大鼠伏膈核生后发育期形态学的变化

目的：探讨大鼠伏膈核的发育变化的形态学规律。方法：选取出生后一个月内不同时期的大鼠，常规切片染色后应用计算机辅助图像分析系统分别测量不同组别大鼠伏膈核细胞的密度及直径。结果：细胞直径随年龄呈增长趋势，至PD30已基本接近成年水平。细胞密度在PD0-PD15呈下降趋势，之后基本不再变化。结论：表明出生后一段时间内伏膈核细胞在形态上仍要经过一个不断成熟和完善的过程。     

Freeze-fracture study of the vestibular hair cell surface during development

Summary The ciliary arrangement and external surface of the vestibular receptor cell were studied in their immature stages. In the first stages, immature stereocilia are similar to the microvilli of the adjacent supporting cells. Later, when the cilia grow, a geometric arrangement of the stereocilia occurs, while the peripheral microvilli disappear. The size of the hair cell apex increases. The biggest and more bulbous cells correspond to type I hair cells, but in the newborn cat it is difficult to distinguish accurately type II flat hair cells from immature hair cells.This project was supported by INSERM and the Medical Research Council (Grant n^o 00720-X)     

Morphological changes of the myenteric plexus during early postnatal development of the rat

The enteric nervous system needs to adapt itself constantly to the postnatal changes of the developing gut. The aim of this study was to examine the morphological changes between the distal and proximal segments of the gastrointestinal (GI) tract during the first two postnatal weeks. Myenteric plexus from the duodenum, proximal and distal colon of 1‐, 7‐ and 14‐day‐old rat pups was dissected and examined under the scanning electron microscope. Wholemounts from the same regions and postnatal stages were stained with cuprolinic blue. Neuronal numbers per ganglionic area were counted and neuronal sizes were measured. Furthermore, segments of the above‐mentioned areas were embedded in resin and semithin sections were cut. The thickness of the circular and longitudinal muscle layers was measured. The morphology of the myenteric plexus depends on localization as well as on the age of the animal. While in younger animals the myenteric plexus is usually densely packed, the network expands with increasing age. Similarly, the thickness of the circular and the longitudinal muscle layers increases. Nerve cell numbers per ganglionic area increase from duodenum to distal colon and decrease from the 1‐day (P1) to the 14‐day‐old (P14) animal. The longest diameters and the area of the nerve cells decrease from duodenum to distal colon and increase with age of the animal. The intensity of the cuprolinic blue staining varies also according to age and segment of the gut. During the first two postnatal weeks the three‐dimensional architecture of the myenteric plexus as well as the size and densities of the enteric neurons change according to the increasing gut length and the thickness of the muscle layer. The differences between duodenum and colon might reflect the different physiological properties of the proximal and distal gut as well as a varying grade of maturity, which is also supported by a variation in the cuprolinic blue staining intensity. Anat Rec 256:20–28, 1999. © 1999 Wiley‐Liss, Inc.     

Morphological changes of the myenteric plexus during early postnatal development of the rat.

The enteric nervous system needs to adapt itself constantly to the postnatal changes of the developing gut. The aim of this study was to examine the morphological changes between the distal and proximal segments of the gastrointestinal (GI) tract during the first two postnatal weeks. Myenteric plexus from the duodenum, proximal and distal colon of 1-, 7- and 14-day-old rat pups was dissected and examined under the scanning electron microscope. Wholemounts from the same regions and postnatal stages were stained with cuprolinic blue. Neuronal numbers per ganglionic area were counted and neuronal sizes were measured. Furthermore, segments of the above-mentioned areas were embedded in resin and semithin sections were cut. The thickness of the circular and longitudinal muscle layers was measured. The morphology of the myenteric plexus depends on localization as well as on the age of the animal. While in younger animals the myenteric plexus is usually densely packed, the network expands with increasing age. Similarly, the thickness of the circular and the longitudinal muscle layers increases. Nerve cell numbers per ganglionic area increase from duodenum to distal colon and decrease from the 1-day (P1) to the 14-day-old (P14) animal. The longest diameters and the area of the nerve cells decrease from duodenum to distal colon and increase with age of the animal. The intensity of the cuprolinic blue staining varies also according to age and segment of the gut. During the first two postnatal weeks the three-dimensional architecture of the myenteric plexus as well as the size and densities of the enteric neurons change according to the increasing gut length and the thickness of the muscle layer. The differences between duodenum and colon might reflect the different physiological properties of the proximal and distal gut as well as a varying grade of maturity, which is also supported by a variation in the cuprolinic blue staining intensity.     

