DIFFERENCES IN RHYTHMS OF ENZYMATIC ACTIVITY OF MATERNAL AND FETAL BLOOD

Blood specimens were obtained at different daily times from the umbilical cord and brachial vein from 53 women within 10 minutes after delivery. Enzyme activities were measured in the white blood cells (WBCs) and serum of each sample. For each enzyme, the results were grouped according to sampling (delivery) times and arrayed to form a 24h time series. Separate time series were generated for maternal and fetal enzymes. Analysis of variance (ANOVA) and cosine best-fit analyses were applied to elucidate significant differences between enzyme activity patterns of mothers and fetuses with regard to time dependency, number of peaks, and acrophases. These and previously documented results indicate that not all mothers and fetuses have rhythms that are concordant. For some enzymes of fetuses, the activity rhythms differ in phase and shape of the time series pattern from those of the mothers; for other enzymes, the activity rhythms develop after birth. (Chronobiology International, 17(2), 221–228, 2000)     

Lack of circadian rhythmicity of rat fetal intestinal enzymes as compared to the dams

The 24-hr activity patterns of intestinal maltase, lactase, leucylnaphthylamine hydrolyzing activity, γ-glutamyltransferase, and alkaline phosphatase were determined in pregnant rats maintained on a 12-12 light-dark cycle, with feeding during the dark period (1800-0600 hr, EST). The activities of these enzymes plus those of lysosomal maltase and lactase were followed during the same time period in 19- to 20-day-old fetuses. The activity patterns in the dams followed circadian rhythms, with peak activities occurring during the feeding-dark period. These rhythms are similar to the feeding schedule-cued rhythms observed in male rats and, therefore, are assumed to be feeding schedule cued also. In the fetuses, which obtained nutrients through the placenta, the activities increased in a somewhat nonlinear manner throughout the entire 24-hr period, but did not display a defined rhythm. It is concluded that endogenous intestinal enzyme rhythms do not exist in utero, and that oral and/or intermittent feeding is necessary for these rhythms to occur.     

Effects of maternal diabetes on the development of carbohydrate metabolizing enzymes in fetal rat liver

Effects of streptozotocin-induced maternal diabetes on fetal hepatic carbohydrate-metabolizing enzyme development and hormonal status has been explored in the rat. Hepatic glycogen synthase a activity of the normal fetus rose to a maximum at 20 days of gestation, then fell prior to parturition. In fetuses of diabetic mothers, this prepartum decline was curtailed, resulting in enhanced synthase a activity and increased glycogen content in fetal livers at term. Elevation in hepatic synthase a in fetuses of diabetic mothers was due, not to altered interconversion between existing synthase a and b, but to equivalent increases in both forms of the enzyme. Both hepatic and free plasma corticosterone levels were elevated in fetuses of diabetic mothers and may be responsible for the enhanced development of total glycogen synthase observed in these fetuses. In normal fetuses hepatic phosphofructokinase and pyruvate kinase activities also rose to maxima at 20 days, then declined prior to term. In fetuses of diabetic mothers pyruvate kinase activity attained higher than normal maximal levels and phosphofructokinase activity fell more gradually, thus resulting in elevations in both enzyme activities at term. Augmentations in these glycolytic enzymes are compatible with hyperinsulinemia observed in fetuses of diabetic mothers. The following conclusions may be drawn from these findings. During late fetal life developmental patterns of rate-limiting hepatic glycogen-synthesizing and glycolytic enzymes are adapted to glucose utilization. In the normal fetus these patterns reverse at term, thereby promoting glucose mobilization, which prepares the fetus for abrupt deprivation of maternal glucose at birth. Maternal diabetes results in retardation of these reversal processes, presumably due to elevations in fetal glucocorticoid and insulin levels. Glycogenolytic and glucogenic capacities are thereby impaired in these fetuses.     

Circadian Rhythms of Enzymes' Activity in Mice Tissues During Exposure to Continuous Illumination

Activity rhythms of enzymes were determined in various tissues of C57BL/6J male mice. The determinations were carried out on mice which were kept in 14 hr light: 10 hr dark regimen, and on day 2, day 5 and day 21 during exposure to continuous illumination. Locomotor activity rhythms were followed in light: dark and up to the seventh day in constant light. All the activities exhibited a significant circadian rhythm in the light: dark regimen. During the exposure to continuous illumination, the locomotor activity exhibit a free running circadian rhythm with a consistent 24 hr and 40 min, major period component. At the same time recording the rhythms of enzyme activity; enzymes exhibited various formats of response which differed from those of the locomotor activity. The results suggest that rhythms of enzyme activity, as well as the desynchronization of the rhythms, are not enzyme specific.     

