34 lsa subramaniyan.pdf

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Life Science Archives (LSA)

ISSN: 2454-1354

Volume – 1; Issue - 4; Year – 2015; Page: 233 - 239

©2015 Published by JPS Scientific Publications Ltd. All rights reserved

Research Article

NEUROMUSCULAR MODULATORY ACTIVITY OF LION FISH

Pterois russelii VENOM IN MICE

R. Saravanamurugan and A. Subramaniyan*

Department of Zoology, Annamalai University, Annamalai Nagar – 608 002, Tamil Nadu, India.

Abstract

The present study was aimed to examine the impact of crude extract as well as partially purified

protein of lionfish venom on the neuronal system of mice. The treated mice were showed various behavioral

changes including miner shivering in fore limbs, loss of balance, opaque eyes, convulsions forming from

mouth and exophthalmia. It is possibly correlated with CNS and PNS dysfunction. The protein was found to

be modulate the functioning of the sodium pump (Na+, K

+ and ATPase) at the presynaptic complex of neuro-

transmission, there by affecting ATP hydrolysis. They increase cholinesterase activity at the post synaptic

complex by modulating the activity of the enzyme Acetyl cholinesterase (AChE) and the modulation was

occurred in dose dependent manner. Histopathological observations revealed that the toxins were affected

the anatomy of the mice brain.

Article History Received : 20.08.2015

Revised : 27.08.2015

Accepted : 06.09.2015

Key words: Lion fish, P. russelii, Proteins,

Sodium pump, Acetylcholine, Neurotoxins and

Enzyme.

1. Introduction

The activity of neuromuscular system is

vital for normal behavior and muscular function

and it represents a prime target system on which

some toxicants can produce a detrimental effect.

The excitability of sensory cells, neurons and

myocytes depends on ion channel and signal

transducers that provide a regulated path for the

movement of inorganic ions across the plasma

membrane in response to various stimuli. Ion

channels in plasma membrane are primary targets

of marine toxins. These channels are important

regulators of cell physiology and many of the

pathological effects of toxins result from the

modulation of ion channels.

* Corresponding author: A. Subramaniyan,

Department of Zoology, Annamalai University,

Annamalai Nagar.

Nerve cells are different from other cells

that they conduct bioelectrical signals for long

distance and possess intercellular connections with

neurons and other tissues. For proper functioning

of the neuron, it is essential that neurotransmission

should be maintained undisturbed. Drugs and

toxins that alter the normal processes of the

neurotransmission and are known to be

neuromodulators yielding beneficial or deleterious

effects on neuronal function. Therefore studies on

neuromodulatory effects of the venom are

worthful because of their profound usefulness in

pharmacological and neurophysiological studies.

ATPase activity is associated with the

active transport system, which is necessary for the

extrusion of Na+ from cells and the accumulation

of K+ within these cells. This enzyme is essential

for such function as the generator of cell volume

and electrolyte composition. Acetylcholine (ACh)

Subramaniyan / Life Science Archives (LSA), Volume – 1, Issue – 4, Page: 233 - 239, 2015 234


is a neurotransmitter that functions in conveying

nerve impulses across synaptic clefts within the

CNS (Tripathi and Sritvastava, 2008). Following

the transmission of an impulse across the synapse

by ACh, AChE is released into the synaptic cleft

(Horton et al., 2006). This enzyme hydrolyzes

ACh to choline and acetate and transmission of the

nerve impulse is terminated (Liesener et al.,

2007). Venoms and toxins represent useful tools to

investigate muscle degeneration and regeneration

since synchronic lesion can be induced by these

agents (Harris, 1992). The aim of the present study

is to know the modulation in ATP hydrolysis by

Na+, K

+ and ATPase (sodium pump) and ACh

hydrolysis by acetylcholinesterase in

neuromuscular junction and nerve ending by the

influence of toxins extracted from lionfish

P. russelii venom.

2. Materials and methods

Venom preparation

Live specimens of the venomous lionfish

P. russelii were collected from Mandapam coast

and brought to the laboratory. The fishes were

killed (by cooling) and the venomous spines were

removed and stored in 10% glycerol solution at -

800C and the venom was extracted as described by

Church and Hodgson (2002). The venom protein

content was estimated by the method of Lowery et

al. (1951).

Behavioral study

Adult male Swiss albino mice (Mus

musculus) of 10 to 12 weeks old (22 ± 2 g) were

obtained from the Central Animal House, Rajah

Muthiah Medical College, Annamalai University.