Morphological changes and cellular proliferation in mouse colon during fetal and postnatal development

To document regional structural and cellular proliferation changes in the developing mouse colon, tissues from fetal, sukling, and weanling mice were analyzed by light microscopy (LM), transmission electron microscopy (TEM), scanning electron microscopy (SEM), [³H]-thymidine incorporation studies, and radioautography. The proximal and distal colon were studied independently at all ages. At 17–18 days of gestation, the mouse proximal colonic mucosa was projected into high and low longitudinal folds disposed in a V-shaped pattern. From birth up to 9 days, the mucosal folds observed by SEM can easily be misinterpreted as being a succession of high and low villus-like structures at LM level. TEM study confirmed the presence of highly specialized absorptive cells in the upper halves of the mucosal folds during this period. No recognizable crypts were noted at birth. Instead, LM and radioautography showed the presence of cell aggregates developing at the base of the epithelium at all levels of the mucosal folds. These cell aggregates evolved into rudimentary crypts giving fully differentiated crypts by day 16 with radiolabeled cells located in the midcrypt portion. As opposed to the proximal segment, a flat mucosa interspersed with well defined short crypts at birth was observed in the distal colon. During the following days, crypts further developed and by 16 days, the radiolabeled epithelial cells were still exclusively located at the base of the crypt. TEM observations illustrated that specialized cells as those found in the proximal segment did not differentiate in this segment. From birth up to 30 days, the labeling indices continuously decreased in the external muscle layer while increasing in the crypt epithelium at different time intervals in both colonic segments. The results show that true villus structures do not develop in proximal colonic mucosa and document regionally related morphological and cellular proliferation changes during mouse colonic maturation. © 1994 Wiley-Liss, Inc.     

Freeze-fracture morphology of the vestibular hair cell-primary afferent synapse in the chick

Summary The intramembrane specializations at vestibular hair cell-primary afferent synapses have been identified and characterized in complementary freeze-fracrure replicas from prehatch and hatchling chick cristae and maculae. Hair cell protoplasmic (P) faces at sites where presynaptic bodies are present exhibit small, tightly packed arrays of 9 nm particles. Hair cell external (E) faces have corresponding arrays of pits. Multiple arrays are often observed in contiguity. Opposite the presynaptic bodies, postsynaptic afferent boutons and calyces exhibit a more extensive array of scattered, irregular E-face particles. Corresponding P-fracture faces of afferent boutons and calyces display little topographical specialization opposite these E-face arrays, which are presumed to be the intramembrane correlate of the postsynaptic density. Examination of complementary replicas has allowed identification of the intramembrane synaptic specializations for all membrane faces at the synaptic apposition.     

Morphometric analysis of thymic medullary non-lymphoid cell changes during postnatal development

Thymic non-lymphoid cells have been shown to influence the differentiation of T-lymphocytes. Large numbers of non-lymphoid cells are concentrated in the thymic medullary zone. A morphometric analysis of these cells in the medulla identified little change in their absolute number and volume density during thymic development in the rat. Expansion of thymic medullary volume with age was documented by planimetry. Several subtypes of medullary non-lymphoid cell were identified by electron microscopy, including both squamous and cystic epithelial cells, interdigitating cells and macrophages. Changes in their relative proportion during development were determined with the percentage of squamous epithelial cells declining, interdigitating cells increasing and the other cell types exhibiting little change. Subcellular evidence of secretory activity was sought by analysis of changes in the volume density of organelles in each of the non-lymphoid classes. Cystic enlargement in epithelial cells and a decrease in the volume density of macrophage inclusion bodies were noted but no clear morphometric trend could be correlated with increased secretory activity expected in the developing endocrine thymus.     

