Neonatal Period In Rat Research Articles

Spleen Morphogenesis during the Neonatal Period in Rats Exposed to Endocrine Disruptor DDT

Spleen morphogenesis during the neonatal period in rats exposed in prenatal and postnatal development to low doses of dichlorodiphenyltrichloroethane (DDT), a persistent universal pollutant with endocrine disrupting properties, was studied. More intensive formation of periarterial lymphoid sheaths and marginal zone and simultaneously decreased rate of B-cell differentiation in the spleen were revealed. A higher content of differentiating T-cells and a lower number of cytotoxic T-lymphocytes by the end of the first week of life indicates a decrease in the differentiation of the latter. A lower content of neutrophils in the marginal zone also indicates a delay in the rate of functional development of lymphoid tissue, as opposed to morphological, in rats developing under exposure to low doses of DDT.

Morphogenesis of the Spleen during the Neonatal Period in Rats Exposed to the Endocrine Disruptor DDT

Morphogenesis of the Spleen during the Neonatal Period in Rats Exposed to the Endocrine Disruptor DDT

Distribution of GABAergic Interneurons and the GABAergic Transporter GAT1 in the Layers of the Neocortex during the Neonatal Period in Rats

Distribution of GABAergic Interneurons and the GABAergic Transporter GAT1 in the Layers of the Neocortex during the Neonatal Period in Rats

Neonatal kaempferol exposure attenuates impact of cerebral palsy model on neuromotor development, cell proliferation, microglia activation, and antioxidant enzyme expression in the hippocampus of rats

ABSTRACT Objectives This study aims to assess the effect of neonatal treatment with kaempferol on neuromotor development, proliferation of neural precursor cells, the microglia profile, and antioxidant enzyme gene expression in the hippocampus. Methods A rat model of cerebral palsy was established using perinatal anoxia and sensorimotor restriction of hindlimbs during infancy. Kaempferol (1 mg/ kg) was intraperitoneally administered during the neonatal period. Results Neonatal treatment with kaempferol reduces the impact of the cerebral palsy model on reflex ontogeny and on the maturation of physical features. Impairment of locomotor activity development and motor coordination was found to be attenuated by kaempferol treatment during the neonatal period in rats exposed to cerebral palsy. Neonatal treatment of kaempferol in cerebral palsy rats prevents a substantial reduction in the number of neural precursor cells in the dentate gyrus of the hippocampus, an activated microglia profile, and increased proliferation of microglia in the sub-granular zone and in the granular cell layer. Neonatal treatment with kaempferol increases gene expression of superoxide dismutase and catalase in the hippocampus of rats submitted to the cerebral palsy model. Discussion Kaempferol attenuates the impact of cerebral palsy on neuromotor behavior development, preventing altered hippocampal microglia activation and mitigating impaired cell proliferation in a neurogenic niche in these rats. Neonatal treatment with kaempferol also increases antioxidant defense gene expression in the hippocampus of rats submitted to the cerebral palsy model.

Expression of Parvalbumin and β-III-Tubulin in Cells of the Subventricular Zone during the Neonatal Period in Rats

Expression of Parvalbumin and β-III-Tubulin in Cells of the Subventricular Zone during the Neonatal Period in Rats

Immunohistochemical features of expression and distribution of antibodies to thyroglobulin in the thyroid glands of newborn rats after prenatal exposure of dexamethasone

