Cellular and Molecular Aspects of Implantation Cellular and Molecular Aspects of Implantation Edited by Stanley R. Glasser and David W. Bullock Baylor College of Medicine Houston, Texas PLENUM PRESS . NEW YORK AND LONDOIV Library of Congress Cataloging in Publication Data Main entry under title: Cellular and molecular aspects of implantation. Proceedings of a conference held in Houston, Tex., Sept. 17-19, 1979, which was sponsored by the Center for Population Research, National Institute of Child Health and Human Devel opment, and other bodies. Includes index. 1. Ovum implantation-Congresses. I. Glasser, Stanley. II. Bullock, David W. II I. United States. National Institute of Child Health and Human Development. Center for Population Research. QP275.C44 599.01'6 80-20471 ISBN-13: 978-1-4613-3182-7 e-ISBN-13: 978-1-4613-3180-3 DOl: 10.1007/978-1-4613-3180-3 © 1981 Plenum Press, New York Softcover reprint of the hardcover 1s t edition 1981 A Division of Plenum Publishing Corporation 227 West 17th Street, New York, N.Y. 10011 All rights reserved No part of this book may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise, without written permission from the Publisher "It is characteristic of eggs and early embryos of lower animals that they are prepared to develop without shelter and nutriment from the mother. When the mammals evolved the phenomenon of utero-gestation, the chosen place of shelter, the uterus, was developed from the part of the oviduct, a channel that had for its purpose the efficient transportation and discharge of eggs, not their retention and maintenance. To fit it for gestational functions, the endocrine mechanism of the corpus luteum was evolved. In the light of this thought it is not surprising that the uterine chamber is actually a less favorable place for early embryos than, say, the anterior chamber of the eye, except when the hormones of the ovary act upon it and change it to a place of superior efficiency for its new function." George W. Corner The Hormones in Human Reproduction, 1947 Preface Long was I hugg'd close-long and long. Immense have been the preparations for me, Faithful and friendly the arms that have help'd me. Cycles ferried my cradle, rowing and rowing like friendly boatmen. For room to me stars kept aside in their own rings, They sent influences to look after what was to hold me. Before I was born out of my mother, generations guided me, My embryo has never been torpid, nothing could overlay it. -Walt Whitman, "Song of Myself" The womb is the seat of all mammalian life. In pregnancy, the uterus acquires this impor tance with the arrival of the fertilized egg, which takes up residence for periods ranging from about 2 weeks in the opossum to about 2 years in the elephant. The arrival of the embryo signals a crucial time for the establishment of pregnancy. For several days the blas tocyst remains free in the uterine lumen, where it depends on uterine secretions for its sur vival and differentiation. During this time, essential changes in the endometrium take place in preparation for attachment of the blastocyst and implantation. Early embryonic loss is an economic problem of global proportions in animal husbandry, where, in pigs and cattle for example, some 30% of all fertilizations fail to result in a pregnancy. In humans this figure may be even higher, and estimates of early spontaneous abortions range from 40 to 60% of all conceptions. Because the time of implantation is before the end of the menstrual cycle, failure of the embryo to implant could occur with little or no delay in a woman's menses. Thus many women engaging in unprotected intercourse may initiate a pregnancy without ever being aware of the fact. From the point of view of human fertility and of animal food production, therefore, knowledge of the mechanisms of implantation and the factors gov erning this process assumes considerable importance. Research in the past has provided much information about the developmental steps related to implantation in many species. While the results have been of immense value, the potential of the techniques commonly employed has been exhausted, and research on im plantation has stagnated as a consequence. Our understanding of the interaction between the embryo and the uterus and of its regulation remained primitive. In recent years power ful new tools have become available from advances in the fields of cellular and molecular biology. Few investigators in implantation had access to these new approaches and the peo- vii viii Preface pIe working in molecular biology were removed from reproductive and developmental biol ogy. We were fortunate that our concern about this state of affairs was sympathetically received by others. With the encouragement of Dr. Bert 0 'Malley, Chairman of the Department of Cell Biology and Director of the Center for Population Research and Repro ductive Biology at Baylor College of Medicine, we sought to organize a conference on the cellular and molecular aspects of implantation. Dr. William A. Sadler, Chief, Population and Reproduction Branch, CPR, NICHD, was enthusiastic about the idea and invited us to plan such a meeting under the auspices of the National Institute of Child Health and Human Development. The intent was to provide a compendium of information on the new ap proaches and their