Modification of spontaneous activity in primary vestibular neurons during development in the cat

Summary The spontaneous activity of primary vestibular neurons was studied during postnatal development in the cat. Activities were categorized as regular, intermediate and irregular on the basis of the coefficient of variation. At birth, few regularly firing units were found while the percentage of intermediate and irregular units was high. During development, the percentage of units meeting the criterion of regularity increased steadily with age. At the same time the number of intermediate and irregular units decreased. The average resting rate of all categories of unit showed an increase in firing from birth up to the adult stage, i. e., around the second postnatal month. The mean firing rate of regularly firing units was always higher than the two other categories throughout all the stages of development. These results were compared with similar work performed in the rat.Supported by grants from the C.N.R.S. (E.R.A.-187) and D.G.R.S.T. (no. 79-7-1073)     

Biomechanical changes in the rabbit masticatory system during postnatal development

Using dissection, biometry, and two three-dimensional mechanical models, the postnatal changes of the rabbit masticatory muscles were studied by analyzing their three-dimensional orientation, their strength and fiber lengths, and certain functional consequences of these changes. The first mechanical model uses length-tension relationships of the muscles and predicts the maximum bite force as a function of mandibular position. It shows that young rabbits are able to generate large bite forces at a wider gape than adult animals and that the forces are directed more vertically. In spite of the postnatal changes the mechanical advantage of the system remains about equal. However, the muscles are reoriented so that they exert a larger degree of parallel action, suggesting a larger bite force magnitude but a smaller range of bite force directions. The second model-predicts this range. It shows that during postnatal development a relative gain occurs in the possibilities for the system to exert forces directed rostrodorsally. In all other directions the capability to exert force decreases. The results suggest that during development the possibility of the system to generate large bite forces is increased at the cost of a restriction in the range of jaw excursion and that a restriction takes place in the range of possible force directions that can be exerted at the molars.     

Progenitor cell cycling during hair cell regeneration in the vestibular and auditory epithelia of the chick

We investigated nucleotide-labeling patterns during ongoing hair cell regeneration in the avian vestibular epithelium and during drug-induced regeneration in the avian auditory epithelium. For utricle experiments, post-hatch chicks received an injection of bromodeoxyuridine (BrdU) and were allowed to survive from 2 hours to 110 days after the injection. Utricles were fixed and immunoreacted to detect BrdU. The number of BrdU-labeled nuclei in the hair cell and support cell layers of the utricular sensory epithelium changes significantly between 2 hours and 110 days post-BrdU. At 2 hours, most labeled cells are isolated, while by 5-10 days, the majority of labeled cells are organized in pairs that are most frequently composed of a hair cell and a support cell. Pairs of labeled cells are seen as late as 110 days. Clusters of more than 3 labeled cells are uncommon at all time-points. The total number of labeled cells increases approximately 1.5-fold between 5 and 60 days post-BrdU. This increase is due primarily to a rise in the number of labeled support cells, and it is likely that it represents additional rounds of division by a subset of cells that were labeled at the time of the BrdU injection. There is a significant decrease in labeled nuclei in the hair cell layer between 60 and 110 days post-BrdU, suggesting that hair cells die during this period. To investigate support cell recycling in the drug-damaged auditory epithelium, we examined nucleotide double labeling after separate injections of BrdU and tritiated thymidine. A small number of support cells that incorporate BrdU administered at 3 days post-gentamicin treatment also label with tritiated thymidine administered between 17 and 38 hours later. We conclude that a small population of support cells recycles during regeneration in both the normal utricle and the drug-damaged basilar papilla.     