Environmental stress alters the developmental pattern of delta 5-3 beta-hydroxysteroid dehydrogenase activity in Leydig cells of fetal rats: a quantitative cytochemical study

Quantitative cytochemistry was used to determine the effect of subjecting pregnant rats to environmental stress on the activity of delta 5-3 beta hydroxysteroid dehydrogenase (3 beta-HSD) in Leydig cells of their fetuses. Enzyme activity was measured by microspectrophotometry in individual Leydig cells in cryostat sections of fetal testes on Days 16-21 postconception. Fetuses of stressed mothers lacked the peak of enzyme activity on Days 18 and 19 of gestation that is characteristic of Leydig cells of normal fetuses at this time. In addition, both before and after these 2 days, 3 beta-HSD activity in Leydig cells of stressed fetuses was significantly higher than normal. The altered developmental pattern of 3 beta-HSD activity in the stressed fetuses largely corresponds to the changes in plasma testosterone found previously in male fetuses of mothers exposed to the same regimen of stress. Thus, in the fetal Leydig cell, the activity of 3 beta-HSD, a key steroidogenic enzyme, can be modified by environmental stress, and provides an index of steroidogenic activity of the fetal testes and of the titers of circulating testosterone.     

Changes in carnitine-palmitoyl-transferase and carnitine-acetyl-transferase activity in rat kidney during development; effects of fasting

Developmental changes in the activities of two enzymes catalysing transfer of fatty acid across the mitochondrial membrane (carnitine-palmitoyl-transferase and carnitine-acetyl-transferase) were studied in the kidneys of developing rats from late fetal life to 10 days post-partum and were compared to cortical adult value. The activities of carnitine-palmitoyl-transferase and carnitine-acetyl-transferase increased after birth to reach a maximal value on day 5. Thereafter both activities decreased to reach adult cortical value. The cytochrome c oxidase activity (index of mitochondrial activity) increases continuously from late fetal age to adult. In kidneys of fetuses from starved mothers the carnitine-palmitoyl-transferase activity is higher than that of controls while carnitine-acetyl-transferase activity is not changed. In postmature fetuses (23 days post-co?tum) carnitine-palmitoyl-transferase activity is the same as in 21 days post-co?tum old fetuses. These results are discussed in relation to variations in nutritional and hormonal changes occurring during the perinatal period.     

Circadian and Ultradian Time Patterns in Human Behaviour: Part 1: Activity Monitoring of Families from Prepartum to Postpartum

Time patterns of activity-rest rhythms during and after pregnancy are increasingly recognised as important factors for the well-being and health of young families. This longitudinal study examined activity-rest patterns of couples during late pregnancy and subsequently the alterations in the periodic structure of parental and neonatal time patterns during the first four months after birth. Part I concentrates on the effects of late pregnancy and birth to the mother's rest-activity patterns and those of the father and, after birth, what time pattern the infant developed. Part II attempts to clarify how activity patterns of the entire family agree or disagree with each other and investigates how the infant synchronises with the environment that includes the process of parent-infant interaction. Activity data of, so far, seven families (father, mother and child) were continuously recorded using non-invasive Actiwatch units. Recordings of parental activity started at the beginning of the 37th week of gestation, and were continued in parallel with the infants' recordings in three series of three weeks each until four months after birth: 1st to 3rd week, 7th to 9th week and 13th to 15th week of life. In a standardised diary, record was kept of household routines, parental activities, type of feeding, initiation of sleep or waking up. Activity data of seven non-pregnant women were collected and used as a control. Irregular nocturnal activity epochs occurred frequently in pregnant women and were absent in non-pregnant women. Period lengthenings and shortenings of the circadian rhythms appeared in both parents from prepartum to postpartum. Activity at night increased from prepartum to postpartum in mothers and fathers. Three infants showed a marked circadian rhythm between day 3 and 14 after birth. All seven infants showed a predominant circadian rhythm between day 8 and 19 after birth. The onset of daytime activity of mothers and their infants corresponded well to each other. Postpartum frequency spectra of parents and child always had some ultradian components in common. Time patterns of activity-rest rhythms of couples and parents are shown to be altered during and after pregnancy and we suggest that the infants' adaptation to the environment begins during the first week that includes the process of mother-infant interaction.     

