REGENRATION OF VENTRAL NERVE CORD IN Eisenia

fetida
PROJECT A

Submitted in partial fulfilment of the requirements for the degree

BACHELOR OF ENGINEERING

BY

VIJAY IYER

HUMAIRA SHAH

Under the Guidance Of

Ms. Reshma Sathian

Assistant Professor

Thadomal Shahani Engineering College

DEPARTMENT OF BIOTECHNOLOGY

THADOMAL SHAHANI ENGINEERING COLLEGE

UNIVERSITY OF MUMBAI

2017-2018
 CERTIFICATE
This is to certify that students whose names are given below have submitted their B.E.
Project in the partial fulfilment of degree of B.E. Biotechnology Engineering.

PROJECT ENTITLED:

REGENERATION OF VENTRAL NERVE CORD OF Eisenia fetida.

YEAR: 2017-2018

NAME OF STUDENTS: 1.VIJAY IYER

2. HUMAIRA SHAH

___________________ _____________________

External Examiner Project Guide

___________________ ______________________

Internal Examiner Head of Department

 ACKNOWLEDGEMENT

We would like to express our sincere gratitude to Dr. MC Arunan (TIFR) for his guidance
and constant support to help us reach this point. We would also like to thank Ms. Reshma
Sathian, our project guide to be our pillar of strength always and to help us shape our project .

We are thankful to Mr. Chandrashekhar Kulkarni, HOD, Department Of Biotechnology

without whom we would have not been able to come upto this project idea. He was always
available to guide us whenever we needed his help in any form.

We are thankful to the entire faculty of Biotechnology Department as well as the non-
teaching staff for all their support.
 CONTENTS

1. Introduction.................................................................................................................1

2. Literature Review........................................................................................................3

2.1 Regeneration introduction.............................................................................3

2.2 Factors Responsible For Regeneration..........................................................4

2.2.1 Role of Stem Cells in Regeneration.4

2.2.2 Reprogramming By Defined Factors...................................................5

2.2.3 The Four Factors.................................................................................5

2.2.3.1 Oct 3/4 ...........................................................................5

2.2.3.2 SOX2 .............................................................................6

2.2.3.3 c-Myc..............................................................................6

2.2.3.4 Klf4.................................................................................6

2.3 Role of SOX in Neurogenesis.......................................................................7

2.4 WNT pathway..8

2.4.1 WNT signalling ..9

2.4.2 Canonical and non canonical pathway9

2.4.2.1 Canonical pathway...9

2.4.2.2 Non canonical pathway10

2.4.2.3 Other pathways11

2.4.3 Regulation12

2.4.4 Induced cell responses..12

2.4.4.1 Embryonic development....12

2.4.4.2 Axis patterning..13

2.4.4.3 Cell fate specification13

2.4.4.4 Cell proliferation14

2.4.4.5 Cell migration.15

 2.4.5 WNT signalling in HSCs.15

2.4.6 WNT & Regeneration..15

2.4.7 Sox & WNT interactions.17

2.5 Nervous System..........................................................................................18

2.5.1 Of Invertebrates...........................................................................18

2.5.2 Of Vertebrates..............................................................................18

2.5.3 Ganglion.......................................................................................20

2.5 Faults in Signal Transmission......................................................................20

2.6 Earthworm as a Model System....................................................................20

2.6.1 Why Eisenia fetida?.....................................................................21

2.6.2 Physiology.....................................................................................22

2.6.3 Nutrition........................................................................................23

2.6.4 Ventral Nerve Cord (VNC)...........................................................23

2.6.5 Neuronal Control of Locomotion..................................................25

2.7 Methods to Alter Regeneration.....................................................................25

2.7.1 Use of Valproic Acid 25

2.7.2 WNT inhibitors..............................................................................25

2.7.3 Lithium chloride ...27

2.7.4 Alpha lipoic Acid..28

2.7.5 Quercetin ..28

2.7.6 Retinoic Acid 28

3. Aim and Objective .....................................................................................................29

4. Materials and Methods...............................................................................................30

4.1 Organisms Used...........................................................................................30

4.2 Rapid Escape Response Assay.....................................................................30

4.3 Lesion Protocol.............................................................................................31

 4.4 Altering Regeneration of Ventral Nerve Cord................................................31

4.4.1 Use of VALPROIC Acid.................................................................31

4.4.2 Use of lithium chloride 31

4.4.3 Use of Quercetin...32

4.4.4 Use of Alpha lipoic Acid..32

4.4.5 Use of Retinoic Acid33

5. Future Prospectus...........................................................................................................34

6. References......................................................................................................................35