Manipulating the mouse embryo: a laboratory manual
Genetics and embryology of the mouse: past, present, and future -- Summary of mouse development -- A mouse colony for the production of transgenic and chimeric animals -- Recovery and in vitro culture of preimplantation embryos -- Isolation, culture, and manipulation of postimplantation embryos -- S...
Saved in:
| Format: | Book |
|---|---|
| Language: | English |
| Published: |
Cold Spring Harbor, NY
Cold Spring Harbor Laboratory Press
2014
|
| Edition: | 4. ed. |
| Subjects: | |
| Online Access: | Inhaltsverzeichnis Klappentext |
| Summary: | Genetics and embryology of the mouse: past, present, and future -- Summary of mouse development -- A mouse colony for the production of transgenic and chimeric animals -- Recovery and in vitro culture of preimplantation embryos -- Isolation, culture, and manipulation of postimplantation embryos -- Surgical procedures -- Production of transgenic mice by pronuclear microinjection -- Embryo-derived stem cell lines -- Germ line-competent stem cells derived from adult mice -- Vector designs for pluripotent stem cell-based transgenesis and genome alterations -- Introduction of foreign DNA into embryonic stem cells -- Production of chimeras -- Genotyping -- Parthenogenesis, pronuclear transfer, and mouse cloning -- Assisted reproduction: ovary transplantation, in vitro fertilization, artificial insemination, and intracytoplasmic sperm injection -- Cryopreservation, rederivation, and transport of mouse strains -- Techniques for visualizing gene products, cells, tissues, and organ systems -- Setting up a micromanipulation lab |
| Item Description: | Includes bibliographical references and index |
| Physical Description: | XXII, 814 S. Ill., graph. Darst., Kt. 29 cm |
| ISBN: | 9781936113002 1936113007 9781936113019 1936113015 0879695749 0879695919 |
Staff View
MARC
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| 245 | 1 | 0 | |a Manipulating the mouse embryo |b a laboratory manual |c Richard Behringer ... |
| 250 | |a 4. ed. | ||
| 264 | 1 | |a Cold Spring Harbor, NY |b Cold Spring Harbor Laboratory Press |c 2014 | |
| 300 | |a XXII, 814 S. |b Ill., graph. Darst., Kt. |c 29 cm | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 500 | |a Includes bibliographical references and index | ||
| 520 | |a Genetics and embryology of the mouse: past, present, and future -- Summary of mouse development -- A mouse colony for the production of transgenic and chimeric animals -- Recovery and in vitro culture of preimplantation embryos -- Isolation, culture, and manipulation of postimplantation embryos -- Surgical procedures -- Production of transgenic mice by pronuclear microinjection -- Embryo-derived stem cell lines -- Germ line-competent stem cells derived from adult mice -- Vector designs for pluripotent stem cell-based transgenesis and genome alterations -- Introduction of foreign DNA into embryonic stem cells -- Production of chimeras -- Genotyping -- Parthenogenesis, pronuclear transfer, and mouse cloning -- Assisted reproduction: ovary transplantation, in vitro fertilization, artificial insemination, and intracytoplasmic sperm injection -- Cryopreservation, rederivation, and transport of mouse strains -- Techniques for visualizing gene products, cells, tissues, and organ systems -- Setting up a micromanipulation lab | ||
| 650 | 7 | |a Animais de laboratório |2 larpcal | |
| 650 | 7 | |a Embriologia comparada |2 larpcal | |
| 650 | 7 | |a Ratos (embriologia) |2 larpcal | |
| 650 | 4 | |a Genetic Engineering |v Laboratory Manuals | |
| 650 | 4 | |a Mice as laboratory animals | |
| 650 | 4 | |a Mice |x Embryos |v Laboratory manuals | |
| 650 | 4 | |a Mice |x Genetic engineering |v Laboratory manuals | |
| 650 | 4 | |a Mice |x embryology |v Laboratory Manuals | |
| 650 | 4 | |a Mice, Transgenic |x genetics |v Laboratory Manuals | |
| 650 | 4 | |a Transgenic mice |v Laboratory manuals | |
| 650 | 4 | |a Mice / Embryos / Laboratory manuals | |
| 650 | 4 | |a Mice / Genetic engineering / Laboratory manuals | |
| 650 | 4 | |a Transgenic mice / Laboratory manuals | |
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| 700 | 1 | |a Behringer, Richard |e Sonstige |4 oth | |
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Record in the Search Index
| _version_ | 1804152159326961664 |
|---|---|
| adam_text | Contents
Preface
xxi
CHAPTER
1
Genetics and Embryology of the Mouse: Past, Present, and Future
