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Intracellular free calcium is a important intracellular second
messager. It controls many very important physiological function of the
living cell. Under normal conditions, the intracellular free calcium concentration
are mediated by many mechanisms and sustain at a relatively stable level.
In order to understand how those mechanism mediate intracellular calcium
and how outside chemical substance affect the calcium signal transduction,
we compare whole cell patch clamp with single cell fluorescence [Ca2+]i
measurement as a mean of detection. Starting at the neuroblastoma cell SH-SY5Y
cell, we study the dynamic process of the intracellular [Ca2+]i.
In the first chapter , the necessary background of the filed discussed
is introduced. Also we present background of the technique used in this
section.
In chapter 2, the detail system setup and the various solutions used in the experiments are given and how to perform experiment are illustrated. Then in the next chapter, calcium channel data in SH-SY5Y cell are demonstrated.
The opioid substance, as a important drugs for pain-curing and addiction, its suppressive effects on the calcium channels are discussed. (Press here to see)
The chapter 5 show the muscarine-triggered calcium oscillation which was reported in SH-SY5Y cells for the first time, and also the mechanism of it. (Press here to see)
Opioid 's effects on the intracellular calcium store of the SH-SY5Y cell are studied in chapter 6, and its excitatory effects are show here.(Press here to see)
In the last chapter, a conclusion of the work we've done and a looking forward to the future works are made.
Key Words: Intracellular free calcium concentration, calcium channel, calcium store, opioid substance, SH-SY5Y cell line, patch clamp, single cell microfluorescence [Ca2+]i measurement