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Carbamoyl-phosphate synthase gene expression is found to be primarily regulated by conditions that enhance hepatic glucocorticosteroid levels (hormone injections) and cyclic AMP levels (induction of diabetes). After birth, changes in the level of carbamoyl-phosphate synthase protein follow changes in the level of carbamoylphosphate synthase mRNA, suggesting a pretranslational control mechanism. In fetal rats, carbamoyl-phosphate synthase gene expression is regulated by the same factors as in adults. However, both the level to which carbamoyl-phosphate synthase mRNA can accumulate and the extent to which mRNA can be translated appear to be limited, indicating control mechanisms at the pretranslational and translational level. Finally, in the immediate postnatal period, a transient but pronounced decrease in the rate of degradation of carbamoyl-phosphate synthase protein may play a role in the accumulation of the enzyme.

Rat (Rattus norvegicus) and spiny mouse (Acomys cahirinus) are closely related murine species that, due to their altricial (rat) and precocial (spiny mouse) modes of development, differ in the developmental timing of birth. A comparison between the developmental profiles of plasma glucagon, insulin, thyroxine, triiodothyronine, and glucocorticosteroid hormone was carried out to elucidate the question to what extent these hormonal profiles were related to the timing of birth. Although corticosterone is the major circulating glucocorticosteroid in rat, only cortisol was found in the spiny mouse. The onset of increases in glucocorticosteroid and thyroid hormone levels occurred at the same developmental time points in both species. A neonatal increase in triiodothyronine levels was observed in the spiny mouse only. In both species the immediate perinatal period was characterized by decreases in the ratio of insulin and glucagon levels and the level of glucocorticosteroids. The observed developmental patterns of hormonal levels were found to be consistent with the observed developmental pattern of enzymic maturation in the respiratory and gastrointestinal tract, which play a critical role in the adaptation to the extrauterine environment.

Rat (Raitus norvegicus)and spiny mouse (Acomys cahirinus)are closely related species that mainly differ in the developmental timing of birth. A comparison between the developmental profiles of some characteristic enzymes of the small intestine (lactase and sucrase) and of the pancreas (amylase) of both species was carried out to elucidate the question to what extent these enzymic profiles and hence the maturation of these organs was related to the process of birth. It was found that these organ-specific enzymes become first detectable at the same developmental stage in both species. Likewise, the weaning phase of the enzymic profiles occurred at the same developmental time point in both species. It is argued that both the first appearance and the weaning increase in enzyme activity follow an inherent biological program that can only be modulated by hormones. In contrast, the perinatal phase of the enzymic profile is completely dependent on the developmental timing of birth, and therefore appears not to be anchored to a particular developmental time point but rather to be dependent on birth-associated (hormonal) adaptation. In accordance with this hypothesis it was found that the development of the microscopic anatomy of the small intestine proceeded independently of the functional adaptation of the intestine to the process of birth.

Rat (Rattus norvegicus) and spiny mouse (Acomys cahirinus) are closely related murinoid species that mainly differ in the developmental timing of birth. A comparison between the developmental profiles of some characteristic enzymes of the liver of both species was carried out to elucidate the question to what extent are these enzymic profiles and hence the maturation of the liver related to the timing of birth? It was found that these organotypic enzymes first become detectable at the same developmental stage in both species. Likewise, the weaning phase of the enzymic profiles occurs at the same developmental time point in both species. It is argued that both the first appearance and the weaning increase in enzyme activity levels occur at endogenously programmed timepoints with only superimposed effects of hormones. In contrast, the perinatal phase of the enzymic profile is completely dependent on the developmental timing of birth and therefore appears not to be anchored to a particular developmental timepoint, but rather to be dependent on birth-associated (hormonal) adaptation. In accordance with this hypothesis it was found that the morphological development of the liver proceeded independent of the timing of birth. Furthermore, the hormonal regulation of the investigated enzymes was found to be the same in both species. Despite the more advanced state of morphological development of the liver in the spiny mouse at birth, it was found that the inducibility of organotypic gene expression by hormones in spiny mouse fetuses was as limited as in rat fetuses. This observation therefore suggests that the intra-uterine environment is responsible for the limited inducibility of enzymes before birth.

A method has been developed to establish the degree of cross-reactivity of an antiserum raised against purified carbamoyl-phosphate synthase (ammonia) from adult rat liver, toward a homologous enzyme from another species without purification of the latter enzyme. For that purpose the ratio between enzyme activity and enzyme protein, i.e., the molecular specific activity in crude liver extracts, was determined by two independent methods. When the molecular specific activity was determined by means of radioimmunoassay using a specific antiserum raised against rat liver carbamoyl-phosphate synthase this ratio was a factor four higher for the axolotl than for the rat. Both axolotl and rat liver carbamoyl-phosphate synthase appear as a very prominent band with an apparant molecular weight of 165,000 after sodium dodecyl sulphate-polyacrylamide gelelectrophoresis. Therefore, the amount of enzyme protein could be determined by means of densitometry of this band using purified rat liver carbamoyl-phosphate synthase as a standard. The ratio between enzyme activity and enzyme protein calculated from this method appeared to be the same for axolotl and rat. From these results it can be deduced that the degree of cross-reactivity between rat and axolotl liver carbamoyl-phosphate synthase is approximately 25% when using the antiserum raised against the rat liver antigen.