1
Introduction
2
Mendelian Inheritance and Linkage: The Beginnings of Mouse Genetics
3
Origins of the Laboratory Mouse
4
Creation of Inbred Strains and Other Resources of Mouse Genetics
4
Origins of Developmental Genetics of the Mouse
9
A Heritage of Experimental Mouse Embryology
11
Manipulating the Mouse Genome
14
Systematic Mutagenesis and Phenotyping of the Mouse
15
CHAPTER
2
Summary of Mouse Development
23
INTRODUCTION
24
Gestation and Embryo Staging
24
Oocyte Maturation and
Ovulation
27
Spermatogenesis
32
Fertilization
34
Cleavage:
Zygote
to Eight-Cell Uncompacted
Morula
35
Morula
Compaction and Blastocyst Formation: The First
Differentiation Events
35
Implantation in the Uterus
37
Trophectoderm and Its Derivatives
39
Formation of the Primitive Endoderm and Epiblast: The Second Round
of Differentiation Events
41
The Epiblast Lineage
41
Epiblast Cells Divide Rapidly
42
The Epiblast Is a Pluripotent Tissue
44
The Pregastrula Embryo
45
Gastrulation: Formation of Mesoderm and Definitive Endoderm
45
viii
/ Contents
Fate Maps of Early
Postimplantation
Embryos
49
Origin of the Germline 49
Generation of Regional Diversity in the Mesoderm
51
The Anterior Visceral Endoderm
52
The Node
52
The Tail Bud
54
Embryonic Turning
54
Somites and Their Derivatives
55
Lateral Plate and Intermediate Mesoderm and Their Derivatives
58
Sex Determination and Differentiation
60
Limb Formation
61
Neurulation: Formation of the Nervous System
61
Regional Diversification in the Neural Tube
62
Neural Crest
65
Formation of the Branchial Arches and the Pharyngeal Region
66
Gut Development
67
Extra-Embryonic Tissues
68
Visceral Endoderm
70
Parietal Endoderm
71
Differentiation of the Extra-Embryonic Mesoderm
72
The Structure and Function of the Chorioallantoic Placenta
72
The Adult Mouse
74
Mouse Coat Color and Its Genetics
74
Morphological and Behavioral Mutants
78
CHAPTER
3
A Mouse Colony for the Production of Transgenic and Chimeric Animals
85
INTRODUCTION
85
Pathogen Control in Experimental Mouse Colonies
86
Location of the Embryo Manipulation Laboratory
87
Establishing a Breeding Colony to Supply Embryo Donor
and Recipient Mice
88
Embryo Donors
89
Female Embryo Donor Mice for Pronuclear
Microinjection
89
Female Embryo Donor Mice for Generation of
ES
Cell Chimeras
90
Fedite
Stud Male Mice
91
Production of Embryo Donors
91
Embryo Recipients
92
Female Mice to Serve as Pseudopregnant Recipients
92
Sterile Male Mice to Induce Pseudopregnancy in Females
93
Production of Embryo Recipients
93
Pregnancy with Manipulated Embryos
94
Setting up Natural Matings
94
Contents
Ι ¡Χ
Inducing
Superovulation
95
Strain Background
%
Age and Weight
96
Dose of Gonadotropins
97
Time of Administration of the Gonadotropins
97
Reproductive Performance of Stud Males
97
Animal Identification and Tissue Biopsies
97
Ear Punching or Notching
98
ľoe
Clipping
98
Tattooing
98
Ear Tags
99
Microchip Implants
100
Tail-Tip Excision
100
PROTOCOLS
1
Selecting Females in Estrus and Checking Plugs
103
2
Preparation of Gonadotropins and
Superovulation
105
3
Intraperitoneal (i.p.) Injection
107
CHAPTER
4
Recovery and In Vitro Culture of Preimplantation Embryos
109
INTRODUCTION
109
Culture Media for Preimplantation Embryos
109
History
109
Two-Cell Block
110
Media for the Development from
Zygotes
to Blastocysts
112
Critical Components of Preimplantation Embryo Culture
113
Glucose
113
Amino
Acids
114
Osmolarity
115
Oxygen Tension, CO
2,
pH,
Light, and Temperature
115
Embryo Incubation Volume and the Role of Growth Factors
117
Effect of Embryo Culture on Gene Expression
117
Important Considerations for Successful In Vitro Embryo Culture
118
Medium Choice and Experimental Outcome
118
Reagents
118
Embryo Collection and Culture
119
Quality Control
119
Preparation of Embryo Culture Media
120
Pipettes for Embryo Handling
121
Collection of Preimplantation Embryos
123
PROTOCOLS
1
Preparation of Media for Embryo Handling and Culture
126
χ
/ Contents
2
Setting Up
Microdrops
Culture 132
3
Making Embryo-Handling Pipettes from Hard Glass Capillaries
134
4
Preparation of Siliconized Pipettes
136
5
Opening the Abdominal Cavity and Locating Female Reproductive Organs
137
6
Collection of
Zygotes
and Removal of Cumulus Ceils with Hyaluronidase
139
7
Collection of Two-Cell- to Morula-Stage Embryos
143
8
Collection of Blastocysts
146
CHAPTER
5
Isolation, Culture, and Manipulation of
Postimplantation
Embryos
149
INTRODUCTION
149
Isolating
Postimplantation
