Fetal Endocrinology and Metabolism: Current Topics in Experimental Endocrinology, Vol. 5

Fetal Endocrinology and Metabolism

Current Topics in Experimental Endocrinology

L. Martini

Department of Endocrinology, University of Milan, Milan, Italy

V.H.T. James

St. Mary’s Hospital, Medical School, University of London, London, England

ISSN  0091-7397

Volume 5 • Number Suppl. (PB) • 1983

Table of Contents

Cover image

Title page

Contributors

Editorial Board

Copyright page

Contributors

Preface

The Fetal Neuroendocrine Axis

Publisher Summary

I Introduction

II The Development of the Hypothalamus and Adenohypophysis

III The Secretion and Function of Fetal Adenohypophyseal Hormones (see Fig. 1)

IV Autonomy of the Fetal Neuroendocrine Unit

Acknowledgments

In Vitro Fertilization

Publisher Summary

I The Natural Ovulatory Cycle and IVF

II The Stimulated or Controlled Ovulatory Cycle

III Maturation of the Oocyte

IV The Technique of in Vitro Fertilization

V The Use of IVF in Male and Idiopathic Infertility

Factors Controlling Placental Endocrine Function in Domestic Animals

Publisher Summary

I Introduction

II Control of Placental Steroid Secretion by Fetal Cortisol in Domestic Animals

III Control of Secretion of Placental Lactogens

IV Effects of β-Catecholaminergic Drugs on the Placenta

V Conclusions

The Fetal Thyroid

Publisher Summary

I Introduction

II The Choice of Preparation

III The Morphological and Secretory Development of the Gland

IV The Placental Barrier

V The Role of the Thyroid in Fetal Development

VI Fetal Thyroid Hormone Kinetics

VII Perinatal Thyroid Hormone Concentrations

VIII Conclusion

Acknowledgment

Carbohydrate Metabolism

Publisher Summary

I Introduction

II Maternal Hormonal and Metabolic Adaptations

III Early Embryonic Development

IV Placental Transfer of Carbohydrates

V Fetal Hormones and Carbohydrate Utilization

VI Fetal Metabolic Pathways

VII Summary

Regulation of Partition of Protein During Pregnancy

Publisher Summary

I Introduction

II Placental Protein

III Fetal Protein

IV Free Amino Acid Pools

V Maternal Protein

Mineral Needs of the Fetus

Publisher Summary

I Introduction

II Calcium, Phosphate, Magnesium

III Parathyroid Hormone (PTH)

IV Vitamin D

V Calcitonin

VI Hormone Interrelations

VII Conclusion

Fetal Metabolism of Cortisol

Publisher Summary

I Introduction

II Circulating Levels of Cortisol in the Fetus

III Sources of Cortisol in the Fetal Circulation

IV Fate of Fetal Cortisol

V Significance of Cortisol in the Fetus

VI Summary

Sexual Differentiation: Normal and Abnormal

Publisher Summary

I Embryology of Sexual Differentiation

II Determinants of Phenotypic Differentiation

III Genetic Control of Sexual Differentiation

IV Sexual Differentiation of the Brain

V Male Pseudohermaphroditism

VI XX Males and True Hermaphroditism

VII Female Pseudohermaphroditism

Metabolic Errors of Adrenal Steroidogenesis

Publisher Summary

I Introduction

II Steroidogenesis and Enzymatic Conversions of Adrenal Steroid Hormones

III Fetal Sexual Development

IV Enzyme Defects in Congenital Adrenal Hyperplasia

V The Zona Fasciculata and Zona Glomerulosa as Separate Glands

VI Treatment of Congenital Adrenal Hyperplasia

VII Genetics of Congenital Adrenal Hyperplasia

VIII Prenatal Diagnosis of Congenital Adrenal Hyperplasia

IX Summary

Acknowledgments

Index

Contributors

Editorial Board

Copyright page

COPYRIGHT © 1983, BY ACADEMIC PRESS, INC.

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Contributors

Charlotte T. Laplante Branchaud(197),     Montreal General Hospital and Montreal Children’s Hospital Research Institutes and McGill University, Montreal, Canada

Bo Dupont(309),     Human Immunogenetics Section, Memorial Sloan-Kettering Cancer Center, New York, New York 10021

A.P. Flint(75),     Institute of Animal Physiology, Agricultural Research Council, Babraham, Cambridge CB2 4AT, England

Peter David Gluckman(1),     Department of Pediatrics, University of Auckland, Auckland, New Zealand

Julianne Imperato-Mcginley(231),     Department of Medicine, Division of Endocrinology, Cornell University Medical College, New York, New York 10021

Lenore S. Levine(309),     Division of Pediatric Endocrinology, Department of Pediatrics, The New York Hospital-Cornell Medical Center, New York, New York 10021