The animals were maintained under controlled

conditions of temperature (23 ± 2 oC), humidity

(50 ± 5%), and light (10 and 14 hrs of light and

dark cycles, respectively) and were fed with

commercial standard pellet and provided water ad

libitum. Animal handling and experimental

procedures were approved by the Institutional

Animal Ethics Committee, Annamalai University

(Registration Number: 953/2012/CPCSEA) and

the animals were cared in accordance with the

“Guide for the care and use of laboratory animals”

and “Committee for the purpose of control and

supervision on experimental animals”. The

lionfish venom at concentrations of 10 µg/ml, 20

µg/ml, 30 µg/ml and 40 µg/ml was administered

to experimental animals through single

intraperitoneal injection and control group was

maintained in each case by injecting an equal

volume of sterile saline. The time of injection and

death, in addition to behavioral changes before

death were recorded.

Neuromodulatory activity

In vitro studies

The effect of lionfish venom on mice brain Na+,

K+ and ATPase enzyme

Na+, K

+ and ATPase were assayed in mice

brain by the method of Shalom and Katyare

(1985) and the inorganic phosphate released was

measured by the method of Fiske and Subbarow

(1925). The assay mixture consisting of 1ml

buffer, 0.2 ml magnesium sulphate, 0.2 ml

potassium chloride, 0.2 ml sodium chloride, 0.2

ml EDTA and 0.2 ml of tissue homogenate was

prepared. The reaction was started by the addition

of 0.2 ml ATP and was incubated. Then, the

contents were incubated at 37 °C for 15 min. At

the end of 15 min, 1 ml of 10% TCA was added to

arrest the reaction. The enzyme activity was

expressed as moles phosphorous liberated / min/

mg/ protein / hour.

The effect of lionfish venom on mice brain

AChE (Acetylcholinesterase) enzyme

AChE activity was assayed in mice brain

using the method of Ellman et al. (1961). Brain

isolated from the albino mice was homogenized in

0.25 M ice cold sucrose and 2 % (W/V) tissue

homogenate was prepared in the same sucrose

solution and kept aside as enzyme source. Three

ml phosphate buffer (pH 8.0) was taken in each

test tube, to which 0.1 ml of 2 % homogenate was

added and stirred. A volume of 100 µl of 0.01 M

DTNB was added and the initial color was

measured at 412 nm. The test solution, 20 µl of

acetylthiocholineiodide (0.075 M) was allowed to

continue for 15 minutes at room temperature. The



color developed was measured at 412 nm in

spectrophotometer.

𝐴𝐶ℎ𝐸 =

𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 × 𝐴𝑚𝑜𝑢𝑛𝑡 𝑜𝑓 𝐴𝐶ℎ 𝑝𝑟𝑒𝑠𝑒𝑛𝑡 𝑖𝑛 𝑡ℎ𝑒 𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑 ×

100Absorbance of the standard ×

Amount of protein in the sample ×incubation time in minutes

The enzyme activity was expressed as moles

of AChE hydrolyzed per mg protein per hour.

Histopathological analyses

The histopathological changes were

observed in mice brain by the standard

histological methods of Luna, 1968. The brain was

removed carefully after the animals died and

immersed in 10% buffered formalin at room

temperature and then they were dehydrated in a

graded series of alcohol and xylene, embedded in

paraffin wax and then sectioned transversely into

3 – 4 μm slices. Multiple slices were made and

stained by hematoxylin and eosin stains.

Histopathological changes were observed under a

light microscope.

Statistical Analysis

The experimental data were expressed as

means ± Standard Deviations. Differences in

means were estimated by using SPSS 16.0. One-

way analysis of variance (ANOVA) followed by

Duncan’s Multiple Range Test (DMRT) were

done. In all cases statistical significance is

indicated by P < 0.05.

3. Results

Behavioral changes in mice

After treating the mice with lionfish

venom, they showed the following behavioral

changes like lying on belly with forelimbs spread

wide, running around the cage in an exited

manner, escape reaction, prolonged palpitation,

closed eyes, grooming, shivering of forelimbs,

loss of balance, opaque eyes, squeaking, tonic

convulsions, gasping for breath, arching of body

backwards, paralysis, micturiction, flexing of

muscles, prodding (insensitive to stimuli),

diarrhea, lethargy, dragging of hind limbs, rolling

of tail, foaming from mouth and exophthalmia.

