Preterm Rats Research Articles

Effects of oxygen-induced lung damage on megakaryocytopoiesis and platelet homeostasis in a rat model.

The association between lung injury and thrombocytopenia was investigated by comparing the megakaryocyte and platelet counts, and platelet activation using P-selectin as a marker, between the prepulmonary (right atrial) and postpulmonary (left atrial) blood in adult and neonatal (preterm and term) rats with and without hyperoxic lung injury. In the healthy controls, the postpulmonary blood had lower megakaryocyte count (prepulmonary versus postpulmonary: Preterm: 8.7[0.6] versus 3.9[0.3] per ml, p < 0.001; Term: 8.7[1.1] versus 2.6[0.4] per ml, p < 0.001; Adult: median [interquartile ranges]: 2.5[1.0, 5.0] versus 1.0[0, 3.0] per ml, p < 0.001), higher platelet count (prepulmonary versus postpulmonary: Preterm: 491.2[11.1] x 10(9)/L versus 595.1[10.2] x 10(9)/L, p < 0.001; Term: 472.5[19.9] x 10(9)/L versus 579.3[26.2] x 10(9)/L, p < 0.001; Adult: 513.9[31.5] x 10(9)/L versus 664.7[28.8] x 10(9)/L, p < 0.001), but similar P-selectin expression. In contrast, the lung-damaged animals did not show any such differences in either megakaryocyte or platelet count, but P-selectin expression was greater in the postpulmonary blood (prepulmonary versus postpulmonary: Preterm: 38.7[3.9] versus 56.4[4.9]% platelets, p = 0.02; Term: 40.9[2.0] versus 54.0[4.2]% platelets, p = 0.002; Adults: 30.0[3.6] versus 49.1[4.7]% platelets, p = 0.003). Peripheral platelet and intra-pulmonary megakaryocyte counts in the lung-damaged rats were significantly lower than those in their respective controls. Intra-pulmonary thrombi or platelet aggregation were detected in the lung-damaged rats but not in the controls. These findings showed that hyperoxic lung damage reduced circulating platelets through (1) failure of the lungs to retain and fragment megakaryocytes to release platelets, and (2) platelet activation leading to platelet aggregation, thrombi formation and platelet consumption. The magnitude of platelet reduction was physiologically significant, as demonstrated by higher counts of megakaryocyte colony forming units in the bone marrow culture of the animals in the hyperoxia group when compared with the controls.

Inhibition of in vivo constriction of fetal ductus arteriosus by endothelin receptor blockade in rats.

The fetal ductus can be constricted by drugs, including cyclooxygenase inhibitors (indomethacin), nitric oxide synthesis antagonists [N-nitro-L-arginine monomethyl ester (L-NAME)], and glucocorticoid hormones (dexamethasone). Constriction of the fetal ductus by endothelin (ET) 1 was reported in an in vitro study. We studied the preventive effect of a dual ET receptor antagonist (bosentan) and a selective ET-A blocker (CI-1020) on pharmacologic fetal ductal constriction in rats. Near-term pregnant Wistar rats at d 21 and preterm rats at d 19 were used. The fetal ductus was constricted by four medications: orogastric administration of indomethacin (10 mg/kg) on fetal d 21, orogastric indomethacin 1 mg/kg combined with muscular injection of L-NAME (10 mg/kg) on fetal d 21, and muscular injection of L-NAME or dexamethasone (1 mg/kg) on fetal d 19. Bosentan (0.1, 1, 10, or 100 mg/kg) was injected intraperitoneally either simultaneously with indomethacin, L-NAME, or dexamethasone, or 4 h after administration of 10 mg/kg indomethacin. CI-1020 (0.01, 0.1, 1, or 10 mg/kg) was injected intraperitoneally simultaneously with indomethacin. After maternal atlas dislocation, cesarean section, fetal whole-body freezing, and cutting on the freezing microtome, measurements were made of the inner diameters of the ductus, main pulmonary artery, and ascending aorta. Bosentan blocked fetal ductal constriction by indomethacin, indomethacin plus L-NAME in the near-term rats, and constriction by L-NAME and dexamethasone in the preterm rats dose dependently. Fetal ductal constriction was nearly completely blocked by simultaneously administered 100 mg/kg of bosentan or 10 mg/kg of CI-1020. Dual ET receptor antagonist (bosentan) and selective ET-A blocker (CI-1020) prevent constriction of the fetal ductus arteriosus induced by ductus-constricting agents in rats, indicating that ET and ET-A receptors are essential in fetal ductal constriction.

