Simple spectroscopic Methods for estimating Brain Neurotransmitters, Antioxidant Enzymes of Laboratory animals like Mice: A review

About Authors:
Habibur Rahman1, M.C Eswaraiah2
1Department of Pharmacology,
2Dept. of Pharmacognosy,
Anurag Pharmacy College,
Ananthagiri (V), Kodad (M),
Nalgonda (Dt.), A.P-508206, India

ABSTRACT
Brain neurotransmitters like Dopamine, serotonin, Adrenaline, Glutamate and enzymes like Acetyl cholinesterase (AchE), Mono amino-oxidase (MAO-A & MAO-B) levels and antioxidant enzymes like Superoxide dismutase, catalase, peroxidase etc play important roles in different complex neurological disorders like Alzheimer’s disease(AD), Parkinsonism, depression etc. But biochemical estimation of these parameters is very difficult to conduct in simple laboratory. This review is a collection of simple spectroscopic methods to determine the common brain neurotransmitters and antioxidant enzymes.

Reference Id: PHARMATUTOR-ART-1244

Introduction
Neurotransmittersare endogenous chemical that transmit signals from a neuron to a target cell across a synapse. Glutamate is the major excitatory neurotransmitter in the brain and spinal cord and it involved in different disease like anxiety, epilepsy, psychosis, schizophrenia, Alzheimer’s disease and others common neurological disorders [1].  Excessive glutamate release can lead to excitotoxicity causing cell death.  GABA is used at the great majority of fast inhibitory synapses in virtually every part of the brain. Many sedative/tranquilizing drugs act by enhancing the effects of GABA [2].   Acetylcholine is distinguished as the transmitter at the neuromuscular junction connecting motor nerves to muscles.  Acetylcholine involved in memory, paralysis and others disorders.  Dopamine has a number of important functions in the brain. It plays a critical role in the reward system, but dysfunction of the dopamine system is also implicated in Parkinson's disease and schizophrenia.[3] Serotonin is a monoamine neurotransmitter to regulate appetite, sleep, memory and learning, temperature, mood, behavior, muscle contraction, and function of the cardiovascular system and endocrine system. It is speculated to have a role in depression, as some. The brain is uniquely vulnerable to oxidative injury, due to its high metabolic rate and elevated levels of polyunsaturated lipids, the target of lipid per oxidation.  Antioxidants appear to prevent oxidative stress in neurons and prevent apoptosis and neurological damage. Antioxidants are also being investigated as possible treatments for neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis [4, 5].

Estimation of Brain Neurotransmitters
Acetylcholineserase (AChE) enzyme determination

Acetylcholineserase (AChE) enzyme activity was estimated by Elman method [6].

Reagents
1. 0.1M Phosphate buffer
Solution A:  5.22g of K2HPO4 and 4.68g of NaH2PO4 are dissolved in 150 ml of distilled water.
Solution B: 6.2g NaOH is dissolved in 150ml of distilled water.
Solution B is added to solution A to get the desired pH (pH 8.0 or 7.0) and then finally the volume is made up to 300ml with distilled water.

2. DTNB Reagent
39.6 mg of DTNB with 15 mg NaHCO3 is dissolved in 10 ml of 0.1M phosphate buffer (pH 7.0).

3. Acetylthiocholine (ATC)
21.67 mg of acetylthiocholine is dissolved in 1 ml of distilled water.

Procedure
The mice were decapitated; brains are removed quickly and placed in ice-cold saline. Frontal cortex, hippocampus and septum are quickly dissected out on a Petri dish chilled on crushed ice. The tissues are we


Comments

I am a student of M.Sc. Semester IV and i am doing my dissertation work on activity of neurotransmitters on chicken brain intoxicated by fugal food. in that my one protocol is study of MAO-A and B , i have fulfill my this protocol by the method of Charles, M. and J. McEwen, 1977. Mao Activity in Rabbit Serum. In: Methods in Enzymology, XVIIB, Tabor, H. and C.W. Tabor (Eds.). Academic Press, New York and London, pp: 692-698. in this method i have got my readings but i do not know the formula to convert it into nmoles/mg protein. so, if u will email me formula for that and related articals it will help me in my dissertation work & i will be thank full to you for that.

Calculations are the biggest headache for rookie researcher. Your instructions are easy to comprehend. Thanks a lot...

dpsingh's picture

Please send the calculation of MAO-A and MAO-B activity in nmol/mg protein.
I have absorbance for the same.
 
Thanks.

SirI AM DOING mpharm in trivandrum medical college. This protocol is very useful for my study and i got the results. but i dont have the formula for calculating the amount of neurotransmittors. I request u to kindly send me the calculations for determination of neurotransmittors. if u send this as soon as possible It will definetly help me to complete my work and am will be very thankful to you.
Thanks
Athira

 

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