Table Of ContentMonographs on Endocrinology
Volume 31
Edited by
F. Gross (t), Heidelberg· M. M. Grumbach, San Francisco
A. Labhart, Zurich· M. B. Lipsett (t), Bethesda
T. Mann, Cambridge· L. T. Samuels (t), Salt Lake City
J. Zander, Munchen
Fred A. Kine!
Hormone Toxicity
in the Newborn
With 26 illustrations
Springer¥erlag
Berlin Heidelberg New York London
Paris Tokyo Hong Kong Barcelona
Fred A. Kind, Ph.D., DSc., FRSC (Deceased)
Professor Emeritus
The College of Staten Island of the
City University of New York
Staten Island, New York, USA
ISBN -13: 978-3-642-83796-8 e-ISBN -13: 978-3-642-83794-4
DOl: 10.1007/978-3-642-83794-4
Library of Congress Cataloging in Publication Data.
Kincl, Fred A.
Hormone toxicity in the newborn 1 Fred A. Kincl.
(Monographs on endocrinology; v. 31)
Includes bibliographical references.
ISBN -13: 978-3-642-83796-8 (U. S.)
1. Fetus--Effect of drugs on. 2. Infants (Newborn)--Effect of
drugs on. 3. Hormones--Toxicology. 4. Sex (Biology)
5. Endocrinology, Experimental. I. Title. II. Series.
[DNLM: 1. Fetus--drug effects. 2. Hormones--adverse effects.
3. Infant, Newborn. 4. Steroids--adverse effects. WI M057 v. 311
WS 420 K51h]
RG627.6.D79K56 1989
618.3'268--dc20
DNLM/DLC
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Preface
The account of "neonatal sterilization" is the story of the advocates of
direct effect of steroids on the gonads and those who believed in the
indirect influence, mediated through the hypothalamus and/or the pituitary
gland. As often happens in biology, both convictions represent the same
image seen from different perspectives.
Prof DC Johnson (Kansas City, KS) reminisced the beginning of the story
in a letter to me. I am paraphrasing parts of the letter with his permission.
"As a starting point we could pick the life-long research of Emil
Steinach ... " Steinach recognized the influence of testes on the develop
ment of accessory sex organs in 1894, described virilization of females and
feminization of males in 1913, and identified the controlling influence of
the hypophysis on the gonads in 1928.
He reviewed his work in a book Sex and Life, Forty Years of Biological and Medical
Experience (E Steinach and L Loebe!; Faber and Faber, London, 1940). He got on the wrong
road in later years and that is the reason everybody seems to have forgotten him. He presented
his hypothesis that estrogen has a direct effect upon the testes, i. e. hormone antagonism, at the
1st International Congress on Sex Research in 1926.
Carl Moore went to Chicago to study embryology under Frank Lillie. Lillie kept Moore
after graduation for the faculty and convinced him that he could make a name. for himself
if he could answer the riddle of the freemartin. Lillie thought that the female was "sterilized"
by the hormones of the male; I think he was influenced by Steinach. However, Moore could
not confirm the direct antagonism Steinach was talking about and found that androgens
were as damaging to the male as wcre estrogens, but that chorionic gonadotropin could
reverse the action of both. He presented his research at the 2nd International Congress on
Sex Research in 1930 (Price was not a co-author). You will notice there is no mention of
"feedback" in the 1932 paper by Moore and Price, but one of the conclusions reached was
that the conccpt of sex hormone antagonism was wrong. Unfortunately the preoccupation
with the indirect action of steroids prevailed for 40 years and only recently have we gained
respect for the "old" direct action.
The next step in the trail leads to LeRoy Goodman (Ant Rec 59: 233, 1934) who, as a
medical student, did a project involving transplantation of ovaries into the anterior chamber
of the eye of female and male rats. He noted that they did not ovulate nor luteinize if in
males. Witschi was interested in finding differences between male and female gonadotropins
and saw the importance of Goodman's experiments. Remember that at this time there was
a controversy over the number of gonadotropins. Witschi firmly believed (until 1961) that
there were three gonadotropins; LH, ICSH, and FSH. Males secreted only ICSH and FSH
but females secreted all three. The presence of testis at birth set the pattern for the loss of LH.