Embryos
150
Visualizing Early Embryo Implantation Sites
150
Isolating
Postimplantation Embryos 150
Isolating Extra-Embryonic Membranes
150
Separating
Postimplantation
Germ Layers
151
Germ-Layer
Expiant
Recombination Culture
151
Isolating Germ Cells from the Genital Ridge
151
Culturing
Postimplantation
Embryos
152
Preparing Embryos
152
Roller Culture of
Postimplantation
Embryos
153
Static Culture of
Postimplantation
Embryos
153
Static Culture of
Postimplantation
Embryos for Imaging
154
Introducing Nucleic Acids into
Postimplantation
Embryos by Electroporation
154
PROTOCOLS
1
Visualizing Early Embryo Implantation Sites by Dye Injection
157
2
Isolating
Postimplantation
Embryos
159
3
Isolating Extra-Embryonic Membranes
167
4
Separating
Postimplantation
Germ Layers
170
5
Germ-Layer
Expiant
Recombination Culture
173
6
Isolating Germ Cells from the Genital Ridge
175
7
Roller Culture of
Postimplantation
Embryos
178
8
Static Culture of
Postimplantation
Embryos
182
9
Static Culture of
Postimplantation
Embryos for Imaging
184
10
Electroporation of
Postimplantation
Embryos In Vitro
187
11
Electroporation of
Postimplantation
Embryos In
Utero
190
CHAPTER
6
Surgical Procedures
195
INTRODUCTION 195
General Guidelines for Mouse Surgeries
196
Anesthesia and Analgesia I97
Anesthetics -J97
Contents
I
xi
Analgesics
198
Vasecto
my
199
Embryo Transfer
199
Background
199
Role of
Zona Pellucida in
Embryo Transfer
200
Number of Embryos to Transfer
200
Recipient Females
201
Technical Aspects
202
Caesarean Section and Fostering
203
Tissue Transplantation
203
Tissue Biopsy
203
Ovariectomy and Castration
203
PROTOCOLS
1
Preparation of Tribromoethanol (Avertin)
206
2
Vasectomy of Mice
208
3
Oviduct Transfer of Mouse Embryos
211
4
Uterine Transfer of Mouse Embryos
216
5
Caesarean Section and Fostering of Mice
220
6
Transplantation of Adult or Embryonic Tissues under the Kidney Capsule of Mice
222
7
Subcutaneous Injection of Pluripotent Stem Cells in Mice
225
8
Tissue Biopsies in Mice
227
9
Blood Collection by Tail Bleeding in Mice
230
10
Ovariectomy of Mice
232
11
Castration of Mice
234
CHAPTER
7
Production of Transgenic Mice by Pronuclear Microinjection
237
INTRODUCTION
238
Applications of Transgenic Mouse Technology
238
Gene Transfer into the Mouse Genome by Pronuclear Microinjection
240
Designing
Transgenes
242
Effects of Prokaryotic Vector Sequences
242
Length of
DNA
Construct
242
Coinjection of Two (or More)
Transgenes
243
Distinguishing Expression of
Transgenes
and Endogenous Genes
243
Reporter Genes
244
Using Previously Identified Tissue-Specific Regulatory Sequences in
Transgene
Constructs
244
Expression of cDNAs and the Role of
Introns
in
Transgene
Expression
245
Using Housekeeping Gene Promoters to Direct Ubiquitous Expression
245
Factors Affecting the Efficiency of Gene Transfer
245
General Considerations about Large
DNA
Constructs
247
isolation and Purification of
DNA 247
General Considerations
247
Contents
Contaminants and Methods for Their Removal 247
Commercially Available Kits for Plasmid Preparation and
DNA
Purification
249
Preparation of Standard-Sized Plasmid
DNA
from Bacterial Cultures
249
Digestion of Plasmid
DNA
for Release of Genetic Construct Fragment
251
Gel Separation 251
Recovery of
DNA
Fragments from
Agarose
Gels
Fragment Purification
Preparing
DNA
for Microinjection 252
Determining and Maintaining High
DNA
Quality
Determining Concentration of
DNA
for Injection
Filtering the
DNA
Solution 254
Storage of Prepared
DNA 254
Preparation of Technical Equipment and
Zygotes
254
Choice of Mouse Strain
254
Preparation of
Zygotes
255
Making Holding Pipettes
256
Making Injection Pipettes
257
Microinjection Setup
258
Microinjection
262
Timing of Pronuclear Injection
262
Microinjection of Mouse
Zygotes
262
After Microinjection
263
Establishment and Maintenance of Transgenic Mouse Lines
263
PROTOCOLS
1
Simple, Reliable Steps for
DNA
Fragment Isolation and Purification
269
2
Isolating
ВАС
DNA
from Bacterial Cultures Using the NucleoBond System
271
3
Purification of
ВАС
DNA
Prepared with the NucleoBond System
273
4
Large-Scale Preparation of Yeast
Agarose
Plugs to Isolate
YAC
DNA 274
5
Purification of
YAC
DNA
Using a Two-Gel Electrophoresis Procedure
279
6
Purification of
YAC
DNA
Using PFGE and
Ultrafiltration 286
7
Preparing Injection Buffer for Standard-Sized
DNA 289
8
Preparing Injection Buffer for
ВАС
/YAC
DNA 290
9
Preparation of In Vitro-Translated
RNA
for Microinjection
292
10
Making Holding Pipettes