Regulation of matrix metalloproteinases (type IV collagenases) and their inhibitors in the virgin, timed pregnant, and postpartum rat uterus and cervix by prostaglandin E2-cyclic adenosine monophosphate

Objective: Our purpose was to examine whether the type IV collagenases (metalloproteinase [MMP]-2 and MMP-9) and their inhibitors (TIMP-1 and TIMP-2) are regulated by a prostaglandin E2 (PGE2)-cyclic adenosine monophosphate (cAMP) mechanism in nonpregnant virgin, preterm and term pregnant and postpartum rats. Study Design: Sprague-Dawley rats were infused with either saline solution or PGE2 over 24 hours or were noninfused. Plasma and tissue were analyzed for cAMP, MMP-2 and MMP-9, and TIMP-1 and TIMP-2. Results: PGE2 evoked elevations in plasma and tissue levels of cAMP and MMP-2 in the preterm and term pregnant rats. MMP-9 levels were elevated in the preterm plasma and uterus, whereas in the term pregnant and postpartum rats, MMP-9 levels were increased in the cervix. Conclusions: MMP levels in the pregnant and postpartum rat uterus and cervix are regulated in part by a PGE2-cAMP mechanism. Specifically, MMP-9 may be involved in preterm labor, cervical maturity, and involution of the postpartum uterus. (Am J Obstet Gynecol 2002;187:202-8.)

Phospholipase A2 activity in rat embryonic brain and in primary cultures of cortical neurons.

We investigated, with the aid of a radio-enzymatic method, the developmental changes in the activity of 85 kDa calcium-dependent cytosolic phospholipase A(2) (cPLA(2)) in pre-term embryonic rat brain and in primary cultures of cortical neurons. PLA(2) activity was highest in the brains of 18 day embryos (E18) and gradually decreased toward birth (E18 through E21). No significant differences were found regarding cPLA(2) activity in different topographies, namely hippocampus, cortex, and total brain. In primary cultures of E18/E19 cortical neurons, cPLA(2) activity was higher than the respective embryonic tissue activity, straight from early development. In vitro cPLA(2) activity peaked on the 4th day in culture, and decreased after the six days of incubation, when the cultures became mature. This result reinforces previous evidence that cPLA(2) plays an important role in the early development of the nervous system, and further suggests that it may be implicated in neurodevelopmental processes and in in vitro neuronal survival. We believe that we have produced a useful model for the tissue culture study of PLA(2) metabolism and biochemistry, which can be further addressed to the investigation of the biology of neurodegenerative and neuropsychiatric disorders.

Oxidative stress in preterm rat brain is due to mitochondrial dysfunction.

Prematurity-mediated cerebral damage has been associated with oxidative stress. The aim of the present work was to study the possible role played by free oxygen radicals generated by mitochondrial respiratory function in cerebral injury in preterm neonates. Our results show that whereas total glutathione concentrations are similar in term and preterm neonates, the GSH/GSSG ratio decreases sharply in preterm neonates immediately after birth. This effect is not due to a lack of enzymes involved in GSH regeneration, such as glutathione reductase and glucose-6-phosphate dehydrogenase, but to a significant increase in free-radical generation in preterm rat brain as shown by the increase in lipoperoxidation. Because the mitochondrion is the main source of free radicals in the cell, mitochondrial respiratory function was studied in the brain of preterm neonates. Our results show that prematurity prevented the postnatal increases in complex II-III activity and ATP concentrations that occur in term neonates at 5 min after delivery. All these effects were counteracted by the oxygen supply, suggesting that the inhibition of mitochondrial function is caused by restricted oxygen availability. Consequently, cerebral damage associated with prematurity may be mediated by mitochondrial free-radical generation as a consequence of hypoxia undergone by preterm neonates at birth.

