Hepatocyte differentiation during early fetal development in the rat

Summary Rat hepatocyte differentiation between day 12 and 19 of fetal life was studied by electron microscopy. The cytoplasmic structures involved in synthetic and secretory function, i.e., rough endoplasmic reticulum and Golgi apparatus, appear to be the first to differentiate, and their development is probably related to the secretion of different kinds of plasma proteins. The cytoplasmic organelles involved in other hepatic functions develop later: lysosomes from day 15, peroxysomes, glycogen rosettes and smooth endoplasmic reticulum still later. However, the morphological differentiation of bile canaliculi begins from day 12.This work was supported in part by a grant from C.N.R. n. 78.02265.04     

Water-soluble hippocampal proteins of the rat in early postnatal ontogeny

Using ion exchange chromatography on DEAE-cellulose, studies have been made on protein extracts obtained at low ionic strength from hippocampal slices of rats within the 1st month of their postnatal life. Significant changes in fractional composition were found. To the end of the 2nd week, a reliable decrease in the relative content of low-acidic fractions was observed together with the increase in the content of those fractions which were eluted by a buffer with a higher molarity. In 3-week animals, the relative content of weak-acidic fractions decreases by 1 1/2 times, the difference being even more significant in adult animals. It is suggested that the observed redistribution of fractions within protein spectra is due to the appearance of the brain specific proteins.     

Normal and abnormal epithelial differentiation in the female reproductive tract

In mammals, the female reproductive tract (FRT) develops from a pair of paramesonephric or Müllerian ducts (MDs), which arise from coelomic epithelial cells of mesodermal origin. During development, the MDs undergo a dynamic morphogenetic transformation from simple tubes consisting of homogeneous epithelium and surrounding mesenchyme into several distinct organs namely the oviduct, uterus, cervix and vagina. Following the formation of anatomically distinctive organs, the uniform MD epithelium (MDE) differentiates into diverse epithelial cell types with unique morphology and functions in each organ. Classic tissue recombination studies, in which the epithelium and mesenchyme isolated from the newborn mouse FRT were recombined, have established that the organ specific epithelial cell fate of MDE is dictated by the underlying mesenchyme. The tissue recombination studies have also demonstrated that there is a narrow developmental window for the epithelial cell fate determination in MD-derived organs. Accordingly, the developmental plasticity of epithelial cells is mostly lost in mature FRT. If the signaling that controls epithelial differentiation is disrupted at the critical developmental stage, the cell fate of MD-derived epithelial tissues will be permanently altered and can result in epithelial lesions in adult life. A disruption of signaling that maintains epithelial cell fate can also cause epithelial lesions in the FRT. In this review, the pathogenesis of cervical/vaginal adenoses and uterine squamous metaplasia is discussed as examples of such incidences.     

Changes in the renal corpuscles during early postnatal ontogeny of the rat (stereologic analysis)

Kidneys of 60 white rats at the age of 1-45 days of postnatal life have been investigated. Increase in size of the renal corpuscles during the developmental process is accompanied with certain changes in the form and variability of volumes in the structures studied. During early postnatal period, against the background of linear increase in the size of the renal corpuscles, the parameters of variability in volumes, form and in the total number of the renal corpuscles in the organ change non-linear. Stabilization moment of the total number (11-14 days) on the 8th-10th day outgoes that for the parameters of the form and variability of the renal corpuscle volumes.     

Myo-inositol in the reproductive tract of the female rat

• 1.1. myo-Inositol concentrations in oviduct, ovary and uterus were many-fold those of blood serum during all four stages of the estrous cycle of the female rat.
• 2.2. Inositol concentration was higher in oviduct than in ovary or uterus and was lower in uterine fluid than in uterus.
• 3.3. Estrus uteri had higher inositol concentrations than uteri in other phases of the cycle.
• 4.4. In order to measure dynamic aspects of the distribution of inositol. the distribution of radioactivity among organs of the reproductive tract of mature female rats was measured 45 min after i.p, injection of [2-³H]myo-inositol.
• 5.5. These organs concentrated inositol from the blood, and the tissue radioactivity (expressed as dpm/mg wet wt of tissue) increased in the sequence: vagina < cervix < uterus < ovary < oviduct.
• 6.6. The uterus and ovary concentrated myo-inositol more strongly during proestrus than during metestrus. diestrus or estrus.
• 7.7. The contents of proestrus follicles were more highly radioactive than was the ovary itself, whereas proestrus uterine fluid was less radioactive than the uterine tissue.

