Postnatal development of the rabbit pineal gland. A light- and electron-microscopic study.

The development of the rabbit pineal gland has been studied by light and electron microscopy from the 1st to the 120th postnatal day. After 24 h of postnatal life, the pineal parenchyma is highly cellular, showing two identifiable cell types: pinealocytes I and II. Immature type II pinealocytes arrange either in cellular cords or clusters or forme rosette-like structures. At the 5th postnatal day, corticomedullar differentiation is established. Rosette-like structures and cellular cords are absent from the cortex. Along the postnatal period, nuclei of pinealocytes are set apart due to cytoplasmic widening and development of cell processes. These structures pervade the cellular cords and rosette-like structures formed by immature type II pinealocytes. Rosette-like structures are no longer seen beyond the 30th postnatal day, and cords of type II pinealocytes from the 90th postnatal day on. At this time, the rabbit pineal gland is considered to be histologically mature.     

Diaphragm muscle fatigue resistance during postnatal development.

Changes in the contractile and fatigue properties of the cat diaphragm muscle were examined during the first 6 wk of postnatal development. Both twitch contraction time and half-relaxation time decreased progressively with age. Correspondingly, the force-frequency curve was shifted to the left early in development compared with adults. The ratio of peak twitch force to maximum tetanic force decreased with age. Fatigue resistance of the diaphragm was highest at birth and then progressively decreased with age. At birth, most diaphragm muscle fibers stained darkly for myofibrillar adenosinetriphosphatase after alkaline preincubation and thus would be classified histochemically as type II. During subsequent postnatal development, the proportion of type I fibers (lightly stained for adenosinetriphosphatase) increased while the number of type II fibers declined. At birth, type I fibers were larger than type II fibers. The size of both fiber types increased with age, but the increase in cross-sectional area was greater for type II fibers. On the basis of fiber type proportions and mean cross-sectional areas, type I fibers contributed 15% of total muscle mass at birth and 25% in adults. Thus postnatal changes in diaphragm contractile and fatigue properties cannot be attributed to changes in the relative contribution of histochemically classified type I and II fibers. However, the possibility that these developmental changes in diaphragm contractile and fatigue properties correlated with the varying contractile protein composition of muscle fibers was discussed.     

Postnatal development of the corticothalamic projections from the visual association cortex to the lateralis posterior complex in the cat

The postnatal development of the corticothalamic projection from the lateral suprasylvian cortex (LS) to the lateral medialis-suprageniculate nucleus (LM-Sg) of the cat thalamus was assessed by means of the anterograde tracer biocytin. In the adult, two types of corticothalamic fibers were found: type I established a network of fine fibers present throughout the LM-Sg, it was characterized by a linear sequence of small (less than 0.5 microm in diameter), single terminal boutons making contact mainly with thin dendrites and/or dendritic spines. Type II, found less frequently, gave off short, side branches near axon terminals and formed clusters of 5-10 large terminal boutons (0.5-1.5 microm in diameter), making contact predominately with medium-sized dendrites and/or vesicle-containing profiles, forming a synaptic glomerulus. At birth (P0), anterogradely-labeled fibers were found in the LM-Sg as in adults. In the early postnatal period (until P6) as well as around the time of eye-opening (P7-P10) to P21, neonatal fibers were largely unbranched many of them having axons tipped with growth cones. Axon terminals containing synaptic vesicles were rarely observed but when present these exhibited considerable variation in their morphological appearance of synapses. Thus, it was not possible to categorize them into the two types of axons which characterize the adult. After P25, terminal swellings bearing a close resemblance to those of type II fibers begin to appear. In this way, the main two corticothalamic fiber types could be identified. These findings demonstrate that significant postnatal changes occur in the synaptology of corticothalamic fibers in the LM-Sg, particularly with the maturation of type II fibers.     