Evaluation of the tocolytic effect of a selective cyclooxygenase-2 inhibitor in a mouse model of lipopolysaccharide-induced preterm delivery.

The inflammatory process is known to cause preterm delivery. Recently, a cyclooxygenase (COX)-2 inhibitor has been developed as an anti-inflammatory drug with few side-effects. We evaluated the COX-2 inhibitor, Celecoxib, for its tocolytic effects and side-effects on dams and pups using a lipopolysaccharide (LPS)-induced preterm delivery mouse model (preterm delivery rates; 95%). With administration of Celecoxib (50, 10, 1 and 0.3 mg/kg), the preterm labour rate was significantly reduced to 18, 30, 36 and 60% respectively. The prostaglandin F(2alpha)(PGF(2alpha)) and PGE(2) concentrations in murine uterine tissue 4 and 10 h after LPS treatment with Celecoxib (10 and 1 mg/kg) were significantly lower than those in the LPS-treated group without CELECOXIB: With administration of 10 or 100 mg/kg Celecoxib, the fetal ductus arteriosus was constricted significantly in preterm and near-term rats, although constriction rates in preterm rats were significantly lower than those in near-term rats. Reproductive and renal functions in offspring whose mothers were treated with LPS and Celecoxib were normal. These data demonstrate that Celecoxib could be used as a new therapy for preterm labour. However, careful attention to constriction of the fetal ductus arteriosus should be given.

The role of nitric oxide in hyperoxic lung injury in premature rats.

To investigate the role of nitric oxide (NO) in hyperoxic lung injury, the 3-day-old preterm rats were randomly assigned to four groups: group I (hyperoxia group), group II (hyperoxia + Nw-nitro-L-arginine methyl ester (L-NAME) group), group III (air group), and group IV (air + L-NAME) group. Group I and II were exposed to > or = 90% O2 for 3 or 7 days. Group II and IV received subcutaneous L-NAMEy on daily basis (20 mg/kg). After 3 day or 7 day exposure, the lung wet weight/dry weight ratio (W/D), total protein and malondialdehyde (MDA) in bronchoalveolar lavage fluid (BALF) and lung pathology were examined in all groups. NO content, expression of endothelial NOS (eNOS) and inducible NOS (iNOS) in lungs were measured in group I and III. Our results showed that after 3 day exposure, group I appeared acute lung injury characterized by the increase of MDA content (P < 0.01) and the presence of hyperaemia, red cell extravasation and inflammatory infiltration; after 7 day exposure, except MDA, total protein and W/D were also increased in comparison with group III (P < 0.01, 0.05), pathological changes were more severe than those after 3 day exposure. After 3 and 7 day exposure, total protein in group II was significantly increased as compared with group I (P < 0.01 for both). The pulmonary acute inflammatory changes were more obvious in group II than in group I. Occasionally, mild hemorrhage was detected in the lungs of group IV. BALF protein content in group IV was higher than that in group III after 7 day exposure (P < 0.01). After 3 and 7 day exposure, NO content in BALF were all significantly elevated in group I as compared with group III (P < 0.01 for all). In the lungs of group I, strong immunostaining for iNOS was observed in airway and alveolar epithelia, inflammatory cells, which were stronger than those in group III. Expression of iNOS in rats after 7 day hyperoxic exposure was stronger than that after 3 day exposure. Shortly after 7 day exposure, stronger immunostaining for eNOS in airway epithelia in group I than that in group III was seen. Our study suggested that treatment with L-NAME worsened acute hyperoxic lung injury in preterm rats and also had a deleterious effect on the rats exposed to air, indicating that endogenous nitric oxide may play a protective role in rats under both physiological and hyperoxic status. Hyperoxia can significantly upregulate the expression of iNOS and eNOS in inflammatory cells, epithelia in the lungs of preterm rats, promote NO generation, which suggests that endogenous NO may mediate the hyperoxic pulmonary damage. Over-stimulation of iNOS may contribute to the pathogenesis of hyperoxic lung injury. NO may have dual roles in pulmonary oxygen toxicity.