Witschi gave Pfeiffer the project, but apparently he was aided by RT Hill who was at the time
working on parabiosis. Upon graduation Hill went to Wisconsin and started RK Meyer on a
VI Preface
long trail of parabiotic research while Pfeiffer went to Yale. He met up with Hamilton, Young
and Wilson who were studying the effects of repeated injections of steroids on rats. They
had noted that injections started soon after birth caused permanent damage and mentioned
the similarity to the effects produced by testicular transplants as shown by Pfeiffer. However,
they mentioned that personal communication with Pfeiffer indicated that androgen did not have
the same effect as testes; Pfeiffer had used testosterone and not the propionate. Pfeiffer did
not follow up on the research which bothered Witschi a great deal but he felt that he should
not "steal" a project from one of this students.
Little attention is given to the tremendous potency of the testes for "androgenization".
Even in cases of ovo-testis when the androgen level is too low to stimulate the Wolffian
ducts, and the production of MUllerian inhibiting substance is too low to prevent production
of uteri, the animal is still "androgenized". Gynandromorphic mice, which have very poor
testicular development, are always anovulatory. Apparently the presence of a Y chromosome
is the key to "masculinization" and "androgenization" is something else entirely.
Finally, let me get back to the original idea which started the research to begin with; aging.
You did not take this up in your review but it is a pet of mine. I think that exposure of the
neonatal female rat, and maybe other species too, causes "damage" to the CNS which results
in a premature aging. Swanson and Van der Werff ten Bosch proposed that the delayed
anovulatory syndrome was aging and there have been several others who have picked up
on the idea. The androgenized female rat has several of the features of the aged female.
Furthermore, fetal anoxia or X-irradiation on the fetus produces premature aging with loss
of estrous cycles (constant estrus). The big question is what do these agents (steroids, DDT,
caffeine, etc.) do to the CNS? I think that their main effect is upon rhythms. Barraclough,
and particularly Gorski, have concentrated upon "tonic" gonadotropin release in males and
"cyclic" function in females. However, this may be backwards. Males "cycle" on a daily
basis-lots of data support this at least for ACTH, corticosterone, LH, prolactin, but not FSH.
Females seem to cycle only at 4-5 day periods, but they can be made to cycle daily after
ovariectomy. Jerry Yochim has shown that the rat behaves as if it has two clocks, one set
internally with a 20 hour day and the other entrained to the 24 hour light-dark cycle. Interactions
between the two produce a cycle of about 5 days. He can manipulate the exogenous light cycle
to various lengths and change the ratio of internal to external time so that he can have 7 or 10 day
cycles. When both clocks are set for a 20 hour day the animal loses cyclicity; the estrus or diestrus
phase will be about 150+ days long. Yochim believes that aging of the CNS gradually lengthens
the internal clock so that eventually the two are phased so that acyclicity is established. Note
that "old" female rats are locked into either pseudopregnant diestrus, or estrus.
Although we can produce a female rat with the androgenization syndrome by such diverse
treatment as constant light or rather mild diabetes mellitus, we do not consider them
"masculinized". Both of these treatments cause loss of daily cyclic pituitary function and again
focus attention upon loss of this as a causative factor ...
I became interested in the sterilizing effects of steroid hormones in the
late 1950's when I met, during a congress in Miami, a young Hungarian
scientist, Bela Flerk6. He lectured in part on the effect of androgens in
the neonate. The company I worked for was developing oral contraceptives,
and I found the subject fascinating. To achieve a permanent sterilization
by a single injection looked the up-and-coming method to have. I had
thought that we could employ this particular activity in veterinary medicine
to castrate male piglets "chemically". We tested synthetic estrogens in rats
and injected several newborn piglets. The results were encouraging, but it
was more convenient, and easier, to castrate the piglet surgically. Next,
we turned to rodents. The indigenous rat population in the macadamia nut
plantations in Hawaii were causing great crop damage. We reasoned that if we
could sterilize a large proportion the damage might become acceptable.
Preface VII
We were using the most effective synthetic estrogen, mestranol, with
encouraging results. However, the management decided against the trials.
They reasoned that it would not look good to use the same estrogen as
a constituent of an oral contraceptive for humans and also as a "rat
poison".