     

Cell proliferation and differentiation in the fetal and early postnatal mouse thymus

The relationships between cell proliferation and cell differentiation during thymus ontogeny were studied by labeling DNA-synthesizing thymocytes with bromodeoxyuridine and staining with antibodies against CD4, CD8, J11d, phagocytic glycoprotein 1, TCR V beta 8 chain, Thy-1, and IL-2R surface proteins. The development of the thymus was discontinuous, with two well defined growth periods from 13 days to 18 days of fetal life and from 3 days to 6 days after birth, and more progressive growth from day 8 to 2 wk. Cell proliferation started on fetal day 12, 1 day after the arrival of hemopoietic stem cells in the third branchial pouch. These cells were phagocytic glycoprotein 1-positive but IL-2R and Thy-1 negative. Thus, cell proliferation preceded IL-2R expression. Until day 15, CD4-8- thymocytes expanded without differentiation. Then CD4-8+ and CD4+8+ cells appeared; this induction was proliferation dependent and occurred on cells which had already lost IL-2R, but just after maximum expression of this receptor. During several days, the thymus remained of constant size (around 10(7) cells) and behaved like the steady state thymus. On day 3 after birth, expansion started again and was correlated with an increase in CD4-8- proliferation index and IL-2R expression. At the same time, the thymic subset capable of expansion without differentiation was again, transiently, detectable. These results suggest that the inflow of precursor cells into the thymus is permanent but transiently increased at several times during ontogeny. Moreover, the behavior of fetal CD4-8- cells does not appear radically different from that of adult precursors, but the actual difference resides in the variation of the relative proportion of CD4-8- cells at different maturation stages, as revealed by striking variations of IL-2R expression by cycling cells.     

Cyto- and synaptogenesis in the arcuate nucleus of the rat hypothalamus during fetal and early postnatal life

Summary Morphogenesis of the arcuate nucleus of the rat from the 15th fetal day to the 6th postnatal day was investigated light and electron microscopically. The arcuate neurons exhibit a gradual development after the 15th fetal day. All cytoplasmic constituents are present in these nerve cells already during the last days of gestation. Nevertheless, they are not fully differentiated at birth. The first synapse-like structures (presynapses) were observed in 17 day-old, the first synapses in 18 day-old fetuses. During the early postnatal period the number of presynapses decreases, but at the same time there is a gradual increase in the number of the relatively mature synapses. This process starts already during the last days of prenatal life. Although all structural elements of the arcuate nucleus of the adult rat appear to be present at birth, the extent of the neuropil area and the number of the presynapses indicate that the arcuate nucleus is still in a fairly undeveloped stage during the first postnatal days.     

Expression and localization of luteinizing hormone receptor in the female mouse reproductive tract

The presence of the LH receptor (LHR) in nongonadal tissues of the reproductive tract has been reported, but localization studies have not been performed. Our objectives were to demonstrate the presence of LHR in the reproductive tract and to localize receptor expression. Reproductive age rats and mice were obtained and (125)I-hCG binding assays were performed on membrane preparations from the uterus, ovary, liver, and testis. In situ hybridizations were performed using (35)S-labeled antisense and sense RNA probes prepared from nucleotides 1-591 of the mouse LHR cDNA. Specific hCG binding was detected in membrane preparations from the ovary, uterus, and testis but not in the liver in both the rat and mouse. In the ovary, LHR mRNA was localized in theca cells, large follicles, and corpora lutea as expected. In the uterus, LHR mRNA was expressed in stromal cells of the endometrium and in the uterine serosa. Uterine smooth muscle cells had low levels of expression, and the endometrial epithelium was negative. In the oviduct, high levels of LHR expression were noted on the serosa and in subepithelial cells. Oviductal smooth muscle had low expression, and the epithelium was negative. We conclude that functional, nongonadal LHR are expressed in the mouse reproductive tract. The presence and localization of LHR expression in the mouse reproductive tract lay the foundation for transgenic models to address the physiologic role of these receptors.     