C. Lowy(117),     Department of Medicine, St. Thomas’s Hospital Medical School, London SE1 7EH, England

Beverley E. Pearson Murphy(197),     Montreal General Hospital and Montreal Children’s Hospital Research Institutes and McGill University, Montreal, Canada

P.W. Nathanielsz(97),     Department of Obstetrics and Gynecology, Laboratory of Fetal Physiology, Harbor-UCLA Medical Center, Torance, California 90509

Maria I. New(309),     Division of Pediatric Endocrinology, Department of Pediatrics, The New York Hospital-Cornell Medical Center, New York, New York 10021

Songya Pang(309),     Division of Pediatric Endocrinology, Department of Pediatrics, The New York Hospital-Cornell Medical Center, New York, New York 10021

Marilyn S. Pollack(309),     Human Immunogenetics Section, Memorial Sloan-Kettering Cancer Center, New York, New York 10021

John C. Stevenson(177),     Endocrine Unit, Royal Postgraduate Medical School, Hammersmith Hospital, London W12 OHS, England

A.L. Thomas(97),     Department of Physiology, University of Southampton, Southampton S09 3TU, England

C.R. Thomas(117),     Department of Medicine, St. Thomas’s Hospital Medical School, London SE1 7EH, England

Alan Trounson(43),     Department of Obstetrics and Gynaecology, Monash University, Queen Victoria Medical Centre, Melbourne 3000, Australia

Maureen Young(145),     Department of Gynaecology, St. Thomas’s Hospital Medical School, London SE1 7EH, England

Numbers in parentheses indicate the pages on which the authors’ contributions begin.

Preface

L. Martini and V.H.T. James

Volume 3 of this series, published in 1978, marked a change in policy by the editorial board, and aimed at bringing together a number of contributors to discuss a single topic in endocrinology. For the reader, it was felt that this approach offered a more useful perspective, enabling him to review fairly extensively a major area in what is now a rapidly growing and rather wide discipline. This policy has been retained, and Volumes 4 and 5 are written by a number of contributors who have dealt with various aspects of one of the most important areas of endocrinology—pregnancy and parturition. The two volumes are to some extent complementary because of the inevitable overlap of these topics.

This volume contains contributions from a group of expert investigators, who review a wide area of fetal endocrinology.

Gluckman deals in detail with the maturation and functions of fetal neuroendocrine structures, and Trounson addresses himself to the topic of in vitro fertilization—an area in which experimental progress is rapid, with major clinical and social implications. Flint, using a comparative approach, has reviewed the controversial issue of the factors controlling placental function in domestic animals. A rapidly growing field is the study of fetal thyroid function, dealt with by Thomas and Nathanielsz. Fetal metabolism has wide and major implications, both for research and for clinical practice; Lowy and Thomas have reviewed carbohydrate metabolism, Young has dealt with protein metabolism, and Stevenson discusses the mineral needs of the fetus. The breadth of these articles illustrates the growing interest and importance of these areas of human fetal research.

Finally, three articles are devoted more specifically to steroid endocrinology. Pearson Murphy and Branchaud discuss cortisol metabolism in the fetus, a field which continues to offer new perspectives as techniques improve. Imperato-McGinley reviews the fascinating area of sexual differentiation. Like the preceding articles this is an area in which recent research has widened our understanding of both normal and abnormal developmental endocrine physiology, and which has important clinical implications. Finally, New and her colleagues deal extensively with errors of adrenal steroidogenesis, an area which is important because it offers insights into normal physiological mechanisms and also has major clinical implications for the practicing physician.

It is a feature of the series that all the authors are practicing investigators, and all are of international standing. We are grateful that they have given up their time to preparing what the editors believe is a valuable oversight of an important and growing area of endocrinology.

The Fetal Neuroendocrine Axis

Peter David Gluckman,     DEPARTMENT OF PAEDIATRICS, UNIVERSITY OF AUCKLAND, AUCKLAND, NEW ZEALAND

Publisher Summary

The mature endocrine system is characterized by tight regulatory loops that subserve the homeostatic requirements of the organism; however, the fetal endocrine system cannot be considered solely in those terms. Not all units of the fetal endocrine system mature at the same rate and consequently, the regulation of fetal hormone secretion is not constant through development. While there is a considerable degree of autonomy from maternal influences, the fetal endocrine system may be influenced by extrinsic signals, particularly of placental origin. The action of fetal hormones may be limited by the immaturity of hormone receptors. Further, the actions of hormones in the fetus may differ from those observed after birth. Due to the ethical and practical limitations, which have restricted the ability to investigate endocrine function in the human fetus, most data in this chapter is derived from animal studies. The interpretation of studies in a variety of species is confounded by the great variation between species in the rate of neural maturation and in the degree of maturation present at birth.