Effect of lipopolysaccharide on uterine contractions and prostaglandin production in pregnant rats

Objective: Our aim was to evaluate the effect of lipopolysaccharide on prostaglandin production and on contraction of isolated myometrial strips from preterm pregnant rats. Study Design: Pregnant Wistar rats on day 17 of gestation were killed 3 hours after intraperitoneal injection of lipopolysaccharide (1.5 mg/kg) or vehicle, with or without pretreatment with indomethacin (5 mg/kg administered intraperitoneally) 1 hour beforehand. Concentrations of endotoxin in maternal serum and amniotic fluid, prostaglandin F2α and prostaglandin E2 in amniotic fluid, and progesterone in maternal serum were determined. Longitudinal uterine strips were prepared, placed in organ chambers with Krebs-Ringer solution, aerated with 95% oxygen and 5% carbon dioxide (37°C, pH ~7.4), and equilibrated at 1g passive tension. Concentration-contraction relationships to oxytocin were determined. Samples of bathing solution were collected 10 minutes after the concentration of oxytocin was maximal. Prostaglandins and progesterone were measured by radioimmunoassay and endotoxin was measured by the Endospecy (Seikagaku Kogyo, Tokyo, Japan) kit. Results: Lipopolysaccharide treatment significantly increased the levels of prostaglandin F2α and prostaglandin E2 in amniotic fluid. Treatment with lipopolysaccharide inhibited the production and release of prostaglandin F2α and prostaglandin E2 that were activated by oxytocin in uterine strips and increased the sensitivity of strips to the contractile effect of oxytocin. Indomethacin did not affect the basal or the lipopolysaccharide-activated levels of endotoxin in serum and amniotic fluid and exerted a counteraction on lipopolysaccharide-induced increases in concentrations of prostaglandin F2α and prostaglandin E2 in amniotic fluid. Indomethacin counteracted oxytocin-activated production and release of prostaglandin F2α and prostaglandin E2 in uterine tissues after lipopolysaccharide administration without changing the sensitivity of uterine strips to oxytocin. Concentrations of progesterone were not changed after lipopolysaccharide, indomethacin, or their combined application, which suggests that the changes described were not associated with alterations in the levels of the hormone. Conclusions: The activation of the uterine contractile system by prostaglandin and oxytocin during intra-amniotic infection may be one of the causes of preterm delivery. A combination of an oxytocin receptor antagonist and an inhibitor of cyclooxygenase may be beneficial in prevention or treatment of preterm labor. (Am J Obstet Gynecol 2001;184:84-9.)

The role of nitric oxide in dilating the fetal ductus arteriosus in rats.

