During wake/sleep transitions in animal/birds, various circuitry in the brain undergoes change/modification, citing the role of various types of neurotransmitters in the regulation of sleep-wake transition. Nucleus coeruleus (LC), a small nuclei in the pons of the brain stem region, has been shown to regulate the above transition as it is involved in maintaining waking and alertness by supplying norepinephrine to various parts of the brain like the pre-frontal cortex, hypothalamus etc. Increases in NE concentration in the brain drastically activate the cortical EEG and thus keep the animal awake. It has been shown that during sleep, LC firing subsides and in REM sleep it almost ceases its firing. I want to monitor the in-vivo changes of the NE concentration in the rat brain during the EEG recording.
You can directly analyse different monoamine level using Microdylasis assisted HPLC-FD/ED Method. That will give you real time analysis for the exact status of monoamines in the CSF.
If you need analysis of monoamines in different parts of brain like hippocampus, cortex, cebellum, midbrain, brain stem, entorhinal cortex, piriform cortex or stiatum, amygdala. Just separate these parts from brain and homogenize it and get it analyse from HPLC-FD/ECD method. For this you can follow my recently published article in Epilepsy and Behavior (Effect of saponin fraction from Ficus religiosa on memory deficit, and behavioral and biochemical impairments in pentylenetetrazol kindled mice. Epilepsy & Behavior 2013 In press)
I hope this will serve you best for the monoamine analysis.
If you wish to study the role of different brain locus in this change.. you can guide your steriotaxic apparatus to administer the specific agonist or antagonist at that locus and then confirm the administration by histological study. Once the agonism or antagonism is done then you can follow the monoamine changes in the discrete brain parts.
If still have some quarry you can ask again.
You could use the head-restrained rat technique (check on Gervasoni et al 1998). By recording EEG activity and measuring NE with microdialysis in the region of interest simultaneously, you should be able to see changes in NE concentration depending of the sleep-wake cycle.
To the best of my knowledge, the only way to measure NE concentration in-vivo in the brain is microdialisys. See: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3054889/
Wise answers below. I do recommend the combined EEG/EMG and microdialysis technique in freely moving animals, and not head restraint due to a non-stressed recording of NE in the different sleep stages. Good luck!
Absolutely, if you are not interested in doing single-cell recordings and local pharmacological manipulations, then go for the polygraphic recordings and microdialysis in freely moving animals! Good luck.
Well, there are various options depending on what facilities you have in/around lab. The common method is to use Microdylasis assisted HPLC-FD/ED Method. The other one would be to use in vivo-NMR spectroscopy. You can also use laser imaging if you can manage to get some suitable fluorescent tag for infrared imaging using IVIS system. I hope that now you have got many options available.
Use a telemetric biosensor with fixed potential amperometry . You will be able to record changes in nor epinephrine on a second scale. It has a better temporal resolution than microdyaisis, you have the advantage that it can be recorded by telemetry, which means no cables attached. Just insert the biosensor and start recording .
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