I Introduction

II The Development of the Hypothalamus and Adenohypophysis

III The Secretion and Function of Fetal Adenohypophyseal Hormones

A Growth Hormone

B Prolactin

C Thyrotropin

D Gonadotropins

E Proopiocortin-Related Peptides

IV Autonomy of the Fetal Neuroendocrine Unit

References

I Introduction

In recent years there has been a rapid expansion of our knowledge of fetal physiology. This has largely been a consequence of the development of techniques of studying the fetus in utero by the chronic implantation of vascular catheters into the fetal sheep, rhesus monkey, and other species. This technique has enabled the fetus to be studied in a relatively undisturbed state and in the absence of the marked changes in the physiological status of the fetus observed following exteriorization or delivery and separation from the placental circulation (Nathanielsz, 1976, 1980). Studies of the fetal endocrine system have been of increasing interest. This interest arises in part from the pioneering studies of Jost on the role of fetal gonads in sexual differentiation (Jost 1954) and more recently from the studies of Liggins et al. (1968, 1973) and others on the role of the fetal hypothalamic–pituitary–adrenal axis in the regulation of parturition in the sheep and the effects of glucocorticoids on fetal lung development (Liggins and Howie, 1972; Liggins 1969).

While the mature endocrine system is characterized by tight regulatory loops which subserve the homeostatic requirements of the organism, the fetal endocrine system cannot be considered solely in these terms. Not all units of the fetal endocrine system mature at the same rate and consequently regulation of fetal hormone secretion is not constant through development. While there is a considerable degree of autonomy from maternal influences the fetal endocrine system may be influenced by extrinsic signals, particularly of placental origin. The action of fetal hormones may be limited by immaturity of hormone receptors. Further the actions of hormones in the fetus may differ from those observed after birth. For example, fetal hormones may have important functions in regulating tissue differentiation. The fetal neuroendocrine unit has been the subject of several reviews (Jost, 1966; Jost et al., 1970, 1974; Gluckman, 1981a; Jenkin et al., 1979; Dussault and Fisher, 1979; Fisher et al., 1977; Kaplan et al., 1976; Cara, 1978; Grumbach and Kaplan, 1974; Goodyer et al., 1979; Aubert, 1979; Gluckman et al., 1980a) and this article shall focus on recent studies of the hypothalamic–pituitary unit which emphasize these unique aspects of the fetal endocrine system.

Because of the ethical and practical limitations which have restricted the ability to investigate endocrine function in the human fetus, most data have been derived from animal studies. Interpretation of studies in a variety of species is confounded by the great variation between species in the rate of neural maturation and in the degree of maturation present at birth (Dobbing and Sands, 1979). For example, the rat has only achieved 12% of adult brain weight at birth and most brain growth occurs after birth. In contrast, the sheep is far more mature at birth, brain growth is primarily prenatal, and the neonatal sheep brain weighs 53% of adult brain weight. The maturation of the hypothalamic–pituitary axis is similarly advanced in species such as the sheep and delayed in species such as the rat. These species differences are not determined by gestational length, body size, or evolutionary rank (see Table I).

Table I

Species Differences in the Development of the Brain and Hypophyseal Portal Vascular System

aRanked in approximate order of neural and neuroendocrine maturation at birth.

bTiming of maximum increase in brain weight (from Dobbing and Sands, 1979).

cExcept where indicated as postnatal age (pn), age in days of fetal life.

II The Development of the Hypothalamus and Adenohypophysis

Until recently it has generally been accepted that the adenohypophysis arises as a diverticulum from the developing oral cavity (stomadeum) and is therefore of ectodermal origin. Recent evidence suggests that this is not so and that the hypothalamus and adenohypophysis are both derived from a common neuroectodermal analage. In careful histological studies in the avian embryo, Takor-Takor and Pearse (1975) have provided evidence that Rathke’s pouch, the progenitor of the adenohypophysis, arises from the ventral neural ridges. This portion of the neural tube also gives rise to the diencephalon. Subsequently, following dispersion of cells from the proliferating neural ridges into the surrounding mesoderm the developing adenohypophysis comes to lie in contact with the developing oral cavity and takes on the classical form of Rathke’s pouch. This observation would provide an explanation of reports that cells of the anterior pituitary have characteristics of cells of the APUD series (Pearse and Takor-Takor, 1976) and that cultures of Rathke’s pouch epithelia do not, in the absence of attached mesenchyme containing neural elements, secrete hormones (Ferrand et al., 1974). This hypothesis is an important revision of earlier concepts of the embryogenesis of the pituitary and warrants further investigation. A common origin of these two components of the neuroendocrine axis would suggest that there may be important interactions between the developing hypothalamus and pituitary from the earliest phases of development.

