Zebrafish Model for Biomedical Research

✍ Scribed by Prasan R. Bhandari (editor), Kala Kumar Bharani (editor), Amit Khurana (editor)

• Publisher: Springer
• Year: 2022
• Tongue: English
• Leaves: 493
• Category: Library

✦ Synopsis

This book provides the latest information on the significance of zebrafish as an ideal model for researching the biomedical field, with references. This book also focused on the evidence of zebrafish as a model in cardiovascular, neurologic, psychiatric and metabolic research. In addition, the book also includes the research carried out on zebrafish in hepatic, renal, ophthalmic, and ENT related areas. Contributed chapters come from the most prominent laboratories working in this field, which provides a unique perspective on zebrafish models from a wide spectrum of the research community. In addition, the book offers a detailed analysis of the most current research in the area for specific zebrafish models including specific research in the area of skin disorders, endocrine diseases, nutritional disorders, gastrointestinal, hematological disorders and cancer. The compilation of chapters in the volume culminates into a comprehensive and definitive text on zebrafish and its suitability for modeling various diseases, providing a critical resource on the potential attributes of the zebrafish as a pharmacological model. In terms of scope, this book is a useful tool for young researchers, professors and pharmaceutical scientists for understanding the significance of zebrafish as an emerging pharmacological model that can significantly aid in the process of drug discovery and development.

✦ Table of Contents

Preface
Contents
Editors and Contributors
1: ABCD of Zebrafish Culture
1.1 Zebrafish: A Brief Background
1.2 Zebrafish: A Vertebrate-Animal Model
1.2.1 Pros
1.2.2 Cons
1.3 Steps in Zebrafish Culture
1.3.1 Acquisition and Maintenance of Zebrafish
1.3.1.1 Static System
1.3.1.2 Flow-Through System
1.3.1.3 Recirculating System
Major Components of the Recirculating System
1.3.2 Breeding of Zebrafish in Lab Conditions
1.3.3 Care of Zebrafish
1.3.4 Diet for Zebrafish Developmental Stages
1.3.4.1 Live, Natural Lab-Grown Diet
1.3.4.2 Commercially Available Formulated Diet
Feeding Regime for Zebrafish
1.4 Conclusion
References
2: Zebrafish: A Model Organism to Understand Tumor Angiogenesis Mechanism
2.1 Introduction
2.2 Tumor Induction in Zebrafish by Chemical or Carcinogen Exposure
2.3 Zebrafish Xenograft Model of Tumor Development
2.4 Choice of Injection: Embryo vs. Juvenile vs. Adult
2.5 Tumor-Induced Angiogenesis in Zebrafish Model
2.6 Potential Screening of Anti-angiogenic Drug in Zebrafish Tumor Model
2.7 Challenges in Tumor Models to Study Anti-angiogenic Therapy
2.8 Conclusion
References
3: Zebrafish as a Xenotransplantation Model for Studying Cancer Biology and Cancer Drug Discovery
