almost everything you've wanted to know about elucidating mouse development

2
BOOK LOOK Almost Everything You’ve Wanted to Know About Elucidating Mouse Development MANIPULATING THE MOUSE EMBRYO: A LABORATORY MANUAL, THIRD EDI- TION by A. Nagy, M. Gertsenstein, K. Vintersten, and R. Behringer Cold Spring Harbor Laboratory Press, New York, NY, 2003, 764 p. ISBN: 0-87969-591-9 $125.00 As an alumnus of the 1988 Cold Spring Harbor Course in Molecular Mouse Embryology, it was a plea- sure to be asked to review the third edition of the companion course book, entitled “Manipulating the Mouse Embryo: A Laboratory Man- ual.” The “Mouse Course” was es- tablished in the 1980s as part of the Cold Spring Harbor Laboratory’s ac- ademic program to communicate new discoveries, concepts, and methodologies in the manipulation of the mouse embryo and its ge- nome to small groups of students. For many participants, including my- self, the instruction of Rosa Bedding- ton, Liz Robertson, Andy McMahon, and Robin Lovell-Badge was so in- sightful and inspirational that entire careers were re-charted. The mouse course is typically reor- ganized every few years by scientists who not only have considerable ex- pertise in transgenesis but boundless energy as well. Because the last edi- tion was published in 1994, both the mouse course and the field of mouse transgenesis have under- gone significant changes. A recent team, led by Andras Nagy, has put together an exhilarating and superb book whose content is based squarely on its two predecessor edi- tions. The new edition, which is both textbook and laboratory manual, is easy to understand, richly illustrated, and full of lively anecdotes. On the whole, it reflects the swift currents of progress made over the past de- cade, many of which were contrib- uted to the field by the present au- thors. Most of the protocols contained within this manual are aimed at un- derstanding mammalian develop- ment through production of geneti- cally altered mice. The manual contains 17 chapters, beginning with a brief and highly entertaining history of mouse genetics. The sec- ond chapter provides a discussion of mouse development. Subsequent chapters are protocol based. Here, the authors have maintained and expanded previous details on the creation of transgenic and chimeric mice, and embryonic stem cell iso- lation and manipulation. Most of the remainder of the manual reflects the amazing breadth of technological expansion concerning genetically altered mice since the last edition, including reporter gene expression, parthenogenesis, cloning, and as- sisted reproduction. Included is a satisfying compendium of tech- niques of animal husbandry, hu- mane sacrifice, general surgery, and set up of the transgenic facility. Most of the major protocols are introduced within an historical con- text and provided with a rationale, as well as general ideas for their use. In addition, many of the protocols contain invaluable step-by-step photographic images and how to evaluate, also with the aid of care- fully chosen photographs, starting material and results. The overall im- pression created by these authors is one of authority: they are so experi- enced in handling questions that they have headed the reader off at the pass by including the minutest of details for success. For example, the authors instruct on everything from keywords during searches to sticking plasticine onto the microscope stage to secure micropipettes dur- ing intricate procedures. They have also provided the sources for mate- rials and reagents, both commercial and in house: it is no exaggeration to admit sheer relief in locating, after several years of desperate search- ing and disappointment, a new dis- tributor of flat mouthpieces to at- tach to aspirator assemblies for manipulating embryonic tissue. In addition, other chapters include techniques relevant to the manipu- lation of nongenetically altered mice, such as in vitro culture of pre- and postimplantation conceptuses, isolation of RNA and estimated re- coveries from the entire gamut of embryonic stages, as well as visual- ization of gene products by both whole-mount and sectional analy- ses. There is a new section on cryo- preservation, rederivation, and trans- port of mice, all of which will be very useful in maintaining valuable mouse *Correspondence to: Dr. Karen M. Downs, Department of Anatomy, University of Wisconsin, 329 Service Memorial Institute, 1300 University Avenue, Madison, WI 53706-1532. E-mail: [email protected] DOI 10.1002/dvdy.20031 Published online 31 March 2004 in Wiley InterScience (www.interscience.wiley.com). DEVELOPMENTAL DYNAMICS 230:199 –200, 2004 © 2004 Wiley-Liss, Inc.