The timing of hypothalamic development in relationship to birth correlates with the rate of neural development in different species as exemplified by the following examples. In the sheep fetus, the supraoptic nuclei are present by the 24 mm state (35 days), the paraventricular nuclei by 27 mm, and by 35 mm most hypothalamic cellular proliferation is complete. By 61 mm (45 days) the hypothalamus is fully differentiated (Diepen, 1941). In the human fetus, the hypothalamus can be identified as the ventral layer of the developing diencephalon by 5 weeks and at this stage primitive fiber tracts are apparent. The arcuate and supraoptic nuclei are present by 7 weeks and the median eminence by 8 weeks. By 13 weeks all the hypothalamic nuclei have differentiated (Gilbert, 1934; Kuhlenbeck, 1954; Weill and Bernfeld, 1954; Papez, 1940). Extensive data are available on the morphogenesis of the hypothalamus in the rat. By the twelfth day of embryonic life, the hypothalamic region can be distinguished (Coggeshall, 1964). The lateral nuclei develop earliest at about day 12 followed by the appearance of the paraventricular and supraoptic nuclei. The periventricular, suprachiasmatic, and arcuate nuclei and the median eminence are among the last regions to differentiate (Altman and Bayer, 1978; Anderson, 1978; Ifft, 1972). The arcuate nucleus is not fully developed until after birth (Koritsansky, 1979; Matsumoto and Arai, 1976). The median eminence initially differentiates between fetal days 13 and 15 but is not layered in the adult manner until 2 weeks after birth (Bitsch and Schiebler, 1979; Kobayshi et al., 1968; Paull, 1978; Munroe and Paull, 1974; Daikoku et al., 1977; Rutzel and Schiebler, 1980). A similar ordering of the development of hypothalamic nuclei is apparent in the mouse (Angevine, 1970; Shimada and Nakamura, 1973).

The neurohormones somatostatin (SRIF), gonadotropin-releasing factor (LRF), and thyrotropin-releasing factor (TRF) are present in the fetal hypothalamus from very early phases of development. In man TRF and LRF are present in brain extracts by at least 5 weeks postconception (Winters et al., 1974a) and SRIF by 11 weeks postconception (Aubert et al., 1977). In the sheep TRF, LRF, and SRIF are all detectable in the median eminence by 58 days gestation (D.M. Styne, P.D. Gluckman, P.L. Mueller, S.L. Kaplan, and M.M. Grumbach, unpublished observations). In the fetal rat LRF is detectable in the hypothalamus by 16 days (Daikoku et al., 1978; Paull, 1978), SRIF by 17 days (Mueller et al., 1978) and TRF by 16 days (Oliver et al., 1980) of gestation. By 18 days, fetal rat hypothalamic cell cultures secrete vasopresin, CRF, β-endorphin, and ACTH (Denizeau et al., 1981). Mouse hypothalamic cells obtained from 16 day fetuses, cultured in serum-free medium, secrete TRF (Faivre-Bauman et al., 1981).

The recent development of techniques to examine presynaptic and postsynaptic markers of developing neuronal systems has lead to a rapid increase in our knowledge of the ontogeny of central neurotransmitter systems, particularly in the rodent. This aspect of neural development has been the subject of several reviews either of a general nature or relating to specific neurotransmitter systems (Baker and Quay, 1969; Lanier et al., 1976; Coyle, 1973, 1977; Filogamo and Marchisio, 1971; Johnston and Coyle, 1981).

In general development of synaptic units proceeds in a caudal to rostral direction and it is likely that hypothalamic systems are assembled and are functional prior to neocortical systems. The synaptic groupings for each neurotransmitter follow an individual timetable of development. Moreover not all the elements of a synaptic unit necessarily develop synchronously and the order of maturation is not constant between different neurotransmitter systems. For example the synthetic enzyme for γ-amino-butyric acid (GABA) appears roughly at the same time that GABA receptors develop but the presynaptic reuptake mechanism for GABA matures more rapidly (Coyle and Enna, 1976). In contrast, dopaminergic receptors generally develop in advance of presynaptic markers of the dopamine synapse. The studies in the rat generally show that cholinergic systems develop later than catecholamine and GABA-ergic neuronal systems (Johnston and Coyle, 1981). Thus the functional status of the developing hypothalamus will depend on the degree of maturation of each neuronal grouping and the degree of maturation of the various components of the synaptic unit as well as the progressive development of interactions between each neuronal grouping by increasing synaptogenesis.

There remains a paucity of data in other species with prenatal or perinatal brain growth patterns with regards the development of neurotransmitter systems. Serotonin, dopamine, and norepinephrine are all present in the human fetal hypothalamus by 12 weeks gestation (Hyyppa, 1972; Bertler, 1961; Nobin and Bjorklund, 1973).