294
11
Making Injection Pipettes
296
12
Microinjection Setup
298
13
Microinjection of Mouse
Zygotes
301
14
Microinjection of
RNA
into Mouse
Zygotes
306
15
Microinjecting Lentivirus into Mouse Embryos
308
16
Infecting Mouse Embryos by Coculturing with Lentivirus
312
DISCUSSION
Troubleshooting Guide for Microinjection of Mouse
Zygotes
and Production
of
Tran
sgen ic
M
ice
315
Contents
I
xiii
CHAPTER
8
Embryo-Derived Stem Cell Lines
321
INTRODUCTION
321
Derivation and Culture of Mouse Embryonic Stem Cells
322
Historic Background and New Developments
322
General Considerations for Optimal
ES
Cell Culture
325
Media and Reagents
327
Serum and Serum Replacement
328
Leukemia Inhibitory Factor (LIF)
329
Trypsin/EDTA
330
Feeder Cells
330
Primary Mouse Embryonic
Fibroblasts
330
STO Fibroblasts
330
Mycoplasma and Pathogen Testing
331
Karyotype
332
In Vitro Differentiation of
ES
Cells
333
Trophoblast and Extra-Embryonic Endoderm Stem Cells
333
Epiblast Stem Cells
334
Progenitor Cells from the Central Nervous System
334
PROTOCOLS
1
Preparation of Primary Mouse Embryonic
Fibroblasts
(MEFs)
338
2
Preparation of Feeder Cell Layers from
STO
or MEF Cells
341
3
Passage of
ES
Cells
343
4
Cryopreservation of
ES
Cells
345
5
Testing Serum Batches
348
6
Derivation of
ES
Cell Lines Using Serum-Containing Culture Conditions
350
7
Derivation of
ES
Cell Lines Using Small-Molecule Inhibitors of Erk and Gsk3 Signaling (2i)
355
8
Chromosome Counting
363
9
Differentiating
ES
Cells into Embryoid Bodies by Suspension Culture
366
10
Differentiating
ES
Cells into Embryoid Bodies by Hanging-Drop Cultures
368
11
Differentiating
ES
Cells into Embryoid Bodies in AggreWell Plates
370
12
Derivation and Culture of TS Cell Lines
373
13
Derivation and Culture of XEN Cell Lines
379
14
Derivation and Culture of EpiSC Lines
382
15
Isolation, Culture, and Differentiation of Progenitor Cells from the Central Nervous System
389
16
Isolation, Propagation, and Differentiation of Radial Clia-Like Neural
Progenitor Cells in Adherent Cultures
395
CHAPTER
9
Germline-Competent Stem Cells Derived from Adult Mice
403
INTRODUCTION
403
Spermatogonial Stem Cells
404
Culture of Spermatogonial Stem Cells
404
XIV
/ Contents
Transplantation
of
Spermatogonial Stern
Cells
405
Induced
Pluripotent Stern
Cells
406
Reprogramming
Vectors
407
Testing
Pluripotency and Germline
Competence of Established
¡PS
Cell
Lines 408
PROTOCOLS
1 Isolation
of the Spermatogonial Stem Cell-Containing Fraction from
Testes 410
2
Culture and Expansion of Primary Undifferentiated Spermatogonial Stem Cells
413
3
Cryopreserving and Thawing Spermatogonial Stem Cells
416
4
Spermatogonial Stem Cell Transplantation to the
Testis
418
5
Integrating piggyBac Transposon
Transgenes
into Mouse
Fibroblasts
by Electroporation
424
6
Integrating piggyBac Transposon
Transgenes
into Mouse
Fibroblasts
Using Chemical Methods
426
7
Reprogramming Mouse
Fibroblasts
with piggyBac
Transposons
428
CHAPTER
10
Vector Designs for Pluripotent Stem Cell-Based
Transgenesis
and Genome Alterations
431
INTRODUCTION
431
Glossary of Elements
432
Cenomic and Gene Elements
432
Selectable Markers
434
Reporters
436
Site-Specific Recombinases
437
Inducible Systems
439
Reporters to Detect
Recombinase
Action
440
ES
Cell-Mediated
Transgenesis 440
Gene Targeting
442
Gene and Promoter Trap
445
Homologous Recombination-Replaceables
446
Site-Specific Recombination-Mediated Insertions
448
Induced Site-Specific Chromosomal Aberrations
449
Isolating
ES
Cells Homozygous for Specific Mutations
451
PROTOCOLS
1
In Vitro Screen for Widespread
Transgene
Expression
455
2
In Vitro Screen to Identify Silent but Activatable (S/A) Integration Sites
for a Tet-lnducible
Transgene
458
CHAPTER
11
Introduction of Foreign
DNA
into Embryonic Stem Cells
461
INTRODUCTION
461
General Considerations
462
Culture Conditions
462
Contents
I XV
Genotyping
Assays
462
Transfection
Methods
462
Targeting
Approaches
463
Purifying
DNA
for Introduction into the
ES
Cell Genome
464
Electroporating
DNA
into
ES
Cells and Selection Methods
464
Isolating Individual
ES
Cell Colonies by Picking, Replica-Plating, and Freezing
ES
Cell Lines
465
PROTOCOLS
1
Electroporating
DNA
into
ES
Cells
468
2
Isolating Individual
ES
Cell Colonies by Picking
472
3
Passaging and Freezing
ES
Cells in 96-Well Plates
475
4
Thawing
ES
Cells from a 96-Well Plate
478
5
Rapid Preparation of
DNA
from Cells in 96-Well Tissue Culture Plates
480
6
Preparation of
DNA
