Signal transduction Research Project You are working in a molecular biology labo

Question

Signal transduction Research Project You are working in a molecular biology laboratory to study Dopamine (DA) signal transduction pathway in spiny lobster. According to previous research, DA alters rhythmic behaviors (food digestion) of spiny lobsters by modulating neuron synaptic and intrinsic firing properties in pylorie network. The 14 pyloric neurons fall into six cell types, and DA actions are cell type specific. The transient potassium current mediated in most cell types. DA changes in opposing directions in pyloric dilator (PD) versus lateral pyloric (LP) neurons. The mechanism (s) underpinning cell-type specific DA modulation ofAs by shal channels (a) is a common target of DA modulation In the canonical pathways (pathways that have been discovered in other organisms), DA acts on several highly conserved DA receptors (DARs) that belong to the G-protein-coupled receptor (GPCR) superfamily. DA receptors (DARs) can be classified as type 1 (DIR) or type 2 (D2R). DIRs and D2Rs are known to increase and decrease intracellular cAMP concentrations respectively. We hypothesized that the opposing DA effects on PD and LP lA were due to differences in DAR expression patterns. Our preliminary data showed that PD neurons express D2Rs, but not DIRs, whereas LP expressed DIRs that were concentrated near synapses but did not express D2Rs. According to the canonical pathways, DIRs couple with Gs to increase adenylyl cyclase (AC) activity and D2Rs couple with Gi/o to decrease AC activity, thereby increasing and decreasing cAMP levels, respectively. Therefore, we will test if DA changes la in the lobster neuron by activating DIRs-AC-cAMP-PKA pathway in this specific project. An adenylyl cyclase (AC) activator, forskolin; 1,9-dideoxyforskolin (dd-forskolin), a structural analogue of forskolin that does not activate AC The cAMP analogue, 8-Br-cAMP: The protein kinase A (PKA) blockers, H-89 and Rp-CAMP . * . How could you use the above drugs to test if DA changes Ia in the lobster neuron by activating DIRS-AC-CAMP-PKA pathway, show the expected results in graphs/figures?

Explanation / Answer

Answer:

If we add adenylcyclase activator , will enhance the dopamine action on lobsters neuron through changes in LP Ia which indicate that D1R-AC-cAMP- protein kinase A is a signal transduction pathway that changes Ia in lobsters neuron.

Addition of AC analogue , decrese the action of adenyl cyclase means decrease cAMP lead to the dopamine effect oppositely on PD Ia.

cAMP analogue also inhibit the action produce the pathway clearly indicate that the above mentioned signal direction pathway responsible for DA changes Ia in lobster neuron.

Protein kinase block the downstream pathway lead to no change in LP Ia but opposite change occur PD Ia.

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