Regulation of adenosine transporter and AMPA receptor subunit localization by protein kinase CK2 in rat hippocampus

The control of extracellular adenosine is crucial to the regulation of synaptic transmission and neuroprotection. Equilibrative nucleoside transporters (ENTs) are highly expressed in the hippocampus and widely accepted as critical regulators of adenosine tone. However, the mechanisms regulating the surface distribution and transport function of ENTs are largely unknown. Since ENT1 and ENT2 contain consensus sequences for phosphorylation by protein kinase CK2, and because this protein has been reported to regulate synaptic plasticity and ENT function in non-neuronal systems, the present thesis outlines the hypothesis that CK2-induced phosphorylation of ENTs is important for their cellular localization and thus the regulation of adenosine tone and synaptic transmission. Here, a functional interaction between adenosine CK2, ENTs and AMPA receptors in the hippocampus is reported. Western blot analysis shows that a variety of CK2 inhibitors (DMAT, TBB and DRB) significantly reduced the density of ENT1 and ENT2 proteins in hippocampal membrane fractions, suggesting that CK2-mediated phosphorylation of ENTs promotes their surface localization. In contrast, it was found that the ENT1 inhibitor NBTI significantly increased in the membrane localization of ENT1, relative to the control. Moreover, ENTs were found to immunoprecipitate with GluR1 and GluR2-containing AMPA receptors; and CK2 inhibitors caused a decrease in the membrane localization of GluR2 and GluR1 AMPA receptors. These results suggest a novel signaling complex linking CK2-regulated adenosine transport to AMPA receptor trafficking in the rat hippocampus. Although the physiological significance of these findings requires further investigation, this thesis provides insight into an adenosine regulation pathway that may be important for the regulation of synaptic transmission and neuroprotection in the rat hippocampus

Analysis of somitogenesis using multiphoton laser scanning microscopy (MPLSM)

In order to study complex cellular interactions in the developing somite and nervous system, we have been refining techniques for labeling and imaging individual cells within the living vertebrate embryo. Most recently, we have been using MPLSM to analyze cellular behaviors, such as cell migration, filopodial extension, cell process collapse, and neuron pathfinding using time-lapse microscopy in 3-dimensions (3-d). To enhance the efficiency of two-photon excitation in these samples, we have been using a Zeiss LSM 510 NLO fiber delivery system with a Grating Dispersion Compensator (GDC). This system not only offers the convenience of fiber delivery for coupling our Ti:Sapphire laser to the microscope, but also affords us precise control over the pulsewidth of the mode- locked beam. In addition, we have developed a novel peptide/non-cationic lipid gene delivery system to introduce GFP plasmid into somite cells. This approach has allowed us to generate detailed 3-d images of somite cell morphologies at various stages of somite development in a way that best preserves the vitality of the cells being imaged

Bright Facet Sign and its Association with Demographic and Clinical Variables

Low back pain has a significant impact on global public health and economics. The bright facet sign (BFS), a common finding on magnetic resonance imaging (MRI) of the lumbar spine, is associated with low back pain. While degenerative joint disease (DJD) affects low back pain, its presence appears independent of the BFS at the disc and facet joints at the same spinal level. Increased BMI, considered a risk factor for DJD, has an inverse association with the BFS. The independent relationship of DJD and the BFS is poorly understood and may represent a previously unreported pain pathway. In this nested case-control quantitative study, based on an accepted conceptual framework, 350 lumbar MRI studies on symptomatic patients with historic and anthropomorphic data related to low back pain were analyzed using Spearman\u27s Rho and Multivariate Logistic Regression to examine any associations between the BFS at 3 spinal levels and the independent variables age, race/ethnicity, physical activity, BMI, trauma, low back pain, and DJD. The findings revealed significant associations between the BFS and the duration of pain, age, and gender at 1 or more spinal levels, the BFS and BMI and degenerative facet disease (DFD) at all 3 spinal levels, and no association between the BFS and degenerative disc disease (DDD). These results, contrary to current medical constructs where BMI, DFD, and DDD are considered predictive of low back pain, facilitate an improved understanding of joint function and contribute to the current body of knowledge related to low back pain. An understanding of the BFS as it relates to DJD and low back pain will assist clinicians with the early detection of spinal degeneration and the mitigation of pain and suffering, contributing to positive social change

Using dual eye tracking to uncover personal gaze patterns during social interaction

We report the personal eye gaze patterns of people engaged in face-to-face getting acquainted conversation. Considerable differences between individuals are underscored by a stability of eye gaze patterns within individuals. Results suggest the existence of an eye-mouth gaze continuum. This continuum includes some people showing a strong preference for eye gaze, some with a strong preference for mouth gaze, and others distributing their gaze between the eyes and mouth to varying extents. Additionally, we found evidence of within-participant consistency not just for location preference but also for the duration of fixations upon the eye and mouth regions. We also estimate that during a 4-minute getting acquainted conversation mutual face gaze constitutes about 60% of conversation that occurs via typically brief instances of 2.2 seconds. Mutual eye contact ranged from 0-45% of conversation, via very brief instances. This was despite participants subjectively perceiving eye contact occurring for about 70% of conversation. We argue that the subjective perception of eye contact is a product of mutual face gaze instead of actual mutual eye contact. We also outline the fast activity of gaze movements upon various locations both on and off face during a typical face-to-face conversation

Studies on limb blood flow in normal and abnormal pregnancy

Abstract Not Provided

Characterization of Kupffer cells in livers of developing mice

Abstract Background Kupffer cells are well known macrophages of the liver, however, the developmental characteristics of Kupffer cells in mice are not well understood. To clarify this matter, the characteristics of Kupffer macrophages in normal developing mouse liver were studied using light microscopy and immunocytochemistry. Methods Sections of liver tissue from early postnatal mice were prepared using immunocytochemical techniques. The Kupffer cells were identified by their immunoreactivity to the F4/80 antibody, whereas endothelial cells were labelled with the CD-34 antibody. In addition, Kupffer cells and endothelial cells were labelled by systemically injected fluorescently labelled latex microspheres. Tissue slices were examined by fluorescence microscopy. Results Intravenous or intraperitonal injections of microspheres yielded similar patterns of liver cell labelling. The F4/80 positive Kupffer cells were labelled with both large (0.2 μm) and small (0.02 μm) diameter microspheres, while endothelial cells were labelled only with the smaller diameter microspheres. Microsphere labelling of Kupffer cells appeared stable for at least 6 weeks. Cells immunoreactive for F4/80 were identified as early as postnatal day 0, and these cells also displayed uptake of microspheres. Numbers of F4/80 Kupffer cells, relative to numbers of albumin positive hepatocytes, did not show a significant trend over the first 2 postnatal weeks. Conclusions Kupffer cells of the developing mouse liver appear quite similar to those of other mammalian species, confirming that the mouse presents a useful animal model for studies of liver macrophage developmental structure and function