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BOOK LOOK

Almost Everything You’ve Wanted to Know AboutElucidating Mouse Development

MANIPULATING THE MOUSE EMBRYO:A LABORATORY MANUAL, THIRD EDI-TION

by A. Nagy, M. Gertsenstein, K.Vintersten, and R. Behringer

Cold Spring Harbor LaboratoryPress, New York, NY, 2003, 764 p.ISBN: 0-87969-591-9 $125.00

As an alumnus of the 1988 ColdSpring Harbor Course in MolecularMouse Embryology, it was a plea-sure to be asked to review the thirdedition of the companion coursebook, entitled “Manipulating theMouse Embryo: A Laboratory Man-ual.” The “Mouse Course” was es-tablished in the 1980s as part of theCold Spring Harbor Laboratory’s ac-ademic program to communicatenew discoveries, concepts, andmethodologies in the manipulationof the mouse embryo and its ge-nome to small groups of students.For many participants, including my-self, the instruction of Rosa Bedding-ton, Liz Robertson, Andy McMahon,and Robin Lovell-Badge was so in-sightful and inspirational that entirecareers were re-charted.

The mouse course is typically reor-ganized every few years by scientistswho not only have considerable ex-pertise in transgenesis but boundlessenergy as well. Because the last edi-tion was published in 1994, both themouse course and the field ofmouse transgenesis have under-gone significant changes. A recentteam, led by Andras Nagy, has puttogether an exhilarating and superb

book whose content is basedsquarely on its two predecessor edi-tions. The new edition, which is bothtextbook and laboratory manual, iseasy to understand, richly illustrated,and full of lively anecdotes. On thewhole, it reflects the swift currents ofprogress made over the past de-cade, many of which were contrib-uted to the field by the present au-thors.

Most of the protocols containedwithin this manual are aimed at un-derstanding mammalian develop-ment through production of geneti-cally altered mice. The manualcontains 17 chapters, beginningwith a brief and highly entertaininghistory of mouse genetics. The sec-ond chapter provides a discussion ofmouse development. Subsequentchapters are protocol based. Here,the authors have maintained andexpanded previous details on thecreation of transgenic and chimericmice, and embryonic stem cell iso-lation and manipulation. Most of theremainder of the manual reflects theamazing breadth of technologicalexpansion concerning geneticallyaltered mice since the last edition,including reporter gene expression,parthenogenesis, cloning, and as-sisted reproduction. Included is asatisfying compendium of tech-niques of animal husbandry, hu-mane sacrifice, general surgery, andset up of the transgenic facility.

Most of the major protocols areintroduced within an historical con-text and provided with a rationale,

as well as general ideas for their use.In addition, many of the protocolscontain invaluable step-by-stepphotographic images and how toevaluate, also with the aid of care-fully chosen photographs, startingmaterial and results. The overall im-pression created by these authors isone of authority: they are so experi-enced in handling questions thatthey have headed the reader off atthe pass by including the minutest ofdetails for success. For example, theauthors instruct on everything fromkeywords during searches to stickingplasticine onto the microscopestage to secure micropipettes dur-ing intricate procedures. They havealso provided the sources for mate-rials and reagents, both commercialand in house: it is no exaggeration toadmit sheer relief in locating, afterseveral years of desperate search-ing and disappointment, a new dis-tributor of flat mouthpieces to at-tach to aspirator assemblies formanipulating embryonic tissue.

In addition, other chapters includetechniques relevant to the manipu-lation of nongenetically alteredmice, such as in vitro culture of pre-and postimplantation conceptuses,isolation of RNA and estimated re-coveries from the entire gamut ofembryonic stages, as well as visual-ization of gene products by bothwhole-mount and sectional analy-ses. There is a new section on cryo-preservation, rederivation, and trans-port of mice, all of which will be veryuseful in maintaining valuable mouse

*Correspondence to: Dr. Karen M. Downs, Department of Anatomy, University of Wisconsin, 329 Service Memorial Institute, 1300University Avenue, Madison, WI 53706-1532. E-mail: [email protected]

DOI 10.1002/dvdy.20031Published online 31 March 2004 in Wiley InterScience (www.interscience.wiley.com).

