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Neuropeptides: A methodology (IBRO handbook series: Methods in the neurosciences vol. 11)
Facets o f Vision is indispensable the newly growing shoots. The in compound eye research I for all those interested in a book is well illustrated and its would guess that, after so many thorough and authoritative pres- printing is of superb quality. brilliant achievements, the multientation of the present frontiers Unfortunately, due to its high tude of variations found among and up-to-date developments of price, quite a few of the inter- arthropod eyes and their adapcompound eye research. It will be ested students and researchers tations to particular behavioural particularly welcome to those will have to go to their library to tasks in a particular specieswho believe that knowledge of read it instead of having it in specific environment will remain the roots of a field of research is a their private collection. the most reliable source of new Speculating on future surprises questions and discoveries. prerequisite for understanding
Neuropeptides: A Methodology (IBRO Handbook Series: Methods in the Neurosciences Vol. 11) £ Cam/e edited by G. Fink and A. J. Harmar, TheUniversityof ManchesterInstitute of Scienceand Technology,PO Box No. 88, Manchester M60 1QD,UK.
116
John Wiley & Sons, 1989. £45.00 (ix + 345 pages) ISBN 0 471 92072 X
The publication of a book with a broad coverage of the techniques for studying the role of peptides in the function of the nervous system is timely, in view of the rapidly expanding interest in all areas of neuroscience. There are many texts available for those wishing to learn about the chemistry and function of peptides, but relatively few are specifically directed at methods applicable to neuropeptides. The stated intention of the book is to provide a compendium of techniques applicable to the study of neuropeptide function, with sufficient detail to permit its use as a laboratory manual wherever possible. With some techniques, the aim is to discuss general strategies and guide the reader to references that supply detailed methodology. Some chapters are excellent, but others fail in varying degrees to fulfil these expectations. An excellent account of neuropeptide-degrading enzymes is presented in chapter 8. A concise introduction is followed by sections on the various peptidases involved. Each section gives information on the properties of the enzymes, detailed assay methods with appropriate references, and helpful comments on important points to be considered. The ultrastructural study of peptide-containing neurones is
thoroughly dealt with in chapter 4. Strategies as well as techniques are rightly emphasized since individual peptides require specific approaches. The careful choice of figures and the first-class reference section will be welcomed by both graduates and more experienced researchers for whom this is an unfamiliar field. Since cells employ several second messenger systems to elicit responses to stimulation, the effect of peptides on these systems in neural tissues is a subject of increasing interest and study. A concise but comprehensive coverage of second messenger systems is provided in chapter 10. The treatment includes assays for the measurement of the compounds involved, and for the enzymes that are activated or inhibited, supported by an up-to-date, highly relevant bibliography. The account of current methods of in situ hybridization for the localization of mRNA for peptide precursors shows excellent attention to detail with indications of points that the researcher might overlook. A description of the theory behind the methods is given, providing information about the reactions involved, the inclusion of controls and the drawbacks of certain methods. Particularly useful is the discussion of the pitfalls associated with some procedures. The discussion of the construction and screening of cDNA libraries is essentially a sound introduction to the principles involved, with references given to the detailed methodology. An appendix contains a useful protocol for the primary screening of cDNA expression libraries with
antibody probes. Workers wishing to apply the methods associated with cDNA libraries will benefit from consulting Methods in Enzymology Vol. 152 (Academic Press, 1987) after reading this chapter. Moreover, the methods discussed have to some extent been superseded by the recently developed polymerase chain reaction (PCR), although this perhaps came too late for inclusion in the text. Some aspects of the book are less satisfactory than those considered above. Although the chapter on neuropeptide release in vivo is an excellent manual for studying the measurement of peptides in hypophysial portal blood, one wonders why release from other tissues is not covered. The treatment of neuropeptide release in vitro merely furnishes a short introduction to the methods available in an important area. The account of the role of peptides in behaviour is readable and informative, although those with experience in handling and administering peptides may feel that some of the difficulties are not sufficiently stressed. The solubility of peptides (p. 312) often presents problems, as does their adherence to reaction vessels. Moreover, the successful intracerebral administration of peptides requires considerable skill and experience. The book begins with a discussion of the Fmoc method for the chemical synthesis of peptides, although many consider tBoc methodology to be superior for longer peptides. It cannot be used as a manual to produce peptides; this would have required a prohibitively long account not relevant to this text. Many peptides TINS, Vol. 13, No. 3, 1990
techniques using tritiated and iodinated peptides for determining receptor numbers and the affinity of receptors for peptides would have been useful, since many scientists working with neuropeptides will, at some time, need to measure these parameters. The several chapters in this book that provide detailed
methodology will be welcomed by experienced scientists wishing to use techniques that are new to them. They will also benefit graduate and postdoctoral workers. For the graduate student certain chapters provide a sound introduction to theory and techniques, but will require supplementation from other sources for a complete methodology.