The lipid peroxidation system in the retina and brain of rats during early postnatal ontogeny

The content of the lipid peroxidation products in the rat retina and brain tissues during the early postnatal period (20-45 day of life) was estimated. It was shown during this period the content of conjugated dienes was decreased: the content of malondialdehyde and Schiff bases was without changes in the rat retina. At the same time the content of the conjugated dienes and Schiff bases in the brain tissue was proportionally increased. The activity of glutathioneperoxidase and glutathionereductase was decreased in the retina and not changed in brain. In the same period of the life the content of alpha-tocopherol in the retina was increased. We observed also the enhance of the rate of the induced lipid peroxidation in brain and retina of older animals.     

Effect of neonatal androgenization on the septal neurons of the brain in female rats in early postnatal ontogeny

Newborn female rats were androgenized, and the reaction of neurons of brain septum on excessive quantity of exogenous androgens, introduced during so-called "crucial" period of formation of centers of gonadotropic regulation of sexual cycles, has been studied in 3, 5, 7, 10, 20, 30, and 60 days old animals. Morphometry of brain septum cell nuclei revealed that most neuron nuclei shrink after androgenization. Monoamine content was significantly increased in septum nuclei of experimental animals. Neonatal androgenization led to the increased capacity of septal complex neurons to bind 3H-estradiol and to the decreased 3H-testosterone binding. The data obtained suggest that the brain septum neurons of female rats depend on sex steroids, particularly during "crucial" period of development.     

Polysomes during early postnatal development of brain in the rat

We have attempted to show eventual modifications in the brain protein synthesis apparatus of rat during the first three weeks after birth. Through this time we noted a steady decrease (about 60%) in the free polysomes, when expressed relative to tissue weight. This decrease does not correlate with changes in the polysome profile, indicating that no loss in the efficiency of protein synthesis was involved. Translation in a reticulocyte lysate also failed to reveal differences.     

The development of the corticoliberin- and vasopressinergic elements of the rat hypothalamus during postnatal ontogeny

In 10--81-day and adult Wistar rats, neurosecretory cells were revealed which react with antisera to corticoliberin and vasopressin. Morphometric analysis of these cells in the supraoptic, paraventricular and anterior commissural nuclei shows that in the latter vasopressinergic cells develop somewhat later than in the supraoptic and paraventricular nuclei. Complete differentiation of neurosecretory cells in all the centres investigated is observed in 2-month animals. Studies were also made on the amount of corticoliberin- and vasopressinergic terminals in the external zone of the median eminence. Vasopressin-immunoreactive fibers are more numerous in young rats than in adult ones. Corticoliberin-positive neurosecretory fibers are more abundant in adult animals. Earlier development of vasopressinergic elements corresponds to a hypothesis of a more ancient origin of nonapeptidergic structures as compared to those producing liberins and statins.     

DNA synthesis in various sections of the brain and in the liver of the rat in early postnatal ontogeny

The intensity of 3H-thymidine incorporation in DNA, as well as DNA content of the liver, brain stem, cerebellum, and neocortical tissues have been investigated in 1-32 days old rats. It was shown that the kinetics of the studied parameters is principally different in the tissues investigated.     

Distribution of P2X receptor subtypes in the rat female reproductive tract at late pro-oestrus/early oestrus

Using immunohistochemistry techniques, we have examined the female reproductive organs of the rat in late pro-oestrus/early oestrus for the presence of purine nucleotide P2X1-7 receptors. In contrast to the male genital organs and the urinary tract, P2X1 receptors were present weakly, if at all, on smooth muscle membranes, except in blood vessels, whereas P2X2 immunoreactivity in smooth muscle was present in ovary and uterus as well as in blood vessels. Neither P2X1 nor P2X2 receptors were present in fallopian tubes. P2X5 receptors were seen in the differentiating cell layers of the stratified squamous vaginal epithelium and also in the very early stages of ovarian follicular development; P2X6 receptors were present in secondary follicles. P2X7 receptors, markers for programmed cell death, were present in the keratinised vaginal epithelium and also in the exfoliating superficial endometrial cells. The possible biological significance of these signalling molecules in the female reproductive tract is discussed.