The pineal gland of the Indian palm squirrel, Funambulus pennanti (Wroughton)

In the adult palm squirrel, F. pennanti the pineal is a club shaped, elongated structure with a connective tissue capsule. It consists of various types of pinealocytes, glial cells, neurons, nerve fibres, blood vessels and connective tissue. Two types of pinealocytes could be identified by light microscopy. They are large rounded with centrally placed nucleus, and small rounded pinealocytes. They have medium sized processes stainable with Alcian blue, periodic acid Schiff and Nissl methods. The pinealocytes are not stainable with bromophenol blue. However, they are moderately stainable with PAS, Sudan black and Baker's acid hematin. Neurons are seen either singly or in groups with axonal processes. Cystic cavities often lined by cells are a normal feature of adult squirrel pineal, and the lining cells are both pinealocytes and glial cells. Often neuronal endings are seen terminating on these lining cells. PAS positive globules were also seen inside the cysts. In some squirrel pineals, fibrous cysts with an inner core of cells are also seen. Occasionally groups of lymphocytes were also encountered in the pineal. In the fetal pineal, the cells are both larger and smaller ones and arranged in a cortex and medulla pattern and no cystic cavities are seen. The third ventricle enters the base of the pineal as pineal recess.     

Immunohistochemical study of the postnatal development of pituitary thyrotrophs in the rat,with special reference to cluster formation

Summary The postnatal development of rat pituitary thyrotrophs was investigated immunohistochemically on days 1, 3, 5, 10, 15 and 25. Fetal thyrotrophs are strongly immunoreactive. In the postnatal period, however, weakly immunoreactive thyrotrophs increase in number to constitute clusters on days 3–5. The numbers and dimensions of the clusters reach a maximum on day 10. Thereafter the clusters break down to give rise to single, scattered neogenic thyrotrophs. Thyrotrophs in clusters on day 10 were investigated by electron microscopy in adjacent sections. They can be characterized as an immature type of basophil, according to the classification of Yoshimura et al. (1977): 1) Type I basophils, which are irregularly shaped with elongate processes, and characterized by rows of secretory granules about 100 nm in diameter. 2) Type I/II basophils, i.e., forms intermediate between Types I and II, containing less numerous secretory granules about 100–150 nm in diameter. Type II basophils which correspond to the classical thyrotrophs are not fully developed on day 10. Thus, most thyrotrophs develop from the clusters in the neonatal period. Such neogenic thyrotrophs retain the immature characteristics of Type I and I/II cells and may develop into Type II cells during subsequent maturation.     

Effect of destruction of the medial forebrain bundle on synaptic organization of the supraoptic nucleus of the rat hypothalamus

An electron-microscopic investigation was made of the synaptic organization of the supraoptic nucleus (SON) of the albino rat hypothalamus before and after electrolytic destruction of fibers of the medial forebrain bundle (MFB). Two types of axon terminals, tentatively of cholinergic (type I) and monoaminergic (type II) nature, are described in the nucleus. In intact animals type I terminals account for 30% and type II for 70%. Both types of terminals form axo-dendritic, axo-somatic, and axo-axonal contacts. Five days after bilateral electrolytic destruction of MFB 13.5% of terminals degenerated in SON, as shown by swelling of the synaptic vesicles, their redistribution in the terminal, and vacuolation of the terminal. Quantitative analysis of preserved endings and comparison with results obtained on intact animals show that mainly type II axon terminals, presumably monoaminergic in nature, degenerate. The decrease in the number of these terminals after destruction of MFB is evidence of the mainly monoaminergic nature of fibers running in this bundle to SON, and it enables the pattern of distribution of different types of axon terminals in the nucleus to be characterized quantitatively.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 14, No. 3, pp. 227–232, May–June, 1982.     

Type II epithelial cells are critical target for hyperoxia-mediated impairment of postnatal lung development

Type II epithelial cells are essential for lung development and remodeling, as they are precursors for type I cells and can produce vascular mitogens. Although type II cell proliferation takes place after hyperoxia, it is unclear why alveolar remodeling occurs normally in adults whereas it is permanently disrupted in newborns. Using a line of transgenic mice whose type II cells could be identified by their expression of enhanced green fluorescent protein and endogenous expression of surfactant proteins, we investigated the age-dependent effects of hyperoxia on type II cell proliferation and alveolar repair. In adult mice, type II cell proliferation was low during room air and hyperoxia exposure but increased during recovery in room air and then declined to control levels by day 7. Eight weeks later, type II cell number and alveolar compliance were indistinguishable from those in room air controls. In newborn mice, type II cell proliferation markedly increased between birth and postnatal day 7 before declining by postnatal day 14. Exposure to hyperoxia between postnatal days 1 and 4 inhibited type II cell proliferation, which resumed during recovery and was aberrantly elevated on postnatal day 14. Eight weeks later, recovered mice had 70% fewer type II cells and 30% increased lung compliance compared with control animals. Recovered mice also had higher levels of T1alpha, a protein expressed by type I cells, with minimal changes detected in genes expressed by vascular cells. These data suggest that perinatal hyperoxia adversely affects alveolar development by disrupting the proper timing of type II cell proliferation and differentiation into type I cells.     