In clinical practice, synthetic glucocorticoids, such as dexamethasone, are used to accelerate fetal maturation in pregnant women at risk of preterm birth. Increasing the concentration of cortisol in humans and other mammals often causes structural and functional changes in fetal tissues, preparing it for childbirth and extrauterine life, but they can have long-term consequences in the structural organization of organs postnatally. Despite the large number of studies on the effect of glucocorticoids on the fetus, there are almost no data on the prenatal effect of dexamethasone on the processes of synthesis and resorption of thyroglobulin by thyroid thyrocytes in the postnatal period of life. Therefore, the aim of the study was to determine the immunohistochemical features of expression and distribution of antibodies to thyroglobulin in the thyroid glands of newborn rats in normal and after prenatal exposure to dexamethasone. The study material was the thyroid gland of Wistar rats aged 1 to 7 days of postnatal development (54 animals): I group - intact animals (norm); ІІ group - control, animals which were injected with 0.9% NaCl solution at a dose of 0.05 ml to each fetus on the 18th day of dated pregnancy; III group - experimental animals, which were administered a solution of dexamethasone at a dilution of 1:40 at a dose of 0.05 ml to each fetus on the 18th day of the date of pregnancy operatively during laparotomy, by intrauterine, transdermal subcutaneous injection into the interscapular area (Ukrainian patent №112288). Thyroglobulin Antibody (2H11) monoclonal antibodies: sc-51708 from Santa Cruz Biotechnology, Inc. were used for immunohistochemical study. Photo documentation of the studied objects was performed using a “Primo Star” microscope (Carl Zeiss, Germany) using an AxioCam camera using the Zeiss Zen program (2011). Analysis of micropreparations of thyroid glands of intact and control rats showed invariance of thyroglobulin synthesis and its accumulation, which was expressed by sufficient immunohistochemical expression of antibodies to thyroglobulin (ТgAb+). Prenatal administration of dexamethasone leads to intensification of the processes of morphological development of hormone-producing structures (follicles and follicular epithelium), production, resorption and iodination of thyroglobulin. This is evidenced by immunohistochemical studies found in 1-3 days of the neonatal period. It should be noted that on the 7th day of life the newborn was found intense changes in the structure of the parenchyma of the thyroid gland of animals of the experimental group: increased relative percentage of follicle cavity due to increased number of large and medium, some follicles had no resorption vacuoles which was accompanied by a slowdown in the excretion of hormones into the bloodstream and led to overstretching of the follicles and, as a consequence, to the flattening of the thyroid epithelium. Intrauterine administration of dexamethasone leads to prenatal acceleration of structure formation, folliculogenesis and enhancement of hormone-producing function, which is confirmed by the peculiarities of immunohistochemical expression of TgAb. By the end of the neonatal period in rats prenatally exposed to dexamethasone, the thyroid gland is depleted of compensatory-reactive internal reserves, which is morphologically and immunohistochemically manifested by signs of hypofunction and hypertrophy. Thus, detected in the thyroid glands of animals prenatally exposed to dexamethasone, aberration of cytoplasmic expression of ТgAb+, intensification of colloidal expression of ТgAb+, flattening of thyroid epithelium, and the absence of resorption vacuoles are signs of impaired hormone-forming function, which is the morphological basis for the development of hypofunctional states and requires further study.

The suppressive effect of immune stress on LH secretion is absent in the early neonatal period in rats

Some physiological functions display weak responses to stress in the early neonatal period; i.e., they exhibit stress hyporesponse periods. In this study, we evaluated whether gonadotropin regulatory factors exhibit stress hyporesponsive periods in male and female rats. Rats were intraperitoneally injected with lipopolysaccharide (100μg/kg) (LPS group) or saline (control group) on postnatal day (PND) 5, 10, 15, or 25. Then, their serum luteinizing hormone (LH) concentrations and hypothalamic mRNA levels of gonadotropin regulatory factors; i.e., kisspeptin (Kiss1), the kisspeptin receptor (Kiss1r), and gonadotropin-releasing hormone (GnRH), were measured at 2h after the injection. The hypothalamic mRNA levels of pro-inflammatory cytokines were also measured because they suppress gonadotropin secretion. The serum LH concentration of the LPS group was lower than that of the control group at PND25 in both sexes, but no such difference was seen at PND5, 10, or 15 in either sex. In both sexes, the hypothalamic tumor necrosis factor (TNF)α and interleukin (IL)-6 mRNA expression levels of the LPS group were higher than those of the control group at PND25, but not at PND5 or 10. The hypothalamic IL-1β mRNA expression level of the LPS group was higher than that of the control group at all time points. The hypothalamic Kiss1, Kiss1r, and GnRH mRNA expression levels of the LPS and control groups did not differ at any time point in either sex. These findings suggest that gonadotropin regulatory factors exhibit stress hyporesponse periods. The hypothalamic–pituitary–gonadal axis (HPG) might become responsive to immune stress between PND15 and 25, which could be related to enhanced hypothalamic cytokine expression. The avoidance of infectious stress during the early neonatal period might be important for normal development of the HPG axis.