From very early stages of development the primoidal pituitary has the capacity to secrete pituitary hormones in culture. By 5 weeks of gestation, the human fetal pituitary in culture secretes GH, PRL, LH, FSH, TSH, and ACTH (Pierson et al., 1973; Siler-Khodr et al., 1974). Similarly by day 12, the fetal rat pituitary in culture secretes all the adenohypophyseal hormones (Begeot et al., 1979; Watanabe and Daikoku, 1976).

It remains uncertain as to whether neural influences play a role in the onset of pituitary hormone secretion. While cultures of Rathke’s pouch cells prior to differentiation into cell types (16 days in rat fetus) secrete all the adenohypophysial hormones (Watanabe and Daikoku, 1976; Watanabe et al., 1973; Shiino et al., 1978; Begeot et al., 1979), these studies are confounded by the use of undefined culture media containing serum and the possibility of neural contamination of the cultures. In general these studies and those in the encephalectomized rat fetus (Daikoku et al., 1973; Chatelain et al., 1976, 1979) suggest that in the absence of hypothalamic stimuli, differentiation into cell types will occur and hormonal secretion can be initiated. Further, these studies suggest that complete maturation of pituicytes and, in particular, the full development of secretory granules require hypothalamic influences. However, Ishikawa et al. (1977) suggest that factors present in fetal brain extracts are necessary for the differentiation in culture of anterior pituitary cell lines from Rathke’s pouch percursors. The subsequent development of Rathke’s pouch into the anterior and intermediate lobes has been reviewed recently (Gluckman et al., 1980a; Gluckman, 1981a).

The earliest interactions between the developing hypothalamus and pituitary are probably by a paracrine route, with simple diffusion of hypothalamic factors from the hypothalamus to Rathke’s pouch (Kaplan et al., 1976; Gluckman, 1981a).

The primary plexus of the portal vascular system arises from an extensive capillary network in the mesenchyme surrounding the developing pituitary (Wislocki, 1937). Initially a well-defined capillary network develops on the ventral surface of the hypothalamus but there is no penetration of capillary loops into the median eminence (Nieminerva, 1950; Rinne, 1962). This plexus has been termed a mantle plexus and persists into adulthood in some reptiles and birds (Enemar, 1960). It therefore seems likely that neurohormones can be secreted into the mantle plexus and that penetration of capillary loops into the median eminence is not essential for a functional portal vascular system.

Subsequently capillary loops from the mantle plexus invade the median eminence to form the true primary plexus. A recent report suggests that the primary plexus is formed by 12 weeks of gestation in the human fetus (Thliveris and Currie, 1980), somewhat earlier than previously accepted (Nieminerva, 1950; Rinne, 1963). SRIF and LRF-containing axonal terminals are in contact with capillaries of the developing primary plexus by 16 weeks of gestation (Bugnon et al., 1976, 1977a,b, 1978). The primary plexus is formed in the horse by 50 days and in the guinea pig by 40 days of gestation (see Table I). In contrast, in the rat and mouse, formation of the primary plexus occurs relatively later in the period surrounding parturition, again demonstrating the differences in rates of development between species. Portal venous trunks appear at about the time that the true primary plexus appears (see Table I).

III The Secretion and Function of Fetal Adenohypophyseal Hormones (see Fig. 1)

A Growth Hormone

1 Secretion

The essential feature of fetal GH secretion is that, in all species studied, plasma GH concentrations are elevated at some phase of fetal life. Despite these very high circulating concentrations there is no evidence that GH plays a major role in the regulation of fetal growth (see Section III,A,2) and it does not appear that these high fetal GH concentrations subserve an essential role in fetal development. Consequently it has been proposed that these high plasma concentrations are a consequence of immaturity of hypothalamic control of fetal GH secretion (Kaplan et al., 1972, 1976; Grumbach and Kaplan, 1973, 1974).

Fig. 1 Diagramatic representation of the pattern of plasma pituitary hormone concentrations in the perinatal period in sheep, man, and rats. The timing of maximum brain growth is indicated by the arrow. Gestational age is expressed in days before or after birth (B). See text for references.

