CSB353H1 Lecture Notes - Lecture 8: Cyclic Adenosine Monophosphate, Green Fluorescent Protein, Reverse Genetics
28 Apr 2016
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How can we measure changes in ca2+ concentration in vivo: extraction and measurement from tissues. Doesn"t give real time info: ca2+ sensitive fluorescent dyes. Real time info; however, disruptive, gets into tissues: genetically-encoded ca2+ indicators. Transgenic cells express endogenous ca2+ sensitive fluorescent protein. Allows us to observe ca2+ signals in response to diverse stimuli. Amplitude and duration of calcium signal varies between cell types and tissues, means there"s variation between channels and receptors. If treat with channel blocker (lacl3) or chelator (egta), calcium signals are dampened. Different types of ca2+ signals are associated with different immune responses. Cyclic nucleotide-regulated ion channels (cngcs) play a key role in animal sensory perception neurons. Cylic nucleotide monophospates (i. e. camp or cgmp) and calmodulin (cam) regulate channel gating. 3000 bp deletion, which results in cngc 11 and 12 being fused expression of chimeric channel. No cpr22 phenotypes were observed in cngc 11 or 12 ko; however, cngc11/12 displayed phenotype.