Reactions of neural elements of neocortex to action of hypoxia at the early neonatal period in rats

In this work we studied reactions of neural elements of various neocortex areas (sensomotor, visual, auditory) on action of acute normobaric hypoxia. The study is performed on the model of human premature pregnancy (action of normobaric hypoxia on rat pups at the 2nd postnatal day). There are revealed monotypical and monodirected structural reconstructions in all studied neocortex parts. The chosen parameters of hypoxic action initiate several direct reactions as early as at the next day: a decrease in sizes of cell bodies and in volume of the cytoplasm, as well as an enhancement, as compared with control, of the apoptotic cell death. By the end of the neonatal period (5 days), several ultrastructural alterations indicating deceleration of processes of differentiation of nerve cells are revealed: arrest of processes of complication of smooth and rough endoplasmic reticulum and of Golgi apparatus, a small number of single ribosomes and polysomes in the cytoplasm, a decrease of the number of growth cones of axons and dendrites in neuropil, delay and disturbance of myelination processes in nerve fibers. The detected morphofunctional reconstructions can serve the structural ground for development of neonatal encephalopathies.

Maternal Transfer of the Cyanobacterial Neurotoxin β-N-Methylamino-L-Alanine (BMAA) via Milk to Suckling Offspring

The cyanobacterial neurotoxin β-N-methylamino-L-alanine (BMAA) has been implicated in the etiology of neurodegenerative disease and proposed to be biomagnified in terrestrial and aquatic food chains. We have previously shown that the neonatal period in rats, which in humans corresponds to the last trimester of pregnancy and the first few years of age, is a particularly sensitive period for exposure to BMAA. The present study aimed to examine the secretion of 14C-labeled L- and D-BMAA into milk in lactating mice and the subsequent transfer of BMAA into the developing brain. The results suggest that secretion into milk is an important elimination pathway of BMAA in lactating mothers and an efficient exposure route predominantly for L-BMAA but also for D-BMAA in suckling mice. Following secretion of [14C]L-BMAA into milk, the levels of [14C]L-BMAA in the brains of the suckling neonatal mice significantly exceeded the levels in the maternal brains. In vitro studies using the mouse mammary epithelial HC11 cell line confirmed a more efficient influx and efflux of L-BMAA than of D-BMAA in cells, suggesting enantiomer-selective transport. Competition experiments with other amino acids and a low sodium dependency of the influx suggests that the amino acid transporters LAT1 and LAT2 are involved in the transport of L-BMAA into milk. Given the persistent neurodevelopmental toxicity following injection of L-BMAA to neonatal rodent pups, the current results highlight the need to determine whether BMAA is enriched mother's and cow's milk.

Noxious stimuli in the neonatal period in rats can cause important peripheral nervous system alterations that persist on adults in a sex-dependent manner

Advances inmedicine have increased the survival of pretermneonates, that are constantly submitted to invasive procedures, such as heelsticks, insertion of indwelling venous and arterial catheters, causing repetitive painful experiences in the neonate period. However, the consequence of early painful experiences in the peripheral nervous system remains unexplored. We aimed to evaluate the possible alterations in the sural nerves of male and female adult Wistar rats, after painful stimulation in the neonatal period.Wistar ratswere followed birth to 180 days of life, separated in 4 groups: 1) Control-male group; 2) Control-female group; 3) Pain-male group; 4) Pain-female group. Pain groups received repetitive needle insertion in plantar and lateral area of the right paw, twice a day for 15 days starting at birth. Control groups were stimulated with a cotton swab, twice per day for 15 days starting at birth. When animals completed 180 days of life, they were killed, and the sural nerves were dissected, and prepared for light microscopy. Visual morphometry was performed with the aid of computer software tomeasure the fascicular andmyelinated fiber parameters (fascicular area, number and density of myelinated fibers and Schwann cell nuclei, area and diameter of myelinated fibers and respective axons and g ratio). Female rats from the pain group show smaller sural nerve, myelinated fibers, and myelinated axon areas compared to female controls. Furthermore, the myelinated fibers diameter distribution shows that the female pain group present larger number of smaller fiber. Males showed no significant difference between groups. These results suggest that male and females can respond on different ways to neonatal injury with females showing changes in peripheral sensory nerve morphometry. In conclusion, noxious stimuli in the neonatal period in rats can cause important peripheral nervous system alterations that persist on adults in a sex-dependent manner.

