Zebrafish: A Model for Marine Peptide Based Drug Screening

82 Pages·2019·3.982 MB·English

by  Saravanan Ramachandran, Senthilkumar Rajagopal

Saravanan Ramachandran 
Senthilkumar Rajagopal
Zebrafi sh:
A Model for
Marine Peptide
Based Drug
Screening
Zebrafish: A Model for Marine Peptide Based
Drug Screening
Saravanan Ramachandran
Senthilkumar Rajagopal
Zebrafish: A Model for
Marine Peptide Based
Drug Screening
Saravanan Ramachandran Senthilkumar Rajagopal
Native Medicine and Marine Pharmacology Department of Biochemistry
Laboratory, Faculty of Allied Health Rayalaseema University
Sciences Kurnool, Andhra Pradesh, India
Chettinad Hospital and Research Institute,
Chettinad Academy of Research and
Education (Deemed to be a University)
Kelambakkam, Tamil Nadu, India
ISBN 978-981-13-7843-0 ISBN 978-981-13-7844-7 (eBook)
https://doi.org/10.1007/978-981-13-7844-7
© Springer Nature Singapore Pte Ltd. 2019
This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of
the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation,
broadcasting, reproduction on microfilms or in any other physical way, and transmission or information
storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology
now known or hereafter developed.
The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication
does not imply, even in the absence of a specific statement, that such names are exempt from the relevant
protective laws and regulations and therefore free for general use.
The publisher, the authors, and the editors are safe to assume that the advice and information in this book
are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the
editors give a warranty, express or implied, with respect to the material contained herein or for any errors
or omissions that may have been made. The publisher remains neutral with regard to jurisdictional claims
in published maps and institutional affiliations.
This Springer imprint is published by the registered company Springer Nature Singapore Pte Ltd.
The registered company address is: 152 Beach Road, #21-01/04 Gateway East, Singapore 189721,
Singapore
Foreword by Balasubramanian
The field of marine peptide research is vast and evolving rapidly in both basic
research and clinical therapeutics. When I was asked to write a foreword to this
book, my immediate thought was that there are many monographs and comprehen-
sive textbooks focusing on zebrafish model for utilizing drug discovery from marine
peptides that cover a wide variety of topics at various levels of detail, so why the
need for another book in this increasingly crowded field? Although quite a lot of
information is available on the zebrafish model for drug discovery, the present
monograph is different from the earlier as the authors provide a comprehensive
information on the basic mechanisms of peptides/toxins and how these mechanisms
are linked with the emergence of common, devastating pathological disorders. The
authors build on current knowledge to detail the link between zebrafish models for
discovery of more drugs. They explain these complex concepts with straightforward
language that allows greater accessibility to a wide audience. The succinct text will
assist the novice in understanding marine peptide complexity research, while the
up-to-date information on the current state of this work and pathophysiology will be
of interest to the experts in the field.
The authors provided all the relevant information in an update fashion in five
different chapters. Chapter 1 highlights the introduction, classification, and biomed-
ical importance of peptide, toxins, alkaloids, polysaccharides, and phenolic com-
pounds of macro- and microorganisms and other marine organisms for various
human diseases and disorders. Chapter 2 emphasizes the isolation, structural
v
vi Foreword by Balasubramanian
characterization techniques, and de novo sequencing of peptide from ascidians and
its teratogenic activity in the zebrafish embryo.
Chapter 3 accentuates the separation, purification, and sequencing techniques of
toxin from the posterior salivary gland of cuttlefish—furthermore, the maximum
tolerated dose and teratogenic activity of toxin by using the embryo of the zebrafish.
Chapter 4 draws the attention to the anticancer properties of cuttlefish toxin and the
development of xenotransplantation model of zebrafish for anticancer property
marine drug screening. Chapter 5 represents the protective effect of marine peptides
for cardiovascular diseases in zebrafish model; each chapter of this book contains an
insight that will be useful to the scientists at all levels.
I congratulate the authors on producing a straightforward text that can be useful to
the researchers with different levels of expertise. I hope that this work will essentially
help to expand interest in the field of the marine peptide using drug discovery.