Neuromodulatory activity

The effect of lionfish venom on mice brain Na+,

K+ and ATPase enzyme activity

The Table - 1 show the changes in the

activity of Na+, K

+ and ATPase in mice brain. It

was revealed that with the increase in

concentration of P. russelii venom an increase in

Na+, K

+and ATPase activity was observed. At 10

µg/ml concentration of P. russelii venom, the

value was 12.46 ± 0.37 µ moles /min/mg of

protein. The value increased to 12.86 ± 0.46 at 20

µg/ml concentration of P. russelii venom and

14.34 ± 0.55 at 30 µg/ml concentration of P.

russelii venom. At 40 µg/ml concentration of P.

russelii venom the value was high 15.26 ± 0.45 µ

moles/min/mg protein.

Table –1: The effect of lionfish venom on mice brain

Na+, K

+ and ATPase enzyme

Concentration

of venom

(µg/ml)

Na+, K

+, ATPase activity

µM Pi liberated min /mg

/protein

Control 8.16±0.40

10 12.46±0.37*

20 12.86±0.46*

30 14.34±0.55*

40 15.26±0.45* (Values are mean ±SD, n=6, * P < 0.05, control and venom

treated animal)

The effect of lionfish venom on mice brain

AChE (Acetylcholinesterase) enzyme activity

An elevated level of AChE was observed

in brain of mice treated with different doses of

crude P. russelii venom. The increase in activity

observed was dose dependent (Table – 2).

The histopathological changes in the

venom treated mice revealed that the main organ

brain was affected by lionfish venom. The

histological section of the control mice brain

tissue showed a regular and compact configuration

with blood sinusoids. But, in the treated mice they

were highly congested with haemolyzed blood and

haemorrhage. Glial nodule formation was

observed in some areas of the cerebrum. The mild



congestion of capillaries and pycnotic nuclei, a

condition formed by the condensation of

chromatin in the nucleus of cell undergoing

necrosis were found in the cerebellum (Fig - 1).

Table –2: The effect of lionfish venom on mice brain

AChE (Acetylcholinesterase) enzyme

Concentration

of venom

(µg/ml)

AChE activity in Brain

µM Ach hydrolyzed /mg

/protein/hour

Control 0.28±0.02

10 0.31±0.03*

20 0.40±0.03*

30 0.63±0.01*

40 0.71±0.05*

(Values are mean ±SD, n=6, * P < 0.05, control and venom

treated animal)

Normal brain

Venom treated brain

A.Congested blood sinusoids,

B.Pycnotic nuclei,

C.Vacoulation,

D.Necrosis.

Fig- 1: Histopathological effects of P. russelii venom

on mice brain.

4. Discussion

The present study reveals that the toxin of

the lionfish altered the hydrolysis of ATP by Na+,

K+

and ATPase in the pre-synaptic region there by

possibly affecting signaling to intercellular

organelles. Our study is similarly with the study of

Xie and Askari (2002) and electrolyte movement

across epithelial cells (Ewart and Klip, 1995).

Some behavioral changes enlisted above such as

flexing of muscles, tonic convulsions, arching of

body backwards, paralysis might be due the

accumulation of AChE and total termination of

neuronal signaling (Zhang et al. ,2005) and also

increase in AChE was corroborated with

neuromuscular paralysis and death by

asphyxiation (Silman et al.,1988).

The membrane-bound enzyme ATPase

plays an important role in the maintenance of

membrane potentials and it has been estimated

that it accounts for upto 50 % of oxidative

metabolism in the brain and was deeply involved

in cellular function (Ratnakumari et al., 1995).

Membrane Na+, K

+and ATPase plays an important

role in active transport of Na+ and K

+ ions across

the plasma membrane. The enzyme was present in

high concentration in brain and muscle. Na+, K

+

and ATPase is ubiquitous in nature in the

mammalian central nervous system and it was

found predominantly in glial and nerve terminals

(Chen et al., 2005). The sodium gradient is

important for the uptake of neurotransmitters into

nerve cells and glia, which suggests that the

changes in Na+, K

+ and ATPase activity result in

the modulation of neurotransmission (Fighera et

al., 2006; Sivan, 2007). This enzyme activity has

been used as a potential indicator for membrane

toxicity (Engelke et al., 1992).

