The progress report summarizes research on the neuroprotective effects of antioxidants against endosulfan-induced neurotoxicity in mice. Experiments were conducted on mice to analyze the effects of endosulfan and antioxidants (resveratrol, alpha-lipoic acid, vitamin E) on levels of acetylcholinesterase, butyrylcholinesterase, and ATPase in the brain. Preliminary results found that endosulfan inhibited these enzyme activities, while antioxidant supplementation protected against this neurotoxicity. Further biochemical analysis and statistical evaluation are planned for the next year to complete the study.

1. A progress report
on
“Evaluation of role of certain antioxidants on Endosulfan induced
neurotoxicity in the brain of Swiss mice (Mus musculus)”
By
HABIBA BANO
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2. INTRODUCTION
One of agricultural poisons.
It is a neurotoxic organochlorine insecticide.
It is an endocrine disruptor & is highly toxic.
TOXICITY : Neurotoxic
COLOUR : Brown
STATE / FORM : Solid, Crystals
ODOUR : Slight Sulphur Dioxide
NATURE : CNS Stimulant
GROUP : Chlorinated Cyclodiene Insecticide
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3. APPLICATIONS
Effective against a wide range of insects like beetles, caterpillars,
woolly apple aphid, colorado beetle, leaf hoppers & cabbage worms -by
contact & stomach action.
Used as a pre-harvest insecticide on fruits, nuts, vegetables, field &
grain crops including rice & ornamentals.
Also used on non-food crops such as tobacco & cotton.
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4. NEUROLOGICAL PROBLEMS
Directly affects CNS & prevents it from working properly.
Causes nausea, dizziness, headache, convulsions.
Learning disabilities, low IQ, scholastic backwardness.
Severe poisoning may result in death within hours or days.
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5. OBJECTIVES OF THE STUDY
To study the neurotoxicity induced by endosulfan in the brain
 To investigate the neuroprotective role of certain antioxidants on
endosulfan induced neurotoxicity.
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6. Materials and Methods
Dose Selection of endosulfan: - Sub chronic dose was determined
experimentally keeping in mind the LD 50 values.
Antioxidants: - Antioxidants Resveratrol, Alpha- lipoic acid and
Vitamin E were used which are very effective in preventing
neurotoxicity.
Animals for experiments: - Male Swiss mice (Mus musculus), 7-8
weeks old, weighing 22 ± 3 gms were used in the study.
Route of Exposure: Oral administration was followed for all the doses.
Administration of Antioxidants: Pre treatment of antioxidants was
given prior to endosulfan administration.
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7. GROUPS
Group 1: Control
Group 2: Endosulfan
Group 3: Resveratrol
Group 4: Endosulfan + Resveratrol
Group 5: Lipoic Acid
Group 6: Endosulfan + Lipoic Acid
Group 7: Vitamin E
Group 8: Endosulfan + Vitamin E
Group 9: Resveratrol + Lipoic Acid + Vitamin E
 Group 10: Endosulfan + Resveratrol+ Lipoic Acid+ Vitamin E
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8. ANTIOXIDANTS USED IN THE STUDY
3.RESVERATROL:
Naturally occurring phytoalexin
Found in the skins of certain red grapes, in peanuts, blueberries.
Act as antiviral. Anti cancer, anti aging, anti inflammatory agent.
Beneficial against number of brain disorders.
It directly scavenges superoxide and inhibits its generation.
Has the capacity to crossn Blood brain barrier.
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9. 2. ALPHA- LIPOIC ACID:
 Potent antioxidant
 Found inn potatoes and red meat.
 Has protective effects on brain and nervous tissue.
 Neutralizes free radicals.
 Prevents cancer, heart diseases, neuropathy, cataract, diabetes and
AIDS also.
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10. 3. VITAMIN E:
Lipid soluble antioxidant
Found in almonds, nuts, mangoes and other foods.
Important in the formation of red blood cells.
Prevents dementia, heart disease, cataract etc.
 Play role in immune function, in DNA repair and other metabolic
processes.
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11. PARAMETERS TAKEN
Histoenzymological experiments for the distribution of
following neurotransmitter enzymes was done on the brain of mice.
 Acetylcholinesterase (Karnowsky and Roots Method)
Butyrylcholinesterase ( Karnowsky and Roots Method)
Adenosine triphosphatase ( Wachstein Method)
Biochemical estimation of Serotonin levels in the brain of
mice.
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12. Summary of research activities done during the period of
04-07-2010 to 03-07-2011
 Experiments for the distribution of AChE, BChE and ATPase of all
the
ten groups under study has been performed on the whole brain of mice
along with their photography.
Estimation of serotonin levels in the brain of mice in all the groups has
been conducted and completed successfully.
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13. Histochemical localisation of AChE activity in different layers of
olfactory lobe
1 2
3
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4 5

