The ontogenetic development of serotonin (5-HT1) receptors in various cortical regions of the rat brain

Summary The distribution of serotonin (5-HT₁) receptors in various cortical regions of the rat brain has been examined during ontogenesis by quantitative autoradiography.An increase in binding site density between the first postnatal day and adult age was observed and could be approximated by a sigmoid shaped (logistic) growth curve. A marked heterochrony in the increase of binding site density is found in the 13 analyzed cortical regions. Binding sites develop earlier in neocortex than in allocortical areas. Fifty pereent of the binding site density of adult age is reached in the motor cortex at the 9th postnatal day, followed by the primary somatosensory cortex one day later, by the medial prefrontal cortex on the 12th day, by the fascia dentata on the 14th day and by the CA1-region on the 20th day. A detailed analysis of the frontal, medial prefrontal and hippocampal regions also shows a heterochrony within these regions. Adult values of binding site densities are also reached at different ages in the various cortical regions. The highest receptor densities were observed in the dorsal subiculum, the lowest in the primary somatosensory cortex.     

Decrease in the response to ADH of the rat kidney as a result of early postnatal treatment with cortisol

Wistar rats were injected just once, intraperitoneally with cortisol (1 g/g) or saline at the age of 5 days. The cortisol-treated rats did not differ significantly in the (U/P)osm ratio from the saline-treated controls before 15 days of life. Their response to ADH was distinct but weaker than in the saline controls aged 30 days. This reduced response persisted to 60 days of life. In the collecting tubule fragments, (³H)AVP specific binding was lower in the cortisol-treated rats than in the controls at the age of 20 and 60 days. There was no (³H)AVP specific binding in the proximal convoluted tubules in the cortisol-and saline-injected rats of both ages. The ontogenetic patterns of cAMP specific binding in the papillary cytosolic fraction were different: the early increase in cAMP binding was protracted in the cortisol-treated rats, and no peak appeared at the age of 25 days. Cytosolic protein kinase activity was lower, no peak appeared at 30 days, no activation of protein kinase occurred to the end of weaning in the cortisol-treated rats. The difference between the cortisol and saline groups was abolished by day 30. The interference of cortisol with the ontogenetic changes in AVP binding capacity and cAMP-dependent protein kinase appears to be a plausible cause of the altered development of the response to ADH.     

Proliferation and differentiation of pituitary corticotrophs during the fetal and postnatal period: a quantitative immunocytochemical study

 To study the proliferation and differentiation of pituitary corticotrophs, we administered bromodeoxyuridine (BrdU) to pregnant rats at 15.5–21.5 days of gestation and to rat pups at 3, 7, and 28 days after birth. The pituitary sections of fetuses and pups were consecutively immunostained with anti-BrdU and anti-adrenocorticotropic hormone (ACTH) to detect proliferating cells and corticotrophs, respectively. The number of cells labeled with BrdU, ACTH, or both were counted. The diameters of their nuclei and the volume of the pituitary were measured. The BrdU-positive cells were around 76,000–96,000/mm³ during the period studied. The corticotrophs were first detected in the fetus at 15.5 days and they increased during the fetal and postnatal periods. The double-labeled cells were first detected in the 17.5-day fetus. They increased markedly at 19.5 days and comprised about one-quarter of the corticotrophs that increased in 24 h at this stage. These results indicate that: (1) at 15.5–18.5 days the corticotrophs were derived almost exclusively from undifferentiated cells; (2) during the later fetal and early postnatal periods, the proliferation of existing corticotrophs contributed, at least in part, to their increase; (3) About 1/20 of proliferating cells differentiated to corticotrophs when their increase was required. Accepted: 5 October 1999     