Maternal entrainment of tau mutant hamsters.

Maternal entrainment of the circadian wheel-running activity rhythm was examined in Syrian hamsters heterozygous for a single gene mutation (tau) that affects the free-running period of circadian rhythms. Heterozygous tau pups were born to and raised by wild-type mothers under constant dim light. The pups' wheel-running activity was recorded after weaning on postnatal day 18 or 24. Pups weaned on day 18 had an average free-running period of 21.70 hr, demonstrating that the tau phenotype was fully expressed at this age. Using the activity onset of the postnatal free-running rhythms as a phase reference, we estimated the phase relationships between the pups and their mothers on days 18 and 24. In contrast to results with wild-type pups, the activity rhythms of tau pups were not in phase with the rhythms of their wild-type mothers; that is, activity onsets of mothers and pups did not coincide. The pups did, however, show synchrony among themselves, indicating that they had been exposed to a synchronizing signal sometime during development. It is likely that this synchronizing signal was provided by the mothers, since pups from different litters showed phase relationships similar to those of their mothers. Thus the mothers provided a signal that was sufficient to cause entrainment, despite the 2-hr difference in free-running period between the mothers and pups. Although the pups' activity rhythms appeared to have been entrained by the mothers, they were clearly free-running by postnatal day 18. The mechanism for entrainment is lost during the course of development, despite continued interaction between the mothers and pups.     

Circadian and Ultradian Time Patterns in Human Behaviour: Part 1: Activity Monitoring of Families from Prepartum to Postpartum

Time patterns of activity-rest rhythms during and after pregnancy are increasingly recognised as important factors for the well-being and health of young families. This longitudinal study examined activity-rest patterns of couples during late pregnancy and subsequently the alterations in the periodic structure of parental and neonatal time patterns during the first four months after birth. Part I concentrates on the effects of late pregnancy and birth to the mother's rest-activity patterns and those of the father and, after birth, what time pattern the infant developed. Part II attempts to clarify how activity patterns of the entire family agree or disagree with each other and investigates how the infant synchronises with the environment that includes the process of parent-infant interaction. Activity data of, so far, seven families (father, mother and child) were continuously recorded using non-invasive Actiwatch units. Recordings of parental activity started at the beginning of the 37th week of gestation, and were continued in parallel with the infants' recordings in three series of three weeks each until four months after birth: 1st to 3rd week, 7th to 9th week and 13th to 15th week of life. In a standardised diary, record was kept of household routines, parental activities, type of feeding, initiation of sleep or waking up. Activity data of seven non-pregnant women were collected and used as a control. Irregular nocturnal activity epochs occurred frequently in pregnant women and were absent in non-pregnant women. Period lengthenings and shortenings of the circadian rhythms appeared in both parents from prepartum to postpartum. Activity at night increased from prepartum to postpartum in mothers and fathers. Three infants showed a marked circadian rhythm between day 3 and 14 after birth. All seven infants showed a predominant circadian rhythm between day 8 and 19 after birth. The onset of daytime activity of mothers and their infants corresponded well to each other. Postpartum frequency spectra of parents and child always had some ultradian components in common. Time patterns of activity-rest rhythms of couples and parents are shown to be altered during and after pregnancy and we suggest that the infants' adaptation to the environment begins during the first week that includes the process of mother-infant interaction.     

Characterization of coelenterazine analogs for measurements of Renilla luciferase activity in live cells and living animals

In vivo imaging of bioluminescent reporters relies on expression of light-emitting enzymes, luciferases, and delivery of chemical substrates to expressing cells. Coelenterazine (CLZN) is the substrate for a group of bioluminescent enzymes obtained from marine organisms. At present, there are more than 10 commercially available CLZN analogs. To determine which analog is most suitable for activity measurements in live cells and living animals, we characterized 10 CLZN analogs using Renilla luciferase (Rluc) as the reporter enzyme. For each analog, we monitored enzyme activity, auto-oxidation, and efficiency of cellular uptake. All CLZN analogs tested showed higher auto-oxidation signals in serum than was observed in phosphate buffer or medium, mainly as a result of auto-oxidation by binding to albumin. CLZN-f, -h, and -e analogs showed 4- to 8-fold greater Rluc activity, relative to CLZN-native, in cells expressing the enzyme from a stable integrant. In studies using living mice expressing Rluc in hepatocytes, administration of CLZN-e and -native produced the highest signal. Furthermore, distinct temporal differences in signal for each analog were revealed following intravenous or intraperitoneal delivery. We conclude that the CLZN analogs that are presently available vary with respect to hRluc utilization in culture and in vivo, and that the effective use of CLZN-utilizing enzymes in living animals depends on the selection of an appropriate substrate.     