DEVELOPMENTAL DYNAMICS 230:199–200, 2004

© 2004 Wiley-Liss, Inc.

strains and keeping down costs of in-creasingly expensive mouse colonies.

Thus, this new edition should figureprominently on the bookshelves ofall transgenic mouse facilities andserious students of embryology. Inscope and detail, it supersedes any-thing currently available. Neverthe-less, having said that, the currentmanual is not without its flaws.

The authors’ hope, as stated intheir Preface, is to train the futureleaders and innovators of the fieldsof mouse developmental geneticsand molecular embryology. A majorconcern of this reviewer towardachieving this noble goal is the em-bryo-centric manner in which theauthors have presented mouse de-velopment in the second chapter;just a few pages deal with extraem-bryonic tissues and little note is takenof the complex placenta. In the ab-sence of balanced treatment of theentire conceptus, the frustrating butcommon practice of investigatorsdiscarding extraembryonic mem-branes in favor of the embryo willundoubtedly be perpetuated. Theconsequences of this practice arefar-reaching, not only in financialterms, but in terms of hampering ho-listic progress in understanding de-velopment of viviparous mammals.

A second concern is that, for themost part, the current edition reliesalmost entirely on manipulating thegenome as a means of elucidatingmammalian development. Whilstfew would deny the profound rele-vance of this approach for address-ing many questions, it is not sufficientfor addressing all of them. The impor-tance of imaginatively designedand carefully controlled experi-ments involving microsurgical pertur-

bations must not be overlooked.These strategies have underpinneda great deal of what is known aboutthe embryo not only in other classicorganisms, but in the mouse, as well.Directives for fate mapping, deter-mination of developmental po-tency, embryonic polarity, tissuerecombination and deletion experi-ments, use of impregnated beads,and injection of compounds intocavities are only a few of the ap-proaches that should be recognizedas necessary, valuable, and less ex-pensive for unraveling genetic ac-tion in both wild-type and mutatedsettings.

Of the chapters, the one on in vitroculture of preimplantation embryosis well intentioned and highly usefulbut could use some pruning and re-organization to eliminate the repeti-tive nature of much of the text andprotocols. In contrast, the chaptercontaining methods for postimplan-tation whole embryo culture is sobrief as to be almost cavalier. In par-ticular, no advice is provided onwhat to look for after culture. Theauthors observe that various culturemethods can lead to a significantlyhigh frequency of embryonic de-fects, but what these defects are isnot discussed. This is an especiallyimportant concern when one be-gins to experimentally perturb em-bryos. Presentation of the Brown andFabro scoring system (1981) shouldbe mandatory. Also, ether has forsome years now been unavailableas an anesthetic, at least at this re-viewer’s institution, for obtainingfrom rats the serum used in postim-plantation embryo culture. Hence, adiscussion of viable alternativeswould be welcome.

Aside from these intellectual con-siderations, presentation of the man-ual itself could be improved for userfriendliness. The list includes, but is byno means exhaustive, a more com-prehensive index which, as it cur-rently stands, is very scanty and in-adequately cross-referenced. Inaddition, an alphabetical list of pro-tocols, tables, figures, and a list ofWeb sites, which are currently buriedthroughout the text, should be in-cluded somewhere. Finally, the firstpage of the manual indicates theavailability of a companion Web sitein early 2003. A potentially great re-source though, at the time of writing,now a year later, the site is not upand running.

Thus, although nothing is perfect,concerning the methodologies foraltering the mouse genome, thisnew manual comes very close. Allinvestigators active in the field, nov-ice and experienced alike, are en-couraged to run to their local book-store and add this truly outstandingresource to their libraries. However,the student of mouse embryologyshould take care to recognize thatthe genetic approach is just one ofmany ways to understand normalmammalian development.

REFERENCE

Brown NA, Fabro S. 1981. Quantitation ofrat embryonic development in vitro: amorphological scoring system. Teratol-ogy 24:65-78.

Karen M. Downs*Department of Anatomy

University of WisconsinMadison, Wisconsin

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