In the human fetus plasma GH concentrations reach peak concentrations of 131 ± 22 ng/ml at 20 to 24 weeks and fall in late gestation to be 34 ± 4 ng/ml in cord blood. There is a further decrease in plasma GH concentrations over the first 2 weeks of postnatal life (Kaplan et al., 1972; Cornblath et al., 1965; Matsuzaki et al., 1971; Von Muhlendahl et al., 1976). Umbilical cord concentrations are higher in premature infants than in term infants and the postnatal fall is more prolonged (Cornblath et al., 1965; Ballard et al., 1980). Even in the youngest fetuses studied at 70 days fetal GH concentrations are markedly elevated compared to postnatal concentrations. Similarly in the monkey (Holland et al., 1979), sheep (Gluckman et al., 1979a; Bassett et al., 1970), and pig (Atinmo et al., 1979a), fetal plasma GH concentrations are markedly elevated compared to adult values in mid and late gestation and decrease in the neonatal period. In the sheep the fall in plasma GH concentrations commences two to three days prior to birth (Gluckman et al., 1979a; Bassett et al., 1970) and has reached low adult values within 24 hours of birth (Bassett and Alexander, 1971). Limited data suggest that plasma GH concentrations rise to very high concentrations after 20 days in the fetal rabbit (Jost et al., 1979). In the rat and mouse, fetal plasma GH concentrations rise very late gestation, are markedly elevated at birth, and fall in the first 2 weeks after birth to adult values (Rieutort, 1974; Blasquez et al., 1974; Sinha et al., 1972).

The metabolic clearance of GH in the late gestation ovine fetus is similar to that of the infant lamb (Wallace et al., 1973). In the human neonate the metabolic clearance of GH is faster than in the adult (Cornblath et al., 1965). There are no data available in either species regarding the disposition of circulating GH earlier in gestation. Growth hormone does not pass the human placenta (Gitlin et al., 1965) and, as plasma GH concentrations are undetectable in the hypophysectomized sheep fetus (Bassett et al., 1970) and decapitated rat fetus (Rieutort, 1972), it seems unlikely that GH passes the placental barrier in these species either.

There is ample clinical evidence that the regulation of GH release in the human neonate is immature (Gluckman et al., 1980a, Gluckman, 1981a). Sleep-associated GH release is not demonstrable until 3 months after birth (Finkelstein et al., 1971; Vigneri and Agnata, 1971; Shaywitz et al., 1971). Paradoxical elevations of GH concentrations are observed following glucose infusion in the human neonate (Reitano et al., 1971, 1978; Cornblath et al., 1965). Similarly L-Dopa and pyridoxine are reported to inhibit rather than stimulate neonatal GH release (Delitata et al., 1978a). On the other hand, arginine (Reitano et al., 1971; Stubbe and Wolf, 1970) and insulin-induced hypoglycemia (Cornblath et al., 1965) stimulate neonatal GH release.

If the high fetal GH concentrations are a consequence of immaturity of control of GH release then this immaturity could involve several potential mechanisms. The possibilities include autonomous pituitary GH secretion, deficient SRIF release, excessive GH-releasing factor (GRF) release, deficient negative feedback, or stimulation of GH release by extinsic influences.

The GH concentrations in the umbilical cord blood of human anencephalic fetuses are low (Kaplan et al., 1976). Anencephalic infants generally have a small adenohypophysis but have absent diencephalic tissue (see Gluckman et al., 1980a). This is evidence that the fetal hypothalamus tonically stimulates fetal GH release in late gestation. Similarly following fetal stalk section of the ovine fetus, between 110 and 130 days, GH concentrations generally fall to low levels (Gluckman et al., 1982). The highest fetal plasma GH concentrations in the human fetus occur at the time of full maturation of the portal vascular system, again suggesting the role of GRF in generating the high fetal GH concentrations.

Immunoreactive somatostatin is present in the human fetal hypothalamus by 11 weeks and its concentration increases between 10 and 22 weeks (Aubert et al., 1977). Immunohistochemical studies demonstrate that by 16 weeks somatostatin-staining neurons arising from the periventricular areas terminate in the median eminence in conjunction with capillaries of the developing primary plexus (Bugnon et al., 1978; Paulin et al., 1976). In the sheep, somatostatin is present in the fetal hypothalamus by 60 days gestation and the concentration increases after 100 days (D.M. Styne, C. Marti-Henneberg, P.D. Gluckman, S.L. Kaplan, and M.M. Grumbach, unpublished data; Fisher et al., 1977). SRIF is first detected in the fetal rat hypothalamus by day 17 (Mueller et al., 1978). Fetal mouse hypothalamic neurons in culture secrete somatostatin (De Vitry et al., 1979). By day 19 in the mouse, SRIF is present in nerve terminals in the median eminence in proximity to capillaries of the mantle plexus (Gross and Longer, 1979). Thus in each species the fetal hypothalamus synthesizes and contains SRIF at the time when fetal GH concentrations are very high.

SRIF inhibits GH release in the normal human neonate (Delitata et al., 1978a) and SRIF inhibits GH release by midgestation human fetal pituitaries in organ culture (Goodyer et al., 1977). Exogenous SRIF suppresses fetal GH release in the sheep fetus by 80 days gestation (Gluckman et al., 1979b; McMillan et al., 1978) with no ontogenic change in the magnitude of the response with advancing gestation. These data suggest the presence of SRIF receptors on the fetal somatrope.