I have surveyed in the monograph Hormone Toxicity in the Newborn the
teratological effects of hormones in the neonate and in the fetus. The first
chapter is a historical introduction. I describe in the second chapter the
control of reproduction as we understand it today; the purpose is to recount
the great complexity which controls the process. The third chapter surveys
the effects of steroid hormones on the embryo and the fourth chapter their
effects in the neonate. Nonsteroidal hormones are discussed in Chapter 5.
Changes in behavior and possible effects on psychosexual development in
humans are covered in Chapter 6. I have summarized the evidence in the
last chapter.
The appendix includes abbreviations used in the text, structures and
biological activity of the principal hormones mentioned, and a brief outline
of biological tests used to assay steroid hormones.
I wish to thank the many friends and colleagues who gave me considerable
help, read part of the manuscript and offered valuable suggestions: S Cekan
(Stockholm, Sweden); IJ Clarke (Melbourne, Australia); G Dorner (Berlin,
DDR); RA Gorski (Los Angeles, CA); JL Hill (Bethesda, MD); T Iguchi
(Yokohama, Japan); DC Johnson (Kansas City, KS); S Kawashima (Hiro
shima, Japan); J Kinsel (Bethesda, MD); CR Kramer (Staten Island, NY);
RD Lisk (Princeton, NJ); L Macho (Bratislava, Czechoslovakia); RA
Maurer (Iowa City, IA); FL Meyer-Bahlburg (New York, NY); T Mori
(Tokyo, Japan); S Nilsson (Falun, Sweden); WH Rooks II (Palo Alto;
CA); DM Sheehan (Jefferson, AZ); A Raynaud (Vabre, France); JM
Reinisch (Bloomington, IN); HW Rudel (Elizabeth, NJ); P Sodersten
(Goteborg, Sweden); J Schreiber (Praha, Czechoslovakia); J Vreeburg
(Rotterdam, The Netherlands); J Weisz (Hershey, PA). S Kawashima
(Hiroshima, Japan) further helped by sending reprints of Japanese publi
cations. T Iguchi, T Mori and A Raynaud sent material for illustrations.
M Gregory (Staten Island, NY) reproduced aged slides into presentable
microphotographs and PA Kincl (New York, NY) helped with illustrations.
My wife, Lada Kincl, proofread all the references and helped to correlate
these with text. B Bevan, P Cosumano and F Pollutri typed parts of text,
references and tables.
Staten Island, NY Fred A. Kincl
October 1989
Springer-Verlag regrets to announce that the author died shortly before
publication of his book.
Contents
1 Introduction................... 1
2 Control of Reproductive Function in the Adult . 5
2.1 The Reproductive Cycle - An Overview 6
2.1.1 Marsupials ....... . 6
2.1.2 Eutherians ........ . 7
2.1.2.1 Cyclic Breeders 7
2.1.2.2 Seasonal Breeders 11
2.1.2.3 Induced Ovulators 12
2.2 Patterns of Hormone Releasc . 13
2.2.1 Males ..... . 14
2.2.2 Females . . . . . 15
2.2.2.1 Rodents 16
2.2.2.2 Primates 16
2.2.3 Seasonal Breeders 17
2.2.4 Annual Cycles . . 17
2.3 Neuroendocrine Control of Reproduction 18
2.3.1 Neural Control of Gonadotropin Release 19
2.3.1.1 Regulation of Prolactin Function 21
2.3.1.2 Endorphins and Enkephalins 22
2.3.1.3 Steroid Hormones ....... . 23
2.3.1.4 Other Peptide Hormones ... . 23
2.3.2 Neural Pathways Regulating Reproduction 23
