Animal Biotechnology : Models in Discovery and Translation. Animal biotechnology : models in discovery and translation / edited by Prof. Ashish S. Verma, PH.D. (Zoology), Jadavpier University, Kolkata, 700032, W.B., India, Dr. Anchal Singh, PH.D. (Biochemistry), Department of Biochemistry, Institute of Science, Banaras Hindu University, Varanasi, 221005, U.P. India.

Front Cover
Animal Biotechnology
Copyright Page
Dedication
Contents
List of Contributors
Preface
Acknowledgments
I. Human diseases: in vivo and in vitro models
1 Drosophila: a model for biotechnologist
Summary
What you can expect to know
Introduction
Classical aspects of Drosophila melanogaster
Physical appearance
Life cycle
Drosophila development
Embryogenesis in Drosophila
Pattern formation in Drosophila
Homeotic genes in Drosophila
Drosophila genome
History
Historical perspective of Drosophila contributions to biotechnology
Principle
Methodology
Culturing of Drosophila
Preparation of Drosophila food medium
Materials required
Handling of flies
Fly disposal
Egg collection
Dechorination of eggs
Preparation of DNA for injection
Protocols
Protocol for germ-line transformation in Drosophila
Materials required
Procedure
Ethical issues
Translational significance
Clinical significance
Drosophila-based models for understanding human neurodegenerative diseases
Drosophila as a model for understanding human metabolic disorders
Drosophila as a model for understanding nephrolithiasis (kidney stones)
Drosophila-based model for understanding the human immunodeficiency virus pathology
Drosophila-based therapeutic peptide production
Turning point
World Wide Web resources
Acknowledgments
References
Further reading
Glossary
Abbreviations
Long answer questions
Short answer questions
Answers to short answer questions
Yes/no type questions
Answers to yes/no type questions
2 Animal models of tuberculosis
Summary
Introduction
Comparative pathology of tuberculosis in humans and animals
Characteristics of a model for tuberculosis with respect to infection and pathogenesis.

Pathogen diversity: crossing species barriers
Host diversity: fundamental processes and fine-tuning
Animal models of tuberculosis: limits and lessons
Animal models: contributions in tuberculosis vaccine testing
Various animal models
Mouse model
Guinea pig model
Rabbit model
Nonhuman primate model
Cattle model
Protocols
Preparing M. tuberculosis inoculum for aerosol exposure
Aerosol infection of mice using the middlebrook apparatus
Aerosol infection of guinea pigs using a madison chamber
Bacteria loading
Intravenous infection of mice with M. tuberculosis
Isolation of samples for determining M. tuberculosis load by real-time-PCR
Determination of bacterial loads in target organs
Preparation of lungs or other tissues for histology
Preparation of lung cell suspension
Ethical issues
Translational significance
World wide web resources
Safety considerations
References
Further reading
Glossary
Abbreviations
Long-answer questions
Short-answer questions
Answers to short-answer questions
Yes/no-type questions
Answers for yes/no type questions
3 Animal models for neurodegenerative disorders
Summary
What you can expect to know
History and methods
Introduction
Neurodegenerative diseases
Amyotrophic lateral sclerosis
Spinal muscular atrophy
Spinal and bulbar muscular atrophy
Principles
Genetics of amyotrophic lateral sclerosis
Superoxide dismutase 1-amyotrophic lateral sclerosis
Amyotrophic lateral sclerosis: genes implicated in RNA metabolism
TAR-DNA-binding protein 43
Fused-in sarcoma/translocated in liposarcoma protein
C9orf72
Genetics of spinal muscular atrophy
Genetics of spinal and bulbar muscular atrophy
Methodology
Generation of transgenic mice.