In the sheep, dopaminergic agonists inhibit GH release and this response is developed by 80 days gestation (Marti-Henneberg et al., 1981). In the rat dopamine inhibits GH release by stimulating SRIF release (Chihara et al., 1979). Assuming a similar mechanism in the sheep, these data suggest that hypothalamic somatostatin release can be stimulated by dopamine in the midgestation ovin fetus.

While the capacity of the fetal hypothalamus to synthesize and secrete SRIF is established, there remains no direct evidence as to whether hypothalamic SRIF is secreted by the fetus and deficient SRIF release may contribute to the high fetal GH concentrations. In the rat the postnatal fall in plasma GH concentrations has been correlated with increasing hypothalamic somatostatin concentrations (Walker et al., 1977).

The ontogeny of neurotransmitter-mediated control of GH release has been primarily studied in the chronically catheterized sheep. These studies have used a neuropharmacological approach with the administration of agonists or antagonists intravenously to the fetus. There are limitations to the interpretation of these studies as a consequence of the experimental approach—it is not known whether the pharmacokinetics of the drugs used are constant during development and there are clearly problems relating to the specificity of the drug action observed. A number of laboratories are actively pursuing the problem of overcoming the considerable problems of more sophisticated neurophysiological approaches to the fetus. Limited progress has been made in developing techniques for placing catheters in the fetal CSF. Konda et al. (1979) have reported a technique of sterotactic placement of electrodes in the fetal pig brain.

It is clear from neuropharmacological studies in the sheep fetus that the potential for various neurotransmitter systems to influence fetal GH release does not develop simultaneously but is spread over a wide gestational range. Even in a precocial species such as the sheep these systems are not fully integrated until after birth.

The serotoninergic agonists, 5-hydroxytryptophan (5HTP) and fluoxetine stimulated fetal GH secretion by 80 days gestation (Marti-Henneberg et al., 1980). The response to 5HTP is abolished by fetal pituitary stalk-section suggesting that serotonin is acting at the hypothalamic level (Gluckman et al., 1982). As fluoxetine acts by inhibiting serotonin reuptake, the data further suggest that endogenous hypothalamic serotonin can modulate neurohormone secretion leading to fetal GH release. The GH response to serotonin agonists decreases markedly after 100 days. The reason for this is speculative. There may be a decrease in the concentration of serotonin receptors as has been described in the infant rat brain (Uzbekov et al., 1979). Alternatively, the decrease in response to serotonin agonists after 100 days may be a consequence of the development of counteracting neuroinhibitory mechanisms which lead to a dampening of the response to the trophic stimulus.

Similarly, the administration of β-endorphin intravenously to the ovine fetus stimulates GH release markedly prior to 100 days gestation and the response decreases late in gestation. However β-endorphin administered via the fetal CSF markedly stimulates GH release in late gestation suggesting that the development of the fetal blood-brain barrier is the prime reason for the reduction in the response to β-endorphin in late gestation (Gluckman, 1981b). The development of the blood-brain barrier is discussed in a separate section of this article.

Dopaminergic agonists inhibit GH release in the fetus by 80 days gestation (Marti-Henneberg et al., 1981) with no developmental change in the magnitude of the response. The GABA agonist muscimol inhibits GH release in the sheep fetus by 100 days (Gluckman, 1981c). In contrast neither the α2 agonist clonidine (Goldsmith et al., 1980) nor the cholinergic agonist physostigmine (Gluckman, 1981d) stimulates GH release until after birth. Both drugs stimulate GH release in the neonatal lamb and the response is blocked by pretreatment with phentolamine and atropine, respectively. The wide scatter in the timing of the development of the ability of these various neurotransmitters to affect fetal GH release in the sheep presumably reflects differing rates of maturation of each neurotransmitter system in the fetal hypothalamus.

Limited data suggest that there may be some tonic inhibitory influences on fetal GH secretion in the sheep fetus. The administration of picrotoxin, a GABA antagonist, to the sheep fetus after 90 days of gestation is associated with an increase in plasma GH concentrations (Gluckman, 1981c). This suggests that there may be partial restraint of fetal GH release in the ovine fetus by a GABA-mediated mechanism.

In the adult, GH exerts negative feedback on its own secretion (Katz et al., 1969). Immaturity of this feedback loop, perhaps as a consequence of immature GH receptors in the hypothalamus, would lead to high fetal GH concentrations. While there is no direct evidence of such immaturity in the fetus, recent evidence suggests that somatogenic receptors are immature in the ovine fetal liver (Gluckman, 1981b).

Finally the possibility that fetal GH secretion may be influenced by extraneural factors must also be considered. The very rapid fall in plasma GH concentrations at parturition in the sheep fetus raises the possibility that the placenta may secrete a tropic substance. As will be discussed subsequently, the placenta has been demonstrated to contain a number of neurohormones and biogenic amines, the function of which remains speculative. Such a hypothesis may explain the observation that plasma GH concentrations do not always fall following pituitary stalk section in the sheep fetus, particularly in late gestation (Nathanielsz et al., 1977; Gluckman et al., 1981).