3.1 Introduction
3.2 Advantages of Zebrafish Xenotransplantation Model for Cancer Research
3.3 Stages of Zebrafish for Xenotransplantation
3.3.1 Embryo Stage
3.3.2 Juvenile Stage
3.3.3 Adult Stage
3.4 Applications of the Zebrafish Xenotransplantation Model in Cancer Biology
3.4.1 Assessing the Tumor Growth, Metastasis and Angiogenesis
3.4.2 Investigating Mechanisms of Cancer in Search for Therapeutic Targets
3.4.3 Assessment of Cancer Treatment Drugs (Drug Screening)
3.5 The Potential of Zebrafish Xenotransplantation for Personalized Cancer Therapy
3.6 Current Limitations of the Zebrafish as a Xenotransplantation Model
References
4: Zebrafish Models for Screening of Metabolic Diseases
4.1 Epidemiology of Metabolic Diseases
4.2 Need of Drug Discovery
4.3 Need of Alternative Animal Models
4.4 Zebrafish Models for Obesity
4.4.1 Larval Models of Obesity
4.4.2 Transgenic and Mutants as Model of Obesity
4.4.3 Diet Models of Obesity
4.5 Zebrafish Models for Diabetes Mellitus
4.5.1 Zebrafish Models of Type 1 Diabetes Mellitus
4.5.2 Zebrafish Models of Type 2 Diabetes Mellitus
4.5.3 Zebrafish Models of Maturity-Onset Diabetes of the Young
4.5.4 Transgenic Zebrafish Lines
4.5.5 Zebrafish Models for Complications of Diabetes Mellitus
4.6 Zebrafish Models of Hyperlipidemia
4.6.1 High-Cholesterol Diet (HCD) Paradigm
4.6.2 Zebrafish APOC2 Deficiency
4.6.3 Zebrafish Liver X Receptor (LXR) Deletion
4.7 Zebrafish Models of Atherosclerosis
4.8 Advantages, Limitations, and Conclusion
4.8.1 Advantages
4.8.2 Limitations
4.9 Conclusion
Reference
Bibliography
5: Role of Zebrafish as an Experimental Model for Renal Disorders
5.1 Introduction
5.2 Zebrafish Models of Acute Kidney Disorders (AKI and AKD)
5.3 Genetic Models of Renal Diseases
5.4 Conclusion
References
6: Evolution of Zebrafish as a Novel Pharmacological Model in Endocrine Research
6.1 Introduction
6.2 Zebrafish as a Preclinical Model for Diabetes Drug Discovery and Development Research
6.2.1 Type 1 Diabetes Mellitus Models of Zebrafish
6.2.2 Type 2 Diabetes Mellitus (T2DM) Models of Zebrafish
6.2.3 Zebrafish Diabetes Models to Study Glucose Homeostasis
6.3 Study of Pancreas Development
6.4 Zebrafish as a Tool for Research and Development in Obesity
6.4.1 Nutritional Modulatory Models of Obesity in Zebrafish
6.4.2 Mutant and Transgenic Model for Obesity
6.4.3 Methods to Study Adiposity in Zebrafish
6.5 Zebrafish as a Tool to Study Osteoporosis
6.5.1 Zebrafish as a Model to Study Impaired Iron Homeostasis
6.5.2 Ectonucleotide Pyrophosphatase/Phosphodiesterase 1 (Enpp1) Gene Mutation and Generalized Arterial Calcification
6.5.3 Zebrafish as a Model to Study Collagenopathies, Osteogenesis Imperfecta, and Scoliosis
6.5.4 Genetic Manipulation in Zebrafish for Studying Osteoporosis
6.5.5 Model to Study Bone Development and Skeleton Mineralization
6.5.6 As a Model to Study Bone Matrix Formation with Emphasis on Bone Regeneration and Fracture Repair
6.6 Zebrafish as a Tool to Study Reproductive Biology
6.6.1 The Hormonal Signaling in Zebrafish and its Similarities to the Human Hormone Axis
6.6.2 Hypothalamic-Pituitary-Gonadal (HPG) Axis
6.6.3 Hypothalamic-Pituitary-Thyroid (HPT) Axis
6.6.4 Hypothalamic-Pituitary-Adrenal (HPA) Axis
6.6.5 Reproductive Biology of Zebrafish
6.6.6 Zebrafish as a Tool to Study Infertility
6.6.7 Genetic and Transgenic Model of Infertility
6.6.8 Stress-Induced Infertility
6.6.9 Chemical- and Environment-Induced Infertility
6.7 Advantages and Limitations of Zebrafish as a Tool for Endocrinology Research
6.8 Future Perspectives and Summary
References
7: Zebrafish as a Versatile Model for Cancer Research
7.1 Introduction to Zebrafish in Cancer Discovery
7.2 Various Cancer Models Developed in Zebrafish