Alveolar differentiation in rabbits

Lung tissue of the white New Zealand rabbit was examined by transmission and scanning electron microscopy from the 23rd to the 30th day post conception (p.c.). The following results were obtained: 1. About day 23 p.c., when the development of capillaries increases, the "canalicular period" starts. This is followed by the "terminal sac period" characterized by the beginning of alveolarisation. On day 28 p.c. typical alveoles can be found. 2. The Pneumoplast is the stem cell of the pneumocyte type I as well as type II. They differentiate parallel in either one or the other. This stem cell of the entodermal origin has a single cilium. During the period of single cilia growth the cell is not mitotic. 3. The maturation of the lamellar body, typical of the pneumocyte type II, can be accomplished in a direct as well as in a indirect way of synthesis. Transitions between both are possible. 4. The most important factors of differentiation are collagenic induction substances beside nerval and humoral factors. Those humoral factors can be transported easier into the cells with advancing capillarisation as a result of the shortened distance of diffusion.     

Transitional expression of OX-2 and GAP-43 glycoproteins in developing rat cochlear nerve fibers

The OX-2 and GAP-43 glycoproteins are two proteins involved in neuronal cell-to-cell interaction and/or growing of dendrites and axons. Therefore, for the auditory receptor the expression of these proteins could provide information on the afferent and efferent nerve fiber organization. The expression and distribution of OX-2 and GAP-43 were analyzed during the auditory receptor development and maturation (from embryonic day E13 to postnatal day P22). Both glycoproteins were early recognized in the cochleae of E13 rats. Then, they slowly but progressively disappeared, being absent when the animals reached the P22 postnatal day. At E13, a weak OX-2 expression was restricted to the perikaryon of the spiral ganglion neurons, while in the same period a strong GAP-43 immunostaining was found in both the neuronal perikaryon and the neurites. During the rat embryonic period (E13 to birth) the expression of both glycoproteins appeared progressively restricted to the neurites. During the rat postnatal period (P0 to P22), OX-2 and GAP-43 exhibited a dissimilar distribution pattern. The OX-2 glycoprotein appeared in the afferent, efferent and fibers of the auditory nerve, while the GAP-43 glycoprotein only appeared in the efferent nerve fibers. Present data suggest that OX-2 and GAP-43 could act as two complementary glycoproteins during the development, organization, and maturation of the cochlear nerve fibers. While both glycoproteins could participate in axonal growing and orientation, OX-2 could also be involved in a similar process for auditory dendrites.     

The pineal gland of nocturnal mammals

Summary In the pineal gland of the pipistrelle bat two different populations of pinealocytes and glial cells were observed electron microscopically. The pinealocytes of populations I and II differ in their content of metabolically active cell organelles. In the pinealocytes of population I, granular vesicles originating from the Golgi apparatus were found in the perikaryon and especially in the endings of the pinealocyte processes. Granular vesicles appeared to be more numerous in hibernating nulliparous females. The pinealocytes of population II are characterized by the presence of small cytoplasmic vacuoles, probably originating from cisternae of the granular endoplasmic reticulum and containing flocculent material of moderate electron density. The classification of the pinealocytes belonging to population II is discussed.This collaboration was initiated with the aid of an SRC European short visit grant to P.A.R.The study was supported by the Foundation for Medical Research, the Netherlands (FUNGO, 13-35-33)     

Postnatal development of "synaptic" ribbons and spherules in the guinea pig pineal gland