application to research on implantation. Two immediate benefits of the meeting were expected to be the opportunity for interaction between scientists from dif ferent fields who had never previously been given a common forum, and the stimulation arising from such interaction that might rekindle interest in research on implantation among scientists and granting agencies. With the generous support of additional sponsors and the aid of our scientific col leagues, the conference was convened in Houston in September 1979 and proved to be a great success, to the pleasant surprise of some who had argued against its premise and to the gratification of its organizers and sponsors. This volume contains the edited proceed ings of the conference, at which some of the leading participants in the "new implanta tion" shared their findings and thoughts with an appreciative audience that engaged in lively discussions. We hope that this book will propagate this impetus to a wider sphere' and will be of use not only to reproductive and developmental biologists but also to gradu ate and medical students and to practitioners in the fields of animal and human fertility. We are grateful to The Rockefeller Foundation, The National Science Foundation, The Upjohn Company, Ortho Pharmaceutical Corporation, G.D. Searle & Co., Schering Corporation, Merck & Co., Inc., Abbott Laboratories, and to the Population Program and the Department of Cell Biology of Baylor College of Medicine for their supplementary financial support, to Ms. Joanne Julian, Ms. Linda Hall, Mr. Tony May, and Mr. James Hawkins for their help, to Mr. Richard Cunningham of Houston, Texas for designing and donating the implantation logo, and to Messrs. Kirk Jensen and Richard Jannaccio of Plenum Publishing Corporation for their help and guidance. Stanley R. Glasser David W. Ballock Houston. Texas Contents Part I: Perspectives Chapter 1 Viviparity E. C. Amoroso I. Introduction ...................................................... 3 II. Viviparity as a Reproductive Mechanism .............................. 3 III. Adaptations for Ovoviviparity and Viviparity in Invertebrates .............. 4 IV. Adaptations for Viviparity in Fishes .................................. 6 A. Elasmobranchs ................................................. 6 B. Teleosts ...................................................... 7 C. Latimeria: The Living Coelacanth ................................. 9 V. Amphibian Viviparity .............................................. 9 VI. Reptilian Viviparity ................................................ 10 VII. Adaptations for Viviparity in Mammals ............................... 12 A. Monotremes (Prototheria) ........................................ 12 B. Marsupials (Metatheria) ......................................... 14 C. Eutheria ...................................................... 16 VIII. The Corpus Luteum and the Establishment of Pregnancy ................. 17 IX. The Maternal Recognition of Pregnancy ............................... 18 X. Immunological Transactions in Early Pregnancy ........................ 19 References ............................................................ 21 Chapter 2 My Life with Mammalian Eggs M. C. Chang I. Introduction ....................................................... 27 II. ln Vitro Fertilization of Mammalian Eggs ............................... 27 III. Storage and Transfer of Eggs ......................................... 29 ix x Contents IV. Interspecific Fertilization and Egg Transfer .............................. 30 V. Effects of Steroids and Other Compounds on the Transportation and Development of Eggs ............................................... 31 VI. Summary ......................................................... 32 References ............................................................ 33 Part II: Cell Biology of the Developing Egg Introduction. . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . .. . . . . . .. 39 John D. Biggers Chapter 3 The Origin of Trophoblast and Its Role in Implantation J. Rossant and W. I. Frels I. Introduction ....................................................... 43 II. The Trophectoderm Cell Lineage ............................. , . .. . . . .. 43 A. What Is Trophoblast? ............................................ 43 B. How and When Is the Trophectoderm Cell Lineage Established? ......... 46 III. The Role of the Trophectoderm Cell Lineage in Implantation ............... 47 A. Uterus-Trophectoderm-ICM Interactions ............................ 47 B. Species Specificity of the Trophoblast-Uterine Interaction .............. 48 IV. Conclusions ....................................................... 50 Discussion ............................................................ 50 References ............................................................ 52 Chapter 4 The Generation and Recognition of Positional Information in the Preimplantation Mouse Embryo M. H. Johnson, H. P. M. Pratt, and A. H. Handyside I. Introduction ....................................................... 55 II. Asymmetries in the Oocyte .......................................... 55 III. Asymmetries at Fertilization .......................................... 56 IV. Asymmetries in Cleavage ............................................ 