Maternal Depression Model: Long-Lasting Effects on the Mother Following Separation from Pups

This study was carried out to ascertain the effects of maternal separation (3 h per day) of mothers from their pups in the neonatal period in rats, which has been suggested to induce a depressive-like state, would have long lasting effects on different parameters including hippocampal Na(+), K(+)-ATPase activity, NO production, free radical production and antioxidant enzymes activities in dams. Fourty-eight Wistar rats were divided into 3 groups: control, brief separation (10 min) and long separation (3 h). The neonatal interventions were done on postpartum days 1-10. At 35 days post-partum the dams were killed and the hippocampal Na(+), K(+)-ATPase activity were measured, as well as the activity of the antioxidant enzymes catalase, glutathione peroxidase, superoxide dismutase, free radicals production, and the production of nitric oxide. Hippocampal Na(+), K(+)-ATPase activity was decreased in the brief separated group and in dams subjected to 3 h separation from their pups. A reduction in nitric oxide levels in the hippocampus in dams of the long separated group was also observed. It is concluded that the withdrawal of pups from their mothers make the mothers more susceptible to the development of neurochemical alterations that could be related to depressive features.

Rescue of social behavior impairment by clozapine and alterations in the expression of neuronal receptors in a rat model of neurodevelopmental impairment induced by GRPR blockade

We have previously shown that pharmacological blockade of the gastrin-releasing peptide receptor (GRPR) during the neonatal period in rats produces behavioral features of developmental neuropsychiatric disorders. Here, we show that social interaction deficits in this model are reversed by the atypical antipsychotic clozapine given in the adulthood. In addition, we analyzed the mRNA expression of three neuronal receptors potentially involved in the etiology of disorders of the autism spectrum. Rats were injected with the GRPR antagonist RC-3095 or saline (SAL) from postnatal days 1-10, and tested for social behavior and recognition memory in the adulthood. One hour prior to the behavioral testing, rats were given a systemic injection of clozapine or saline. The mRNA expression of the NR1 subunit of the N-methyl-D-aspartate (NMDA) receptor, the epidermal growth factor receptor (EGFR), and GRPR was measured in the hippocampus and cortex of a separate set of rats given RC-3095 or SAL neonatally. Rats given neonatal RC-3095 showed decreased social interaction and impaired object recognition memory. Clozapine rescued the social interaction impairment. Neonatal treatment with RC-3095 also resulted in dose-dependent decreases in the expression of GRPR, NR1, and EGFR in the cortex, whereas all three receptor mRNAs were increased in the hippocampus in rats treated with the lower dose of RC-3095. The results contribute to further validate the novel rat model of neurodevelopmental disorders induced by GRPR blockade, and shows alterations in the expression of neuronal receptors in this model.

Low Concentrations of Alcohol Inhibit BDNF-Dependent GABAergic Plasticity via L-type Ca2+Channel Inhibition in Developing CA3 Hippocampal Pyramidal Neurons

Fetal alcohol spectrum disorder (FASD) is associated with learning and memory alterations that could be, in part, a consequence of hippocampal damage. The CA3 hippocampal subfield is one of the regions affected by ethanol (EtOH), including exposure during the third trimester-equivalent (i.e., neonatal period in rats). However, the mechanism of action of EtOH is poorly understood. In CA3 pyramidal neurons from neonatal rats, dendritic BDNF release causes long-term potentiation of the frequency of GABAA receptor-mediated spontaneous postsynaptic currents (LTP-GABAA) and this mechanism is thought to play a role in GABAergic synapse maturation. Here, we show that short- and long-term exposure of neonatal male rats to low EtOH concentrations abolishes LTP-GABAA by inhibiting L-type voltage-gated Ca2+ channels. These findings support the recommendation that even light drinking should be avoided during pregnancy.

A behavioural characterization of neonatal infection-facilitated memory impairment in adult rats

We have reported that exposure to bacteria ( Escherichia coli) during the neonatal period in rats is associated with impaired memory for a novel context in adulthood. However, impairment is only observed if a peripheral immune challenge (bacterial lipopolysaccharide (LPS)) is administered immediately following context exposure. The goal of the current study was to more fully characterize this phenomenon. In Experiment 1, memory impairment as a result of neonatal infection and subsequent LPS challenge was observed in juvenile rats, indicating that the changes induced by infection occur early on and are then manifest throughout the lifespan. In Experiment 2, infection in juvenile rats did not lead to LPS-induced memory impairment in adulthood, suggesting there is a critical period for early infection-induced alterations. In Experiments 3 and 4, memory for a novel context was impaired in neonatally infected rats, a task that is dependent on the hippocampus, whereas cued memory for a tone, which does not depend on the hippocampus, was not impaired. Furthermore, long-term, but not short-term contextual memory was impaired in adult rats infected as neonates following an LPS challenge either 24 h before or immediately after conditioning. Finally, in Experiment 5, no neonatal group differences were observed in corticosterone or open field behaviour, suggesting that decreased freezing to a conditioned context reflects impaired memory, and not simply hyperactivity or altered stress reactivity. Taken together, we have demonstrated that neonatal infection results in robust hippocampal-dependent memory impairment following an immune challenge in adulthood using a number of conditioning paradigms.

