Hi,
I am going to investigate on my research drug regarding influence on neurotransmitter.
Kindly give your valuable suggestions regarding ideal biological sample like brain homogenate or serum or any other to get fruitful result from my research.
In my view, the ideal sample for neurotransmitter is the sample extract by microdialysis followed by HPLC/Mass etc. in conscious animals. This will provide the exact idea about the neurotransmitter release. However, region specific homogenate are also useful to detect the total quantity of neurotransmitter. But in this way we also need to check for its metabolite. Whole brain homogenate is not useful because region specific effect of neurotransmitter. Sometime it may compensate one region with another. Serum will never be a good choice to detect neurotransmitter.
Other alternative ist determine enzymatic activity of enzyme involved in synthesis of neurotransmitters in homogenates.
Irwin test is the gold behavioral test for finding first clues for suspected effects of a drug action on neurotransmitters. Once you have some evidences about any putative effect, the next step could be the in vivo microdialysis (e.g., HPLC detection) for neurotransmitters and/or neurotransmitter metabolites in order to establish more accurate hypothesis. You can start with ventricular microdialysis before going to brain specific regions.
Hi Pravin,
I will suggest you go for brain micordialysis, where you will perfuse specific brain regin with aCSF and collect dialysed CSF. In collected CSF you can measure different neurotransmitters by employing sensitive analytical method (LC-MS/MS or HPLC). Advantage with this method is that you can collect CSF at different intervals (0 to 360 mins) and you can correlate with plasma drug levels. Where as with brain homogenate technique you have to sac several animals to collect relevant result. Mainly handling brain tissue for measuring neurotransmitter levels is tricky. If you have stereotaxic apparatus and hands oon experience go for in vivo brain microdialysis.
Regards
Ramanatha
Depending on how you intend to perform the research, and which neurotransmitters you're looking for, you can analytically measure the NTs using LC-MS. It will likely be faster and even less expensive to perform your work using primary neurons in vitro. This will likely only work for GABA and glutamate. The numerous other NTs can be looked at as other researchers have suggested above using in vivo microdialysis.
If the target is brain I prefer microdialysis wşth HPLC. Serum samples of course analysed by HPLC using relevant standards, columns, analytical procedures available in literature.
Thank you to all researcher your guidance really help me a lot in my research.
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