Synapsin I expression in the rat retina during postnatal development

Summary The expression of the synapsin I gene was studied during postnatal development of the rat retina at the mRNA and protein levels. In situ hybridization histochemistry showed that synapsin I mRNA was expressed already in nerve cells in the ganglion cell layer of the neonatal retina, while it appeared in neurons of the inner nuclear layer from postnatal day 4 onward. Maximal expression of synapsin I mRNA was observed at P12 in ganglion cells and in neurons of the inner nuclear layer followed by moderate expression in the adult. At the protein level a shift of synapsin I appearance was observed from cytoplasmic to terminal localization during retinal development by immunohistochemistry. In early stages (P4 and P8), synapsin I was seen in neurons of the ganglion cell layer and in neurons of the developing inner nuclear layer as well as in the developing inner plexiform layer. In the developing outer plexiform layer synapsin I was localized only in horizontal cells and in their processes. Its early appearance at P4 indicated the early maturation of this cell type. A shift and strong increase of labelling to the plexiform layers at P12 indicated the localization of synapsin I in synaptic terminals. The inner plexiform layer exhibited a characteristic stratified pattern. Photoreceptor cells never exhibited synapsin I mRNA or synapsin I protein throughout development.Abbreviations GCL ganglion cell layer - INB inner neuroblast layer - INL inner nuclear layer - IPL inner plexiform layer - ONB outer neuroblast layer - ONL outer nuclear layer - OPL outer plexiform layer     

Expression of preproenkephalin mRNA in rat superior cervical ganglion during postnatal development

The number of principal neurons in the rat superior cervical ganglion (SCG) exhibiting enkephalin-peptide immunoreactivity is reported to be limited. To better determine the degree of enkephalinergic phenotype in sympathetic neurons, sections of SCGs from rats aged newborn to adult were processed for in situ hybridization histochemistry, using a [³⁵S]cRNA probe directed against preproenkephalin (PPENK). > 50% of principal ganglion neurons express mRNA for PPENK in adults. Striking variability in labeling intensity is observed. PPENK mRNA is detected in developing ganglia beginning at postnatal days 4–7. Both the number of cells and intensity of labeling increases with postnatal development. These results indicate that expression of PPENK mRNA is more widespread than expression of enkephalin peptides and develops postnatally.     

Amiloride-sensitive sodium transport of the rat distal colon during early postnatal development

To evaluate developmental changes in colonic sodium transport, the sensitivity of the transepithelial potential and short-circuit current to amiloride was investigated. The amiloride-sensitive short-circuit current (I _sc ^Na ), which represents the electrogenic sodium transport through Na⁺ channels, rose significantly from day 5, reached a peak on day 10, and entirely disappeared after weaning. The maximum rate of electrogenic, amiloride-sensitive sodium transport was 12.0 Eq/cm² · h. TheI _sc ^Na was suppressed by adrenalectomy and/or premature weaning but not by a mineralocorticoid antagonist, spironolactone. On the contrary, treatments which increase aldosterone levels in vivo (low-sodium diet, furosemide-induced natriuresis, high dietary intake of potassium) stimulated theI _sc ^Na . The effect of adrenalectomy increased during postnatal development. The sensitivity ofI _sc ^Na to aldosterone was highest at the end of the weaning period. High-sodium diet, which causes a decrease in circulating aldosterone, was associated with a partial inhibition ofI _sc ^Na (P<0.016). These data suggest that the distal colon of neonatal rats can transport sodium via an electrogenic, amiloride-sensitive mechanism and that adrenocortical hormones exert the main regulatory control of this pathway.Parts of this study have been presented at the 6th Symposium on Developmental Pharmacology, Schloss Reinhardsbrunn 1986     

The postnatal development of rat dentate gyrus and the effect of early thyroid hormone treatment

Summary The postnatal development of the dentate gyrus and the effect of 5 g/day triiodothyronine treatment was examined by means of tritiated thymidine autoradiography.The polymorph layer of the dentate gyrus is a secondary germinal layer and forms cells for the granular layer. The cell formation in the dentate gyrus was found to be nearly completed on the 21st day, but development ended only in adulthood.Thyroid-hormone treatment did not affect significantly the postnatal cell formation of the dentate gyrus, but resulted in the appearance of a great number of pycnotic cells in the granular layer.     

