castaneum. The pre-mRNA of Tcdsx is sex-specifically spliced into three female (Tcdsxf1, Tcdsxf2 and Tcdsxf3) and one male-specific (Tcdsxm) isoforms. Cis-regulatory elements potentially involved in sex-specific splicing of the Tcdsx pre-mRNA were identified in the female-specific exon and the adjoining intronic sequences. All the three female-specific TcDsx proteins share common OD1 and OD2 domains and differ in their C-terminal sequences. Knockdown of Tcdsx resulted in a reduction in the oocyte development, egg production and hatching of eggs laid ...

Video Codec Performance (Excel Spreadsheet), George Mcnamara

George McNamara

Video codec performance (Excel spreadsheet). Movie was made in 2005-2006 when I worked at City of Hope National Medical Center. VTLF refers to Video Timelapse Light Facility. Videos were outputted from MetaMorph as AVI files. Personally, I always recommend uncompressed video files fro scientific uses. I also encourage posting the original scientific data format (ex. .lsm, .zvi, .lif, .stk).

Molecular Mechanisms Regulating Neonatal Oocyte Survival And Primordial Follicle Formation In The Mouse Ovary, Robin L. Jones

Biology - Dissertations

In mammals, formation of the primordial follicle is a complex process involving the breakdown of germ cell cysts, where oocytes must separate from each other and subsequently become surrounded by somatic cells. As cysts separate, a large number of germ cells are lost by apoptosis, however the mechanisms by which cyst breakdown and germ cell death occur are not well understood. We first hypothesized that two anti-apoptotic regulators from the BCL2 family of proteins, BCL2 and MCL1, may be responsible for regulating neonatal oocyte survival. To elucidate the effects of BCL2 in the neonatal ovary, we examined ovaries of both ...

Pubspectra Tattletales, George Mcnamara

George McNamara

Tattletales for Multiplex Fluorescent Reporters in Single Cells for Metabolomics

George McNamara

Analytical Imaging Core Facility, University of Miami, Miller School of Medicine, Miami, FL

As of April 2013: L.J.N. Cooper & D.A. Lee Cellular Immunotherapy Lab, University of Texas M.D. Anderson Cancer Center, Houston, TX

Email: gmcnamara@med.miami.edu, geomcnamara@earthlink.net

Tattletales is my concept for spatial multiplexing many fluorescent protein (FP) biosensors in the same live cell. For example, there are excellent FP biosensors to Ca++ ions, pH, glucose, ribose, glutamine, glutamate, ATP, redox, ROS, pyruvate, cAMP, cGMP, IP3, PI(3,4,5)P3, cell cycle indicators (Fucci2), PKA, PKC, photsphatases, caspase(s) [1, 2]. However, these are typically used one biosensor per experiment, due in part to flooding the cell with soluble biosensor. That is, conventionally, either a metabolite (glucose) reporter or a signal transduction (Ca++) reporter can be imaged. By flooding the cell with the reporter, signal to noise ratio is compromized by autofluorescence.

Tattletales takes advantage of spatial multiplexing to both increase the number of different reporters, and improve signal to noise ratio by localizing each biosensor to a small volume. I started with the observation by Robinett et al [3] who localized 512 GFP-nls-LacI to a 256 LacO array as a 200 nm diameter diffraction limited spot (nuclear background due to overexpression). Many thousands of DNA binding proteins, of known sequence specifities, exist (LacI, TetR, GalR, etc for cell line studies; ZF-FPs, TALE-FPs to STRs, telomere repeat binding factors-FPs, etc for primary cells) and can be fused (as cDNAs) to different fluorescent proteins and FP biosensors.

Many biosensors are available as affinity series [1, 4], now enabling extended dynamic range. I realized that spatial multiplexing of many DNA binding protein-reporters by localizing to different spots in the cell nucleus and distinguished by combinatorial addressing, where N address colors ...

