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Male reproductive toxicology. Methods in toxicology, volume 3, part A
Reproductive Toxicology,Vol. 8, No. 2, pp. 193-194, 1994 Copyright © 1994ElsevierScienceLtd Printed in the USA. All rights reserved 0890-6238/94$6.00 + .00
Pergamon 0890-6238(93)E0003-Z
• Book Review
Male Reproductive Toxicology. Methods in Toxicology, Volume 3, Part A. Chapin RE, Heindel JJ, eds. San Diego, CA: Academic Press; 1993. 389 pages, $69.95 paper, $155 hardcover. The methods in Toxicology series, edited by Charles Tyson and Hanspeter Witschi, is an effort to make standard test protocols available to the laboratory scientist in toxicology. The third volume in the series deals with reproductive toxicology; this first book of volume 3 is concerned with the male. Doctors Chapin and Heindel, the editors of this part and of part B (Female Reproductive Toxicology), are reknowned scientists at the National Institute of Environmental Health Sciences. These editors have joined a group of 38 other original researchers in the field to produce a very practical laboratory guide to toxicology methods. The scope of this manual is best appreciated from the table of contents, which is reproduced in Table 1. It is clear that laboratory animals, particularly the rat and to a lesser extent the mouse, are the focus of this book. This focus makes sense, given the heritage of rodent studies in the male reproductive physiology, histology, and toxicology literature, but I will voice a few reservations about the underrepresentation of other species in a moment. First, it is important to stress the overriding success of this book in laying our laboratory methods as clearly and comprehensively as it does. With admirable consistency, the authors present their material in a format that leads directly to the ability to perform the experiments of interest. The opening few paragraphs of each chapter describe the goal of the procedures that follow, giving a brief account of what ends will be attained and why an investigator might want to attain them. There follows a shopping list of the animals, glassware, reagents, and equipment that need to be gathered. Particularly helpful are the details that so often wreak havoc if unattended: how to sterilize a particular solution, temperatures at which to store things, which materials must be fresh. Finally, there is a detailed, step-by-step explanation of the experimental protocol, often with illustrations to show how things are supposed to look at particular stages of the process. The paperbound version is very obviously meant to sit on the lab bench: the wire-O binding lies flat and the back cover includes a fold-over bookmark for keeping one's place. The print is large and clear, although the margins are not as wide as might be useful for scribbling notes. I have to admit that the reference style leaves me cold: the citations do not include paper titles or inclusive pages, a practice that saves rather little space in exchange for considerable user inconvenience. Still, as a laboratory manual for practical use, the references do little to detract from the overall utility of the book.
Now, with regard to the concentration on rats and mice, it is necessary to admit that these are the primary animals used in laboratory work and as such ought to dominate the book. The inclusion of a chapter on the rabbit is very reasonable, given the interest of some investigators in using this animal for repeatable evaluations of semen parameters. My principal criticism of this concentration is the virtual absence of information on laboratory tests using human material. Make no mistake: Steven Schrader gives a nice overview of techniques used in field studies in the final chapter; however, this is too much material to cover in 9 pages and should have served as an introduction to several subsequent chapters on the methods used in field studies and
Table 1. Contents 1. Introduction 2. Male mouse sexual behavior test 3. In vitro techniques for assessing pituitary secretory function 4. Histological methods for evaluation of the testis 5. Application of testicular sperm head counts in the assessment of male reproductive toxicity. 6. Stage synchronization in rat seminiferous tubules using vitamin A depletion and repletion 7. Spermatogonial stem cells: assessing their survival and ability to produce differentiated cells 8. Dominant lethal tests in male and female mice 9. Transillumination-phase-contrast microscopic techniques for evaluation of male germ cell toxicity and mutagenicity 10. Isolation and culture of Leydig cells from adult rats 11. Isolation and primary culture of Leydig cells 12. Purification and primary culture of Leydig cells 13. Preparation and use of Sertoli cell-enriched cultures from 18-day-old rats 14. Two-compartment cultures of Sertoli cells--applications in testicular toxicology 15. Isolation, separation, and short-term culture of spermatogenic ceils 16. Collection of interstitial fluid and seminiferous tubule fluid from the rat testis 17. Isolation and culture of epidiymal epithelial cells from adult rats 18. Evaluation of protein synthesis by the epididymis 19. Utilizing Cryo Resources CellSoft computer-assisted sperm analysis system for rat sperm motility studies 20. Computer-assisted sperm analysis of rodent epididymal sperm motility using the Hamilton-Thorn Motility Analyzer 21. Methods for evaluation of rat epididymal sperm morphology 22. Semen analysis and fertility assessment in the rabbit 23. General techniques for assessing male reproductive potential in human field studies