2.3.3 Photic Stimulation . . . . . . . . . . . . . 24
2.3.3.1 Location of the Biological Clock. 25
2.3.3.2 Contribution of the Pineal Gland 26
2.3.4 Olfactory Control ......... . 28
2.3.5 Auditory Cues . . . . . . . . . . . . 29
2.4 The Hypothalamus-Pituitary-Gonadal Axis. 29
2.4.1 The Hypothalamus ........ . 30
2.4.1.1 Gonadotropin Releasing Hormone (GnRH) 30
2.4.1.2 Pep tides Similar to GnRH ..... . 34
2.4.2 The Pituitary Gland . . . . . . . . . . . . . . 34
2.4.2.1 Follicle Stimulating Hormone (FSH) 36
2.4.2.2 Luteinizing Hormone (LH) .... . 37
2.4.2.3 Prolactin (PRL) .......... . 39
2.4.2.4 Melanocyte Stimulating Hormone (Melanotropen, MSH) 39
2.4.3 The Gonads .. . 40
2.4.3.1 Males ............... . 40
2.4.3.2 Females ............. . 41
2.4.3.3 Biosynthesis of Steroid Hormones. 45
2.4.4 Nonsteroid Hormones . . . . . . . . . . . . 51
x Contents
2.4.4.1 Inhibin . . . 52
2.4.4.2 Activins 52
2.4.4.3 Follicostatin 53
2.4.4.4 Prostaglandins 53
2.4.4.5 Other Factors 53
2.5 Activity of Steroid Hormones . 54
2.5.1 Stimulatory and Inhibitory Effects 55
2.5.1.1 Androgens . . . 56
2.5.1.2 Anabolic Agents 57
2.5.1.3 Estrogens.. 57
2.5.1.4 Progesterone 60
2.5.2 Feedback 62
2.5.2.1 Males.. 64
2.5.2.2 Females 64
2.5.3 Synergism . . . . 65
2.5.3.1 Gonadotropin Releasing Hormone 65
2.5.3.2 Pituitary Hormones ..... 65
2.5.3.3 Gonadal Hormones . . . . . 66
2.5.3.4 Other Hormones and Factors 68
2.6 Hormone Receptors. . . . . . . . . . 68
2.6.1 Nature of Receptor Molecules 69
2.6.1.1 Receptor Interaction. 71
2.6.1.2 Sex Differences 71
2.6.1.3 Receptors in Neonatal and Aged Animals. 72
2.6.2 Gonadotropin Releasing Hormone . . 73
2.6.3 Pituitary Hormones . . . . . . . . . . 73
2.6.3.1 Follicle Stimulating Hormone 74
2.6.3.2 Luteinizing Hormone .. 74
2.6.4 Androgen Receptors ....... 74
2.6.4.1 Regulation of Formation. 75
2.6.5 Estrogen Receptors . . . . . . . . 75
2.6.5.1 Structure......... 76
2.6.5.2 Specificity of Estrogen Receptors 76
2.6.5.3 Regulation of Formation. 76
2.6.6 Progesterone Receptors . . . . . . 76
2.6.6.1 Structure......... 77
2.6.6.2 Regulation of Formation. 77
2.7 Interactions with Other Hormonal Systems . 77
2.7.1 Adrenal Function 77
2.7.1.1 Males.. 78
2.7.1.2 Females 78
2.7.2 Thyroid ..... 78
2.7.2.1 Hypothyroidism 79
2.7.2.2 Hyperthyroidism 79
2.8 Reproduction and the Immune Response. 80
2.8.1 Sex Dimorphism . . . . . . . . . 80
2.8.2 Thymus ............. 80
2.8.2.1 Effects of Thymectomy 81
2.8.2.2 Effects of Hormones. 81
3 Teratogenicity of Hormones in Utero . 121
3.1 Introduction.............. 121
3.2 Steroid Hormones: Malformation of the External Genitalia and Interference with
the Reproductive Function . . . . . . . . 123
3.2.1 Gonadal Hormones ....... . 123
3.2.1.1 Effects on Genetic Males 126
Contents XI
3.2.1.2 Effects on Genetic Females 128
3.2.2 Synthetic Steroid Hormones .. 137
3.2.2.1 Ethinyl Estradiol (EE) 137
3.2.2.2 Oral Contraceptives 138
3.2.3 Diethylstilbestrol (DES) . . . 140
3.2.3.1 Effects in Animals . 142
3.2.3.2 Effects on Humans. 144
3.2.4 Adrenocortical Hormones and Stress 145
3.2.4.1 Effects in Animals . 145
3.2.4.2 Effects on Humans. 146
3.3 Other Effects of Hormones .... . 146