Hormonal And Morphological Aspects Of Growth And Sexual Maturation In Wild-Caught Male Vervet Monkeys (Chlorocebus Aethiops Pygerythrus), Tegan J. Gaetano

Theses and Dissertations

Knowledge of baseline changes in testosterone (T) and other androgens is central to both investigations of morphological, physiological, and behavioral correlates of inter-individual variation in the timing and shape of key events and transitions over the life course and questions of the evolution of species-specific schedules of maturation in primates. T represents an important determinant of spermatogenesis in male mammals and plays a central role in the expression of male sexual behavior and the development of secondary sex characteristics. This research integrates hormonal and morphometric methods to determine age-related changes in fecal testosterone (fT) metabolites and morphological markers of sexual ...

Nucleoredoxin And Wnt Signaling In F9 Cells, Leanne Sandieson

Electronic Thesis and Dissertation Repository

Nucleoredoxin is a redox sensitive protein recently shown to be involved in the Wnt/β-catenin pathway through binding dishevelled. When Wnt is present, dishevelled prevents a destruction complex from degrading β-catenin. This facilitates the translocation of β-catenin into the nucleus where it binds to TCF-LEF to impart changes in gene expression. This study used mouse F9 cells as a model to examine whether or not nucleoredoxin is involved in augmenting the Wnt/β-catenin signaling required for extraembryonic endoderm differentiation. It was found that nucleoredoxin and three dishevelled isoforms were present in F9 cells. Furthermore, nucleoredoxin was found to bind to ...

Cloning And Analysis Of Reck During Early Xenopus Laevis Development, Jessica Willson

Electronic Thesis and Dissertation Repository

Extracellular matrix (ECM) remodeling is crucial for the development and maintenance of multicellular organisms. Degradation of ECM components occurs through the activity of matrix metalloproteinases (MMPs) and their inhibitors. Reversion-inducing cysteine-rich protein with Kazal motifs (RECK) gene encodes a membrane-anchored protein and plays an important role in mediating ECM remodeling by inhibiting MMPs. To date, few in vivo studies exist examining RECK during development. The present study focuses on cloning and examining the expression of RECK during early Xenopus laevis development. A mature cDNA clone of the RECK gene was generated. RT-PCR, in situ hybridization, and immunohistochemistry were used to ...

Mating System Biology Of The Florida Native Plant: Illicium Parviflorum, Nicholas Earl Buckley

Masters Theses

Self-incompatibility is thought to have played a profound role in the evolution of the angiosperms. However, there is little evidence of self-incompatibility systems in early diverging lineages of flowering plants. Illicium parviflorum, one such early-divergent angiosperm, is an evergreen perennial species endemic to central Florida, particularly within the Ocala National Forest. Although locally abundant, I. parviflorum is currently listed as endangered at the state level due to being under constant threat of habitat disturbance and over-harvesting. Notably, this species had been described as self-incompatible due to its low seed-set. However, low seed set may also be a result of strong ...

Developing Monitoring Methods For Leptasterias Spp. As Sentinel Species In Detecting Local Environmental Changes, Zachary Sturbaum, Kathryn Nuessly, Riley J. Smith, C. Sarah Cohen

STEM Teacher and Researcher (STAR) Program Posters

Leptasterias spp., a six-rayed sea star, is found in rocky intertidal habitats ranging from Alaska to central California. Leptasterias spp. can be monitored on a broad scale throughout their range by a variety of means using timed counts and random plot censusing in order to detect both large-scale and local-level changes in the environment due to climate change, land-based human activity, or other environmental events. Leptasterias brood their young externally until the embryos grow into fully developed juveniles. These juveniles disperse by crawling away, limiting their dispersal potential. This localized dispersal provides an opportunity to use Leptasterias spp. as sentinel ...

Halloween 2012 Jack O'Lanterns Trick Or Treat, George Mcnamara

George McNamara

Halloween 2012 makes trick or treating more visual and interactive than in past years.

the download is a ZIP file containing three files.