The observed increase in the activity of

Na+, K

+ and ATPase by P. russelii venom

suggests the presence of antinociceptive substance

in the venom. The increase in activity was dose

dependent. A similar increase in the enzyme

activity was observed by morphine and P.

valitance venom in the mouse brain (Masocha et

al., 2002; Balasubashini et al., 2006). Acetyl

cholinesterase (AChE) enzyme involved in the

hydrolysis of the neurotransmitter acetylcholine



and contributes to the integrity and permeability of

the synaptic membrane that occurs during

neurotransmission and conduction (Grafius et al.,

1971). This enzyme has been implicated in

cholinergic and non-cholinergic actions, which

may play a role in neurodegenerative diseases

(Cummings, 2000; Law et al., 2001; Habila et al.,

2012). Likewise, puffer fish Arothron hispidus

exhibited positive modulation in Na+, K

+ and

ATPase, Mg++

, ATPase and AChE enzyme

activity (Bragadeeswaran et al., 2010). The effect

of P. russelii venom on the AChE activity in mice

brain revealed that fish venom increases the

activity of enzyme in a dose dependent manner.

This may be either due to presence of

acetlycholine (Garnier et al., 1995) or by the

massive release of acetylcholine from the nerve

terminal that potentiates the activity of the enzyme

in mouse brain (Church and Hodgson, 2002).

Cohen and Olek (1989) have concluded

that the cellular action of toxin is by inducing

massive release and subsequent depletion of

acetylcholine from the nerve terminal.

Experiments with rat synaptosomes revealed that

stonefish venom affects neurotransmission and has

demonstrated the stimulation of release of the

neurotransmitter acetylcholine (Khoo et al., 1992).

The soldier fish G. marmoratus stimulate the

release of AChE to act at muscarinic receptors on

guinea - pig, gastrointestinal smooth muscle

(Hopkins et al., 1997). Trachynilysin (TLY)

isolated from Sirachynisvenom enhances the

release of acetylcholine from atrial cholinergic

nerve terminals (Colasante et al., 1996; Sauviat et

al., 2000; Ouanounou et al., 2000). The crude

venom exhibited neurostimulatory response on

mice brain AChE activity and the inhibitory effect

on AChE activity ranging between 23 % and 39 %

were caused by venom of C. lentiginosus (Pawan

Kumar et al., 2014).

Various lines of evidence strongly suggest

that the P. russelii venom was affected the

neuronal system functioning in mice. The brain of

mice that died upon the lionfish toxin

envenomation was due to the enlargement of

capillaries and glial nodules. The foci of

microglia in degenerating neurons were found

particularly in the cerebrum. Mild congestion of

capillaries and pycnotic nuclei, a chromatin in the

nucleus of a cell undergoing necrosis were found

in the cerebellum. Similar observations were

reported by Ravindran et al. (2010) in the sea

anemone extracts administrated to mice.

The histopathological changes in

P. russelii venom treated mice brain was the

vascular endothelium appeared to be injurious and

caused congestion of blood vessels and cloudy

swellings. Brain tissue showed spongiosis

(oedema) throughout the parenchyma. Congested

blood vessels, spongiosis, pycnotic nuclei, glial

cell accumulation, focal area necrosis state was

due to the neurotoxicity of P. russelii venom. The

similar results were observed in Bungarus

caeruleus envenoming. Considering the site and

the mode of action of this poison on tissue level,

this findings may contribute for the discovery of

antivenom related valuable pharmaceutical

products (Al-Mamun et al., 2015).

5. Conclusion

The present study demonstrated that the

marine lionfish P. russelii venom protein have

potent pharmacological properties without any

series of toxic effects at low dosage level. Hence,

such protein could be used against cholinesterase

inhibitors and neuromodulatory drugs can be

developed from lionfish venom. These finding

strengthen the health care industry for the

development of indigenous medicine and it can be

used as remedies for analgesic and neurological

disorder.

Acknowledgement

The authors thankful the University Grand

commission, New Delhi (India) for financial

assistance.

6. References

1) Al-Mamun M. A., Rahman M. A., Hasan R.,

Rahmann Z., Haque K. M. F., (2015).

Histopathological alterations induced by

common krait Bungarus caeruleus venom on

hepatic, renal and cardiac tissues of albino



mice.Int. J. Pharm and Pharmaceutical

Sciences.Vol 7, Issue 1.