7.2.1 Various Tools and Techniques in Zebrafish as a Model for Cancer Screening
7.2.2 Zebrafish Cancer and Human Cancers: How Much Is the Correlation?
7.2.3 Studying Tumor Angiogenesis and Metastasis in Zebrafish with Emphasis on Tumor Microenvironment
7.2.4 Zebrafish as a Screening Model of Anticancer NCE
7.3 Zebrafish as a Tool for Chemical Genetic Manipulation for Cancer Discovery
7.3.1 Chemical Screen for Identifying Novel Oncogenes
7.3.2 Zebrafish as a Tool to Identify Melanoma Therapies
7.4 Genetic- and Transplantation-Based Cancer Models in Zebrafish
7.4.1 Liver Cancer
7.4.2 Brain Cancer
7.4.3 Skin Cancer
7.4.4 Blood Cancer
7.5 Genetic Predisposition to Cancer in Zebrafish
7.5.1 Reverse Genetics and Insertional Mutagenesis Model
7.5.2 Zebrafish for Chemical Modifier Screen
7.5.3 Zebrafish as a Tool to Study Oncogenomics
7.5.4 Zebrafish as Tool to Study Cancer Drug Resistance and Epigenetic Modifications
7.5.5 Screens Using Developmental Surrogate Markers
7.6 Advantages of Zebrafish as a Tool for Cancer Research: Screen, Imaging, and Chemical Treatment in Search of Drug
7.7 Challenges and Limitations of Zebrafish Model in Cancer Research
7.8 Future Perspectives and Summary
References
8: Zebrafish as an Indispensable Tool for Infectious Diseases and Immune Modulatory Studies
8.1 Suitability of Zebrafish in Infectious Diseases Research
8.1.1 Bacterial Infections
8.1.2 Viral Infections
8.1.3 Fungal Infections
8.2 Immunomodulatory Studies
8.3 Conclusion
References
9: Zebrafish: Promising Model for Cancer Research
9.1 Introduction
9.2 Zebrafish Models Used in Cancer Research
9.2.1 Models for Tumour Angiogenesis
9.2.1.1 Zebrafish Xenograft Model
9.2.2 Zebrafish Models for Neuroendocrine Tumours
9.2.2.1 NET Transplantable Model in Zebrafish Embryos
9.2.2.2 MYCN Model
9.2.3 Zebrafish Melanoma Models
9.2.3.1 The BRAFV600E Zebrafish Melanoma Model
9.2.3.2 The Mitf Zebrafish Melanoma Model
9.2.3.3 Genetic Model of Melanoma in Zebrafish
9.2.4 Models for Leukaemia
9.2.4.1 T-ALL Zebrafish Transgenic Model
9.2.5 Hepatocellular Carcinoma Models
9.2.5.1 Wnt-Activated Hepatocellular Carcinoma Model
9.2.6 Bone Cancer Models
9.2.6.1 Miscode
9.2.6.2 CSC Model
9.2.7 Paediatric Model
9.2.7.1 Rhabdomyosarcoma Model
9.2.8 Techniques Used for Anticancer Research Using Zebrafish Model
9.2.8.1 Cell Xenograft and Quantification
9.2.8.2 Drug Administration by Soaking and Microinjection
9.2.8.3 Anti-Proliferation Determination in Zebrafish Tumour Model
9.2.9 Cancer Imaging in Zebrafish Model
9.2.9.1 Macroscopic Observation of Tumour Growth
9.2.9.2 Stereomicroscopy to Image Tumour Growth
9.2.9.3 LED Fluorescence Macroscope Imaging
9.2.9.4 Microscopic Observation in Tumorigenesis
9.3 Conclusion
References
10: Zebrafish Model and Cardiovascular System for Novel Therapies
10.1 Introduction
10.2 Genetic Approach in Zebrafish Cardiovascular Research
10.2.1 Forward Genetics
10.2.2 Reverse Genetics
10.2.2.1 mRNA Overexpression
10.2.2.2 Transgenesis
10.2.2.3 Morpholino-Mediated Knockdown
10.2.2.4 Genome Editing Techniques
10.3 Similarities of Human and Zebrafish Cardiovascular Development and Role in Research
10.3.1 Cardiomyocyte Differentiation
10.3.2 Vasculogenesis and Angiogenesis
10.3.3 Valvulogenesis
10.4 Cardiovascular Disease Model
10.4.1 Arrhythmia Model
10.4.2 Cardiomyopathy Model
10.4.3 Cardiovascular Regeneration Model
10.4.4 Complex Congenital Heart Disease Model
10.5 System Pharmacology and Zebrafish Model
10.6 Challenges and Future Perspective of the Zebrafish Model
References
11: Zebrafish Model for Drug Discovery and Screening
11.1 Introduction
11.2 Strengths of Zebrafish
11.3 Genetics and Zebrafish
11.3.1 Identifying Novel Drug Targets
11.3.2 Genes and Polycystic Kidney Disease