ChAT-like immunoreactivity of olivocochlear fibres on rat outer hair cells during the postnatal development

Summary Several studies present a great deal of information about putative efferent neurotransmitters and their distribution in the adult and developing cochlea. Anatomical mapping of outer hair cell efferent fibres during ontogeny is still not available. Using quantitative electron microscopy in combination with immunocytochemistry, the distribution of ChAT-like immunoreactivity in the developing rat was investigated. Adult-like immunoreactivity in the whole cochlea is first observed in 30-day-old rats. We localized the adult-like immunoreactivity in all efferent fibres and synapses of the outer hair cells along the entire cochlear duct. An adult-like reaction in the whole cochlea could be observed on the 25th day after birth in two out of three cases. On the 20th postnatal day, no adult-like ChAT immunoreactivity was found, with the exception of one case where labelling was seen in the basal region only. The adult-like ChAT immunoreactivity on the 30th day, 2–3 weeks after the onset of hearing, is the latest maturation of all features of the organ of Corti so far investigated. Synaptogenesis of the outer hair cell efferents reaches an adult-like appearance already on the 16th day after birth.     

Postnatal development of neuropeptide Y- and calcitonin gene-related peptide-immunoreactive nerves in the rat urinary bladder

The postnatal development of neuropeptide Y- and calcitonin gene-related peptide-immunoreactive (NPY-IR and CGRP-IR) nerve fibers in the rat urinary bladder was investigated using whole-mount preparations and cryostat sections. In newborn and 3-day-old rats, many NPY-IR nerve fibers were observed in the subserous and muscle layers. Many NPY-IR nerve cell bodies clustered at branching points of the subserous nerve bundles. Within 4 weeks after birth, these cell bodies drastically decreased in number and spread along the bundles, although the number of NPY-IR nerve fibers increased moderately. In contrast, CGRP-IR nerve fibers in newborn and 3-day-old rats were less developed, and no CGRP-IR nerve cell body was observed in any rat. However, CGRP-IR nerve fiber distribution in the urinary tissues conspicuously increased within 4 weeks after birth. Especially, an increase of the infraepithelial fibers showing a meshwork appearance was prominent in the fundus and corpus of the bladder. The infra- and intraepithelial CGRP-IR nerve meshwork of the ventral wall was more dense than that of the trigone. At 4 weeks, NPY-IR and CGRP-IR nerves were similar to those of the adult rat (8–12 weeks old). The present study suggests a correlation between the development of the peripheral nervous system in the urinary bladder and maturation of micturition behavior in the rat.     

酒精对大鼠胚胎肝脏酶活性影响的组织化学观察

本文采用酶组织化学方法,研究和观察了酒精对大鼠胚胎肝细胞酶活性的影响。实验结果显示：实验组大鼠胚胎肝细胞琥珀酸脱氢酶（ＳＤＨ）和葡萄糖－６－磷酸酶（Ｇ－６－Ｐ）活性减弱,乳酸脱氢酶（ＬＤＨ）和酸性磷酸酶（ＡＣＰ）活性有所增强。经统计学处理,实验组和对照组两组酶活性有显著性差异（Ｐ〈０．０５）,表明酒精能明显影响胚胎肝细胞的正常代谢功能。     

Quantitative cellular changes during postnatal development of the rat dorsal lateral geniculate nucleus

Summary Quantitative changes in cell number during development of the dorsal lateral geniculate nucleus were determined using semithin serial sections of tissue obtained from 28 rats on postnatal day 0, 5, 8, 10, 20, 30, 90 or 165. Our results show three phases of postnatal development in the rat dorsal lateral geniculate nucleus: phase 1 from birth until eye opening, which occurs around the 12th day in these litters; phase 2 from eye opening through stabilization of neuron number on the 30th postnatal day, and phase 3 from that event until adulthood. During the first period increases in neuron number and in glial cell number are found accompanying a nearly seven-fold increase in dorsal lateral geniculate nucleus volume. Phase 2 includes a high incidence of neuronal cell death and a continuous increase in the number of glial cells. The third phase is characterized by a stabilization in the number of neurons, although the glial cell number continues to increase. Neuronal density decreases exponentially throughout the postnatal life of the rat, while the density of glial cells remains relatively stable over the period of study. The postnatal phenomenon of an initial increase in neuron number followed by a period of neuron death may be related to modulating and plastic functions which occur in the rat dorsal lateral geniculate nucleus before a stable neuronal population is achieved on the 30th postnatal day.     

