Dissertation > Excellent graduate degree dissertation topics show

Calcium Dynamics in Spine by FRET Based-on GFP

Author: LiuXiuLi
Tutor: GongHui
School: Huazhong University of Science and Technology
Course: Biomedical Engineering
Keywords: Hippocampal neurons Dendritic spines Calcium ion concentration Cameleon FRET
CLC: Q421
Type: Master's thesis
Year: 2005
Downloads: 102
Quote: 1
Read: Download Dissertation

Abstract


Intracellular calcium as a second messenger pass information on cell growth and differentiation plays an important role. The synthesis and release of neurotransmitters in the nervous system, the transmission of information between cells, the growth of dendrites, dendritic spine formation and synaptic plasticity and so on intracellular calcium ion has a close relationship. Studies have shown that calcium within dendritic spines involved in the regulation of multiple signaling pathways, and has an important contribution of long-term potentiation (LTP) and long-term depression (LTD) and LTP and LTD is considered to be learning and memory neural basis. Therefore monitoring changes in calcium concentration of single dendritic spines of hippocampal neurons to study of synaptic plasticity molecular and cellular mechanisms critical. The development of laser scanning confocal microscopy and calcium fluorescent probes detected dendritic spines of calcium signaling in the physiological state. Laser scanning confocal microscope with a small scattering background, the color difference is small, high resolution advantage. Gene probe the endogenous macromolecules kinetics of intracellular calcium, Ratio imaging can quantitatively detect the concentration of calcium ions. In this study, using the GFP-based calcium gene probe - Cameleon to study calcium dynamics of dendritic spines. First, the optimization of the cell culture, the calcium phosphate transfection method, using a 458 nm laser in place of the 430 nm laser light to perform the detection of fluorescence resonance energy transfer (FRET). Then, the comparison of the dynamic range of different Cameleon YC2.1, YC6.1, and YC3.60FRET ratio range of variation in the 458 nm excitation, respectively: 33%, 70%, 220%. Finally, preliminary perfected the method of FRET quantitative calcium concentration applied: (1) resting state calcium distribution. In the resting state, the cell bodies of neurons in the hippocampus, a dendritic two dendrites, three dendrites and dendritic spines calcium concentrations were: 104 ± 22 nM, 71 ± 10 nM, 55 ± 6 nM, 40 ± 7 nM, 56 ± 14 nM, dendritic spines of calcium ion concentration is higher than the corresponding dendritic confirmed dendritic neck diffusion barrier function; also found that the resting state distribution of calcium concentration in dendritic spines range of 13 ~ 80 nM, this calcium ion concentration difference may be related to the maturity of dendritic spines, function, and calcium works. (2) physiological stimulation of calcium ion concentration within the cell body and dendritic spines. To study calcium dynamics of the cell bodies of neurons and dendritic spines blowing plus glutamate extracellular concentration of calcium in the body increased from 100 nM to 1 μM, 100 μM glutamate stimulation, dendritic spines within calcium concentration increased from 50 nM to 500 nM. The results show that using Cameleon FRET ratio change the quantitative monitoring calcium dynamics in dendritic spines is feasible, and real-time, non-destructive, long-term monitoring for the study of neurotransmitter action potential to stimulate dendritic spines within calcium signal provides an important research tool.

Related Dissertations

  1. Effect of Chloride Channel Blocker on the Hippocampal Neuronal Apoptosis Induced by No in Culture,R363
  2. The Research on Fluorescence Spectrum Imaging and Photobleaching at Single Molecule Level,O657.3
  3. Intrathecal chloroprocaine neurotoxicity experimental study,R614
  4. Studies on the in Vivo Interaction between MD-2 and TLR4 Based on FRET,R459.7
  5. Studies on the Endocytic Trafficking and furin Protease-induced Cleavage of Several Recombinant Pseudomonas Exotoxin Proteins in Hela Cells,Q93
  6. Fabrication and Properties Research of Novel Fluorescent Nanoparticles for Cupric Ion Detection,TB383.1
  7. Studies on the Synthesis of CD36 Transmembrane Peptides and Mechanism of Transmembrane Domain Interactions,R96
  8. Gold Nanoparticles Quenching Fluorescence Probe for Glucose Detection and Cell Imaging Based on Fluorescence Resonance Energy Transfer (fret),Q503
  9. Inverstigation of Sulfhydryl Oxidases and Their Probes,Q55
  10. Preparation of Fluorescent Quantum Dots Probes and Their Application in Detecting Staphylococcus Aureus,O471.1
  11. Preparation and Characterization of Ratiometric Mercury Ion Sensor Based on FRET,TQ316.334
  12. Preparation and biological analysis of semiconductor nanomaterials,TB383.1
  13. Quantitatively Imaging Induced Activation of Rac1, Cdc42 Signaling Pathways,Q26
  14. Application of Fluorescence Resonance Energy Transfer (FRET) Technique in the Study of Cellular Signal Pathway,Q25
  15. Mechanism Study of the Biological Effects of Laser Irradiation in Living Cells,Q27
  16. Research on Mechemism of Photodynamic Threapy-Induced Apoptosis,R730.5
  17. Interaction between the N-terminal Domains of the AMPA Receptor Analyzed by Fluorescence Detection Methods,R33
  18. Identification of Interaction between Angiogenin and Progranulin,R73-3
  19. Screening for Candidate Molecule Interacts with LINGO-1 in Yeast Two Hybrid System,Q78
  20. Investigation of Nanolabeling Techniques Based on Quantum Dots and Their Toxicity to Cells,R319

CLC: > Biological Sciences > Physiology > Neurophysiology > The structure and function of nerve cells (neurons),glial
© 2012 www.DissertationTopic.Net  Mobile