11.3.3 Genes and Heart Diseases
11.3.4 Morpholino Oligonucleotide Screens
11.3.5 Phenotype-Based Drug Discovery
11.4 Structure-Activity Relationship
11.5 Screening in Adult Zebrafish
11.6 Screening for DNA Toxicity: Mutagenesis
11.7 Target-Based Drug Discovery
11.8 Heart in Drug Development and Drug Toxicity
11.9 Liver: Drug Development and Toxicity
11.9.1 Microsomal Enzymes and Drug Development
11.9.2 Intestines
11.9.3 Pancreas
11.9.4 Gall Bladder
11.10 Kidney: Drug Development and Drug Toxicity
11.11 Nervous System: Drug Development and Drug Toxicity
11.11.1 Pineal/Circadian Rhythm
11.12 Haematopoietic System and Drug Development and Drug Toxicity
11.13 Endocrine System: Drug Development and Drug Toxicity
11.14 Ototoxicity and Drug Development
11.15 Ocular Toxicity and Drug Development
11.16 P-Glycoprotein and Drug Resistance
11.17 Zebrafish and Cancer Drug Development
11.17.1 Chemical Carcinogenesis
11.17.2 Transgenic Models in Zebrafish Neoplasia
11.18 Challenges in Zebrafish as a Model for Drug Development and Drug Toxicity
References
12: Zebrafish as a Novel Pharmacological Screening Model for Drug Discovery and Development Against Hematological Disorders
12.1 Introduction to Zebrafish in Hematology
12.2 The Hematological Features of Zebrafish
12.2.1 Zebrafish and Human Hematology: Similarities and Differences
12.2.2 Zebrafish Biology and Relevance for Hematology Model Development
12.2.2.1 Zebrafish Model for Hematopoietic Development
12.2.2.2 Zebrafish Model for Studying Coagulation Factors and Coagulation-Related Disorders
12.2.2.3 Zebrafish Model for Studying Blood Disease
12.3 Zebrafish as a Tool for Studying Hematopoiesis
12.3.1 Conserved Biology of Hematopoiesis in Zebrafish
12.3.2 In Vivo Models for Investigating Hematopoiesis and High-Throughput Screening in the Zebrafish
12.3.2.1 Malignant Models of Hematopoietic Disorders
12.3.2.2 Non-malignant Models of Hematopoietic Disorders
Primary Immunodeficiencies
Inherited Bone Marrow Failure Syndromes
Anemia
12.3.3 Genetic Mutations to Study Hematopoiesis in the Zebrafish
12.3.3.1 Impact of Mutations on Embryonic Blood Cells in Zebrafish
12.3.3.2 Mutations Affecting Blood Cell Generation (Bloodless Mutants)
12.3.4 Real-Time Studies and Optical Imaging to Study Hematopoiesis in the Zebrafish
12.4 Zebrafish as a Tool to Develop Hemostasis and Thrombosis Models
12.4.1 Models for Studying Coagulation Factors
12.4.2 Models for Studying the Pathology of Human Blood Coagulation Disorders
12.5 Zebrafish as a Tool to Study Leukemia: Lymphoid and Myeloid Origin
12.5.1 Zebrafish Models to Study T-Cell Acute Lymphoblastic Leukemia (T-ALL)
12.5.2 Zebrafish Models to Study B-Cell Acute Lymphoblastic Leukemia (B-ALL)
12.5.3 Models for Studying Acute Myeloid Leukemia (AML), Myeloproliferative Malignancy, and Myelodysplastic Syndrome (MDS)
12.5.3.1 Myeloproliferative Malignancies
12.5.3.2 Myelodysplastic Syndrome (MDS)
12.5.3.3 Acute Myeloid Leukemia (AML)
12.6 Summary
References
13: Zebrafish: A Metamorphosis in Ophthalmological Research-A Literature Review
13.1 Background
13.2 Zebrafish as an Animal Model
13.3 Visual Systems of Zebrafish
13.3.1 Various Parts of the Eye in Zebrafish
13.3.2 Structure of the Retina
13.4 Embryology of the Zebrafish Eye
13.5 Visual Behavioural Assays in Zebrafish
13.5.1 Optokinetic Response (OKR)
13.5.2 Optomotor Response (OMR)
13.5.3 Escape Response
13.6 Startle Response (SR) and Visual Motor Response (VMR)
13.7 Phototactic Behaviour (PTB)
13.8 Limitations
13.9 Zebrafish Models in Human Ophthalmological Disorders
13.9.1 Aniridia
13.9.2 Retinitis Pigmentosa
13.9.3 Usher Syndrome
13.9.4 Leber Congenital Amaurosis
13.10 Diabetic Retinopathy
13.11 Age-Related Macular Degeneration (AMD)
13.12 Corneal Dystrophy and Cornea Plana