Previous studies have shown that the functionally enigmatic pineal "synaptic" ribbons are structurally a heterogeneous group of organelles consisting of rodlike ribbons sensu stricto, spherules, and intermediate forms. As ribbons and spherules react differently under various experimental conditions, these organelles were studied qualitatively and quantitatively during the postnatal period in guinea pigs. It was found that the pinealocytes were highly differentiated at birth and contained all three forms of "synaptic" structures. Ribbons and intermediate forms were more abundant than spherules and exhibited a striking increase in number on postnatal days 1 and 2; this increase was followed by a distinct trough and by a second peak at days 12 and 13, after which their numbers declined to reach adult levels by day 20. The spherules were small in number at birth and did not show the large immediate postnatal increase observed for the ribbons and intermediate forms. Instead there was a steady numerical increase up to day 12 (absolute number) or day 15 (relative numbers), followed by a decrease to adult level by day 20. Whereas during the early postnatal period (days 1 to 3) the majority of pinealocytes were characterized by ribbons and intermediate forms, with increasing age spherule-bearing pinealocytes increased in number. As ribbons and spherules were usually not found in the same pinealocyte, the present findings are interpreted to mean that ribbons and spherules characterize different types of pinealocytes showing an inverse numerical development postnatally. Developmentally intermediate forms behave like ribbons.     

Prenatal and early postnatal development of the glial cells in the median eminence of the rat

Summary The development of the glial cells of the rat median eminence (ME), including the supraependymal cells, was investigated from embryonic day (ED) 14 through postnatal day (PD) 7, and pituicyte development from ED 12 through ED 17. The anlage of the ME and neurohypophysis shows a neuroepithelial-like structure at ED 12. From ED 13 to 15, the cells of both regions start to differentiate. At the ultrastructural level, only one cell type appears. At the beginning of ED 16, glioblasts of the oligodendrocyte and astrocyte series migrate laterally (from the region of the arcuate nucleus) into the ME. Also at this time the first distinctive structural features appear in the neurohypophysial anlage, the cells of which later develop into pituicytes. Starting at ED 18, tanycytes and astrocytic tanycytes arise in the ME from local glial cells, and somewhat later oligodendroblasts and astroblasts are formed from immigrant glioblasts. Due to their common features, the pituicytes, tanycytes and astrocytic tanycytes apparently represent different forms of the same parent cell type. Microglial and supraependymal cells are first seen at ED 12. Initially, they resemble the prenatal phagocytic connective tissue cells and mature in the fetus into typical electron-dense microglia and macrophage-like supraependymal cells. Both cell types are apparently of mesodermal origin. The microglial elements of the ME probably migrate from the mesenchyma through the basement into the nervous tissue. The intraventricular macrophages of the infundibular region may originate from microglia, epiplexal cells and subarachnoid macrophages.Dedicated to Prof. I. Törö, Budapest, on the occasion of his 80th birthday     

A push-pull set of elastic strand stretch receptor neurons for the swimmeret in an isopod Bathynomus doederleini

There coexist two types of neuronal terminal processes attaching to elastic strands at the socket of the swimmeret in Bathynomus doederleini. One of the processes, stretch receptor I is derived from the 1st nerve root of the abdominal ganglion. The other, stretch receptor II is derived from the 2nd nerve root of the ganglion. Both axons of stretch receptors are very thick (30-60 micro m) at sites before the terminal arborization. Cell bodies of the stretch receptors are located in the ganglion of their own segments. The neuronal cell body of the stretch receptor I is located at the anterior half of the hemiganglion ipsilateral to the periphery, and the neuronal cell body of the stretch receptor II at the posterior half of the hemiganglion contralateral to the periphery. Their signaling modalities in response to swimmeret movements were analyzed from intracellular recordings from the cell bodies. Stretch receptor I produced a sustained hyperpolarizing potential in response to protraction of the swimmeret. Stretch receptor II produced a sustained depolarizing potential in response to the protraction, and moreover, generated spike potentials on the rising phase of the depolarizing potential according to its height and steepness. Both the stretch receptors are a push-pull set of elastic strand stretch receptors for the angular position and velocity of swimmeret movements.     