3.3.1 Induction of Cancer .... . 146
3.3.1.1 Reproductive Tract. 146
3.3.1.2 Mammary Glands 148
3.3.2 Other Teratological Effects . 148
3.3.2.1 Mammary Glands . 148
3.3.2.2 Growth and Bone Structure 149
3.3.2.3 Nervous System 151
3.3.2.4 Immune System 151
3.4 Nonsteroidal Agents ..... . 152
3.4.1 Chorionic Gonadotropin 152
3.4.2 Prostaglandins ... 152
3.4.3 Thyroid Hormones 152
3.4.3.1 In Animals 153
3.4.3.2 In Humans 153
3.4.4 Insulin .. 153
3.4.5 Melatonin . . . . 153
3.4.6 Vitamins ..... 154
3.4.7 CNS Active Drugs 154
4 Effects of Steroid Hormones in the Neonate 168
A. Androgens and Estrogens . . . . . . . . . 168
4.1 Effects on Reproductive Function in Rats 168
4.1.1 Testosterone ... 168
4.1.1.1 Females 171
4.1.1.2 Males .. 179
4.1.2 Other Androgens 181
4.1.3 Estradiol ..... 183
4.1.3.1 Females 183
4.1.3.2 Males .. 185
4.1.4 Other Estrogens . 190
4.2 Effects of Estrogens During Lactation 193
4.2.1 Transfer of Steroids to Milk . . 195
4.2.1.1 Effects on Humans .. 195
4.3 Effects of Androgens and Estrogens on Reproduction in Other Species 196
4.3.1 Mice ...... . 196
4.3.1.1 Females 197
4.3.1.2 Males. 199
4.3.2 Guinea Pigs 202
4.3.3 Hamsters. 202
4.3.4 Rabbits . 203
4.3.5 Ferrets .204
4.3.6 Sheep . 204
4.3.7 Pigs .204
4.3.8 Dogs. .204
XII Contents
4.3.9 Bovine .. 204
4.3.10 Primates . 205
4.4 Tissue Responses 205
4.4.1 Hypothalamus-Pituitary Axis 205
4.4.1.1 Androgenized Animals. 205
4.4.2 Restoration of Gonadal Function 206
4.4.2.1 Androgenized Animals. 206
4.4.2.2 Estrogenized Animals . 211
4.4.3 Peripheral Organs - The Carcinogenic Effect 214
4.4.3.1 Androgenized Animals. 214
4.4.3.2 Estrogenized Animals . 215
4.4.4 Uptake in Various Tissues .... 215
4.4.4.1 Androgenized Animals. 216
4.4.4.2 Estrogenized Animals 217
4.4.5 Hormone Receptors . . . . . . . 217
4.4.5.1 Brain Receptors .... 217
4.4.5.2 Receptors in Other Tissues 218
4.4.6 Steroid Biosynthesis and Metabolism 218
4.4.6.1 Androgenized Animals. 219
4.4.6.2 Estrogenized Animals 220
4.4.6.3 Feminized Males 221
4.5 Other than Reproductive Effects . 221
4.5.1 Somatic Growth ..... . 221
4.5.1.1 Growth Hormone 223
4.5.2 Brain Development ... . 224
4.5.3 Mammary Glands .... . 224
4.5.4 Other Endocrine Functions 225
4.5.4.1 The Adrenals .. . 225
4.5.4.2 The Thyroid .. . 226
4.5.5 Effects on Metabolizing Enzymes 227
4.5.5.1 Adrenal Enzymes .. . 227
4.5.5.2 Liver Enzymes .... . 227
4.5.6 Increased Sensitivity to Carcinogens 229
B. Pregnanes ............ . 229
4.6 Adrenocortical Hormones 229
4.6.1 Influences on Reproduction 230
4.6.2 Other than Reproductive Effects 230
4.6.2.1 Toxic Effects ..... . 231
4.6.2.2 Body Growth 232
4.6.2.3 Adrenocortical Steroids in Blood 233
4.6.3 Stress 233
4.6.3.1 Response to Stress . 233
4.7 Progesterone .......... . . 234
4.8 Other Pregnane Derivatives. . . . 234
4.8.1 Estrane Derivatives with Progestational Activity 235
c. Protection Against Steroid Hormone Damage 235
4.9 Females ................... . 236
4.9.1 Progesterone.............. 236
4.9.1.1 Synthetic Progestational Agents 237
4.9.2 Androstanes ..... . 238
4.9.3 Diverse Agents ... . 238
4.9.3.1 Antiestrogens 238
4.9.3.2 Retinoids 239
4.10 Males 239