Sonic hedgehog-patched Gli signaling in the developing rat prostate gland: lobe-specific suppression by neonatal estrogens reduces ductal growth and branching

While prostate gland development is dependent on androgens, other hormones including retinoids and estrogens can influence this process. Brief exposure to high-dose estrogen during the neonatal period in rats leads to permanent, lobe-specific aberrations in the prostate gland, a phenomenon referred to as developmental estrogenization. We have previously shown that this response is mediated through alterations in steroid receptor expression; however, further downstream mechanisms remain unclear. Herein, we examined Sonic hedgehog ( Shh)-patched ( ptc)- gli in the developing rat prostate gland, its role in branching morphogenesis, and the effects of neonatal estrogens on its expression and localization to determine whether a disturbance in this signaling pathway is involved in mediating the estrogenized phenotype. Shh was expressed in epithelial cells at the distal tips of elongating ducts in discreet, heterogeneous foci, while ptc and gli1–3 were expressed in the adjacent mesenchymal cells in the developing gland. The addition of Shh protein to cultured neonatal prostates reduced ductal growth and branching, decreased Fgf10 transcript, and increased Bmp4 expression in the adjacent mesenchyme. Shh-induced growth suppression was reversed by exogenous Fgf10, but not noggin, indicating that Fgf10 suppression is the proximate cause of the growth inhibition. A model is proposed to show how highly localized Shh expression along with regulation of downstream morphogens participates in dichotomous branching during prostate morphogenesis. Neonatal exposure to high-dose estradiol suppressed Shh, ptc, gli1, and gli3 expressions and concomitantly blocked ductal branching in the dorsal and lateral prostate lobes specifically. In contrast, ventral lobe branching and Shh-ptc-gli expression were minimally affected by estrogen exposure. Organ culture studies with lateral prostates confirmed that estradiol suppressed Shh-ptc-gli expression directly at the prostatic level. Taken together, the present findings indicate that lobe-specific decreases in Shh-ptc-gli expression are involved in mediating estradiol-induced suppression of dorsal and lateral lobe ductal growth and branching during prostate morphogenesis.

Behavioral and growth effects induced by low dose methamphetamine administration during the neonatal period in rats

The investigation of methamphetamine exposure during neonatal development in rats has demonstrated that long-term spatial learning deficits are induced. A previous dose–response study showed that administration of 5mg/kg methamphetamine, four times daily from postnatal days 11 to 20 produced these deficits, although the effects were not as severe as at higher doses of 10 or 15mg/kg. This study examined concentrations of methamphetamine at or below 5mg/kg given over the same period of time. Five different concentrations of methamphetamine (i.e., 5, 2.5, 1.25, 0.625, or 0) were administered every 2h four times daily from postnatal days 11 to 20. Body weights, zero maze performance, and Morris water maze learning were examined. A dose-dependent decrease in body weight was observed during the period of methamphetamine administration and these lower weights continued throughout adulthood for the 5, 2.5, and 1.25mg/kg concentrations, although the adult decreases were negligible. No differences were noted in the zero maze. In the Morris water maze during the acquisition period, dose-dependent differences in spatial orientation were seen, however non-dose related deficits were observed for other parameters. During the shifted platform phase (“reversal”), a similar dose-dependent difference in spatial orientation was observed, although no other effects were noted during this phase. Females performed worse than males regardless of treatment or the phase of learning in the Morris water maze. These data suggest that even lower doses of methamphetamine can alter learning and memory in adulthood, although with less consistent results than with doses higher than 5mg/kg/dose. These data would caution against even casual use of methamphetamine by women during pregnancy since even low doses could alter the ability of the child to learn.

