Schizophrenia: Methods and Protocols (Methods in Molecular Biology, 2687)

✍ Scribed by Leyre Urigüen (editor), Rebeca Díez-Alarcia (editor)

• Publisher: Humana
• Year: 2023
• Tongue: English
• Leaves: 160
• Category: Library

✦ Synopsis

This volumes presents current methods used in the research of schizophrenia. Chapters guide readers through molecular techniques, in vivo approaches, cell cultures and heterologous expression systems, and even research methods involving human studies. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Schizophrenia: Methods and Protocols aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.

✦ Table of Contents

Preface
Contents
Contributors
Chapter 1: A Novel Method to Estimate Levels of Receptors Using an Allosteric Modulator: Focus on Muscarinic M1 Receptor
1 Introduction
2 Materials
2.1 Solutions
2.2 Preparation of 1 mM Acetylcholine
2.3 Preparation of Radioligand [3H]NMS for Membrane-Binding Assay
2.4 Preparation of Radioligand [3H]NMS for Autoradiography
2.5 Preparation of BQCA
2.6 Equipment
3 Methods
3.1 BQCA Acetylcholine Displacement Shift Assay: Homogenates
3.1.1 Membrane Homogenates Preparation
3.1.2 Protein Measurement
3.1.3 BQCA Acetylcholine Displacement Shift Assay
3.2 BQCA Acetylcholine Displacement Shift Assay: In Situ Radioligand Binding with Autoradiography
3.2.1 Preparation of Frozen Tissue Sections
3.2.2 BQCA Acetylcholine Displacement Shift Autoradiographic Assay
4 Notes
References
Chapter 2: Enhanced Bystander BRET (ebBRET) Biosensors as Biophysical Tools to Map the Signaling Profile of Neuropsychiatric D...
1 Introduction
2 Materials
2.1 Cell Culture
2.2 BRET Experiment
2.3 Data Analysis
3 Methods
3.1 Cell Culture
3.2 Transfection
3.3 BRET Experiment
3.3.1 Treatment of the Cells
3.3.2 BRET Measurement
3.3.3 Data Analysis
4 Notes
References
Chapter 3: [35S]GTPγS (Guanosine-5′-O-(γ-thio)triphosphate-[35S]) Binding Scintillation Proximity Assay Experiments in Postmor...
1 Introduction
2 Materials
2.1 Samples
2.2 Equipment
2.3 Solutions
2.4 Other Reagents
2.5 Consumables
2.6 Software
3 Methods
3.1 Membrane Preparation
3.2 [35S]GTPγS Binding, Immunoprecipitation, and Detection
3.3 Data Analysis
4 Notes
References
Chapter 4: CRISPR/Cas9 as a Simple Technique for the Generation of Murine Knockout Models for Neuropsychiatric Diseases
1 Introduction
2 Materials
2.1 Animals
2.2 Hormones and Drugs
2.3 TE Buffer
2.4 M2 Medium
2.5 KSOM Medium
2.6 STES Buffer
2.7 LB Agar
2.8 LB Broth
3 Methods
3.1 Designing RNA Guide for Your Gene Disruption with CRISPR
3.2 Microinjection of Fertilized Mouse Oocytes
3.3 Embryo Culture
3.4 Preparing Female Recipient for Embryo Transfer
3.5 Embryo Transfer
3.6 Genotyping
3.7 Allele Sequencing
4 Notes
References
Chapter 5: Evaluation of Sensorimotor Gating Deficits in Mice Through Prepulse Inhibition (PPI) of the Startle Response
1 Introduction
2 Materials
2.1 Animals
2.2 Drugs and Administration Material
2.3 Equipment
3 Methods
3.1 Design of the Protocol
3.2 Administration of DOI
3.3 Prepulse Inhibition Assay
3.4 Analyses of the Data
4 Notes
References
Chapter 6: Automated Detection of Psychedelic-Induced Head-Twitch Response in Mice
1 Introduction
2 Materials
2.1 Ear Tagging
2.2 Behavioral Apparatus and Detection System (Fig. 1)
2.3 Chemicals and Reagents
3 Methods
3.1 Ear Tag Installation (Fig. 3)
3.2 Behavioral Experiment and Data Acquisition
3.3 Data Processing
3.4 Basic Data Analyses
4 Notes
References
Chapter 7: Gastrointestinal and Microbiome Profiling in Rodent Models of Neurological and Psychiatric Disorders
1 Introduction