13.12.1 Cataract
13.12.2 Glaucoma
13.12.3 Coloboma
13.12.4 Microphthalmia/Anophthalmia
13.12.5 Cyclopia
13.13 Ocular Toxicity Test in Zebrafish
13.14 Gene Therapy
13.15 Ocular Drug Discovery in Zebrafish
13.16 Retinal Regeneration
13.17 Summary
References
14: Zebrafish: A Potential Preclinical Model for Neurological Research in Modern Biology
14.1 Introduction
14.2 An Introduction to Zebrafish
14.3 Neurobiology and Genetics of Zebrafish
14.4 The Emerging Utility of Zebrafish in Neuroscience Research
14.5 Zebrafish Models of Neurological Disorders
14.5.1 Huntington´s Disease
14.5.2 Alzheimer´s Disease
14.5.3 Parkinson´s Disease
14.5.4 Epilepsy
14.5.5 Schizophrenia
14.5.6 Anxiety and Depression
14.6 Assays for Assessing Cognition in Zebrafish
14.7 Zebrafish Neuroimaging Models
14.8 Neural Circuits in Zebrafish
14.9 Conclusion
References
15: Zebrafish Screening for Skin Diseases
15.1 Introduction
15.2 Zebrafish Model for Hereditary Pigmentary Disorders
15.3 Melanoma
15.4 Psoriasis
15.5 Model for Selective Pruritus
15.6 Model for Wound Healing Research
15.7 Drug Discovery: Evaluation of Percutaneous Absorption of Drug and Drug Delivery Systems Using Zebrafish
15.8 Conclusion
References
16: Growing Importance of Zebrafish in Translational Neuroscience
16.1 Introduction
16.2 Zebrafish Brain Organization
16.3 Modelling Neural Development.
16.4 Modelling Major Neurological Disorders
16.5 Alzheimer´s Disease (AD)
16.6 Parkinson´s Disease (PD)
16.7 Huntington´s Disease (HD)
16.8 Epilepsy
16.9 Multiple Sclerosis (MS)
16.10 Spinal Cord Injury (SCI)
16.11 Glioma
16.12 Conclusion
References
17: Supporting the Next Generation of Risk Assessment in Toxicology: The Design of AOPs Based on the Alternative Model Zebrafi...
17.1 AOP: Concept, Importance, and Current Framework for Development
17.2 AOPs Developed for Non-mammalian Species
17.2.1 Relevance of Non-mammalian-Based AOPs
17.2.2 Use in Risk Assessment
17.3 Current Zebrafish-Based AOPs
17.3.1 Description of Specific AOPs
17.3.2 Common Events Across AOPs
17.4 Conclusions and Perspectives
References
18: Zebrafish: A Novel Model in Psychopharmacological Research
18.1 Introduction
18.1.1 Rationale for the Use of Zebrafish as a Novel Model in Psychopharmacological Research.
18.1.2 Anxiety and Anxiolytics
18.1.3 Depression and Antidepressants
18.1.4 Psychoses and Antipsychotics
18.1.5 Mood Stabilizers/Lithium
18.1.6 Alcohol Research
18.1.7 Nootropics/Cognitive Enhancers
18.1.8 Epilepsy and Antiepileptics
18.1.9 Autism Spectrum Disorder (ASD)
18.1.10 Drug Abuse and Withdrawal
18.1.11 Pain/Analgesics
18.2 Conclusion
References
19: Pharmacological Modeling of Gastrointestinal Disorders in Zebrafish for Drug Discovery and Development
19.1 Introduction of Zebrafish as a Model for Gastrointestinal Tract Disorders: Similarities and Differences with Humans
19.2 Zebrafish as a Tool for IBD Disease Understanding
19.2.1 Zebrafish as a Tool to Study IBD Genetic Susceptibility
19.2.2 Zebrafish as a Tool to Study IBD Immunology
19.2.3 Zebrafish as a Tool to Study Intestinal Development and GIT Transit
19.2.4 Zebrafish as a Model for Pharmacological Screening of Anti-IBD New Chemical Entities: Chemical Models
19.3 Zebrafish as a Tool to Develop GIT Cancer Models
19.3.1 Zebrafish Model for Tumor Metastasis
19.3.2 Zebrafish Model for Drug Screening
19.3.3 Zebrafish Model for Intestinal Cancer
19.4 Zebrafish as a Tool to Study the Intestinal Microbiome and Host-Microbe Interactions
19.4.1 Intestinal Microbiome
19.4.2 Immune Surveillance of Zebrafish
19.4.2.1 Innate Immunity
19.4.2.2 Adaptive Immunity
19.4.3 Zebrafish as a Model of Infectious Diseases of GIT
19.4.3.1 Bacterial Commensals and Pathogens
Edwardsiella and Aeromonas spp.