Postnatal development of the vascular pattern in the rat telencephalic pia-arachnoid

Summary The postnatal changes in arrangement of the vascular system of the pia-arachnoid of rats are described based on scanning electron microscopy of microcorrosion casts and transmission electron microscopy. At birth, the distal arteries and veins are embedded in a dense plexiform network of immature capillaries. Arteries and veins are interconnected by many small capillary anastomoses. The trunks are located above the pial plexus. The underlying plexiform vessels provide the matrix for the formation of additional collateral and precortical segments during further development. During the first postnatal week, the distal pial arteries and veins become visible as separate channels and emerge from the subjacent capillary plexus. The pattern of anastomosing arterial rings is now clearly visible. The pial arterial tree can be subdivided into conductive, collateral, and precortical distributive segments, according to Jokelainen et al. (1982). Subsequently, passive expansion of the vascular system takes place during the period of rapid brain growth. In young adults the majority of the formerly closed arterial rings are interrupted, possibly by regression of single collateral arterial segments (Fig. 6). The dense venous capillary plexus of the pia is maintained during the first eight days in spite of marked brain growth. The process of reduction of this capillary plexus starts at the arterial side and proceeds from proximal to distal segments of the veins during the second and third week. The capillary segments, which provide anastomosis between arterial and venous vessels, disappear at the same time as the regression of the dense venous capillary network. Two processes may biodify the architecture of the pial vasculature during development: brain growth and maturation on the one side and cytodifferentiation of vascular walls on other.This work was presented in part at the Third World Congress for Microcirculation, Oxford, 9–14 September 1984, and was published in abstract form (1984, Int J Microcircul Clin Exp 3:308)     

Architectural design,fiber-type composition,and innervation of the rat rectus abdominis muscle

The rectus abdominis muscle is architecturally compartmentalized by tendinous intersections and is supplied by multiple thoracic nerves. In this study, the rectus abdominis of the rat has been qualitatively and quantitatively examined with regard to muscle dimensions, fiber organization, fiber-type composition, and innervation. The muscle exhibits architectural heterogeneity and different patterns of innervation among its thoracic, epigastric, and hypogastric parts. The epigastric part, adherent to the rectus sheath via tendinous intersections, represents relatively simple design. It is formed by serially arranged compartments with shorter fibers, compared with the other parts. These compartments are segmentally supplied by thoracic nerves. The hypogastric part is more complex, forms an interdigitation of muscular slips, and has segmental distribution of thoracic nerves in mediolateral direction. The thoracic part much differs from the other parts. It has smaller cross-sectional areas, compartments composed of abundant nonspanning fibers with intrafascicular termination, and non-segmental distribution of thoracic nerves. In addition to these craniocaudal specializations among the three parts, the muscle exhibits mediolateral differences in fiber-type composition. Slow-twitch oxidative fibers are more densely distributed in the medial half region than the lateral, whereas fast-twitch glycolytic fibers follow an inverse pattern. The mediolateral differences in fiber-type composition as well as the craniocaudal specializations in architectural design and innervation imply regionally differentiated recruitments of the muscle in various behaviors.© Willey-Liss, Inc.     