Carbonic anhydrase activity in blood of early sheep fetuses

• 1.1. The level of carbonic anhydrase activity in the red cells was measured in sheep fetuses at different times after conception: 39, 56, 77, 90 and 140 days, the last being close to full term. Measurements were also made on blood from four of the mothers.
• 2.2. There was a low level of the enzyme present in the 39 day fetuses (0.037 enzyme units (E.U.)/100 μg Hb) and its increase up to 90 days of gestation (0.19 E.U./100 μg Hb) had a form approximating exponential.
• 3.3. The earliest levels were only 11% of the full term levels and only 4% of the adult levels previously reported.
• 4.4. Even the earliest samples were of blood that was fetal rather than embryonic but these results are the earliest carbonic anhydrase activities reported in this mammal.

     

Strain dependent response of circadian rhythms during exposure to continuous illumination

Enzymes activities were measured, at three hours intervals, during 30 hours, in various tissues of C57BL/6J and A/J male mice. The measurements, were carried out on mice which were exposed for two, five and twenty one days to continuous illumination. Identical measurements were performed also on mice which were kept in alternating 14 hours light: 10 hours dark. Activity patterns of each group were analysed to test the presence, or absence, of rhythm characteristics. The results of the experiments with C57BL/6J have been previously reported. The comparison of the results, which were obtained from the two strains revealed that under exposure to alternating light: dark conditions all activity patterns exhibited a significant circadian rhythm. Except for one enzyme (thymus GAPD), the times of peak activity (acrophase) were identical for all other examined enzymes, in both strains. On the other hand when the two strains were exposed to continuous illumination they differed in their response to the effect of continuous light. The activity of the same enzyme exhibited different periodicity and/or different acrophase in each of the two strains. This variability reflects the existence of genetic differences, between the strains in the free running behavior of these enzymes' activity rhythms.     

Subcellular redistribution of newly synthesized macrophage lysosomal enzymes. Correlation between delivery to the lysosomes and maturation

Cultured mouse peritoneal macrophages were fractionated by two methods at various times after pulse labeling with [35S]methionine. The lysosomal enzymes beta-glucuronidase and beta-galactosidase were isolated from each fraction by immunoprecipitation and electrophoresis on sodium dodecyl sulfate-acrylamide gels. Two distinct peaks of label were obtained on Percoll density gradients. An early appearing peak of low density, containing the precursor forms of both enzymes, co-sedimented with markers for the endoplasmic reticulum, the Golgi apparatus, and the plasma membrane. With time, immunoprecipitable label cosedimented with the bulk of the lysosomal enzyme activity at high density and corresponded to the mature forms of the lysosomal enzymes. By differential centrifugation, newly synthesized enzymes were found predominantly in small particle fractions, unlike the bulk of the lysosomal enzymic activity which was found in larger particle fractions. With increasing time, newly synthesized enzymes were transferred to assume a distribution similar to that of lysosomal enzymic activity. The results suggest that transport of newly synthesized enzymes to lysosomes and conversion to mature forms are closely linked events. Conversion of lysosomal precursors to mature forms occurs either in a prelysosomal vesicle or shortly after reaching the lysosome. The two enzymes follow similar subcellular pathways at similar rates. Also, the macrophage system appears suitable for direct analysis of newly synthesized lysosomal enzymes during subcellular transport.     