from Cells in 24-Well Tissue Culture Plates
483
7
Genotyping
ES
Cell Colonies before Picking
485
CHAPTER
12
Production of Chimeras
489
INTRODUCTION
489
General Considerations
490
Types of Chimeras
500
Diploidi
Embryos Embryo Aggregation Chimeras
500
ES Cells^Diploid
Embryo Aggregation and Injection Chimeras
501
Diploid^Tetraploid Embryo Aggregation Chimeras
502
ES
Cells^Tetraploid Embryo Aggregation and Injection Chimeras
502
Preparation of Embryos for Injection or Aggregation
503
ES
Cell Injection Chimeras
505
The
Microinjection
Setup
505
Preparation of the Injection and Holding Pipettes
506
Transfer of Chimeric Blastocysts
506
Breeding Chimeric Mice
507
General Troubleshooting for Chimera Production and Breeding
for Germline Transmission
509
PROTOCOLS
1
Making
ES
Cell Injection Needles
514
2
Assembling the
Microinjection
Setup
518
3
Preparation of
ES
Cells for Injection
520
4
Injecting Biastocysts
522
5
Injecting Eight-Cell-Stage Embryos
526
6
Preparation of
ES
Cells for Aggregation
531
7
Preparation of the Aggregation Plate
534
8
Removal of the
Zona Pellucida
536
9
Production of Tetraploid Embryos
539
10
Assembly of the Aggregates
542
XVI
/ Contents
CHAPTER
13
Genotyping
11
Disaggregating Cleavage-Stage Embryos and the Inner Cell Mass
of Blastocysts into Individual Cells
12
Immunosurgery: Isolating the Inner Cell Mass of Blastocysts
547
549
551
INTRODUCTION
Tissue Samples and
DNA
Preparation for Genotyping or Characterization of
Transgenes
Detection and Analysis of
Transgene
Produced by Pronuclear Injection
Detection and Analysis of
ES
Cell-Mediated Gene/Genome Alterations
Cloning
Transgene/Host DNA
Junctions
Identifying Homozygous Transgenic Mice or Embryos
In Situ Hybridization to
Interphase
Nuclei
Test Breeding
Southern Blot Analysis Using a Flanking Probe
PCR Analysis Using a Flanking Primer
PROTOCOLS
1
Isolation of High-Molecular-Weight
DNA
from Mouse Tail Tips
2
Isolation of High-Molecular-Weight
DNA
from Embryonic Tissues, Yolk Sac,
Umbilical Cord, and the Like
3
Preparation of Tissue Lysates for PCR Template
INTRODUCTION
Generating Parthenogenotes
Performing Pronuclear Transfer
Cloning Mice
PROTOCOLS
1
Parthenogenetic Activation of Oocytes
2
Pronuclear Transplantation in the Mouse Embryo
3
Cloning Mice
551
551
552
554
555
555
556
556
556
557
558
560
562
•
Alternative Protocol: Preparation of Template from Embryonic and Adult Tissue Samples
564
•
Alternative Protocol: Preparation of Template from Mouse Tail Tissue
566
4
Polymerase Chain Reaction
567
5
Preparation of
DNA
for Southern Blot Analysis or PCR from
ES
or Other Cultured Cells
569
CHAPTER
14
Parthenogenesis, Pronuclear Transfer, and Mouse Cloning
571
571
571
572
572
574
577
583
CHAPTER
15
Assisted Reproduction: Ovary Transplantation, in Vitro Fertilization, Artificial
Insemination, and Intracytoplasmic Sperm Injection
593
INTRODUCTION
Approaches to Bypass Mouse Infertility
593
593
Contents
I
xvii
Female Infertility
594
Male Infertility
595
Strategies to Rescue the Cermline of Mice
596
Ovary Transplantation
596
In Vitro Fertilization
596
Artificial Insemination
597
Intracytoplasmic Sperm Injection
598
PROTOCOLS
1
Ovary Transplantation
601
2
In Vitro Fertilization
604
3
Nonsurgical Artificial Insemination
615
4
Determining the Stage of the Estrous Cycle by Vaginal Smear
617
5
Surgical Artificial Insemination
619
6
Intracytoplasmic Sperm Injection
623
CHAPTER
16
Cryopreservation, Rederivation, and Transport of Mouse Strains
631
INTRODUCTION
631
Embryo Cryopreservation
632
Factors to Consider in Embryo Cryopreservation
632
Equilibrium Methods of Cryopreservation
633
Nonequilibrium Methods of Cryopreservation
634
Sperm Cryopreservation
635
Oocyte and Ovary Cryopreservation
636
Storage and Records
637
Rederivation of Mouse Strains
637
Transport of Mouse Embryos and Germ Plasm
640
PROTOCOLS
1
Embryo Cryopreservation by Slow Freezing
646
2
Embryo Cryopreservation by Rapid Cooling
652
3
Embryo Cryopreservation by High-Osmolality Vitrification
658
•
Alternative Protocol: High-Osmolality Vitrification of Embryos in Insemination Straws
662
4
Sperm Cryopreservation Using Cryoprotectant Containing Monothioglycerol (MTG)
665
5
Sperm Cryopreservation Using Cryoprotectant Containing L-Glutamine
671
6
Ovary Cryopreservation
676
7
Shipment of Live Preimplantation-Stage Embryos
679
CHAPTER
17
Techniques for Visualizing Gene Products, Cells, Tissues, and Organ Systems
683
INTRODUCTION
684
Visualizing Gene Products
684
Isolating Total
RNA