2) Balasubashini, M., Karthigayan, S.,

Somasundaram, S.T., Balasubramanian, T.,

Viswanathan, P. and Menon, V. P. (2006). In

vivo and in vitro characterization of the

biochemical and pathological changes induced

by lionfish Pterios volitans venom in mice.

Toxicology Mechanisms and Methods. 16: 525

- 531.

3) Bragadeeswaran, S, Therasa D, Prabhu K,

Kathiresan K., (2010). Biomedical and

pharmacological potential of tetrodotoxin-

producing bacteria isolated from marine

pufferfish Arothron hispidus Muller, 1841. J.

Venom. Ani. Toxi. inclu. Tropi. Dis. 16:(3)

421-431.

4) Chen, P. W., Liu, S. H., Hsu, C. J. and Liu-

Shiau, S. V. (2005). Correlation of increased

activities Na+, K

+ ATPase and Ca

+ ATPase

with the reversal of cisplatin ototoxicity

induced by D-methionine in guinea pig.

Hearing Res., 205: 102 - 109.

5) Church, J.E. and Hodgson, W.C. (2002). The

pharmacological activity of fish venoms.

Toxicon., 40:1083 -1093.

6) Cohen, A.S. and Olke, J. (1989). An extract of

lionfish (Pterois valitans) spine tissue contains

acetylcholine and a toxin that affects

neuromuscular transmission. Toxicon, 27 (12):

1367 – 1376.

7) Colasante, C. Meunier, F.A. Kreger, A.S. and

Molgo, J. (1996). Selective depletion of clear

synaptic vesicles and enhanced quantal

transmitter release at frog motor nerve endings

produced by trachynilysin, a protein toxin

isolated from stonefish (Synanceia trachynis)

venom. Eur. J. Neuroscrl, 8: 2149 - 2156.

8) Cummings, J. L. (2000). The role of

cholinergic agents in the management of

behavioral disturbances in AIzheimer’s

disease. Int. J. Neuropsycho pharmacol., 3:21

- 29.

9) Ellman, G.L., Coutney, K.D., Andres, V. and

Featherstone, R.M. (1961). A new and rapid

colorimetric determination of Acetylcholine

esterase activity. Biochem. Pharmacol, 7:

88 - 95.

10) Engelke, M., Diehl, H. and Tahti, H. (1992).

Effects of toluene and dn-hexane on rat

membrane fluidity and integral enzyme

activities. Pharmacol. Toxicol, 71: 343 347.

11) Ewart S.H. and Klip A, (1995). Hormonal

regulation of the Na+, K

+ ATPase:

Mechanisms underlying rapid and sustained

changes in pump activity, Am J Physiol, 269C

– 295.

12) Fighera, M. R., Royes, L. F. F., Furian, A. F.,

Oliviera, M. S., Fiorenza, N. G., Filho, R. F.,

Petry, J. C., Coelho, R. C. and Carlo, F. M.

(2006). GM1 gangliosides prevent seizures,

Na+, K

+ ATPase activity inhibition and

oxidative stress induced by glutaric acid and

pentylenetetrazole. Neurobioi. Dis.ln press.

13) Fiske, C.H. and Subbarow, Y. (1925). The

Colorimetric determination of phosphorus. J.

Biol. Chem, 66: 375 – 400.

14) Garnier, P., Goudey-Perriere, F., Breton, P.,

Dewulf, C., Petek, F. and Perriere, C. (1995).

Enzymatic properties of the stonefish [S.

verrucosa Bloch and Schneider, 1801] venom

and purification of a lethal, hypotensive and

cytolytic factor. Toxicon., 33: 143 - 155.

15) Grafius, M. A., Bond, H. E. and Millar, D. B.

(1971). Acetyl cholinesterase interaction with

a lipoprotein matrix. Eur. J. Biochem, 22: 382

- 390.

16) Habila N., Inuwa H.M., Aimola I.A., Lasisi

O.I., Muhammad A., Okafor A. I., Williams

I.S., (2012). Acetylcholinesterase activity in

the brain and blood of mice infected with Naja

nigricolis venom. Biological Segment: 3(1)

BS/1565.

17) Harris, J.B. (1992). Natural toxins in study of

degeneration and regeneration of skeletal

muscle. in: Methods in Neurosciences, Vol. 8,

Neurotoxins [Ed. P.M.Conn] San Diego.

Academic Press, pp.298 - 310.