Prostaglandin E is a major dilator of the fetal ductus arteriosus (DA), but the role of nitric oxide in fetal ductal dilation has not been established. We studied the effects of a potent nitric oxide synthase inhibitor, Nomega-nitro-L-arginine methyl ester (L-NAME), on the fetal DA in rats. L-NAME was injected into the dorsum of pregnant rats, and fetal DA was studied 4 h later with a rapid whole body freezing method. The inner diameters of the DA and the main pulmonary artery were measured on a freezing microtome. The inner diameter ratio of DA to main pulmonary artery (DA/PA) was 1.02+/-0.03 (mean +/- SEM; number of fetuses [n], 21) in normal near-term fetuses. The effect of prostaglandin synthesis inhibition was studied after orogastric administration of indomethacin to pregnant rats. In near-term rats on the 21st day of gestation (term, 21.5 d), a large dose of L-NAME (100 mg/kg) caused only mild ductal constriction, with DA/PA reduced to 0.83+/-0.05 (n = 20). Indomethacin (1 mg/kg) caused moderate ductal constriction, and DA/PA was decreased to 0.65+/-0.05 (n = 21). Combined administration of L-NAME (10 mg/kg) and indomethacin (1 mg/kg) caused severe ductal constriction, with DA/PA of 0.26+/-0.03 (n = 16). In preterm rats on the 19th day of gestation, a moderate dose of L-NAME (10 mg/kg) caused severe ductal constriction, with a DA/PA of 0.32+/-0.05 (n = 24). Indomethacin (1 mg/kg) alone caused only mild ductal constriction, with DA/PA 0.86+/-0.02 (n = 16). In conclusion, prostaglandin has a major role and nitric oxide has a minor role in dilating the DA in the near-term fetal rat. In contrast, nitric oxide has a major role and prostaglandin has a minor role in dilating the DA in preterm fetal rats.

Spontaneous and Nitrosourea-Induced Primary Tumors of the Central Nervous System in Fischer 344 Rats Chronically Exposed to 836 MHz Modulated Microwaves

We have tested an 836.55 MHz field with North American Digital Cellular (NADC) modulation in a 2-year animal bioassay that included fetal exposure. In offspring of pregnant Fischer 344 rats, we tested both spontaneous tumorigenicity and the incidence of induced central nervous system (CNS) tumors after a single dose of the carcinogen ethylnitrosourea (ENU) in utero, followed by intermittent digital-phone field exposure for 24 months. Far-field exposures began on gestational day 19 and continued until weaning at age 21 days. Near-field exposures began at 35 days and continued for the next 22 months, 4 consecutive days weekly, 2 h/day. SAR levels simulated localized peak brain exposures of a cell phone user. Of the 236 original rats, 182 (77%) survived to the termination of the whole experiment and were sacrificed at age 709-712 days. The 54 rats (23%) that died during the study ("preterm rats") formed a separate group for some statistical analyses. There was no evidence of tumorigenic effects in the CNS from exposure to the TDMA field. However, some evidence of tumor-inhibiting effects of TDMA exposure was apparent. Overall, the TDMA field-exposed animals exhibited trends toward a reduced incidence of spontaneous CNS tumors (P < 0. 16, two-tailed) and ENU-induced CNS tumors (P < 0.16, two-tailed). In preterm rats, where primary neural tumors were determined to be the cause of death, fields decreased the incidence of ENU-induced tumors (P < 0.03, two-tailed). We discuss a possible approach to evaluating with greater certainty the possible inhibitory effects of TDMA-field exposure on tumorigenesis in the CNS.

Accelerated maturation of fetal ductus arteriosus by maternally administered vitamin A in rats.

Maturation of fetal ductus arteriosus is associated with increased constriction in response to maternally administered indomethacin. Recently retinoic acid has been shown to be important in development of the fetal ductus arteriosus. To determine whether retinoid might be of value in the treatment of patent ductus arteriosus in premature infants, we studied the response of fetal ductus arteriosus to indomethacin with and without pretreatment with vitamin A (1 mg (3000 IU)/kg, intramuscular injection) in near-term and preterm rats. Maturation of the ductus arteriosus was studied by measuring the inner diameters of the ductus arteriosus (D) and main pulmonary artery (P) to get D/P ratio 4 h after orogastric administration of 1 mg/kg indomethacin. D/P was 1.0 in the fetus before administration of indomethacin. In near-term fetuses on the 21st d without vitamin A, D/P decreased to 0.54 with indomethacin, whereas it decreased to 0.27 (p < 0.05) in those with pretreatment with vitamin A on the 19th and 20th d. In preterm fetuses on the 20th d without pretreatment with vitamin A, D/P decreased to 0.82 with indomethacin, whereas it decreased to 0.66 (p < 0.05) in those with pretreatment with vitamin A on the 19th d. It is concluded that maternally administered vitamin A accelerates maturation of the ductus arteriosus in fetal rats.