194
Reproductive Toxicology
in fertility evaluations. I would have liked to see the same shopping list approach and detailed protocols as in the animal methods applied to material (nearly always semen) obtained from men. There are a number of methods that are increasingly being used in reproductive toxicology studies of workplace, environmental, and therapeutic exposures, including standard semen analysis, computerassisted motility analysis and morphometry, sperm penetration of zona-free hamster oocytes or of human hemizonas, acrosome assays, chromatin denaturation tests, and karyotyping of decondensed sperm heads, to name a few. To be sure, inclusion of laboratory methods for the evaluation of human material would have lengthened the book by at least 25%, and it is possible that the editors were under a space constraint. After all, the argument could be made that inclusion of all laboratory aspects of reproductive toxicology, including, for example, work with do-
Volume 8, Number 2, 1994 mestic animals of commercial interest, would rapidly enlarge the book beyond the size of a laboratory manual. Investigators who use rats, mice, and rabbits will not, however, find the omission of human methods to be an important detractor. There is a tremendous amount of material in these pages that cannot be easily obtained elsewhere, and Doctors Chapin and Heindel have done a tremendous service to the toxicology community in bringing it together in such a useful manner. At less than $70 for the softcover version, this text is a steal when measured by the hours of library time it will save researchers in the field.
ANTHONY R. SCIALLI, MD
Reproductive Toxicology Center Washington, DC
Pergamon 0890-6238(93)E0003-Z
• Book Review
Male Reproductive Toxicology. Methods in Toxicology, Volume 3, Part A. Chapin RE, Heindel JJ, eds. San Diego, CA: Academic Press; 1993. 389 pages, $69.95 paper, $155 hardcover. The methods in Toxicology series, edited by Charles Tyson and Hanspeter Witschi, is an effort to make standard test protocols available to the laboratory scientist in toxicology. The third volume in the series deals with reproductive toxicology; this first book of volume 3 is concerned with the male. Doctors Chapin and Heindel, the editors of this part and of part B (Female Reproductive Toxicology), are reknowned scientists at the National Institute of Environmental Health Sciences. These editors have joined a group of 38 other original researchers in the field to produce a very practical laboratory guide to toxicology methods. The scope of this manual is best appreciated from the table of contents, which is reproduced in Table 1. It is clear that laboratory animals, particularly the rat and to a lesser extent the mouse, are the focus of this book. This focus makes sense, given the heritage of rodent studies in the male reproductive physiology, histology, and toxicology literature, but I will voice a few reservations about the underrepresentation of other species in a moment. First, it is important to stress the overriding success of this book in laying our laboratory methods as clearly and comprehensively as it does. With admirable consistency, the authors present their material in a format that leads directly to the ability to perform the experiments of interest. The opening few paragraphs of each chapter describe the goal of the procedures that follow, giving a brief account of what ends will be attained and why an investigator might want to attain them. There follows a shopping list of the animals, glassware, reagents, and equipment that need to be gathered. Particularly helpful are the details that so often wreak havoc if unattended: how to sterilize a particular solution, temperatures at which to store things, which materials must be fresh. Finally, there is a detailed, step-by-step explanation of the experimental protocol, often with illustrations to show how things are supposed to look at particular stages of the process. The paperbound version is very obviously meant to sit on the lab bench: the wire-O binding lies flat and the back cover includes a fold-over bookmark for keeping one's place. The print is large and clear, although the margins are not as wide as might be useful for scribbling notes. I have to admit that the reference style leaves me cold: the citations do not include paper titles or inclusive pages, a practice that saves rather little space in exchange for considerable user inconvenience. Still, as a laboratory manual for practical use, the references do little to detract from the overall utility of the book.
