The alveolar pores of Kohn in young postnatal rat lungs and their relation with type II pneumocytes.

In order to obtain more information on the development, morphology and function of the pores of Kohn, the lungs of Wistar rats are studied during their early postnatal period, up to 3 weeks of age, by scanning and transmission electron microscopy. The substantial development of the interalveolar pores on days 14 and 21 coincides with the period of septal rearrangement when secondary interalveolar septa become lengthened and thinner. The high frequency of transseptal type II pneumocytes from day 7 onwards, and their typical localization near the pores of Kohn at this period of lung development especially suggests that type II pneumocytes are engaged in the formation of the pores of Kohn. During early lung development, the pores of Kohn seem to serve as passageways for alveolar macrophages.     

The effect of age on lung structure in male BALB/cNNia inbred mice

Morphometric examination using light, scanning electron, and transmission electron microscopy was performed on the lungs from 32 inbred male BALB/ cNNia mice between 38 days and 28 months of age. Between 38 days and 9 months of age the changes were primarily those of growth. Alveolar multiplication and total elastic-fiber length appeared complete by 38 days of age. The major increase in the number of interalveolar pores occurred by 68 days, but there was a significant further increase from 68 days to 9 months of age. At 9 months, approximately 10% of the alveolar wall was formed by pores. Alveolar ducts, the cylindrical core of air central to alveolar mouths, increased more in diameter than length. Between 9 and 28 months the changes were attributed to aging and were different from those reported in humans and other species. Lung volume, alveolar surface area, and total volume of alveolar wall increased with age; there was no change in mean linear intercept and volume proportion of alveolar and alveolar duct air. Total area of pores increased with age, but their number and area fraction of the alveolar wall did not change. No transmission electron microscopic changes were seen in the alveolar walls. We speculate that the morphometric differences between our animals and those studied in other reports may reflect the fact that our animals were specific-pathogen-free animals and kept under protected circumstances.     

Distribution of pores within alveoli in the human lung

Usually the wall opposite the orifice of alveoli has been used to study interalveolar pores by scanning electron microscopy. To ascertain whether biased results may be obtained from this, the distribution of pores within alveoli was studied in human lungs. By the use of scanning electron photomicrographs, the number, major axes of pores, and proportional area of pores to the alveolar wall were estimated. The alveolar wall seen opposite the orifice was defined as the bottom wall. Average number of pores per alveolus was 13-21, and one-half of them was located in the bottom walls. The average length of major axes was 7-19 micron, and average area fractions were 0.8-5%. The distribution of the numerical density, area fraction, and size of pores was uniform regardless of their location within the alveolus and the size of alveoli. Thus pores can be compared using the bottom walls of alveoli. This will facilitate the study of the effects of age, smoking, and topography on pore size and frequency in humans.     

Pores of Kohn are filled in normal lungs: low-temperature scanning electron microscopy.

Interalveolar pores of Kohn, small uniform-sized epithelium-lined openings in alveolar walls of normal lung, have historically been demonstrated with electron-microscopic techniques that remove water. We show these pores to be present but almost invariably filled with material when water and surfactant are preserved in frozen hydrated lung examined with low-temperature scanning electron microscopy. In the normal mouse, 16 open empty pores per alveolus were found in instillation-fixed dried lung vs. less than 1 per alveolus in frozen hydrated lungs (P less than 0.001). In the normal rat, 13 pores were seen per alveolus in instillation-fixed dried lung vs. less than 1 per alveolus in frozen hydrated lungs (P less than 0.001). We suggest that pores of Kohn 1) function primarily as conduits for interalveolar movement of alveolar liquid, surfactant components, and macrophages, 2) provide distributed sites for tubular myelin storage without increasing gas diffusion pathway thickness in the alveolar subphase itself, and 3) do not function as pathways for collateral ventilation during normal breathing in the absence of atelectasis or obstruction.     

Study by transmission and scanning electron microscopy of the morphogenesis of three types of lingual papillae in the albino rat (Rattus rattus)

El‐Bakry, A.M. 2010. Study by transmission and scanning electron microscopy of the morphogenesis of three types of lingual papillae in the albino rat (Rattus rattus).—Acta Zoologica (Stockholm) 91 : 267–278 Tongues were removed from albino rat foetus on days 12 (E12) and 16 (E16) of gestation and from newborns (P0) and from juvenile rats on days 7 (P7), 14 (P14) and 21 (P21) postnatally for investigation by light, scanning, and transmission electron microscopy. Significant changes appeared during the morphogenesis of the papillae. At E12, two rows of rudiments of fungiform papillae were extended bilaterally on the anterior half of the tongue. At E16, the rudiments of fungiform papillae were regularly arranged in a lattice‐like pattern. A rudiment of circumvallate papillae could be recognized. No rudiment of filiform papillae was visible. No evidence of keratinization was recognizable. At P0, rudiments of filiform papillae were visible but had a more rounded appearance, with keratinization. The fungiform and circumvallate papillae were large and their outlines were somewhat irregular as that found in the adult rat. At P7, the filiform papillae were large and slender. The fungiform papillae became large and the shape of circumvallate papillae was almost similar to that observed in the adult. At P14 and P21, the shape and structure of the three types of papillae were irregular as those found in the adult. In conclusion, the rudiments of the fungiform and circumvallate papillae were visible earlier than those of the filiform papillae. The morphogenesis of filiform papillae advanced in a parallel manner with the keratinization of the lingual epithelium, in the period from just before birth to a few weeks after birth.     