Formation and maturation of axo-glandular synapses and concomitant changes in the target cells of the rat subcommissural organ

Synapse formation and maturation in the subcommissural organ (SCO) of Wistar rats were studied from birth to the end of the first month. Modifications of the secretory ependyma were analyzed over the same period. On the 1st postnatal day, the large varicosities in contact with the SCO ependymocytes appeared immature (absence or low density of vesicular population, no synaptic membrane differentiation). The synaptic contacts were formed from the 3rd postnatal day, near the glandular cell nuclei (0.1 micron distance); progressively, the content of the axonal boutons and the pre- and post-synaptic specializations became similar to those of adults. From the 21st day on, the axo-glandular innervation was considered analogous to that in the adult. Using immunocytochemistry, it was found that the increase in the serotonin-immunoreactive fiber density in the whole organ was time-dependent. Light and electron microscopy demonstrated changes in the morphology of SCO ependymocytes during the first postnatal weeks, notably in the endoplasmic reticulum and content ot apical protrusions. On postnatal day 14, two types of ependymal cells, neonatal-like and adult-like, coexisted. The evolution of SCO ependymocytes coincided with the progressive onset and maturation of axo-glandular innervation taking place after birth.     

Immunolocalization of type IV collagen and laminin during rat gonadal morphogenesis and postnatal development of the testis and epididymis

Summary The distribution of type IV collagen and laminin was studied by immunocytochemistry during rat gonadal morphogenesis and postnatal development of the testis and epididymis. Immunostaining appeared as early as the 12th day of gestation along the basement membranes of the mesonephric-gonadal complex. The connection between some mesonephric tubules and coelomic epithelium was seen between the 12th and 13th day of gestation. Discontinuous immunostained basement membranes delineated the differentiating sexual cords in 13-day-old fetuses; this process probably began in the inner part of the gonadal ridge. The seminiferous cords surrounded by a continuous immunoreactive basement membrane are separated from the coelomic epithelium by the differentiating tunica albuginea in 14-day-old fetuses. During the postnatal maturation of epididymis and testis, the differentiation of peritubular cells is accompanied by a progressive organisation of the extracellular matrix into a continuous basement membrane. This change is associated with a gradual condensation of peritubular cells inducing an increase of immunostaining. In adult animals, the tubular wall of epididymis is thicker than the lamina propria of seminiferous tubules. Both type IV collagen and laminin immunostaining paralleled during ontogenesis at the light-microscope level.     

Changes in the content of the fibrillar collagens and the expression of their mRNAs in the menisci of the rabbit knee joint during development and ageing

Summary The menisci are first seen as triangular aggregations of cells in the 20-day rabbit fetus. At 25-days, a matrix that contains types I, III and V collagens has formed. These collagens are also found in the 1-week neonatal meniscus, but by 3 weeks, type II collagen is present in some regions. By 12 to 14 weeks, typically cartilaginous areas with large cells in lacunae are found and by 2 years, these occupy the central regions of the inner two-thirds of the meniscus. The surface layers of the meniscus contain predominantly type I collagen. From 12 to 14 weeks onwards, there is little overlap between the regions with types I or II collagens, that is, these are discrete regions of type I-containing fibrocartilage and type II-containing cartilage. Types III and V collagens are found throughout the menisci, particularly in the pericellular regions. All the cells in the fetal and early neonatal menisci express the mRNA for type I collagen. At 3 weeks postnatal, cells that express type I collagen mRNA are found throughout the meniscus, but type II collagen mRNA is expressed only in the regions of developing cartilage. At 12- to 14-weeks, only type II collagen mRNA is expressed, except at the periphery next to the ligament where a few cells still express type I collagen mRNA. Rabbit menisci, therefore, undergo profound changes in their content and arrangement of collagens during postnatal development.     