Vice Chancellor T. Balasubramanian
Chettinad Academy of Research and Education
Kelambakkam, Tamil Nadu, India
Foreword by Madeswaran
It is a distinct honor to have been invited by my colleagues in marine peptide-based
drug screening arena to write the foreword to this very articulate and scientifically
state-of-the-art book entitled Zebrafish: A Model for Marine Peptide-Based Drug
Screening. The book covers all the areas required to create a robust category and
perform read-across. I am certain that the readers, including the faculties, research-
ers, and students, will find this book extremely informative, interesting, and inspir-
ing. Hence, I hope that you will find this book possess sufficient disclosure and
adequate utility. This book aims to simplify the revolution and to fortify the
researcher with the information needed to use marine protein/peptides/toxins with
complete confidence and the best compound that can be applied for therapy of the
individual. The book explores in many ways and makes a good sense to investigate
further on the isolation of these peptides from marine species. Moreover, this valu-
able text opens the doors for the progression that occurs when one discovers a fact,
becomes interested, and then begins investigating and discovering the natural
process.
I am a scientist G in the Ministry of Earth Sciences (MoES), who is presently
working at the National Centre for Coastal Research (NCCR), Chennai. Since 1994,
I have handled the following four major research and development programs, viz.,
(i) Development of Potential Drugs from Ocean (Drugs from Sea); (ii) Integrated
Coastal and Marine Area Management (ICMAM); (iii) Marine Living Resources
(MLR); and (iv) Coastal Ocean Monitoring and Prediction System (COMAPS). In
vii
viii Foreword by Madeswaran
addition, I have handled two international programs, i.e., (i) Commission for
Conservation of Antarctic Marine Living Resources (CCAMLR), Hobart, Australia,
and (ii) South Asia Co-operative Environment Programme (SACEP), Colombo, Sri
Lanka.
As a consequence, a large number of books, thick and thin, are being published
continuously on various aspects of beneficial usage of marine peptides. The central
theme of this monograph is giving fundamental mechanisms of marine peptides/
toxins on physiological activities in our body. The authors have made an effort to
unify all the content of scattered research literature in this area of research and tried
to provide thorough information. By and large, this monograph is not just a collec-
tion of papers, but it is an essence of the diverse marine peptides-/toxins-based
drugs screened using the zebrafish model. In all chapters, they have provided the
basic information relevant to the topics, and at the same time, they have described
the perspective knowledge about using the zebrafish model. I believe that this mono-
graph could be an informative resource in the form of condensed handbook for both
fundamental and advanced researchers.
Consequently, this monograph covers information on marine pharmacognosy.
The authors, Saravanan Ramachandran and Senthilkumar Rajagopal, have brought
through understanding the milieu of clinical research, experimental research, marine
omics, and pharmacology which, collected revenue, focused on the most active
areas of cancer research in the zebrafish model.
Best wishes to the authors.

Scientist G, National Centre for Coastal Research P. Madeswaran
Ministry of Earth Sciences, NIOT Campus
Chennai, Tamil Nadu, India
Contents
1 Biomedical Importance of Marine Peptides/Toxins . . . . . . . . . . . . . . . . . . . 1
1.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
1.2 Marine Environment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
1.3 Toxins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
1.4 Toxins and Therapeutics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
1.5 Classification of Toxins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
1.6 Marine Toxins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
1.7 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
2 Teratogenic Activity of Peptides in Zebrafish Model . . . . . . . . . . . . . . . . . 15
2.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
2.2 Ascidians . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
2.3 Morphology of Ascidians . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
2.4 Importance of Ascidians . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
2.5 Results and Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
2.5.1 FT-IR Spectroscopy . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
2.5.2 MALDI-TOF and Mascot Analysis . . . . . . . . . . . . . . . . . . . . 21
2.5.3 Embryo Toxicity of Peptide . . . . . . . . . . . . . . . . . . . . . . . . . . 22
2.6 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
3 Teratogenic Activity of Toxins in Zebrafish Model . . . . . . . . . . . . . . . . . . 27
3.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
3.2 Cephalopods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
3.3 Cephalopod Toxins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
3.4 Results and Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
3.4.1 Structural Characterisation of PSG Toxin . . . . . . . . . . . . . . . 31
3.4.2 CD Spectroscopy of PSG Toxin . . . . . . . . . . . . . . . . . . . . . . . 32
3.4.3 KD Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
2
3.4.4 MALDI-TOF/MS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
ix
x Contents
3.4.5 MASCOT Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
3.4.6 Teratogenicity of PSG Toxin . . . . . . . . . . . . . . . . . . . . . . . . . 39
3.4.7 Zebrafish Embryotoxicity Assay . . . . . . . . . . . . . . . . . . . . . . 39
3.5 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
4 Anticancer Properties of Marine Peptides/Toxins
Using Zebrafish Model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
4.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
4.2 Cancer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
4.3 Causes of Cancer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
4.4 Marine Peptides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
4.5 Bioactive Cyclic Peptides from Marine Sources . . . . . . . . . . . . . . . . . 47
4.6 Bioactive Toxins from Marine Species . . . . . . . . . . . . . . . . . . . . . . . . 47
4.7 Cytotoxicity of PSG Toxin Against MCF-7 Cancer Cell
Line by MTT Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
4.8 Zebrafish as a Model for Cancer Research . . . . . . . . . . . . . . . . . . . . . 49
4.9 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
5 Protective Effect of Marine Peptides/Toxins in CVD
Using Zebrafish Model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
5.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
5.2 Prevention . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
5.2.1 Diet . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
5.2.2 Dietary Supplements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
5.3 Medication . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
5.4 Physical Activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
5.5 Exercise . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
5.6 Adverse Effects of Smoking and Tobacco Usage . . . . . . . . . . . . . . . . 63
5.7 Maintain a Healthy Weight . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
5.8 Quality Sleep . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
5.9 Manage Stress . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
5.10 Health Monitoring . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
5.10.1 Screening of Blood Pressure . . . . . . . . . . . . . . . . . . . . . . . . . 64
5.10.2 Examine the Cholesterol . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
5.10.3 Observation of Diabetes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
5.11 Cardiovascular Disease Treatment . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
5.12 Zebrafish: An Excellent Model for Study of CVD . . . . . . . . . . . . . . . 66
5.13 Summary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70