Expression of glucocorticoid receptors in the hippocampal region of the rat brain during postnatal development

Circulating glucocorticoids, of which their concentration is largely under the control of the hypothalamic-pituitary adrenal (HPA) axis, acting through the glucocorticoid receptors (GR) regulate a large variety of pivotal functions of the organism such as growth, development, immune- and stress-response. The main mechanism of regulation of the HPA axis activity is via negative feedback at all levels of the HPA axis itself as well as at the extra-hypothalamic level, a central part of which is the hippocampus. During neonatal development, the HPA axis of rats undergoes a period of hyporesponsiveness (SHRP)-when most stress stimuli fail to induce stress-response. Here, we describe the pattern of GR proteins expression in the hippocampal area of the rat brain during postnatal development and in adulthood. We demonstrated that the GR protein, of which its expression level is gradually enhanced in the hippocampus during postnatal life, exists in three different molecular sized forms. A larger molecular form was expressed at rather high levels at all studied time periods. A second smaller variant of GR was transiently expressed during the first one and a half weeks that corresponds with SHRP and then appeared again only in the adulthood. By the end of SHRP on PD 13, third smallest protein form of GR started to be detected in the hippocampal area. Thus, it remains to be disclosed in the nearest future, how the hippocampal GR isoforms may be involved in regulation of the neonatal HPA axis hyporesponsiveness as well as in functions of this system during the ensuing period of the brain maturation.     

The pre- and postnatal development and ageing of interscapular brown adipose tissue in the rat

Summary Developmental and ageing changes in interscapular brown adipose tissue have been studied in white Wistar rats by light, fluorescence and electron microscopy.Cellular aggregation was noted in the fetal interscapular area on the 15th day of gestation and vascularised primitive lobules of brown adipose tissue became unequivocally identifiable on the 17th day in utero. Brown adipocyte precursors appeared to be derived directly from mesenchymal cells and were uniquely characterised by larger (0.8–1.7 m diameter) mitochondria. Numbers of these precursor cells (pre-adipocytes) were seen in mitosis during intrauterine life. Pericapillary cells similar in appearance to embryonic pre-adipocytes were regularly observed within brown fat lobules throughout later life.Cardinal features noted in mature brown adipose tissue were parenchymal cells with a multilocular lipid distribution and numerous large mitochondria with distinctive inparallel cristae, as well as an extensive vascular network and a dense catecholaminergic vasomotor and parenchymal innervation.Brown adipocytes generally retained a multilocular lipid distribution into old age, and although the catecholaminergic fluorescence of the nerves supplying the tissue was reduced, a widespread distribution of noradrenergic vasomotor and parenchymal nerves and of nexuses between brown adipocytes continued to be demonstrable by electron microscopy in the brown adipose tissue of senile rats.     

Expression pattern of galectin-1 and galectin-3 in rat testes and epididymis during postnatal development

Galectins are a family of lectins-binding beta-galactosides involved in a variety of extracellular and intracellular processes, thereby contributing to homeostasis, cell adhesion, cellular turnover, and immunity. This study aimed to determine the localization and expression of galectin-1 (Gal-1) and galectin-3 (Gal-3) in the testis and epididymis of rats at postnatal [(prepubertal (day 5), pubertal (day 20), postpubertal (day 50) and mature (day 70)] periods by using immunohistochemistry and Western blotting. Gal-1 and Gal-3 were differentially expressed in different types of cells in the testis and epididymis during postnatal development. While we detected Gal-1 expression in some spermatogenic cells and Leydig cells in the testis, not in the epididymal epithelium, Gal-3 was expressed in Sertoli cells, peritubular myoid cells, Leydig cells, smooth muscles and interstitial CD68-positive macrophages. Epithelial cells of the corpus and cauda epididymis showed an intense Gal-3 expression. Gal-1 expression was higher in the testis than in the epididymis on days 50 and 70. The expression of Gal-3 in the testis increased from the prepubertal to mature period. While the expression difference of Gal-3 was not statistically significant in the testis and epididymis until puberty, Gal-3 expression in the postpubertal and mature periods was higher in the epididymis. The expression of Gal-3 in the corpus and cauda epididymis was higher than that in the caput epididymis. In conclusion, our findings suggest that puberty has potential regulatory effect on the expression of galectins in testis and epididymis of rats. Gal-1 and 3 may play a role in the development of the reproductive system and the preservation of the immune-privileged environment in the testis, due to their pro-apoptotic and anti-apoptotic functions. The presence of intense expression of Gal-3 in the corpus and cauda epididymis may contribute to the maturation and storage of spermatozoa.     