from Mouse Embryos or Fetal Tissues
684
onfenís
Cenerà/
Techniques
for ¡mmunohistochemistry and In Situ Hybridization
of Mouse Embryo Sections
¡mmunohistochemistry of Embryo Sections
686
Immunohistochemistry of Whole-Mount Embryos
686
Fluorescent Analysis of Whole-Mount Embryos and Fetal Organs
686
In Situ Hybridization of Embryo and Tissue Sections with
RNA
Probes
686
In Situ Hybridization of Whole-Mount Embryos with
RNA
Probes
687
Staining for
ß-Galactosidase
Activity
691
Staining for Alkaline Phosphatase Activity
691
Visualizing Cells
692
Visualizing Fluorescent Proteins
692
Visualizing Tissues and Organ Systems
693
Visualizing the Mouse Skeleton Using Histochemical Stains
693
Visualizing the Fetal Vasculature by India Ink Injection
694
Visualizing the Fetal
Creat
Blood Vessels by Plastic Casting
694
PROTOCOLS
1
Isolating Total
RNA
from Mouse Embryos or Fetal Tissues
695
2
Preparing Tissue Fixation Solutions
698
•
Alternative Protocol: Bouin s Fixative
699
3
Handting Blastocysts
for Fixation
700
4
Embedding Samples in Wax
701
•
Additional Protocol: Embedding Small Embryos
703
5
Preparing Glass Slides and
Coverslips
for In Situ Hybridization
704
6
Cutting Wax Sections Using a Microtome
706
•
Alternative Protocol: Transferring Sections onto Water Droplets
708
7
Cutting Thick Sections Using a Vibratome
709
•
Alternative Protocol·. Modifications for Live Specimens
712
8
Dewaxing and Rehydrating Sections before In Situ Hybridization or Staining
714
9
Immunohistochemistry of Embryo Sections
715
10
Immunohistochemistry of Whole-Mount Embryos
719
11
DAPI Staining of Whoie-Mount Embryos or Fetal Organs
722
12
Immunofluorescent Staining of Whole-Mount Embryos
724
13
Mounting Embryos for Microscopic Observation and Imaging
726
14
In Situ Hybridization of Embryo and Tissue Sections with Radioiabeled
RNA
Probes
728
•
Additional Protocol: Autoradiography
733
15
In Situ Hybridization of Whole-Mount Embryos with Nonradiolabeled
RNA
Probes
735
16
Imaging Embryos after Whole-Mount In Situ Hybridization
741
17
Sectioning Embryos after Whole-Mount In Situ Hybridization
742
•
Alternative Protocol: Wax Sections
744
18
Staining Whole Embryos and Sections for
ß-Galactosidase
(LacZ) Activity
745
•
Alternative Protocol: Staining Frozen Sections
748
19
Staining for Alkaline Phosphatase Activity
751
20
Visualization of Fluorescent Protein Expression in Whole-Mount
Postimpiantation-Stage Embryos
753
Contents
I
xix
21
Observation of Fluorescent Proteins in Living Cells
755
22
Intracellular Observation of Fluorescent Proteins in Fixed Cells
756
23
Fixation and Paraffin Embedding for GFP Visualization
757
24
Alcian Blue Staining of the Fetal Cartilaginous Skeleton
759
25
Alcian Blue/Alizarin Red Staining of Cartilage and Bone
760
26
Alizarin Red Staining of Postnatal Bone
762
27
Visualizing the Fetal Vasculature by India Ink Injection
763
28
Visualizing the Fetal Great Blood Vessels by Plastic Casting
765
CHAPTER
18
Setting Up
a
Micromanipulation
Laboratory
767
INTRODUCTION
767
The
Micromanipulation
Laboratory and Its Environment
767
General Considerations
767
Inbred, Outbred, Hybrid, and Genetically Modified Mice
768
General Equipment
769
Refrigerators and Freezers
769
Incubators
769
Laminar Flow Hoods and Biological Safety Cabinets
770
Microscopes
771
Stereomicroscopes
771
Inverted Microscopes
772
Temperature Controllers for Heating and Cooling
Microscope Stages
772
Vibration-Free Tables
773
Micromanipulation 773
Micromanipulators 773
Microinjection
Systems
773
Piezo
Impact and Laser Systems
774
Production of
Microtools
from Glass Capillaries
774
Pullers
775
Microforges, Bevelers,
and Grinders
775
Electrofusion
775
Surgical Instruments
776
Cryopreservation
777
APPENDIX
1
Buffers and Solutions
779
Acidic Tyrode s Solution for Removing Zonae Pellucidae from Preimplantation Embryos
780
Alkaline Phosphatase Buffers (NTMT)
780
Alkaline Phosphatase Staining Solution
780
Chicago Sky Blue 6B, Also Called Pontamine Sky Blue
781
Hyaluronidase
781
XX
/ Contents
Man
η
ito
1 (0.3
M) 781
Methylene
Blue
Solution 7Sri
Pancreatin/Trypsin
Solution
for Separating Germ and Tissue Layers
781
PBSMT for Jmmunohistochemistry of Whole-Mount Embryos
782
PBT 782
PhenolrChloroform Solution for Isolating Total
RNA
from Mouse Embryos or Fetal Tissues
782
Phosphate-Buffered Saline (PBS)
782
Pronase Solution
783
Saline/EDTA Buffer plus Glucose for Isolation of Germ Cells and Tissue Culture
783
SSC
(20
x)
783
SSPE
(20
x)
783
