Experiment :

Anticonvulsant effect of drugs by MES and PTZ method

1. Introduction and Purpose of the experiment

Electroconvulsiometer is used to evaluate the anticonvulsant effects of drugs against

electroshock induced convulsions in experimental animals. It delivers the electric shock
of required intensity for required duration. An electrical stimulus with an intensity that
induces characteristic convulsion is applied in animals like rats or mice through the
electrodes placed in ear pinna. The effect of drug on duration and different phases of
seizures are recorded. Anticonvulsant like Phenytoin reduces the duration of seizures
induced by electrical shocks. The purpose of these experiments is to determine effect of
drug on convulsions in order to identify potential anticonvulsant drugs.
Pentylenetetrazol (PTZ) is a central nervous system stimulant. It produces jerky type
of clonic convulsions in rats and mice. The convulsive effect of this drug is considered to
be analogus to petitmal type of convulsions in man. PTZ act through GABA-
benzodiazepine receptor mechanisms in the brain. The purpose of these experiments is
to determine effect of drug on convulsions and also to identify potential anticonvulsant
drugs against petitmal type of convulsions.

2. Aim and Objectives

 To study the pattern of seizures induced by Maximal Electrocolvulsive Shock (MES)
 To record the duration of each phase of convulsion
 To demonstrate the effects of Phenytoin on MES induce convulsions in mice
 To study the anticonvulsant effect of diazepam against PTZ induced convulsions in
mice

3. Experimental setup
Animals – Mice
Drug – Phenytoin, PTZ and diazepam

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 Apparatus - Electroconvulsiometer
4. Experimental
Procedure

MES Method
 Weigh and number the animals. Divide them into control and test group each
consisting of 6 mice.
 Place the corneal electrodes on the ear pinna of animals and apply current of 30 mA
for 0.2 sec by using the electroconvulsiometer.
 Note the different stages of convulsions i.e, tonic flexion, tonic extensor, clonic
convulsion, stupor, recovery or death.
 Inject Phenytoin (25 mg/kg, i.p) to the animals and after 30 min, subject the animals
to electroshock and observe different stages of convulsions.
 Compare the duration of extension phase for both control and test groups and
analyze the observations.

PTZ Method
 Weigh and mark the animals, divide them into two groups each consisting of 4-6 rats.
Use one group as control and the other for diazepam treatment.

 Inject PTZ to control animals and note the onset of action (indicated by Straub’s tail,
jerky movements of the whole body and convulsions) and severity of convulsions due
to the drug.
 Inject diazepam to second group. After 30 min inject PTZ to these animals which have
received diazepam. Note the onset and severity of convulsions.
 Note either delay or complete abolition of convulsions in mice treated with diazepam.
 Compare the results of both control and test groups and analyze the observations.

5. Presentation of
results MES
Average duration of
Treatment group tonic extensor phase Report
(sec)

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 Control (Vehicle)

Test (Phenytoin)

PTZ

Convulsions

Treatment Onset Recovery/ Report

Nature and severity
(sec) death

PTZ

PTZ+diazepam

6. Analysis and Discussion

Maximal Electro Shock (MES) induced convulsions in animals represent grand mal
type of epilepsy. Electric shock is applied through the corneal electrodes. MES
convulsions are divided into 5 phase’s i.e. tonic flexion, clonic extension, clonic
convulsion, stupor, recovery or death. A substance is known to produce
anticonvulsant effect if it reduce or abolish the extension phase of MES.
Phenytoin is a hydantoin derivative also called as Diphenyl hydantoin or DPH.
Phenytoin particularly targets the Sodium ion channels which are repetitively
opening and closing. By preventing the opening of these channels, Phenytoin inhibits
the development of repetitive action potentials of the neurons (which cause
seizures). The great advantage of Phenytoin is that it can stop epileptic seizures
without producing sedation.
PTZ a CNS stimulant acts through GABA-benzodiazepine receptor mechanism in the
brain and produces jerky clonic convulsions. These effects of PTZ are antagonised by
diazepam. Diazepam enhances the frequency of GABA-mediated Chloride channels
and decreases the excitation. At higher doses they also block use-dependent sodium
channels which contribute to its effectiveness to treat convulsions.

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7. Conclusion

8. Comments
a) Limitations of the experiment: Strength and duration of electric shock alters the
results, may even cause death of animals; dose of PTZ that causes seizures in 80-
90% of the animals varies with strains being used.
b) Limitation of results: time taken for absorption of drugs may alter the results
c) Learning happened: Anticonvulsant activity of Phenytoin against maximum
electric shock induced convulsions
d) Recommendations
 Kulkarni, S.K. (1999) Hand Book of Experimental Pharmacology, 3rd Edition,
Delhi: VallabhPrakashan.
 VrushabendraSwamy, B.M., Jayaveera, K.N. and VenkateshwarReddy, A.(2014),
Experimental Pharmacology and Toxicology, New Delhi: S Chand and Company
Pvt , New Delhi: S. Chand & Company Pvt. Ltd.