Longitudinal data for intrauterine levels of fetal IGF-I and IGF-II

OBJECTIVE: An increasing body of evidence supports a major role for the insulin-like growth factors (IGFs) in the control of human fetal growth. Individual data at various times of pregnancy suggest that IGF-I and IGF-II levels remain stable up to the 33rd week of pregnancy. Thereafter, both increase to reach values 2-3 times higher at term. In order to provide an accurate reflection of fetal IGFs in utero, we sampled fetal blood from the umbilical cord by cordocentesis. METHODS: We measured IGF-I and IGF-II in 12 fetuses longitudinally for up to 5 times between the 21st week of gestation and delivery. RESULTS: All patients showed a progressive increase in IGF-I and IGF-II levels. Data determined during different time intervals (before 29th, 29th to 32nd, after 32nd week) were compared and the main increase was found after the 32nd week. The median for IGF-I before the 29th week was 33.5 ng/ml (range 19-40.5) and increased to 41 ng/ml (32-59) between the 29th to 32nd and further to 54.1 ng/ml (range 17-70) thereafter. During the same time interval, the median for IGF-II increased from 217 ng/ml (86-326) to 349 ng/ml (227-467). In 7 patients, cord blood after delivery was available. For IGF-II a further increase was consistently found after birth (from 282 ng/ml (175-511) to 393 ng/ml (297-513)), whereas only 2 fetuses showed an increase in IGF-I. CONCLUSION: We conclude that in human fetuses, IGF-I and IGF-II levels increase longitudinally throughout pregnancy. Therefore, they may become important markers of healthy fetal development.     

Seasonal changes in the circadian activity rhythm of light-dark (LD) and completely dark (DD) regimen in the mouse submandibular gland in the presence of light-dark (LD) and completely dark (DD) regimen

Influence of seasons on circadian activity changes and the influence of one and six weeks of DD upon these changes of acid phosphatase (AP) and beta-acetylglucosaminidase (AM) was studied in the submandibular gland of sexually mature male mice. Total enzyme activity was determined in tissue homogenates at four-hour intervals in March, June, October, and February under standard LD12/12 conditions and after one and six weeks of the DD regime. The rhythms were analysed according to cosinor method. Under constant lighting conditions the seasonal differences in the AM circadian activity rhythm were found. AP activity was considerably less influenced by seasonal changes. Both enzyme activity changes were independent of each other and each rhythm was differently influenced by DD. In the case of AM the most pronounced circadian activity changes had the highest amplitude and mesor occurred in summer. The strongest influence of DD upon this enzyme activity rhythm was observed in spring and summer especially after the first week, after six weeks the acrophase returned to the LD group value (spring). In autumn and winter the reaction to DD was different to that of summer and spring. For AP the circadian changes of activity were non-rhythmic in spring, whereas in all other seasons the acrophases occurred almost at the same time in the afternoon. In DD the activity rhythm significantly changed after six weeks. In all seasons, except spring the circadian rhythm of activity was not observed after six weeks of DD. An attempt was made to explain the observed results by the certain kind of genetic memory present in laboratory animals the neurohormonal system of which is influenced by seasonal changes.     

Dopamine beta-hydroxylase in rat serum and lymph: changes with age and effect of cold exposure.

The pH optimum for rat serum dopamine beta-hydroxylase (DBH) (3,4-dihydroxyphenylethylamine, ascorbate:oxygen oxidoreductase (beta-hydroxylating)(EC 1.14.17.1) was 4.0 in acetate buffer; other requirements were as reported by others. DBH activity in serum of 20-day-old fetuses is slightly higher than in that of their mothers. Levels of the enzyme in blood a few hours after birth are almost five times greater than in the adult, remain high during the suckling period, then drop rapidly during the 4th week after birth to about three times the adult level, which is then slowly reached over the next few weeks. These fluctuations in serum DBH activity coincide with the period of intense development and maturation of the sympathetic nervous system. There was not significant effect of cold exposure on blood DBH activity when newborn, suckling, weanling or adult warm- and cold-acclimated rats were exposed to cold. Similarly, exposure to cold that elicited two- to three-fold increases in O2 consumption failed to increase DBH activity in thoracic duct lymph. Therefore serum and lymph DBH activities are not sensitive indices of sympathetic secretory activity in the intact rat.     