2 Materials
2.1 Carmine Red Dye Solution
2.2 4 kDa Fluorescein Isothiocyanate (FITC)-Dextran
2.3 Fecal Genomic DNA Extraction
2.4 V4 16S rRNA Library Amplification and Cleanup
2.5 16S rRNA Library Denaturation and Sequencing
3 Methods
3.1 Structural and Functional Phenotyping of the Rodent Gut
3.1.1 Fecal Output and Water Content
3.1.2 Gastrointestinal Transit Time
3.1.3 FITC-Dextran Intestinal Permeability
3.1.4 Macroscopic Measures
3.2 Profiling the Gut Microbiome
3.2.1 Fecal Sample Collection for 16S rRNA Sequencing
3.2.2 Fecal Genomic DNA Extraction
3.2.3 16S rRNA V4 Amplicon Library Preparation
3.2.4 Preparing DNA Amplicon Libraries for Sequencing
3.2.5 Sequencing of Amplicon Libraries on MiSeq Platform
3.2.6 Data Analysis Using QIIME2
4 Notes
References
Chapter 8: Electroencephalography for the Study of the Auditory P300 Evoked Potential and Derived Measurements
1 Introduction
1.1 Electroencephalography
1.2 Event-Related Potentials
1.3 P300
1.4 Time-Frequency Analysis
1.5 Entropy
2 Materials
2.1 Laboratory Conditions
2.2 Recording Equipment
2.3 Stimulation Equipment
2.4 Material for Signal Processing
3 Methods
3.1 Lab and Material Setup
3.2 EEG and EOG Preparation
3.3 Stimulating Presentation Paradigm
3.4 EEG Recording
3.5 Signal Pre-processing
3.6 Evoked Potential Processing
3.7 Calculation of Derived Measures
3.7.1 Noise Power (NP)
3.7.2 Spectral Entropy (SE)
3.7.3 Median Frequency (MF)
3.7.4 Relative Power (RP)
3.7.5 Parameter Baseline Correction
4 Notes
References
Chapter 9: Quantification of Endocannabinoids in Human Plasma
1 Introduction
2 Materials
2.1 Supplies
2.2 Equipment
2.3 Reagents
2.4 Standard Solutions
2.5 Calibration Solutions (See Note 9)
2.6 Quality Control Solutions
2.7 Mobile Phases
3 Methods
3.1 Blood Collection and Plasma Extraction
3.2 Extraction of Endocannabinoids from Plasma
3.3 Analysis with LC-MS/MS
3.4 Procedural Blanks and Quality Controls
4 Notes
References
Chapter 10: Neuropsychological and Functional Assessment of Cognitive Impairments in Schizophrenia
1 Introduction
2 Materials
2.1 Neuropsychological Batteries and Tests for Cognitive Assessment in Schizophrenia
2.1.1 MATRICS Consensus Cognitive Battery (MCCB)
2.1.2 Brief Assessment of Cognition in Schizophrenia (BACS)
2.2 Approach to Main Cognitive-Functional Performance Assessment Scales
2.2.1 Cognition Assessment Scale in Schizophrenia (ScoRS)
2.2.2 Cognitive Assessment Interview (CAI)
2.3 Measurement of Day-to-Day Functionality: The Role of Functional Performance Scales
2.3.1 Global Measures of Functioning in Community
2.3.2 Measurement of the Subject´s Functional Performance: Functional Capacity
UCSD Performance-Based Skills Assessment (UPSA)
The Brief International Functional Capacity Assessment (BIFCA)
3 Methods
3.1 Prior Considerations for the Neuropsychological Assessment of Neurocognitive Deficits
3.1.1 How and When to Assess Cognition in Psychotic Disorders
3.2 Neuropsychological assessment
3.2.1 Before Examination
3.2.2 During Assessment
3.2.3 Major Considerations to Be Taken After a Neuropsychological Assessment
4 Notes
References
Chapter 11: Clinical Management and Assessment of the Early Stages of Psychosis: A Multidimensional Approach
1 Introduction
1.1 Experience from the First-Episode Psychosis Intervention Program: Rationale for the CRUPEP Intervention Program
2 Materials
2.1 Initial Multidisciplinary Assessment
2.1.1 Clinical Assessment
2.1.2 Family and Social Assessment
2.1.3 Psychological Intervention
2.1.4 Occupational and Employment Assessment
3 Clinical Follow-Up Assessments
References
Index