TAE (Tris-Acetate/EDTA) Buffer
(1
χ
Solution)
783
TE (Tris-EDTA)
Buffer
783
Trypsin
(0.25%)
in
Tris-Saline
for Tissue Culture
784
Trypsin-EDTA
Solution
784
Tyrode Ringer s Saline
(pH 7.6-7.7),
Ca24 /Mg24 -Free
784
APPENDIX
2
WWW Resources
785
APPENDIX
3
General Safety and Hazardous Material Information
789
Index
795
Manipulating the Mouse Embryo
A LABORATORY MANUAL
·
FOURTH EDITION
ι
his fourth edition of The Mouse Manual is once again the definitive reference source
for mouse development, transgenesis techniques, and molecular biology. Its preeminent
authors Richard Behringer, Marina Gertsenstein,
Kristina
Nagy,
and
Andras
Nagy
have
reorganized and updated this edition to include new information and protocols on:
•
generation of induced pluripotent stem cells
•
RNA microinjections
•
lentiviral microinjections and infection
•
assisted reproduction techniques for sperm and embryo cryopreservation
•
isolation, generation, and transplantation of spermatogonial stem cell lines
•
in
utero electroporation
of gene constructs into
postimplantation
embryos
•
vibratome sectioning of live and fixed tissues for imaging thick tissue sections
•
whole-mount fluorescent staining methods for three-dimensional visualization
Recombinant
DNA
techniques and methods for studies of mouse embryonic development
have been updated from previous editions, as has the wealth of information on mouse
laboratory strains, mouse housing and breeding, surgical procedures, assisted reproduc¬
tion, handling of embryos, and
micromanipulation
setups. The first edition of this classic
work appeared more than
20
years ago, and authors of the early editions included
Brigid
Hogan, Rosa Beddington, Frank Costantini, and Elizabeth Lacy.The field s technological
sophistication has grown exponentially but the manual remains the essential practical and
theoretical guide for all students, lab technicians, and investigators who work with mice.
|
| any_adam_object | 1 |
| building | Verbundindex |
| bvnumber | BV041826204 |
| callnumber-first | Q - Science |
| callnumber-label | QL737 |
| callnumber-raw | QL737.R6 |
| callnumber-search | QL737.R6 |
| callnumber-sort | QL 3737 R6 |
| callnumber-subject | QL - Zoology |
| classification_rvk | WC 4900 WC 5088 WC 6570 WG 3450 WX 6037 |
| classification_tum | BIO 750f |
| ctrlnum | (OCoLC)870207830 (DE-599)BSZ400204320 |
| dewey-full | 571.861935 616.027333 571.8/61/935 |
| dewey-hundreds | 500 - Natural sciences and mathematics 600 - Technology (Applied sciences) |
| dewey-ones | 571 - Physiology & related subjects 616 - Diseases |
| dewey-raw | 571.861935 616.027333 571.8/61/935 |
| dewey-search | 571.861935 616.027333 571.8/61/935 |
| dewey-sort | 3571.861935 |
| dewey-tens | 570 - Biology 610 - Medicine and health |
| discipline | Biologie Medizin |
| edition | 4. ed. |
| format | Book |
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| id | DE-604.BV041826204 |
| illustrated | Illustrated |
| indexdate | 2024-07-10T01:06:17Z |
| institution | BVB |
| isbn | 9781936113002 1936113007 9781936113019 1936113015 0879695749 0879695919 |
| language | English |
| lccn | 2013023915 |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-027271173 |
| oclc_num | 870207830 |
| open_access_boolean | |
| owner | DE-19 DE-BY-UBM DE-355 DE-BY-UBR DE-29T |
| owner_facet | DE-19 DE-BY-UBM DE-355 DE-BY-UBR DE-29T |
| physical | XXII, 814 S. Ill., graph. Darst., Kt. 29 cm |
| publishDate | 2014 |
| publishDateSearch | 2014 |
| publishDateSort | 2014 |
| publisher | Cold Spring Harbor Laboratory Press |
| record_format | marc |
| spelling | Manipulating the mouse embryo a laboratory manual Richard Behringer ... 4. ed. Cold Spring Harbor, NY Cold Spring Harbor Laboratory Press 2014 XXII, 814 S. Ill., graph. Darst., Kt. 29 cm txt rdacontent n rdamedia nc rdacarrier Includes bibliographical references and index Genetics and embryology of the mouse: past, present, and future -- Summary of mouse development -- A mouse colony for the production of transgenic and chimeric animals -- Recovery and in vitro culture of preimplantation embryos -- Isolation, culture, and manipulation of postimplantation embryos -- Surgical procedures -- Production of transgenic mice by pronuclear microinjection -- Embryo-derived stem cell lines -- Germ line-competent stem cells derived from adult mice -- Vector designs for pluripotent stem cell-based transgenesis and genome alterations -- Introduction of foreign DNA into embryonic