dated. Also uncertain are the roles that opioid peptides might play in the neural mechanisms underedited by R. G. Dyer and R. J. lying seasonal reproductive Bicknell, Oxford University Press, rhythms and in the develop1989. £45.00 (v + 365 pages) ISBN mental processes that lead to 0 19 857694 3 reproductive maturation and The number of investigations on senescence. Perhaps the chief the neuroendocrine, and par- value of this book lies in its ticularly the reproductive neuro- presentation of the diverse endocrine, actions of opioid drugs approaches currently being used is perhaps exceeded only by to resolve these issues, which studies on their roles in neural provides an overview of how the systems modulating pain percep- field now stands. tion. Interest in this issue dates The editors have grouped the from studies by Everett and contributions into five sections Sawyer in the earliest days of covering (1) the interactions bemodern neuroendocrinology and tween opioids and gonadal horwas rekindled soon after the dis- mones in the regulation of goncovery of the endogenous opioid adotrophin secretion, (2) the peptides. This volume contains interactions of opioid systems contributions from a number of with other neuronal networks investigators in this field, and for regulating gonadotrophin sethe most part, serves admirably to cretion, (3) the role of opioids in bring the reader up to date on controlling masculine and femipoints of agreement, as well as on nine sexual behavior, (4) the the areas of uncertainty and con- effects of opioids on oxytocin troversy that still exist in this secretion, and (5) the role of active area of research. As the opioids in human reproduction. editors point out, a general con- At present, these are all major sensus has developed that in rats areas of investigation in neuroand other species, endogenous endocrinology, and their coveropioids largely exert inhibitory age is representative. However, in effects on the secretion of view of the substantial literature gonadotrophins from the anterior attesting to the involvement of pituitary, the release of oxytocin endogenous opioids in the control from the posterior pituitary, and of prolactin secretion, and in the on the expression of both mascu- mediation of gonadal hormone line and feminine mating behav- feedback in the non-human priiors. It is widely held that en- mate, it is somewhat surprising dogenous opioid peptidergic that these topics were not also systems participate in the physio- covered. logical regulation of gonadoSome of the individual essays in trophin secretion and mating be- these sections present new havior by gonadal hormones, and research findings not previously of oxytocin secretion by various published, while others provide afferent stimuli, but many of the reviews of the current state of details concerning sites, mechan- knowledge in a particular area, as isms of action, and physiological well as unpublished or recently significance remain to be eluci- published data. Particularly note-
worthy among the latter are the paper by Wilkinson and Landymore, which critically reviews the controversies surrounding the role of opioids in influencing the onset of puberty, and that by Lincoln and Ssewannyana on possible opiatergic mechanisms involved in seasonal reproduction, both of which provide provocative new data and overviews. Similarly, the papers in the second section by Kalra and co-workers, Grossman and Dyer, and by Hoffman eta/. address a number of critical issues such as the mediation by endogenous opioids of gonadal hormone feedback on the secretion of luteinizing hormone, and the interaction of opioids with noradrenergic systems controlling the release of luteinizing hormone regulatory hormone (LHRH), and with the LHRH systems themselves. Clearcut inhibitory actions of opioids on oxytocin secretion have been repeatedly demonstrated in recent years. Papers by Leng and Russell, Summerlee, Summy-Long, and Bicknell and Zhao go beyond these findings to delve further into the physiological significance of these controls during parturition and lactation. These contributions are particularly welcome for endocrinologists working in this area. Neuroendocrinologists investigating the mechanisms that control pituitary hormone secretion may be less familiar with the nature of the neural controls over mating behavior, and the four papers on opioid regulation of both masculine and feminine reproductive behavior (Herbert, Wiesner and Moss, Sodersten et a/., and Baum) provide comprehensive summaries of this emerging work.
are now commercially available. If a peptide is unavailable or required in large amounts, synthesis is best left to specialists working in large research groups. For those who require it Solid Phase Peptide Synthesis: A Practical Approach
by E. Atherton and R. C. Sheppard (Oxford University Press, 1989) provides a detailed methodology. The inclusion of a chapter on
Brain Opioid Systems in Reproduction
TINS, VoL 13, No. 3, 1990
William R. Crowley Departmentof Pharmacology, Universityof Tennessee,Memphis Collegeof Medicine, Memphis, TN38163, USA.