14. 6 7
8
14
9 10

15. Effect of Endosulfan and Antioxidants on the distribution of AChE in
different layers of olfactory lobe of mice
GROUPS OLFACTORY LOBER LAYERS
PFL GL EPL ML IPL GRL
C
-- ++++ +- +++ ++- +-
E
-- ++- +- ++/-- +- +-
R
+++ +++ +- +++ +- +-
RE
-- ++++ +- +++ +- +-
L
+++ +++ +- +++ +- +-
LE
-- ++++ +- +++ +- +-
++++ = strong V
+++ = intense +++ +++ +- +++ +- +-
VE
++ = less intense
++- = moderate VLR
-- ++++ +- +++ +- +-
+- = mild
+++ +++ +- +++ +- +-
+_ = negligible VLRE
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-- =negative -- ++++ +- +++ +- +-

16. Effect of Endosulfan and Antioxidants on the distribution of BChE in
different layers of olfactory lobe of mice
GROUPS OLFACTORY LOBER LAYERS
PFL GL EPL ML IPL GRL
C
+_/-- +++ +_ ++++ ++- +-
E
-- +- -- ++ +- +_
R
++++ +++ +_ +++ ++- +-
RE
+- +++ +_ +++ ++- +-
L
++++ +++ +_ +++ ++- +-
LE
+- +++ +_ +++ ++- +-
V
++++ +++ +_ +++ ++- +-
VE
+- +++ +_ +++ ++- +-
VLR
++++ +++ +_ +++ ++- +-
VLRE
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+- +++ +_ +++ ++- +-

17. Effect of Endosulfan and Antioxidants on the distribution of ATPase in
different layers of olfactory lobe of mice
GROUPS OLFACTORY LOBER LAYERS
PFL GL EPL ML IPL GRL
C +++ ++ +- ++- +- +-
E +- +_ +_/-- +- +_ +_
R ++++ +++ ++-/+- ++- +- +-
RE +++ ++- +- ++ +- +-
L ++++ +++ ++-/+- ++- +- +-
LE +++ ++- +- ++ +- +-
V ++++ +++ ++-/+- ++- +- +-
VE +++ ++- +- ++ +- +-
VLR ++++ +++ ++-/+- ++- +- +-
VLRE +++ ++- +- ++ +- +-
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18. RESULTS
 Endosulfan treatment exhibited inhibition of ATPase , AChE
and BChE activity in different layers, nuclei and fibre tracts of
different regions of the brain.
 Antioxidants supplementation, prior to endosulfan treatment
provided a protection cover to ATPase, AChE and BChE activity ,
compared to the control group.
 In antioixidant groups marked elevation in enzyme activity was
seen as compared to control group.
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19. Plan of Work for the Next Year
Biochemical studies for AChE and BChE will be done on the brain of
mice for all the groups under study.
Comparative, critical and statistical analysis of biochemical parameters
and discussion for the paradigms undertaken for the study would be
done.
Comparative analysis for all the histoenzymological and biochemical
studies as a whole, publications and written work of thesis require one
more year to accomplish the work.
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