Missing Links in Perinatal Hypoxic Brain Injury in the Preterm Rat Pup. 1756

Missing Links in Perinatal Hypoxic Brain Injury in the Preterm Rat Pup. 1756

Prenatal dexamethasone administration to premature rats exposed to prolonged hyperoxia: A new rat model of pulmonary fibrosis (bronchopulmonary dysplasia)

Objective: To evaluate the postnatal effects of prenatal dexamethasone treatment of preterm rats and to test the hypothesis that prenatal dexamethasone treatment protects against pulmonary oxygen toxicity in the preterm rats and stimulates lung antioxidant enzyme levels in response to hyperoxia. Study design: We administered dexamethasone (0.4 mg/kg, intraperitoneally), or equivolume saline solution to pregnant rats at 48 and 24 hours before premature delivery at gestation day 21. Both groups of prematurely delivered rat pups were randomly assigned to either >95% O 2 or room air immediately after birth and brief resuscitation. Results: The hyperoxic survival rates from day 1 through day 14 were similar in both dexamethasone-treated and control preterm O 2 groups. At 7 days of hyperoxia, the preterm pups demonstrated similar lung antioxidant enzyme activity and surfactant content responses to high O 2 in the dexamethasone-treated and control groups. Lung quantitative morphometry changes were similar (equal degree of inhibition of normal alveolar development) in both groups. Unexpectedly, the lungs of the preterm O 2 control rats showed evidence of septal fibrosis and the pups that received dexamethasone-O 2 showed even greater severity of septal fibrosis and a greater increase (+50%) of lung hydroxyproline compared with the O 2 group control rats. Conclusions: In preterm animals, prenatal dexamethasone administration does not show any of the hypothesized protective effects against hyperoxia or protective biochemical lung changes during prolonged O 2 exposure. However, prenatal dexamethasone administration with prolonged exposure of the preterm rat to hyperoxia results in a pulmonary pathologic picture quite similar to bronchopulmonary dysplasia. (J Pediatr 1997;130:409-16)

PRENATAL DEXAMETHASONE (DEX) ADMINISTRATION TO PREMATURE RATS EXPOSED TO PROLONGED HYPEROXIA: A NEW RAT MODEL OF PULMONARY FIBROSIS[BRONCHOPULMONARY DYSPLASIA (BPD)]. † 1953

Although prenatal DEX treatment is known to improve hyperoxic survival in term newborn rats and to stimulate the responses of the lung antioxidant enzyme (AOE) system to hyperoxia, little is know about the postnatal effects of prenatal DEX on prolonged exposure to hyperoxia in preterm animals. To examine the hypothesis that prenatal DEX treatment would also show protective effects in preterm rats exposed to hyperoxia, we administered DEX(0.4 mg/kg, i.p.), or equivolume saline to pregnant rats at 48 and 24 h prior to premature delivery at gestation d 21 (term=22 d). Both groups of preterm rat pups were randomized to either >95% O2 or room air immediately after birth. The hyperoxic survival rates from d 1 through d 14 turned out to be similar in both DEX- and control-O2 groups. At 7 d of hyperoxia, the preterm pups demonstrated similarly increased lung AOE activity and surfactant content responses to high O2 in the DEX and control groups. Lung quantitative morphometry was changed similarly (equal degree of inhibition of normal alveolar development) in both O2 groups. Unexpectedly, the lungs of preterm control-O2 pups showed evidence of septal fibrosis at 13-14 d in O2, and the DEX-O2 group showed even much greater severity of septal fibrosis and a greater increase of lung hydroxyproline compared with the O2 controls. We conclude that in preterm animals, prenatal DEX administration does not show any of the hypothesized protective effects during prolonged O2 exposure. However, prenatal DEX administration with prolonged exposure of the preterm rat to hyperoxia results in a pulmonary pathologic picture quite similar to BPD. This preterm rat model is a much more practical experimental model than the earlier excellent (but very expensive) baboon/monkey model of BPD. Table

Premature rats treated with propylthiouracil show enhanced pulmonary antioxidant enzyme gene expression and improved survival during prolonged exposure to hyperoxia.