stem cells -- Production of chimeras -- Genotyping -- Parthenogenesis, pronuclear transfer, and mouse cloning -- Assisted reproduction: ovary transplantation, in vitro fertilization, artificial insemination, and intracytoplasmic sperm injection -- Cryopreservation, rederivation, and transport of mouse strains -- Techniques for visualizing gene products, cells, tissues, and organ systems -- Setting up a micromanipulation lab Animais de laboratório larpcal Embriologia comparada larpcal Ratos (embriologia) larpcal Genetic Engineering Laboratory Manuals Mice as laboratory animals Mice Embryos Laboratory manuals Mice Genetic engineering Laboratory manuals Mice embryology Laboratory Manuals Mice, Transgenic genetics Laboratory Manuals Transgenic mice Laboratory manuals Mice / Embryos / Laboratory manuals Mice / Genetic engineering / Laboratory manuals Transgenic mice / Laboratory manuals Maus (DE-588)4169148-9 gnd rswk-swf Embryo (DE-588)4014555-4 gnd rswk-swf Experiment (DE-588)4015999-1 gnd rswk-swf Embryonalentwicklung (DE-588)4070792-1 gnd rswk-swf Maus (DE-588)4169148-9 s Embryo (DE-588)4014555-4 s DE-604 Embryonalentwicklung (DE-588)4070792-1 s Experiment (DE-588)4015999-1 s Behringer, Richard Sonstige oth Digitalisierung UB Regensburg - ADAM Catalogue Enrichment application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027271173&sequence=000003&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis Digitalisierung UB Regensburg - ADAM Catalogue Enrichment application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027271173&sequence=000004&line_number=0002&func_code=DB_RECORDS&service_type=MEDIA Klappentext |
| spellingShingle | Manipulating the mouse embryo a laboratory manual Animais de laboratório larpcal Embriologia comparada larpcal Ratos (embriologia) larpcal Genetic Engineering Laboratory Manuals Mice as laboratory animals Mice Embryos Laboratory manuals Mice Genetic engineering Laboratory manuals Mice embryology Laboratory Manuals Mice, Transgenic genetics Laboratory Manuals Transgenic mice Laboratory manuals Mice / Embryos / Laboratory manuals Mice / Genetic engineering / Laboratory manuals Transgenic mice / Laboratory manuals Maus (DE-588)4169148-9 gnd Embryo (DE-588)4014555-4 gnd Experiment (DE-588)4015999-1 gnd Embryonalentwicklung (DE-588)4070792-1 gnd |
| subject_GND | (DE-588)4169148-9 (DE-588)4014555-4 (DE-588)4015999-1 (DE-588)4070792-1 |
| title | Manipulating the mouse embryo a laboratory manual |
| title_auth | Manipulating the mouse embryo a laboratory manual |
| title_exact_search | Manipulating the mouse embryo a laboratory manual |
| title_full | Manipulating the mouse embryo a laboratory manual Richard Behringer ... |
| title_fullStr | Manipulating the mouse embryo a laboratory manual Richard Behringer ... |
| title_full_unstemmed | Manipulating the mouse embryo a laboratory manual Richard Behringer ... |
| title_short | Manipulating the mouse embryo |
| title_sort | manipulating the mouse embryo a laboratory manual |
| title_sub | a laboratory manual |
| topic | Animais de laboratório larpcal Embriologia comparada larpcal Ratos (embriologia) larpcal Genetic Engineering Laboratory Manuals Mice as laboratory animals Mice Embryos Laboratory manuals Mice Genetic engineering Laboratory manuals Mice embryology Laboratory Manuals Mice, Transgenic genetics Laboratory Manuals Transgenic mice Laboratory manuals Mice / Embryos / Laboratory manuals Mice / Genetic engineering / Laboratory manuals Transgenic mice / Laboratory manuals Maus (DE-588)4169148-9 gnd Embryo (DE-588)4014555-4 gnd Experiment (DE-588)4015999-1 gnd Embryonalentwicklung (DE-588)4070792-1 gnd |
| topic_facet | Animais de laboratório Embriologia comparada Ratos (embriologia) Genetic Engineering Laboratory Manuals Mice as laboratory animals Mice Embryos Laboratory manuals Mice Genetic engineering Laboratory manuals Mice embryology Laboratory Manuals Mice, Transgenic genetics Laboratory Manuals Transgenic mice Laboratory manuals Mice / Embryos / Laboratory manuals Mice / Genetic engineering / Laboratory manuals Transgenic mice / Laboratory manuals Maus Embryo Experiment Embryonalentwicklung |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027271173&sequence=000003&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=027271173&sequence=000004&line_number=0002&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT behringerrichard manipulatingthemouseembryoalaboratorymanual |