117
Neuropeptides: A Methodology (IBRO Handbook Series: Methods in the Neurosciences Vol. 11) £ Cam/e edited by G. Fink and A. J. Harmar, TheUniversityof ManchesterInstitute of Scienceand Technology,PO Box No. 88, Manchester M60 1QD,UK.
116
John Wiley & Sons, 1989. £45.00 (ix + 345 pages) ISBN 0 471 92072 X
The publication of a book with a broad coverage of the techniques for studying the role of peptides in the function of the nervous system is timely, in view of the rapidly expanding interest in all areas of neuroscience. There are many texts available for those wishing to learn about the chemistry and function of peptides, but relatively few are specifically directed at methods applicable to neuropeptides. The stated intention of the book is to provide a compendium of techniques applicable to the study of neuropeptide function, with sufficient detail to permit its use as a laboratory manual wherever possible. With some techniques, the aim is to discuss general strategies and guide the reader to references that supply detailed methodology. Some chapters are excellent, but others fail in varying degrees to fulfil these expectations. An excellent account of neuropeptide-degrading enzymes is presented in chapter 8. A concise introduction is followed by sections on the various peptidases involved. Each section gives information on the properties of the enzymes, detailed assay methods with appropriate references, and helpful comments on important points to be considered. The ultrastructural study of peptide-containing neurones is
thoroughly dealt with in chapter 4. Strategies as well as techniques are rightly emphasized since individual peptides require specific approaches. The careful choice of figures and the first-class reference section will be welcomed by both graduates and more experienced researchers for whom this is an unfamiliar field. Since cells employ several second messenger systems to elicit responses to stimulation, the effect of peptides on these systems in neural tissues is a subject of increasing interest and study. A concise but comprehensive coverage of second messenger systems is provided in chapter 10. The treatment includes assays for the measurement of the compounds involved, and for the enzymes that are activated or inhibited, supported by an up-to-date, highly relevant bibliography. The account of current methods of in situ hybridization for the localization of mRNA for peptide precursors shows excellent attention to detail with indications of points that the researcher might overlook. A description of the theory behind the methods is given, providing information about the reactions involved, the inclusion of controls and the drawbacks of certain methods. Particularly useful is the discussion of the pitfalls associated with some procedures. The discussion of the construction and screening of cDNA libraries is essentially a sound introduction to the principles involved, with references given to the detailed methodology. An appendix contains a useful protocol for the primary screening of cDNA expression libraries with
antibody probes. Workers wishing to apply the methods associated with cDNA libraries will benefit from consulting Methods in Enzymology Vol. 152 (Academic Press, 1987) after reading this chapter. Moreover, the methods discussed have to some extent been superseded by the recently developed polymerase chain reaction (PCR), although this perhaps came too late for inclusion in the text. Some aspects of the book are less satisfactory than those considered above. Although the chapter on neuropeptide release in vivo is an excellent manual for studying the measurement of peptides in hypophysial portal blood, one wonders why release from other tissues is not covered. The treatment of neuropeptide release in vitro merely furnishes a short introduction to the methods available in an important area. The account of the role of peptides in behaviour is readable and informative, although those with experience in handling and administering peptides may feel that some of the difficulties are not sufficiently stressed. The solubility of peptides (p. 312) often presents problems, as does their adherence to reaction vessels. Moreover, the successful intracerebral administration of peptides requires considerable skill and experience. The book begins with a discussion of the Fmoc method for the chemical synthesis of peptides, although many consider tBoc methodology to be superior for longer peptides. It cannot be used as a manual to produce peptides; this would have required a prohibitively long account not relevant to this text. Many peptides TINS, Vol. 13, No. 3, 1990
techniques using tritiated and iodinated peptides for determining receptor numbers and the affinity of receptors for peptides would have been useful, since many scientists working with neuropeptides will, at some time, need to measure these parameters. The several chapters in this book that provide detailed
methodology will be welcomed by experienced scientists wishing to use techniques that are new to them. They will also benefit graduate and postdoctoral workers. For the graduate student certain chapters provide a sound introduction to theory and techniques, but will require supplementation from other sources for a complete methodology.