In full-term newborn rats, propylthiouracil (PTU) treatment has been previously shown to decrease susceptibility to O2-induced lung damage and improve survival during hyperoxic exposure. However, no differences were found in lung antioxidant enzyme (AOE) activity responses to hyperoxia compared with O2-exposed untreated (control) term rats. To further explore possible pulmonary protective effects of PTU treatment in prematurely delivered animals, we administered PTU (0.015%) in drinking water to timed-pregnant rats for the final 10 d of gestation prior to delivery 1 d before term, and during lactation; control pregnant/nursing rats received untreated water. Both groups of 21-d premature rat pups were randomized to either > 95% O2 or room air exposure after birth for up to 14 d. The left lungs of 7-d exposure pups were used to quantitate the concentrations of AOE mRNA by solution hybridization; the right lungs of the same pups were assayed for AOE activities. PTU treatment resulted in survival rates of O2-exposed preterm rat pups that were consistently higher at all time periods in hyperoxia including 7 d [PTU, 67 of 82 (82%) versus control pups, 58 of 113 (51%); p < 0.001] and 14 d [PTU, 31 of 39 (79%) versus control, 15 of 66 (23%); p < 0.001]. Further evidence of increased tolerance to > 95% O2 in PTU pups included a significant decrease in the incidence of microscopic intraalveolar edema and a significant increase in lung tissue surfactant-related phospholipids compared with O2-exposed control pups.(ABSTRACT TRUNCATED AT 250 WORDS)

Mammalian adaptation to extrauterine environment: mitochondrial functional impairment caused by prematurity

In this paper we report that, compared with term rat neonates, both mitochondrial content and function are diminished in liver of preterm neonates (delivered 24 h before full term) compromising cellular energy provision in the postnatal period. In addition, there is a parallel reduction in the content of mRNAs encoding mitochondrial proteins in preterm rats. Also, efficient oxidative phosphorylation is not attained in these pups until 3 h after birth. Although isolated liver mitochondria from preterm neonates show a two-fold increase in F1-ATPase beta-subunit and cytochrome c oxidase activity 1 h after birth, the abnormal coupling efficiency between respiration and oxidative phosphorylation (ADP/O ratio) is due to maintenance of high H(+)-leakage values in the inner mitochondrial membrane. Postnatal reduction of the H+ leak occurs concomitantly with an increase in intra-mitochondrial adenine nucleotide concentration. Accumulation of adenine nucleotides in preterm and term liver mitochondria parallels the postnatal increase in total liver adenine nucleotides. Delayed postnatal induction of adenine biosynthesis most likely accounts for the lower adenine nucleotide pool in the liver of preterm neonates. The delayed postnatal accumulation of adenine nucleotides in mitochondria is thus responsible for the impairment in oxidative phosphorylation displayed by organelles of the preterm liver.

Changes in Mitochondrial Rhodamine-123-Fluorescence Populations of Rat Hepatocytes in Primary Culture

Flow cytometry analysis of mitochondria isolated from rat hepatocytes at different stages of development revealed two different rhodamine-123-stained fluorescence populations distinguishable by their main fluorescence channel. The high-fluorescence population (HFP) was minor, accounting for about 32, 33, and 23% of the total mitochondrial fraction in hepatocytes from pre-term, term, and early (1 h) newborn rats, respectively. The percentage of HFP decreased during the first 20 h of hepatocytes in culture from preterm and term fetuses but not those from early newborns, in which the decrease occurred during the second day of culture. However, after 20 h in culture more than 80% of hepatocytes reached the G0/G1 phase, whatever the stage of development in which they were sown. This suggests that the observed changes in the fluorescence populations are not associated with the cell cycle; instead these changes mimic those observed during postnatal development and are therefore presumably due to mitochondrial differentiation.