18) Horton, H.T., Moran, L.A., Scrimgeour, K.G.,

Perry, M.D., Rawn, D.J., (2006). Principles of

Biochemistry 4th ed. Pearson Int. ed. pp 147.

19) Hopkins, B. J., Hodgson, W.C. and

Sutherland, S.K., (1997). An in vitro

pharmacological examination of venom from

the soldier fish Gymnapistes marmoratus.

Toxicon, 35: 1101 - 1111.



20) Khoo, H.E., Yuen, R., Poh, C.H. and Tan,

C.H. (1992). Biological activities of Synanceia

horrida [Stonefish] venom. Nat. Toxins., 1: 54

- 60.

21) Law, A., Gauthier, S. and Quirion, R. (2001).

Say NO Alzheimer's disease: the putative links

between nitric oxide and dementia of the

Alzheimer’s type. Brain Res. Rev., 35:

73 - 96.

22) Liesener, A., Perclive, A., Schoni, R., Schebb,

N.H., Wilmer, M., Karst, U., (2007).

Screening of acetylcholinesterase inhabitors in

snake venom by electro spraying mass

spectrometry. Pure Appl. Chem. 79, 2339 –

2349.

23) Lowry, O.H., Rosebrough, N.J., Farr, A.L.,

Randal, R.J., (1951). Protein measurement

with Folin-phenol reagent. J. Biol. Chem.,

193:265 - 275.

24) Luna, L.G., 1968. Manu. Histol. Stain. Armed.

Forces. Inst. Pathol. McGraw- Hill Book

Company, New York, NY. p. 258.

25) Masocha, W., Gonzaliz, L. G., Baeyens, J. M.

and Agil, A. (2002). Mechanism involved in

morphine-induced activation of synaptosomal

Na+, K

+ ATPase. Brain Res, 957: 311

- 319.

26) Ouanounou, G., Mattel, C., Meunier, F.A.,

Kreger, A.S. and Molgo, J. (2000).

Trachynilysin, a protein neurotoxin isolated

from stonefish Synanceia trachynis venom,

increases spontaneous quantal release from

Torpedo marmorata neuromuscular junction.

Cybium, 24: 149 -156.

27) Pawan Kumar, K. Venkateshvaran, P. P.

Srivastava, S. K. Nayak, S. M. Shivaprakash,

S. K. Chakraborty, (2014). Pharmacological

studies on the venom of the marine snail

Conus lentiginosus Reeve, 1844. International

Journal of Fisheries and Aquatic Studies.

1(3): 79-85.

28) Ratnakumari, L., Aude, R., Quershi, I. A. and

Butterworth, R. F. (1995). Na+, K

+ ATPase

acitivies are increased in brain both in

congenital and acquired hyperammonemia

syndromes. Neurosci. Lett., 197: 89 - 92.

29) Ravindran, V.S, Kannan L & Venkateshvaran,

K, (2010). Biological activity of sea anemone

protein. I. Toxicology and Histopathology.

Indian J.Exp.Biol, 48: 1225.

30) Sauviat, M.P., Meunier, F.A., Kreger, A. and

Molgo, J. (2000). Effects of trachynilysin, a

protein isolated from stonefish [Synanceia

trachynis] venom on frog atrial heart muscle.

Toxicon., 38: 945 - 959.

31) Shallom, J.M. and S.S. Katyare, (1985).

Altered synaptosomal ATPase activity in rat

brain following prolonged in vivo treatment

nicotine. Biochem. Pharmacol, 34: 3445-3449.

32) Silman I. and Futerman A.H. (1988). Modes of

attachment of acetylcholinesterase to the

surface membrane, Eur. J. Biochem, 170: 11.

33) Sivan, G., Venketesvaran, K. and

Radhakrishnan, C.K. (2007). Biological and

Biochemical properties of Scatophagus argus

venom, Toxicon., 50 : 563-571.

34) Tripathi, A., Srivastava, U.C., (2008).

Acetylcholineasterase: A versatile enzyme of

the nervous system. Annals of Neurosci. 15,

106 – 110.

35) Xie Z. and Askari A, (2002). Na+ K

+ ATPase

as a signal transducter, Eur J. Biochem, 269:

2434.

36) Zhang D. and McCammon J.A, (2005). The

association of tetrameric Acetylcholinesterase

with Co1Q Tail: A Block Normal Mode

Analysis, Plos Comput Biol, 1: 6.

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