Vibriosis
19.5 Summary
References
20: Zebrafish Models for Liver Diseases
20.1 Introduction
20.1.1 Comparative Anatomy and Physiology of Zebrafish Liver
20.1.2 Liver Development and Maturation in Zebrafish
20.2 Liver Disease Models of Zebrafish
20.2.1 Chemical-Induced Liver Cirrhosis and Necrosis Models
20.2.1.1 Fatty Liver Disease Models
Alcohol Induced
Non-alcoholic Fatty Liver Disease (NAFLD)
Overfeeding-Induced NAFLD
High Fructose Diet-Induced NAFLD
High-Fat Diet-Induced NAFLD
High Cholesterol-Induced NAFLD
20.2.1.2 Drug-Induced Liver Injury Model
Paracetamol
Valproic Acid
20.2.1.3 Antibiotic-Induced Liver Injury Model.
Mitoxantrone
Tetracycline Hydrochloride
Erythromycin
20.2.1.4 Preservative-Induced Liver Injury Model
Bisphenol A (BPA)
Bisphenol S (BPS)
20.2.1.5 Pesticide-Induced Liver Injury Models
Tricylazole
20.2.1.6 Carcinogen-Induced Liver Injury Models
Diethylnitrosamine (DEN)
Thioacetamide (TA)
20.2.1.7 Heavy Metal-Induced Liver Injury Models
Arsenic
20.2.1.8 Mycotoxin-Induced Liver Injury Models
Aflatoxin
Ochratoxin
20.2.2 Polycystic Liver Disease Model
20.2.3 Genetically Engineered Model of Liver Disease
20.2.3.1 Transgenic Models
20.2.3.2 Mutant Models
20.2.4 Hepatocellular Carcinoma Model
20.3 Limitation and Advantages of Zebrafish
20.3.1 Advantages
20.3.2 Limitations
20.4 Conclusion and Future Prospects of Zebrafish in Liver Disease Modelling
References
21: Zebrafish Model for Neurotoxic Drug Screening: Methodologies and Protocols
21.1 Introduction
21.2 Limitation of the Zebrafish Model
21.3 Neurotoxicity Assessment of Drugs Using Zebrafish Model
21.4 Experimental Protocols
21.4.1 Mode of Drug Exposure to Zebrafish
21.4.2 Behavioural Model
21.4.2.1 Larvae Behaviour
21.4.2.2 Adult Fish Behaviour
Video Recording
Fish Tracking and Analysis of Data
Tracking Using idTracker
Calculation ofVarious Parameters
Behaviour Parameters
21.4.3 Biochemical Assays
21.4.3.1 Sample Preparations
Embryo and Larvae
Extraction and Storage of the Brain (for Adult Fish)
21.4.3.2 ROS Study of Zebrafish Brain
21.4.3.3 TBARS (MDA)
21.4.3.4 Catalase Activity
21.4.3.5 AChE Activity
21.4.4 Whole-Body Cortisol Measurement
21.4.5 Metabolomics
21.4.5.1 Sample Extraction for LC-MS/MS-Based Metabolomics Study
21.4.5.2 Extracting Solution
21.4.5.3 Internal Standard
21.4.5.4 Sample Preparations for LC-MS/MS Study
21.4.5.5 Analytical Instrument
21.4.5.6 Chromatographic Column
21.4.5.7 Chromatographic Conditions
21.4.6 Proteomics
21.4.6.1 Sample Preparation
21.4.6.2 Data Processing and Analysis
21.4.7 Immunohistochemistry of Brain Tissue
21.4.8 Accumulation of Drugs
21.5 Summary
References