Nitrergic neurons during early postnatal development of the prefrontal cortex in the rat: Histochemical study

The presence of nitrergic cells in the prefrontal cortex has been confirmed, however little is known about the postnatal development of these cells. Nitrergic neurons were studied histochemically by using NADPH-diaphorase staining in the prefrontal cortex of male Wistar rats from postnatal day 7–21 (P7–21). Neuronal NADPH-diaphorase is a nitric oxide synthase that provides a specific histochemical marker for neurons producing nitric oxide (NO). NO acts as a neurotransmitter and intracellular signaling molecule in the nervous system. We observed in 7 day old rats NADPH-d containing neurons that were intensely stained. These neurons were bipolar with a short dendrite with average length of 23 μm. During the second postnatal week, the neurons were mainly bipolar and were rarely multipolar. By P14 the cells were located primarily in cortical layers III–VI. Nitrergic neurons of the 21 day old rats were histochemically identified as multipolar cells with long radial extending dendrites. Dendrites of neurons in 14 and 21 day old rats were a similar length with an average of 57 μm. These results suggest that nitrergic neurons differentiate during a relatively short period of time and reach their structural maturity by the end of the second week of postnatal development.     

Substance P-containing sensory nerves in the rat iris. Normal distribution, ontogeny and innervation of intraocular iris grafts

Trigeminal, substance P-containing nerves have been studied in the stretch-prepared rat iris with immunohistochemical techniques. The normal iris exhibited a slightly irregular plexus of individual fibres in the dilator, intermingled with thin, meandering axon bundles. The sphincter contained more circumferentially oriented fibres. Occasional free nerve endings were present in all parts of the iris; no obvious association with blood vessels was detected. All substance P-positive nerves in the iris disappeared after lesioning the trigeminal nerve. Irides of neonates showed scattered, smooth fibres in a sparse plexus, without visible axon bundles. Over the first two postnatal weeks, the density of innervation developed rapidly, reaching a transiently supranormal level and fluorescence intensity, compared to adulthood. From 3 weeks on, the pattern and density of substance P-containing fibres approached the normal adult appearance. In irides grafted to the anterior eye chamber, the intrinsic substance P nerves degenerated, disappearing completely after 5 days. Reinnervation from the host irides transpired over the next few weeks, approximating normal density after 3 weeks, and organotypic density and distribution from 4 weeks on. No obvious hyperinnervation was encountered after longer postoperative times (3 months). In the host iris, many substance P fibres disappear or exhibit low fluorescence intensity during the first postoperative week, recovering fully during the next 2 weeks. Over longer postoperative periods irregular, moderate hyperinnervation developed with increased numbers of axons in bundles. In conclusion, we show normal distribution and plasticity during ontogeny and maturity of substance P-containing iris nerves in the rat, with a sensitive immunohistochemical technique in iris whole mounts.     

Effects of infant and adult amygdaloid lesions upon acquisition of two-way active avoidance by the adult rat: Influence of rearing conditions

When performed in the adult rat, bilateral and complete amygdalectomy resulted in a clear deficit in the acquisition of a 2-way active avoidance in a shuttle-box. When performed in the 7 day old rat pup, the same complete lesion resulted in no disruption of acquisition of the 2-way AA task by the rats when adult, irrespective of the environmental conditions in which they were reared from weaning. However, isolation-reared rats showed shorter response latencies than did group-reared rats. When the amygdaloid lesion was restricted to the centromedial area of the amygdala, operations carried out at 7 days resulted in a reliable impairment of the 2-way active avoidance in the initial phase of the acquisition, but only when the rats had been group-reared from weaning. The same lesion resulted in a reliable improvement in acquisition when the rats had been reared in isolation. In short, the degree of recovery of function with respect to 2-way AA acquisition following a bilateral amygdaloid lesion depends on the age at which the lesion is carried out (at infant or adult age), on the extent of the lesion (complete or restricted to the CM area) and on the rearing conditions (in groups or in isolation).