Preparation and purification of transgenic construct (step 1)
Harvesting donor eggs (step 2)
Microinjection of transgene to fertilized egg (step 3)
Implantation of microinjected egg to pseudopregnant female mice (step 4)
Screening of founder mice for expression of transgene (step 5)
Establishing stable transgenic line (step 6)
Cre-loxP technology
Amyotrophic lateral sclerosis models
SOD1G37R transgenic mice
SOD1G93A transgenic mice
SOD1WT transgenic mice
Spinal muscular atrophy models
Severe spinal muscular atrophy mice (mSMN−/−
SMN2+/+)
Spinal muscular atrophy type II mice (mSMN−/−
SMN2+/+
SMNΔ7+/+)
Spinal and bulbar muscular atrophy models
AR-97Q and AR-24Q transgenic mice
Examples and their applications
Superoxide dismutase 1-linked amyotrophic lateral sclerosis
Gain of toxicity from mutant superoxide dismutase 1 established as pathomechanisms through engineering mutant superoxide di...
Evaluating phenotype and clinical course of mutant superoxide dismutase 1 transgenic mice
Toxicity from misfolded mutant superoxide dismutase 1 protein
Non-cell autonomous neurodegeneration demonstrated by superoxide dismutase 1 mouse models
Stem cell-derived motor neurons established from mutant superoxide dismutase 1 mice
Other amyotrophic lateral sclerosis
Spinal muscular atrophy
Human SMN2 transgenic mice
Neuron-specific deletion of survival of motor neuron in mice using Cre-loxP systems
Spinal and bulbar muscular atrophy
AR-97Q mice as spinal and bulbar muscular atrophy Model
Androgen hormone and mutant androgen receptor central to spinal and bulbar muscular atrophy pathogenesis
Clinical correlations
Protocols
Ethical issues
Translational significance
World Wide Web resources
Acknowledgment
References
Further reading
Glossary.

Abbreviations
Long answer questions
Short answer questions
Answers to short answer questions
Yes/no type questions
Answers for yes/no type questions
4 Epigenetics and animal models: applications in cancer control and treatment
Summary
What you expect to know
Introduction
History
Principle
Use of mouse models in the epigenetics of cancer
Examples with applications
Brain cancer
Breast cancer
Colorectal cancer
Esophageal cancer
Gastric cancer
Head and neck cancer
Lung cancer
Lymphoma and leukemia
Prostate cancer
Liver cancer
Other approaches
Methodology
Methylation profiling
Histone profiling
Nucleosome mapping
Protocols
Ethical issues
Translation significance
Clinical significance
Web resources
Turning point
Flow chart
World Wide Web resources
Something interesting about this chapter
References
Glossary (terms used in text with examples)
Abbreviations
Long answer questions
Answers to long answer questions
Short answer questions
Answers to short answer questions
Yes/no type questions
Answers to yes/no type questions
5 Development of mouse models for cancer research
Summary
What you can expect to know
Introduction
History
Principle
Institutional Animal Care and Use Committee approval
Institutional Animal Care and Use Committee guidelines
Methodology
Inbred mice
Examples with applications
Immunocompetent mice
Spontaneous tumor models
The genetically engineered mouse models
The Cre/Lox system: a superior genetically engineered mouse model
Immunodeficient mice
Allograft transplants
Xenograft transplants
Humanized mice
Checklist for a successful in vivo experiment
Protocols
An orthotopic mouse model of colorectal cancer
Design and execution.

Interpretation of results
A xenograft model of prostate cancer metastasis
Design and execution
Interpretation of results
Humanized mouse models for tumor xenografts
Design and execution
Ethical issues
Turning point
Translational significance
World Wide Web resources
References
Glossary
Abbreviations
Long answer questions
Short answer questions
Answers to short answer questions
Yes/no type questions
Answers to yes/no type questions
6 The clinico-molecular approaches for detection of human papillomavirus
Summary
What you can expect to know
Introduction
Cancer
Cervical cancer
Historical overview
Mistaken theories of cervical cancer causation
The first breakthrough
zur Hausen
Prevalence and epidemiology of cervical cancer
Global scenario
Symptoms of cervical cancer
Anatomy of female pelvis
Types of cervical cancer
Risk factors for cervical cancer
Human papillomaviruses
Genomic organization of human papillomavirus
Transcriptional regulation of human papillomavirus
Life cycle of human papillomavirus
Functions of human papillomavirus oncoproteins E6 and E7
Inactivation and degradation of p53 through the E6/E6AP complex
Screening and diagnostic methodologies of cervical cancer
Screening
Methods used for screening/diagnosis of cervical cancer
Visual methods
Indications
Other screening tests
Reporting systems terminology
Precancer classification
WHO classification
CIN classification (Bhambhani., 2007)
Bethesda classification
Cancer classification
Colposcopy and biopsy
New technologies
DNA cytometry
Human papillomavirus DNA-based screening methods (protocol)
Urine-based noninvasive human papillomavirus DNA detection method.

Simple "paper smear" method for rapid detection of human papillomavirus infection.