The Pre‐ and Postnatal Development of Na+/K+‐ATPase in Gastrointestinal Organs of the Rat

SummaryDevelopmental changes in Na+/K+‐ATPase enzyme activity have been documented postnatally in a number of organs, but little is known about prenatal levels or the factors affecting them. This study determined the normal developmental patterns of Na+/K+ ‐ATPase activity in the pancreas, liver, small intestine, kidneys, and colon of pre‐ and postnatal rats. In addition the effect of betamethasone administration on enzyme activity was determined both antenatally and postnatally. The individual organs showed a variable pattern of maturational change in enzyme activity. In the pancreas, changes in Na+/K+‐ATPase activity appeared to reflect the changing ratio of ductular to acinar cells. The pattern of enzyme activity in the small intestine was similar to that of the kidney and was inversely related to that of the colon. Enzyme induction by administration of betamethasone was also variable and bore no relationship to the DNA/protein ratio. In the preterm rat, betamethasone increased the Na+/K+‐ATPase activity only in the colon. Use of steroids to increase enzyme activity in the colon at an early stage of development may have therapeutic implications for treating abnormalities of potassium and sodium homeostasis in the preterm infant.

The pre- and postnatal development of Na+/K(+)-ATPase in gastrointestinal organs of the rat: effect of betamethasone treatment.

Developmental changes in Na+/K(+)-ATPase enzyme activity have been documented postnatally in a number of organs, but little is known about prenatal levels or the factors affecting them. This study determined the normal developmental patterns of Na+/K(+)-ATPase activity in the pancreas, liver, small intestine, kidneys, and colon of pre- and postnatal rats. In addition the effect of betamethasone administration on enzyme activity was determined both antenatally and postnatally. The individual organs showed a variable pattern of maturational change in enzyme activity. In the pancreas, changes in Na+/K(+)-ATPase activity appeared to reflect the changing ratio of ductular to acinar cells. The pattern of enzyme activity in the small intestine was similar to that of the kidney and was inversely related to that of the colon. Enzyme induction by administration of betamethasone was also variable and bore no relationship to the DNA/protein ratio. In the preterm rat, betamethasone increased the Na+/K(+)-ATPase activity only in the colon. Use of steroids to increase enzyme activity in the colon at an early stage of development may have therapeutic implications for treating abnormalities of potassium and sodium homeostasis in the preterm infant.

Antioxidant enzyme responses to hyperoxia in preterm and term rats after prenatal dexamethasone administration.

Although prenatal steroid therapy is known to enhance in utero maturation of the surfactant and antioxidant enzyme systems, little is known about the effects of steroids on the antioxidant system after birth. We measured activities of the antioxidant enzymes, catalase, superoxide dismutase, and glutathione peroxidase, in lung homogenates from both preterm and term rat pups after prenatal dexamethasone treatment. Enzyme activities were measured at birth and after exposure to > 98% oxygen. Dexamethasone treatment resulted in significantly higher survival of the preterm pups at 24 h (91.3% for dexamethasone versus 57% for saline). In preterm pups, the activities of catalase and superoxide dismutase at birth were higher after dexamethasone treatment (p < 0.05). However, after 24 h of hyperoxic exposure, there were no differences in activities of any of the antioxidant enzymes between the dexamethasone and control groups of prematurely born pups. In term pups, antioxidant enzyme activities did not differ significantly at birth; nor did they differ after 24 to 72 h of hyperoxic exposure in the dexamethasone and control treatment groups. Our results indicate that although prenatal dexamethasone treatment augments survival and catalase and superoxide dismutase activities at birth in preterm rat pups, dexamethasone does not result in altered early postnatal antioxidant enzyme activities after exposure to hyperoxia.