Expression patterns of bone-related proteins during osteoblastic differentiation in MC3T3-E1 cells

Bone formation involves several tightly regulated gene expression patterns of bone-related proteins. To determine the expression patterns of bone-related proteins during the MC3T3-E1 osteoblast-like cell differentiation, we used Northern blotting, enzymatic assay, and histochemistry. We found that the expression patterns of bone-related proteins were regulated in a temporal manner during the successive developmental stages including proliferation (days 4–10), bone matrix formation/maturation (days 10–16), and mineralization stages (days 16 –30). During the proliferation period (days 4–10), the expression of cell-cycle related genes such as histone H3 and H4, and ribosomal protein S6 was high. During the bone matrix formation/maturation period (days 10–16), type I collagen expression and biosynthesis, fibronectin, TGF-β1 and osteonectin expressions were high and maximal around day 16. During this maturation period, we found that the expression patterns of bone matrix proteins were two types: one is the expression pattern of type I collagen and TGF-β1, which was higher in the maturation period than that in both the proliferation and mineralization periods. The other is the expression pattern of fibronectin and osteonectin, which was higher in the maturation and mineralization periods than in the proliferation period. Alkaline phosphatase activity was high during the early matrix formation/maturation period (day 10) and was followed by a decrease to a level still significantly above the baseline level seen at day 4. During the mineralization period (days 16–30), the number of nodules and the expression of osteocalcin were high. Osteocalcin gene expression was increased up to 28 days. Our results show that the expression patterns of bone-related proteins are temporally regulated during the MC3T3-E1 cell differentiation and their regulations are unique compared with other systems. Thus, this cell line provides a useful in vitro system to study the developmental regulation of bone-related proteins in relation to the different stages during the osteoblast differentiation. © 1996 Wiley-Liss, Inc.     

Juvenile hormone regulation of structural changes and DNA synthesis in the follicular epithelium of Leucophaea maderae

Juvenile hormone (JH I) stimulates specific morphological and biochemical changes in the follicular epithelium surrounding the terminal oöcytes in Leucophaea maderae. These include extracellular and intracellular structural changes, increased rates of follicle cell DNA synthesis, and elevated follicle cell DNA concentrations.Using females decapitated 24 hr after ecdysis, we have shown that JH I injections stimulate the following structural changes in the follicular epithelium: the appearance of channels between adjacent follicle cells and of spaces between the follicular epithelium and the maturing oöcyte; an increase in follicle cell size; the development of an extensive rough endoplasmic reticulum system; and an enlarged nucleus within each follicle cell. No increase in the number of follicle cells surrounding the developing terminal follicles is found in 7-day JH I-treated females, although the terminal follicles are almost twice as long as those in untreated females.In addition, we have demonstrated that JH stimulates the following biochemical events in the ovary: a 3.5 fold increase in thymidine incorporation into follicle cell DNA, with no subsequent transfer of such DNA to the developing oöcyte, and a 1.4 fold increase in ovarian DNA in 7-day JH-treated females. These data indicated that JH stimulates follicle cell DNA synthesis. The absence of any corresponding division of follicle cells suggests that JH I may induce polyploidy in follicle cells.Extended exposure of decapitated females to JH I does not result in complete ovarian maturation. Although fat bodies in the treated insects continue to display an increasing rate of vitellogenin synthesis, DNA synthesis in the terminal follicles declines rapidly after day 9, and the terminal follicles ultimately degenerate.     

Endosperm differentiation in barley wild-type and sex mutants

The cereal endosperm is a storage organ consisting of the central starchy endosperm surrounded by the aleurone layer. In barley, endosperm development is subdivisible into four main stages, i.e. the syncytial (I), the cellularization (II), the differentiation (III) and the maturation stage (IV). During stage I, a multinucleate syncytium is formed, which in stage II develops into the undifferentiated cellular endosperm. During stage III the cells of the endosperm differentiate into two types of aleurone cells (peripheral and modified) and three different starchy endosperm cell types (irregular, prismatic and subaleurone). To elucidate the ontogenetic relationship between the endosperm tissues, the phenotypes of sex (shrunken endosperm mutants expressing xenia) mutant endosperms were studied. These mutants can be classified into two groups, i.e. those in which development is arrested at one of the four wild-type stages described above, and those with abnormal development with new organizational patterns in the endosperm or with novel cell types. Based on these studies, it is suggested that the two endosperm halves represent cell lines derived from the two daughter nuclei of the primary endosperm nucleus, and that the prismatic starchy endosperm cells arise from a peripheral endosperm meristematic activity during stage III. Finally, a model for the main molecular events underlying the morphogenetic processes is discussed.