Blood and liver enzymes in rainbow trout (Salmo gairdneri Rich.) with emphasis on their diagnostic use: Study of CCl4 toxicity and a case of Aeromonas infection

The effect of one intraperitoneal injection of 1.33 ml of CCl₄ per kg of fish was studied. Two experimental series were performed and studied for 10 days (with diluent) and 24 h (pure CCl₄) periods. LDH, GOT, GPT, GR, GDH, CPK, G-6-Pase, and AlkPase were studied. The activity of all enzymes in blood increased: LDH (four times the control), GOT (two times), GPT (three times); they reached a maximal activity 12 h after injection of diluted CCl₄. The levels of some enzymes were also examined in the liver. With pure CCl₄, maximal enzyme activity in blood occurred earlier (6 h). A 6 to 10 times increase was observed for GOT, GPT, LDH, GR, and GDH. Histopathological observations were correlated with these enzymes studies.
An Aeromonas disease characterized by the destruction of the dermis, the exposure of the muscle, and by the presence of numerous petechiae in the liver enabled us to examine the relationships between naturally induced tissue damage and enzyme levels in blood. The levels of seven blood enzymes were determined and the most significant modifications were observed for LDH and CPK. which increased their concentration from 3 to 7 times respectively. A pyruvate saturation test demonstrated that LDH was probably from liver as it was observed after CCl₄ poisoning. The contribution of such biochemical studies in fish research is evaluated.     

(Na, K)ATPase activity in membrane preparations of ouabain-resistant HeLa cells.

Membrane preparations from two independent ouabain-resistant HeLa cell clones, HI-B1 and HI-C1, each appear to contain two species of (Na,K)ATPase. Two-thirds of the total (Na,K)ATPase in each mutant is indistinguishable from the enzyme in preparations of wild type cells with respect to ouabain binding, ouabain inhibition of (Na,K)ATPase activity, and dependence of ATP hydrolysis on Na, Mg, K, and ATP concentration. The remaining (Na,K)ATPase activity in the mutants is up to 1000 and 10 000 times, respectively, more resistant to ouabain than wild type enzyme. Resistance results from a lower affinity of the mutant enzymes for the inhibitor. The presence of Na, K, or Mg has little or no effect on the degree of resistance expressed by the mutant enzymes, although the resistance of the wild type enzyme varies 400-fold in the presence of different ligands. Incubation with 5 X 10(-8) M ouabain abolishes the activity of the wild type enzyme without affecting the activity of the resistant enzymes. Using this procedure we compared the parameters of ATP hydrolysis via the resistant and wild type enzymes. Ouabain-resistant (Na,K)ATPase of HI-C1 has an apparent K0.5 for potassium 3-4 times higher than that of either wild type enzyme or the resistant enzyme of HI-B1.     

Vaginal treatment with a prostaglandin F2 alpha derivative (alfaprostol) for inducing labor in pregnancy at term

The effectiveness and acceptability of Alfaprostol (an analog of PGF2 alpha) in inducing labor were assessed in 20 pregnant women at term. All subjects had no spontaneous uterine activity before treatment and the mean (M +/- SE) Bishop score was 2.45 +/- 0.21. The drug was administered by vaginal route at the dose of 10 mg every 3 hours. Regular uterine contractions appeared in all patients and delivery occurred in 85% of the patients after a mean time of 9h50min +/- 0h55min following the start of treatment. The mean dose of Alfaprostol utilized to achieve delivery was 29.4 +/- 2.0 mg. No major side effects were noted in the mothers and their fetuses at any time during treatment. Two patients exhibited vomiting. The Apgar score of all newborns at birth was 8 or more. These results suggest the usefulness of Alfaprostol to induce labor in pregnant women at term, as it has oxytocic activity without adverse effects on either the mother or the fetus.     

Age-dependent changes in the multiple forms of the soluble 17 beta-hydroxysteroid dehydrogenase of female rabbit liver.

The soluble NADP-dependent 17 beta-hydroxysteroid dehydrogenase activity of female rabbit liver increases with the age of the animal, the specific activity of the enzyme in the 56-day-old rabbit being 3 times that of the 28-day-old animal. The increase in activity is accompanied by a change in the molecular heterogeneity of the enzyme. Three forms (enzymes I, II and III) were identified in the liver cytosol of the 56-day-old female rabbit, whereas only one major form (enzyme IIIY) was present in the 28-day-old animal. Peptide maps of the four purified enzymes showed that there were minor differences in structure. The enzyme present in the liver of the 28-day-old rabbit was distinct from the three enzymes of the 56-day-old animal. All of the enzymes exhibited bifunctional activity, having 17 beta-hydroxysteroid dehydrogenase activity towards androgen and oestrogen substrates and 3 alpha-hydroxysteroid dehydrogenase activity towards androgens of the 5 beta-androstane series. The differences in substrate specificity of the enzymes paralleled their differences in structure. The data suggest that one enzyme (enzyme III) may have a special role in steroid metabolism during development in the female rabbit.