A completely defined medium (EHM-1), which reflects the amino acid composition of fetal rat serum and contains albumin as the sole proteinaceous compound, allows the accumulation of carbamoylphosphate synthetase and phosphoenolpyruvate carboxykinase in the presence of dexamethasone, dibutyryl cyclic AMP, and triiodothyronine to approximately twice the level attained in a standard culture medium (RPMI 1640) supplemented with 10% fetal bovine serum (and hormones). Using the EHM-1 medium we could show that the capacity of hepatocytes to synthesize phosphoenolpyruvate carboxykinase in the presence of hormones is manifest as soon as the cells differentiate from the embryonic foregut (embryonic Day 11). Furthermore we could show that embryonic hepatocytes can become binuclear or polyploid when cultured in the presence of thyroid hormone.

Rat (Rattus norvegicus) and spiny mouse (Acomys cahirinus) are closely related species that mainly differ in the developmental timing of birth. A comparison between the developmental profiles of some characteristic enzymes of the small intestine (lactase and sucrase) and of the pancreas (amylase) of both species was carried out to elucidate the question to what extent these enzymic profiles and hence the maturation of these organs was related to the process of birth. It was found that these organ-specific enzymes become first detectable at the same developmental stage in both species. Likewise, the weaning phase of the enzymic profiles occurred at the same developmental time point in both species. It is argued that both the first appearance and the weaning increase in enzyme activity follow an inherent biological program that can only be modulated by hormones. In contrast, the perinatal phase of the enzymic profile is completely dependent on the developmental timing of birth, and therefore appears not to be anchored to a particular developmental time point but rather to be dependent on birth-associated (hormonal) adaptation. In accordance with this hypothesis it was found that the development of the microscopic anatomy of the small intestine proceeded independently of the functional adaptation of the intestine to the process of birth.

Experimental studies of rat development have shown that the first appearance and possibly part of the preweaning increase of many liver-specific enzymes is due to developmental maturation of the liver itself, but that the major increases in enzyme activity in the perinatal and preweaning periods are due to changing hormone levels. To establish the general nature of this type of developmental regulation we have studied enzyme activity changes during development of the spiny mouse, a closely related murine rodent which has apart from a different developmental timing of the moment of birth, a similar developmental timescale as the rat. Comparison of these altricial (rat) and precocial (spiny mouse) modes of development shows that the first appearance of enzyme activity in the liver (but also in the pancreas and small intestine) is developmental stage-specific, but that the first major increase in enzyme activities awaits the (differently timed) perinatal period. The preweaning increase in enzyme activity appears to be at least partly a developmental stage-specific event. These comparative studies therefore confirm the regulatory model of enzyme accumulation which was derived from experimental studies in the rat. Developmental profiles of human liver enzymes resemble those of the spiny mouse more than they do those of the rat.

The construction and cloning of a cDNA complementary to the mRNA of rat liver carbamoylphosphate synthetase (ammonia) is described. Using this cDNA, the size of the mature, cytosolic carbamoylphosphate synthetase (ammonia) mRNA is estimated to be 6.0 Kb. The levels of carbamoylphosphate synthetase (ammonia) mRNA in liver are shown to be regulated by glucocorticosteroids and cyclic AMP. By studying mRNA levels of carbamoylphosphate synthetase, albumin and phosphoenolpyruvate carboxykinase, using specific cDNA clones, we show that carbamoylphosphate synthetase gene expression, like that of albumin is liver-specific.

Rat and spiny mouse (Acomys cahirinus) are closely related murinoid species that represent altricial (rat) and precocial (spiny mouse) modes of development. The late intrauterine developmental stages of the spiny mouse therefore seem comparable to the early extra-uterine developmental stages of the rat. To elucidate the question to what extent the development of the lung is related to the developmental timing of birth, we have studied some enzymes involved in the de novo synthesis of phosphatidylcholine. Of the enzymes studied, cholinephosphate cytidylyltransferase shows peaks in activity in the perinatal period (rat and spiny mouse) and at the beginning of the 3rd postnatal week (rat only). This enzyme fulfils the requirements for a developmental parameter best as changes in activity of this enzyme can be correlated with phases of cell proliferation and surface expansion in the lung of the rat. The single peak of cholinephosphate cytidylyltransferase activity in the spiny mouse as well as microscopical examination of the lung support the hypothesis that the processes of proliferation and surface expansion, which occur consecutively in the rat, develop concurrently in the spiny mouse.