Immunocytochemical and electrophoretic analyses of changes in myosin gene expression in cat posterior temporalis muscle during postnatal development

Summary Changes in myosin gene expression during the postnatal development of the homogeneously superfast kitten posterior temporalis muscle were examined using immunocytochemical techniques supplemented by pyrophosphate gel electrophoresis and gel electrophoresis-derived enzyme linked immunosorbent assay (GEDELISA) of myosin isoforms. The antibodies used were polyclonals directed against the heavy chains of superfast and foetal myosins and monoclonals against the heavy chains of slow and fast myosins. The fibres of the posterior temporalis in the newborn kitten stained almost uniformly with the anti-foetal myosin antibody and the largest of these fibres stained strongly for superfast myosin. A subpopulation of fibres staining for superfast myosin also stained lightly for slow myosin. These slow staining fibres were evenly distributed in the centres of muscle fibre bundles, reminiscent of primary fibres in limb fast muscle. During subsequent development, slow myosin staining disappeared and superfast myosin replaced foetal myosin so that by 50 days the muscle was virtually homogeneously superfast as in the adult. Fast myosin was never expressed at any stage. It is proposed that fibres staining transiently for slow myosin are superfast primary fibres which are homologous to fast primary fibres recently described in regions of limb muscles devoid of slow fibres in the matured animal. Other jaw-closing muscles have significant populations of slow fibres in the mature animal and it is postulated that there exists in these muscles a second class of jaw primary fibres, the slow primary fibres, in which slow myosin synthesis would be sustained in the adult. It is suggested that the myogenic cells of jaw-closing and limb muscles are of two distinct types preprogrammed to express different muscle genes.     

Sodium current expression during postnatal development of rat outer hair cells

 Outer hair cells of the cultured organ of Corti from newborn rats (0–11 days after birth) were studied in the whole-cell patch-clamp configuration. A voltage-activated sodium current was detected in 97% (n = 109) of the cells at 0–9 days after birth. The properties of this current were: (1) its activation and inactivation kinetics were fast and voltage-dependent, (2) the voltage at half-maximum activation was –45.0 mV, (3) its steady-state inactivation was temperature-sensitive (the half-inactivating voltage was –92.6 mV at 23°C and –84.8 mV at 37°C), (4) the reversal potential (80 mV) was close to the sodium equilibrium potential and currents could be abolished by the removal of extracellular sodium, and (5) tetrodotoxin blocked the current with a K _d of 474 nmol/l. Current amplitudes were up to 1.7 nA at room temperature. Mean current amplitudes showed a developmental time course with a maximum at postnatal days 3 and 7 for outer hair cells from the basal and apical part of the cochlea, respectively. In current-clamp mode cells had membrane potentials of –59.7 ± 11.7 mV (n = 9). When cells were hyperpolarized by constant current injection, depolarizing currents were able to trigger action potentials. At 18 days after birth, sodium currents were greatly reduced and barely detectable. The results show that, unlike adult outer hair cells, immature outer hair cells regularly express voltage-gated sodium channels. However, due to mismatching of the sodium current inactivation range and membrane potential in vitro, a physiological function appears questionable. Received: 13 May 1997 / Received after revision and accepted: 7 July 1997     

Neuropeptide Y immunoreactive axons in the corpus callosum of the cat during postnatal development

Many immunocytochemical studies have identified different types of neurotransmitters localized in the corpus callosum (CC) axons in the adult mammal. Few studies have looked at the development of different neurochemically identified CC systems. Previous studies on the development of cat CC axons have indicated that a large number of transitory CC axons project to the cortex during early postnatal development. The present study focuses on the development of one neurochemically identified group of CC axons in the cat, labeled with an antibody against neuropeptide Y (NPY), to determine if this group participates in transitory CC axonal growth. Cats at specified ages from birth to adulthood were studied with a routine method of immunocytochemistry for antiserum to NPY. NPY-immunoreactive (ir) CC axons were detected at all stages examined, from newborn to adult; the peak density occurred during postnatal weeks (PNW) 3–4. During PNW 1–2, the denisty of NPY-ir CC axons increased gradually; some NPY-ir axons at this age had growth cones located within the CC bundle between the cerebral hemispheres. The density of the NPY-ir CC axons decreased gradually during PNW 5–7, and from PNW 8 to maturity only a few NPY-ir CC axons were observed. These results indicate that at least two types of NPY-ir CC axons (i.e., transitory and permanent) exist during development, and that most of these axons are eliminated or only express NPY-ir for a short period during development. The results also indicate that neurochemical subsets of CC axons participate in the extensive transitory growth observed by means of the membrane tracer DiI but they may follow unique developmental timetables.     

