Medicine and Health Sciences Commons^™

Rna-Guided Rna Cleavage By A Crispr Rna-Cas Protein Complex, Brenton R. Graveley, Sara Olson, Michael O. Duff

UCHC Articles - Research

Compelling evidence indicates that the CRISPR-Cas system protects prokaryotes from viruses and other potential genome invaders. This adaptive prokaryotic immune system arises from the clustered regularly interspaced short palindromic repeats (CRISPRs) found in prokaryotic genomes, which harbor short invader-derived sequences, and the CRISPR-associated (Cas) protein-coding genes. Here we have identified a CRISPR-Cas effector complex that is comprised of small invader-targeting RNAs from the CRISPR loci (termed prokaryotic silencing (psi)RNAs) and the RAMP module (or Cmr) Cas proteins. The psiRNA-Cmr protein complexes cleave complementary target RNAs at a fixed distance from the 3' end of the integral psiRNAs. In Pyrococcus furiosus …

Neuropeptide Y Is Expressed By Osteocytes And Can Inhibit Osteoblastic Activity, John C. Igwe, Xi Jiang, Frane Paic, Li Ma, Douglas J. Adams, Carol C. Pilbeam, Ivo Kalajzic

UCHC Articles - Research

Osteocytes are the most abundant osteoblast lineage cells within the bone matrix. They respond to mechanical stimulation and can participate in the release of regulatory proteins that can modulate the activity of other bone cells. We hypothesize that neuropeptide Y (NPY), a neurotransmitter with regulatory functions in bone formation, is produced by osteocytes and can affect osteoblast activity. To study the expression of NPY by the osteoblast lineage cells, we utilized transgenic mouse models in which we can identify and isolate populations of osteoblasts and osteocytes. The Col2.3GFP transgene is active in osteoblasts and osteocytes, while the DMP1 promoter drives …

Effects Of Estrogen On Muscle Damage In Response To An Acute Resistance Exercise Protocol, Megan R. Wolf

Honors Scholar Theses

Creatine Kinase (CK) is used as a measure of exercise-induced muscle membrane damage. During acute eccentric (muscle lengthening) exercise, muscle sarcolemma, sarcoplasmic reticulum, and Z-lines are damaged, thus causing muscle proteins and enzymes to leak into the interstitial fluid.

Strenuous eccentric exercise produces an elevation of oxygen free radicals, which further increases muscle damage. Muscle soreness and fatigue can be attributed to this membrane damage. Estradiol, however, may preserve membrane stability post-exercise (Brancaccio, Maffulli, & Limongelli, 2007; Carter, Dobridge, & Hackney, 2001; Tiidus, 2001). Because estradiol has a similar structure to Vitamin E, which is known to have antioxidant properties, …

The Role Of Epha4 In Glial Scar Formation Following Injury, Krysti Todd

Honors Scholar Theses

Although many areas of the brain lose their regenerative capacity with age, stem cell niches have been identified in both the subventricular zone (SVZ) along the lateral walls of the lateral ventricles and the subgranular zone (SGZ) of the dentate gyrus (Gage, 2000; Alvarez-Buylla et al., 2001; Alvarez-Buylla and Lim, 2004). The SVZ niche utilizes many mechanisms to determine the migration patterns of neuroblasts along the RMS into the olfactory bulb, one being Eph/ephrin signaling (Conover et al., 2000; Holmberg et al., 2005). EphA4-mediated signaling is necessary for axon guidance during development, and its continued expression in the SVZ niche …

Sphingosine-1-Phosphate/Sphingosine-1-Phosphate Receptor 2 Signaling Induces Cox-2 Expression In Wilms Tumor, Mei-Hong Li, Teresa Sanchez, Timothy Hla, Fernando Ferrer

UCHC Articles - Research

Purpose

Cyclooxygenase-2 (COX-2) has been reported to be ubiquitously expressed in Wilms tumor, the most common malignant renal tumor in children. However, the regulation mechanism of COX-2 expression remains unexplored.

Materials and Methods

Quantitative real-time PCR and western blot analysis were performed to detect COX-2 mRNA and protein expression in WiT49 cells upon the stimulation by sphingosine-1-phosphate (S1P) as well as S1P₂ and COX-2 mRNA expression in 10 fresh frozen Wilms tumor tissues and their matched normal tissues. Overexpression, blockade and downregulation of S1P₂ were performed using adenoviral transduction, S1P₂ antagonist JTE-013 and siRNA transfection, respectively. The …

Regulation Of The Prostaglandin Pathway During Development Of Invasive Bladder Cancer In Mice, John A. Taylor Iii, Benjamin Ristau, Mathilde Bonnemaison, Olga S. Voznesensky, Poornima Hegde, George A. Kuchel, Carol C. Pilbeam

UCHC Articles - Research

Prostaglandin E₂ (PGE₂) is reported to play an important role in tumor development. We explored the differential expression of genes governing production of, and response to, PGE₂ during development of invasive bladder cancer. N-butyl-N-(4-hydroxybutyl)-nitrosamine (BBN) or vehicle-treated mice (n = 4–5) were euthanized after 4–8 weeks (period 1, P1), 12–16 weeks (P2), and 20–23 weeks (P3). Half of each bladder was analyzed histologically and the other half extracted for mRNA analysis by quantitative real-time PCR. Bladders from BBN-treated mice showed progression from submucosal inflammation (P1) to squamous metaplasia/focal CIS (P2) to poorly …

Microinjection Of Follicle-Enclosed Mouse Oocytes, Laurinda A. Jaffe, Rachael P. Norris, Marina Freudzon, William J. Ratzan, Lisa M. Mehlmann

UCHC Articles - Research

The mammalian oocyte develops within a complex of somatic cells known as a follicle, within which signals from the somatic cells regulate the oocyte, and signals from the oocyte regulate the somatic cells. Because isolation of the oocyte from the follicle disrupts these communication pathways, oocyte physiology is best studied within an intact follicle. Here we describe methods for quantitative microinjection of follicle-enclosed mouse oocytes, thus allowing the introduction of signaling molecules as well as optical probes into the oocyte within its physiological environment.