dated. Also uncertain are the roles that opioid peptides might play in the neural mechanisms underedited by R. G. Dyer and R. J. lying seasonal reproductive Bicknell, Oxford University Press, rhythms and in the develop1989. £45.00 (v + 365 pages) ISBN mental processes that lead to 0 19 857694 3 reproductive maturation and The number of investigations on senescence. Perhaps the chief the neuroendocrine, and par- value of this book lies in its ticularly the reproductive neuro- presentation of the diverse endocrine, actions of opioid drugs approaches currently being used is perhaps exceeded only by to resolve these issues, which studies on their roles in neural provides an overview of how the systems modulating pain percep- field now stands. tion. Interest in this issue dates The editors have grouped the from studies by Everett and contributions into five sections Sawyer in the earliest days of covering (1) the interactions bemodern neuroendocrinology and tween opioids and gonadal horwas rekindled soon after the dis- mones in the regulation of goncovery of the endogenous opioid adotrophin secretion, (2) the peptides. This volume contains interactions of opioid systems contributions from a number of with other neuronal networks investigators in this field, and for regulating gonadotrophin sethe most part, serves admirably to cretion, (3) the role of opioids in bring the reader up to date on controlling masculine and femipoints of agreement, as well as on nine sexual behavior, (4) the the areas of uncertainty and con- effects of opioids on oxytocin troversy that still exist in this secretion, and (5) the role of active area of research. As the opioids in human reproduction. editors point out, a general con- At present, these are all major sensus has developed that in rats areas of investigation in neuroand other species, endogenous endocrinology, and their coveropioids largely exert inhibitory age is representative. However, in effects on the secretion of view of the substantial literature gonadotrophins from the anterior attesting to the involvement of pituitary, the release of oxytocin endogenous opioids in the control from the posterior pituitary, and of prolactin secretion, and in the on the expression of both mascu- mediation of gonadal hormone line and feminine mating behav- feedback in the non-human priiors. It is widely held that en- mate, it is somewhat surprising dogenous opioid peptidergic that these topics were not also systems participate in the physio- covered. logical regulation of gonadoSome of the individual essays in trophin secretion and mating be- these sections present new havior by gonadal hormones, and research findings not previously of oxytocin secretion by various published, while others provide afferent stimuli, but many of the reviews of the current state of details concerning sites, mechan- knowledge in a particular area, as isms of action, and physiological well as unpublished or recently significance remain to be eluci- published data. Particularly note-
worthy among the latter are the paper by Wilkinson and Landymore, which critically reviews the controversies surrounding the role of opioids in influencing the onset of puberty, and that by Lincoln and Ssewannyana on possible opiatergic mechanisms involved in seasonal reproduction, both of which provide provocative new data and overviews. Similarly, the papers in the second section by Kalra and co-workers, Grossman and Dyer, and by Hoffman eta/. address a number of critical issues such as the mediation by endogenous opioids of gonadal hormone feedback on the secretion of luteinizing hormone, and the interaction of opioids with noradrenergic systems controlling the release of luteinizing hormone regulatory hormone (LHRH), and with the LHRH systems themselves. Clearcut inhibitory actions of opioids on oxytocin secretion have been repeatedly demonstrated in recent years. Papers by Leng and Russell, Summerlee, Summy-Long, and Bicknell and Zhao go beyond these findings to delve further into the physiological significance of these controls during parturition and lactation. These contributions are particularly welcome for endocrinologists working in this area. Neuroendocrinologists investigating the mechanisms that control pituitary hormone secretion may be less familiar with the nature of the neural controls over mating behavior, and the four papers on opioid regulation of both masculine and feminine reproductive behavior (Herbert, Wiesner and Moss, Sodersten et a/., and Baum) provide comprehensive summaries of this emerging work.
are now commercially available. If a peptide is unavailable or required in large amounts, synthesis is best left to specialists working in large research groups. For those who require it Solid Phase Peptide Synthesis: A Practical Approach
by E. Atherton and R. C. Sheppard (Oxford University Press, 1989) provides a detailed methodology. The inclusion of a chapter on
Brain Opioid Systems in Reproduction
TINS, VoL 13, No. 3, 1990
William R. Crowley Departmentof Pharmacology, Universityof Tennessee,Memphis Collegeof Medicine, Memphis, TN38163, USA.
117