2 Function

It is clear that GH does not play a significant role in the regulation of fetal growth. The evidence for this statement has been extensively reviewed (Gluckman and Liggins, 1983; Cheek and Hill, 1974; Cheek et al., 1977; Jost, 1953, 1954, 1961, 1966, 1977, 1979; Liggins, 1974). In general removal of the influence of fetal GH by fetal hypophysectomy, fetal stalk-section, or fetal decapitation in a variety of species including the pig, rabbit, rat, and mouse does not necessarily lead to fetal growth retardation (Colenbrander et al., 1979; Hill et al., 1979; Jost, 1966; Bearn, 1968; Eguchi, 1961). In the rhesus monkey and sheep fetus, fetal decapitation is associated with growth retardation (McNatty et al., 1973; Kittinger, 1977; Liggins and Kennedy, 1968; Barnes et al., 1977), but in these species, fetal hypothyroidism also causes growth retardation (Kerr et al., 1972; Thorburn, 1974). When fetal stalk-section is performed in these species (so that GH secretion is suppressed but some thyroid function is maintained), normal fetal growth is observed (Kittinger, 1977; Liggins, 1974; Liggins et al., 1973). The human fetus with pituitary agenesis is of normal birth length (Reid, 1970; Sadeghi-Nejad and Senior, 1974; Kauschansky et al., 1979).

Similarly evidence in the fetal rabbit (Hill et al., 1979) and human anencephalic neonate (Gluckman and Brinsmead, 1976; Foley et al., 1980) suggests that fetal somatomedin secretion is independent of GH. However conflicting results are reported in the fetal sheep (Brinsmead and Liggins, 1979; Falconer et al., 1979).

The reason why fetal growth is independent of GH is speculative. It may relate to immaturity of growth hormone receptors, particularly in the liver which is the major site of somatomedin production (D’Ercole et al., 1980). The binding of human GH to hepatic microsomal membranes is low in tissues obtained from fetal rabbits and rats and the binding increases in postnatal tissues (Kelly et al., 1974, 1976). Studies of the binding of hGH to ovine heptic tissues demonstrated low binding in fetal tissues and a sixfold increase in binding after birth. Cross-reaction studies confirmed with studies using ¹²⁵I-labeled ovine PRL and ¹²⁵I-labeled ovine GH suggest that there is a deficiency of somatogenic binding sites in the ovine fetal liver and that they appear in the neonatal period. In contrast lactogenic binding sites are present from at least 90 days of gestation (Gluckman, 1981b). The lack of somatogenic binding sites in the fetal ovine liver is not due to saturation of the receptors by high circulating HG concentrations as the binding is not enhanced by pretreatment in vitro with 5 M MgCl2 to remove endogenously bound hormone from the binding sites. As the binding is not increased in fetuses decapitated 20 days previously, the apparent lack of somatogenic sites cannot be explained by down-regulation by the high fetal GH concentrations (P.D. Gluckman and J. Butler, unpublished observations). However, somatogenic receptors may be present in other fetal tissues and GH may exert affects on fetal development through other receptor systems.

Congenital GH deficiency in man may be associated with hypoglycemia and the hypophysectomized or decapitated ovine, rat, or rabbit fetus has deficient hepatic glycogen deposition in late gestation (Jost and Picon, 1970; Jost et al., 1979; Jost and Jaquot, 1958; Barnes et al., 1977). It has been suggested that GH may play a role in the stimulation of fetal glycogen deposition (Jost et al., 1979). GH may also play a role in the induction of enzymes. Administration of GH to the fetal rat stimulates the precocious appearance of lung phosphorylase a activity (Jost et al., 1979). Any action of GH on glycogen deposition might be mediated by enzyme induction.

GH has a synergistic action with ACTH on adrenal steroidogenesis in adult rates (Colby et al., 1973; Kramer et al., 1977). Human GH has been reported to stimulate dehydroepiandrosterone sulfate production by the human fetal adrenal gland in vitro (Brown et al., 1978). Human growth hormone stimulated corticosteroid production by the fetal sheep adrenal both in vitro and in vivo and by the fetal rabbit in vitro. However, in maternal tissues, no effect on adrenal secretion was observed (Devaskar et al., 1981). These studies raise the possibility that GH may have a steroidogenic function in the fetus.

B Prolactin

1 Secretion

In species demonstrating predominantly prenatal or perinatal brain development, fetal plasma PRL levels are low in midgestation and rise to relatively high concentrations in late gestation. Particularly in the human fetus, plasma PRL concentrations