Ultrastructure of hair follicle afferent fibre terminations in the spinal cord of the cat

Summary In acute electrophysiological experiments on anaesthetized cats, single identified hair follicle afferent fibres were injected with horseradish peroxidase (HRP). The HRP was injected from an intra-axonal microelectrode in the lumbosacral spinal cord. One to six hours after injection the animals were perfused and the tissue prepared for light and electron microscopy (EM). Axon collateral arborizations containing HRP reaction product were identified in thick sections under the light microscope and the same tissue then cut on the ultramicrotome for EM study. The terminal branches of the collaterals kept their myelin sheaths until they were 0.45–l.0 m in diameter, just before they formed synaptic boutons. Synaptic boutons (1.0–4.0 m in diameter) were usually of theen passant variety and made contact with dendrites. The contacts were asymmetrical (Type I) and contained round, clear synaptic vesicles of 35–60 nm diameter. Both the non-myelinated portion of the terminal axon and the synaptic boutons received axo-axonic contacts. These axo-axonic boutons contained clear (agranular) vesicles irregular in profile.MRC research student.     

Differential phosphorylation of MAP1b during postnatal development of the cat brain

Summary Microtubule-associated protein 1b, previously also referred to as microtubule-associated protein 5 or microtubule-associated protein 1x, is a major component of the juvenile cytoskeleton, and is essential during the early differentiation of neurons. It is required for axonal growth and its function is influenced by phosphorylation. The distribution of microtubule-associated protein 1b in kitten cerebellum and cortex during postnatal development was studied with two monoclonal antibodies. Hybridoma clone AA6 detected a non-phosphorylated site, while clone 125 detected a site phosphorylated by casein-kinase II. On blots, both monoclonal antibodies stained the same two proteins of similar molecular weights, also referred to as microtubule-associated protein 5a and 5b. Antibody 125 detected a phosphorylated epitope on both microtubule-associated protein 1b forms; dephosphorylation by alkaline phosphatase abolished the immunological detection. During development of cat cortex and cerebellum, AA6 stained the perikarya and dendrites of neurons during their early differentiation, and especially labelled newly generated axons. The staining decreased during development, and axonal staining was reduced in adult tissue. In contrast to previous reports which demonstrated that antibodies against phosphorylated microtubule-associated protein 1b label exclusively axons, antibody 125 also localized microtubule-associated protein 1b in cell bodies and dendrites, even in adulthood. Some nuclear staining was observed, indicating that a phosphorylated form of microtubuleassociated protein 1b may participate in nuclear function. These results demonstrate that microtubule-associated protein 1b is subject to CK2-type phosphorylation throughout neuronal maturation and suggest that phosphorylation of microtubule-associated protein 1b may participate in juvenile and mature-type microtubule functions throughout development.     

Morphological changes in the pancreas during the development of experimental diabetes

Summary Repeated administration of small doses of alloxan (5 mg 100 g of body weight) caused diabetes to develop gradually in albino rats. A direct relationship could then be observed between the degree of disturbance of the carbohydrate metabolism and the extent of the morphological changes in the pancreas: as the blood sugar concentration in the islets of Langerhans rose, the death rate of the beta-cells also rose, and new gamma-cells were formed which developed from the plasmodium. Most of the medium sized and small islets were found in diabetic rats. Changes were found in the acinous cells of the peri-insular zones, indicating their possible transformation into islets.(Presented by Active Member AMN SSSR S. V. Anichkov) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 54, No. 12, pp. 103–105, December, 1962