Now, with regard to the concentration on rats and mice, it is necessary to admit that these are the primary animals used in laboratory work and as such ought to dominate the book. The inclusion of a chapter on the rabbit is very reasonable, given the interest of some investigators in using this animal for repeatable evaluations of semen parameters. My principal criticism of this concentration is the virtual absence of information on laboratory tests using human material. Make no mistake: Steven Schrader gives a nice overview of techniques used in field studies in the final chapter; however, this is too much material to cover in 9 pages and should have served as an introduction to several subsequent chapters on the methods used in field studies and
Table 1. Contents 1. Introduction 2. Male mouse sexual behavior test 3. In vitro techniques for assessing pituitary secretory function 4. Histological methods for evaluation of the testis 5. Application of testicular sperm head counts in the assessment of male reproductive toxicity. 6. Stage synchronization in rat seminiferous tubules using vitamin A depletion and repletion 7. Spermatogonial stem cells: assessing their survival and ability to produce differentiated cells 8. Dominant lethal tests in male and female mice 9. Transillumination-phase-contrast microscopic techniques for evaluation of male germ cell toxicity and mutagenicity 10. Isolation and culture of Leydig cells from adult rats 11. Isolation and primary culture of Leydig cells 12. Purification and primary culture of Leydig cells 13. Preparation and use of Sertoli cell-enriched cultures from 18-day-old rats 14. Two-compartment cultures of Sertoli cells--applications in testicular toxicology 15. Isolation, separation, and short-term culture of spermatogenic ceils 16. Collection of interstitial fluid and seminiferous tubule fluid from the rat testis 17. Isolation and culture of epidiymal epithelial cells from adult rats 18. Evaluation of protein synthesis by the epididymis 19. Utilizing Cryo Resources CellSoft computer-assisted sperm analysis system for rat sperm motility studies 20. Computer-assisted sperm analysis of rodent epididymal sperm motility using the Hamilton-Thorn Motility Analyzer 21. Methods for evaluation of rat epididymal sperm morphology 22. Semen analysis and fertility assessment in the rabbit 23. General techniques for assessing male reproductive potential in human field studies
194
Reproductive Toxicology
in fertility evaluations. I would have liked to see the same shopping list approach and detailed protocols as in the animal methods applied to material (nearly always semen) obtained from men. There are a number of methods that are increasingly being used in reproductive toxicology studies of workplace, environmental, and therapeutic exposures, including standard semen analysis, computerassisted motility analysis and morphometry, sperm penetration of zona-free hamster oocytes or of human hemizonas, acrosome assays, chromatin denaturation tests, and karyotyping of decondensed sperm heads, to name a few. To be sure, inclusion of laboratory methods for the evaluation of human material would have lengthened the book by at least 25%, and it is possible that the editors were under a space constraint. After all, the argument could be made that inclusion of all laboratory aspects of reproductive toxicology, including, for example, work with do-
Volume 8, Number 2, 1994 mestic animals of commercial interest, would rapidly enlarge the book beyond the size of a laboratory manual. Investigators who use rats, mice, and rabbits will not, however, find the omission of human methods to be an important detractor. There is a tremendous amount of material in these pages that cannot be easily obtained elsewhere, and Doctors Chapin and Heindel have done a tremendous service to the toxicology community in bringing it together in such a useful manner. At less than $70 for the softcover version, this text is a steal when measured by the hours of library time it will save researchers in the field.
ANTHONY R. SCIALLI, MD
Reproductive Toxicology Center Washington, DC