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Full-Text Articles in Physiology

Assembly Of Α4Β2 Nicotinic Acetylcholine Receptors Assessed With Functional Fluorescently Labeled Subunits: Effects Of Localization, Trafficking, And Nicotine-Induced Upregulation In Clonal Mammalian Cells And In Cultured Midbrain Neurons, Raad Nashmi, Mary E. Dickinson, Sheri Mckinney, Mark Jareb, Cesar Labarca, Scott E. Fraser E. Fraser, Henry A. Lester Dec 2003

Assembly Of Α4Β2 Nicotinic Acetylcholine Receptors Assessed With Functional Fluorescently Labeled Subunits: Effects Of Localization, Trafficking, And Nicotine-Induced Upregulation In Clonal Mammalian Cells And In Cultured Midbrain Neurons, Raad Nashmi, Mary E. Dickinson, Sheri Mckinney, Mark Jareb, Cesar Labarca, Scott E. Fraser E. Fraser, Henry A. Lester

Biology Faculty Publications

Fura-2 recording of Ca2+ influx was used to show that incubation in 1 μM nicotine (2–6d) upregulates several pharmacological components of acetylcholine (ACh) responses in ventral midbrain cultures, including a MLA-resistant, DHβE-sensitive component that presumably corresponds to α4β2 receptors. To study changes in α4β2 receptor levels and assembly during this upregulation, we incorporated yellow and cyan fluorescent proteins (YFPs and CFPs) into the α4 or β2 M3–M4 intracellular loops, and these subunits were coexpressed in human embryonic kidney (HEK) 293T cells and cultured ventral midbrain neurons. The fluorescent …


Minus-End Capture Of Preformed Kinetochore Fibers Contributes To Spindle Morphogenesis, Alexey Khodjakov, Lily Copenagle, Michael B. Gordon, Duane A. Compton, Tarun M. Kapoor Mar 2003

Minus-End Capture Of Preformed Kinetochore Fibers Contributes To Spindle Morphogenesis, Alexey Khodjakov, Lily Copenagle, Michael B. Gordon, Duane A. Compton, Tarun M. Kapoor

Dartmouth Scholarship

Near-simultaneous three-dimensional fluorescence/differential interference contrast microscopy was used to follow the behavior of microtubules and chromosomes in living alpha-tubulin/GFP-expressing cells after inhibition of the mitotic kinesin Eg5 with monastrol. Kinetochore fibers (K-fibers) were frequently observed forming in association with chromosomes both during monastrol treatment and after monastrol removal. Surprisingly, these K-fibers were oriented away from, and not directly connected to, centrosomes and incorporated into the spindle by the sliding of their distal ends toward centrosomes via a NuMA-dependent mechanism. Similar preformed K-fibers were also observed during spindle formation in untreated cells. In addition, upon monastrol removal, centrosomes established a transient …


Asymmetry Of The Central Apparatus Defines The Location Of Active Microtubule Sliding In Chlamydomonas Flagella, Matthew J. Wargo, Elizabeth F. Smith Jan 2003

Asymmetry Of The Central Apparatus Defines The Location Of Active Microtubule Sliding In Chlamydomonas Flagella, Matthew J. Wargo, Elizabeth F. Smith

Dartmouth Scholarship

Regulation of ciliary and flagellar motility requires spatial control of dynein-driven microtubule sliding. However, the mechanism for regulating the location and symmetry of dynein activity is not understood. One hypothesis is that the asymmetrically organized central apparatus, through interactions with the radial spokes, transmits a signal to regulate dynein-driven microtubule sliding between subsets of doublet microtubules. Based on this model, we hypothesized that the orientation of the central apparatus defines positions of active microtubule sliding required to control bending in the axoneme. To test this, we induced microtubule sliding in axonemes isolated from wild-type and mutant Chlamydomonas cells, and then …


Exposure Of Nervous System Cells To Polychlorinated Biphenyls (Pcbs) Results In Significant Alterations In Levels Of Expression Of Neurotrophic Factors, Charles F. Ide, Jay C. Means, Trisha M. Basford, John Spitsbergen Jan 2003

Exposure Of Nervous System Cells To Polychlorinated Biphenyls (Pcbs) Results In Significant Alterations In Levels Of Expression Of Neurotrophic Factors, Charles F. Ide, Jay C. Means, Trisha M. Basford, John Spitsbergen

Biological Sciences Faculty and Graduate Student Research

Toxic insult by PCBs results in learning and memory deficits in humans. Alterations in expression of neurotrophic factors and/or their receptors have been linked to changes in cognition. How PCBs affect cognition is not known. We propose that PCBs affect cognition by altering neurotrophic factor expression or effects. We exposed cultured C6 glial cells in medium containing PCB (Aroclor 1254 (10ppm)). Control cells were treated with DMSO or regular medium. Cells were incubated at 37o C for up to 72 hours. Medium samples were taken at 6hr, 24hr, 48hr, and 72hr intervals. Enzyme-linked immunosorbent assays (ELISA) were used to determine …