57 V. Compaction ....................................................... 58 A. Description of Compaction ........................................ 58 B. Control of Compaction ........................................... 61 C. Consequences of Compaction for Blastocyst Formation ................. 64 1. Cytochalasin D Treatment ...................................... 64 2. Treatment with Antiserum to Embryonal Carcinoma Cells ............ 65 Contents xi D. Mechanisms by Which Compaction Might Generate Positional Information during Blastocyst Formation ....................................... 67 VI. Summary and Conclusions ........................................... 70 Discussion ............................................................ 70 References ....................................................•....... 72 Chapter 5 Relationship between the Programs for Implantation and Trophoblast Differentiation Michael I. Sherman, Martin H. Sellens, Sui Bi Atienza-Sarnols, Anna C. Pai, and Joel Schindler I. Introduction ....................................................... 75 II. What Is the Nature of Blastocyst Factors Involved in Implantation? .......... 77 III. What Is the Relationship between Implantation and Other Differentiative Events? . .... . ..... . . ... . ... . ... . .. . . .... ... . .... .... ......... . . . .. 80 IV. When Are Implantation-Related Gene Products Produced? ................. 82 V. Conclusions and Speculations ......................................... 84 Discussion ............................................................ 85 References ............................................................ 87 Chapter 6 Cellular and Genetic Analysis of Mouse Blastocyst Development Roger A. Pedersen and Akiko I. Spindle I. Introduction ....................................................... 91 II. Cell Determination during Early Mouse Embryogenesis ................... 92 A. Cell Determination in the Inner Cell Mass ........................... 92 B. Cell Determination in the Primary Ectoderm ......................... 94 III. Mutations at the a Locus Affecting Implantation ......................... 96 A. Genetics ....................................................... 96 B. Embryology .................................................... 97 1. Histological Observations ...................................... 97 2. Ultrastructural Observations .................................... 99 3. Experimental Observations ..................................... 100 IV. Conclusions ....................................................... 102 Discussion ............................................................ 103 References ............................................................ 105 Part III: Macromolecular Synthesis in the Developing Egg Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 111 Charles J. Epstein xii Contents Chapter 7 Mobilization of Genetic Information in the Early Rabbit Trophoblast Cole Manes, Michael J. Byers, and Andrew S. Carver I. Introduction ....................................................... 113 II. Results and Discussion .............................................. 114 A. Unique-Sequence DNA Transcription ............................... 114 B. Polysome Assembly and Ternary Complex Formation .................. 116 C. Cytoplasmic DNA ............................................... 117 D. RNA-Directed DNA Polymerase ................................... 119 III. Summary ......................................................... 120 Discussion ............................................................ 121 References ............................................................ 123 Chapter 8 Activity of RNA and DNA Polymerases in Delayed-Implanting Mouse Embryos H. M. Weitlauf and A. A. Kiessling I. Introduction ....................................................... 125 II. Materials and Methods .............................................. 126 A. Preparation of Embryo Donors ..................................... 126 B. Radiolabeling of Embryos for the Determination of RNA Polymerase Activity ....................................................... 126 C. Specific Activity of the [3H]-UTP Pools ............................. 126 D. DNA Polymerase Assay .......................................... 127 E. Analysis of Data ................................................ 128 III. Results ........................................................... 128 A. Specific Activities of the [3H]-UTP Pools ............................ 128 B. Rate of Incorporation of [3H]Uridine into RNA ....................... 129 C. RNA Polymerase Activity ........ ,............................... 129 D. DNA Polymerase Activity ........................................ 129 IV. Summary.......................................................... 131 Discussion ............................................................ 132 References ............................................................ 135 Chapter 9 A Reexamination of Messenger RNA Populations in the Preimplantation Mouse Embryo Gilbert A. Schultz, Jeremy R. Clough, Peter R. Braude, Hugh R. B. Pelham, and Mo. H. Johnson I. Introduction ....................................................... 137 II. Materials and Methods .............................................. 138 III. Results and Discussion .............................................. 139