Print out the (unnumbered) image on as large and nice printer paper as possible - I used glossy 44" wide here in Miami (University of Miami, MillerSchool of Medicine, Calder Library, Biomedical Communications dept - I also made another print on "fabric", also 44" wide to take with me to an HHMI Janelia Farm conference on 'turning images into knowledge' that ends on Oct 31 - might stay up for a second conference, "GFP..." that start Nov 4).

The other ...

Transcriptional Regulation Of Profilin Is Required For Drosophila Larval Wound Closure, Amanda Brock

UT GSBS Dissertations and Theses (Open Access)

Injury is an inevitable part of life, making wound healing essential for survival. In postembryonic skin, wound closure requires that epidermal cells recognize the presence of a gap and change their behavior to migrate across it. In Drosophila larvae, wound closure requires two signaling pathways (the Jun N-terminal kinase (JNK) pathway and the Pvr receptor tyrosine kinase signaling pathway) and regulation of the actin cytoskeleton. In this and other systems, it remains unclear how the signaling pathways that initiate wound closure connect to the actin regulators that help execute wound- induced cell migrations. Here we show that chickadee, which encodes ...

Har1 And Ljamp1 Dependent Regulation Of Root Architecture In Lotus Japonicus, Chong Sung Kim

Electronic Thesis and Dissertation Repository

Here, I characterize a root-branching hypermorph of Lotus japonicus, called cluster root-like1 (crl1), which originated from a har1-1 suppressor screen. I have shown that the crl1 root phenotype is determined by two independently segregating recessive mutations, har1-1 and Ljamp1-1, with corresponding HAR1 and LjAMP1 encoding an LRR‑receptor-like kinase and a predicted homologue of the Arabidopsis ALTERED MERISTEM PROGRAM 1 protein, respectively. Unlike har1-1, the Ljamp1 mutation does not affect the symbiotic properties of L. japonicus Gifu but exerts a pleiotropic effect on shoot development. Root architecture, however, is regulated by a synergistic action of HAR1 and LjAMP1, as the ...

Mcnamara 20120831fri-20120904tue Cosmic Ray Particles By Ccd Imaging, George Mcnamara

George McNamara

McNamara 20120831Fri-20120904Tue Cosmic Ray Particles by CCD imaging.zip contains image files in support of a Microscopy Today article - please see

http://www.microscopy-today.com/

Uncovering Dual Roles For Perk Signaling During Experimentally Induced Pancreatitis, Elena Fazio

Electronic Thesis and Dissertation Repository

Pancreatitis is characterized by inappropriate activation of digestive enzyme

precursors, or zymogens, local and systemic inflammation, dysregulation of

cellular calcium (Ca2+), and induction of the unfolded protein response (UPR).

The UPR consists of three distinct pathways all of which are activated during

pancreatitis. However, the molecular roles of each remain unclear. The

protein kinase RNA (PKR)-like ER kinase (PERK) pathway reduces general

protein translation by phosphorylating eIF2!, and is activated within minutes

of initiating pancreatic damage. Microarray analysis carried out by our lab

revealed robust upregulation of the PERK pathways members Activating

Transcription Factor (ATF) 3 and stanniocalcin (STC ...

Genetic Analysis Of The Hippo Pathway In Mouse Liver, Li Lu

UT GSBS Dissertations and Theses (Open Access)

Cancer therapy and tumor treatment remain unsolved puzzles. Genetic screening for tumor suppressor genes in Drosophila revealed the Hippo-signaling pathway as a kinase cascade consisting of five core components. Disrupting the pathway by deleting the main component genes breaks the balance of cell proliferation and apoptosis and results in epithelial tissue tumorigenesis. The pathway is therefore believed to be a tumor suppressor pathway. However, a corresponding role in mammals is yet to be determined. Our lab began to investigate the tumor suppression function of the potent mammalian Hippo pathway by putting floxed alleles into the mouse genome flanking the functional-domain-expressing ...