Effect of food deprivation during early development on cognition and neurogenesis in the rat

Food deprivation has been recognized as having pronounced beneficial effects in adult animals, increasing longevity, reducing seizure susceptibility, and enhancing resistance to neurotoxins. It is not known whether food deprivation in developing animals is neuroprotective or harmful. To evaluate the effects of food deprivation on brain development, we evaluated visual–spatial learning and memory and neurogenesis in the dentate gyrus of the hippocampus in food-deprived (FD) and well-fed (WF) rats. To induce food deprivation, pups were removed from their dams for 12 hours per day from Postnatal Day (P) 2 to P19. FD and WF rat pups were then subjected to status epilepticus (SE) induced by lithium–pilocarpine at P20. After SE, neurogenesis was measured, while in another group of P38 rats, learning and memory were evaluated using the Morris water maze. Food deprivation was found to reduce neurogenesis when assessed after the period of food deprivation. Although SE reduced neurogenesis in the WF animals, it had little effect additional to food deprivation on neurogenesis in the FD rats. Compared with the WF group, FD rats had a mild impairment in memory in the water maze testing after SE. Our study demonstrates that food deprivation during the neonatal period in rats is associated with a decrease in neurogenesis and mild impairment of visual–spatial memory. Although SE decreased neurogenesis in the WF group, in FD animals, SE did not reduce neurogenesis more than what was seen with food deprivation alone. Our results suggest that although food deprivation during early development reduces dentate gyrus neurogenesis, the reduced neurogenesis is not a major factor in cognitive impairment after SE in FD rats.

Developmental changes in the expression of alpha-tocopherol transfer protein during the neonatal period of rat.

It is well known that the cellular concentration of alpha-tocopherol (alpha-toc) is about 10-fold greater than that of gamma-tocopherol (gamma-toc) despite the ingestion of a large amount of gamma-toc in the diet. Recently, this discrimination of tocopherol is considered to be attributed to alpha-tocopherol transfer protein (alpha-TTP) which binds to alpha-toc in preference to the other forms of tocopherol in the liver. In the present study, developmental changes in expression of alpha-TTP after birth were investigated using rats with respect to plasma changes of tocopherols. The expression of alpha-TTP in the neonatal rat liver, which was very low immediately after birth, increased steadily during the two weeks of life before weanling and reached the adult level at four weeks. During the suckling period, the plasma ratio of alpha-toc to gamma-toc linearly increased, because plasma alpha-toc which was low immediately after birth, increased rapidly during the period, while gamma-toc remained unchanged. The increase in the ratio seemed to correlate with the developmental expression of alpha-TTP in the liver during this period.

Temporal determinants of neonatal alcohol-induced cerebellar damage and motor performance deficits

The timing and duration of alcohol exposure was manipulated in neonatal rats by using a “binge” model of alcohol exposure during the “third trimester equivalent.” Groups of Sprague-Dawley rats were exposed to binges via artificial rearing on postnatal days (PD) 4–9, on PD 4–6 or on PD 7–9, which produced peak blood alcohol concentrations representative of human alcohol abusers (~250 mg/dl). Motor performance was assessed using parallel bar traversal on PD 42–44, and total Purkinje cell numbers were determined by using the 3-dimensional stereological optical fractionator method. PD 4–9 binge exposure induced the most severe Purkinje cell loss (to 68% of controls) and PD 4–6 binge exposure also produced significant loss (to 86% of controls), whereas PD 7–9 binge exposure had no significant effect (98% of controls). Unexpectedly, all three alcohol treatments resulted in significant impairments on the parallel bar task. The time of exposure during the early neonatal period in rats strongly influences the degree of Purkinje cell loss, but Purkinje cell loss is not necessary for the alcohol-induced motor performance deficits. Both neuromorphological and neurobehavioral assessments are needed for a full description of alcohol-related neurodevelopmental disorders.

Evidence for the transient expression of P-450 during the neonatal period in rat

A cytochrome P-450 N (N for Neonatal) was resolved and partially purified from 15-day-old newborn rat livers. Antibody raised in rabbit against P-450 N detects two bands in liver microsomes, the major one comigrating with the partially purified P-450 N. This protein is not expressed in lung and kidney and exhibits an unusual ontogenic profile in the liver with maximal levels in 2-week-old animals. Expression of P-450 N is not modified in animals given β-naphthoflavone or pregnenolone 16α-carbonitrile, but is markedly increased by phenobarbital. Its amino-acid composition differs from that of other P-450s and it is not recognized by antibodies directed against CYP1A, CYP2A1, CYP2B, CYP2Cll, CYP3A or epoxide hydrolase. It could thus be considered as an unique P-450 expressed transiently before puberty.