Development of neuroepithelial bodies in intact and cultured lungs of fetal rats

Intact, 14- to 21-day fetal rat lung pairs, neonatal lungs, and cultured 15- and 16-day lung explants were examined in 2-micron-thick glycol methacrylate sections stained by PAS-lead hematoxylin. Selected stages were also studied in histochemical preparations for aliesterase and formaldehyde-induced monoamine fluorescence, as well as by scanning and transmission electron microscopy. Neuroepithelial bodies (NEBs) first appear in pseudoglandular lungs at 15 days in vivo as pyramidal groups of basal, diffusely lead-hematoxylin-positive cells in glycogen-depleted epithelium of main and lobar bronchi. By day 16, primitive NEBs occur within three to four generations of the terminal buds, and older, proximal bodies are larger and more distinctive than at 15 days. Aliesterase activity is first detected in basally located, developing NEBs on day 16. During the canalicular and alveolar sac periods, NEBs appear and mature on a proximal-to-distal gradient along the airway, as they do in developing rabbit and human lungs. As earlier-formed airways elongate, additional NEBs appear and supplement the population already present. By days 20-21, NEBs occur at all airway levels down to the bronchiolo-alveolar junctions, and many of the cells have discrete PAS- and lead-hematoxylin-positive, infranuclear granules. Near term some NEBs exhibit serotonin fluorescence after incubation in 5-hydroxytryptophan and have abundant, ca. 100-nm, electron-dense granules. These are concentrated toward the cell base like the stained granules visualized by light microscopy. Similar results were obtained from lungs placed in organ culture. From 2 days in culture to a time equivalent to term, NEB formation parallels that in vivo, indicating that developmental requirements are met in in vitro. Taken altogether, morphologic and cytochemical evidence suggests that NEBs of rats are functional in late fetal life and that their development is relatively independent of extrapulmonary influences and of the intraepithelial ingrowth of sensory nerve endings.     

Treatment with tacrolimus enhances alveolar bone formation and decreases osteoclast number in the maxillae: a histomorphometric and ultrastructural study in rats

Recent studies have suggested that tacrolimus monotherapy is a beneficial therapeutic alternative for the normalization of cyclosporin-induced bone loss in animal models and humans. The mechanism accounting for this action is unclear at present. In the present study, we attempted to determine the effect of tacrolimus monotherapy on alveolar bone using histological, histomorphometric and transmission electron microscopy (TEM). Groups of rats (n=10 each) were treated with either tacrolimus (1mg/kg/day, s.c.) or drug vehicle for 60 days. Fragments containing maxillary molars were processed for light microscopy to investigate the alveolar bone volume, trabecular separation, number of osteoclasts and osteoblasts, and transmission electron microscopy to investigate their ultrastructural basic phenotype. Treatment with tacrolimus monotherapy during 60 days may induce increases in alveolar bone volume (BV/TV,%; P<0.05) and a non-significant decrease in trabecular separation (Tb.Sp,mm; P>0.05), represented by a decrease in osteoclast number (N.Oc/BS; P<0.05) and maintenance of osteoblast number (N.Ob/BS; P>0.05). Osteoblasts were often observed as a continuous layer of active cells on the bone surface. Osteoclasts appeared to be detached from the resorbed bone surface, which was often filled by active osteoblasts and collagen-rich matrix. Moreover, osteoclasts in the treated group were frequently observed as inactive cells (without ruffled border, clear zone and detached from the bone surface). Within the limits of the present study, we conclude that tacrolimus leads to an increase in alveolar bone formation, which probably exerts action on osteoclasts. Tacrolimus could, therefore, play a crucial role in the control of both early osteoclast differentiations from precursors, as well as in functional activation.     

Localization of cholinesterases in structures of the alveolar portion of the mammary gland

Cholinesterase (ChE) localization in the alveolar compartment of the mammary gland has been studied by light microscopy and ultrahistochemical methods. The product of reaction to ChE is associated with micropinocytotic vesicles of the capillar endothelium and myoepithelial cells, separate nerve terminals in interalveolar connective tissue and perivascular nerve terminals. In secretory cells, the ChE activity is revealed in the sites of fat droplet and on the membranes of endoplasmatic reticulum.     

Dimensions and alveolar surface of the ineralveolar septa of the rat lung]

The dimensions of the alveolar surface of interalveolar septa were studied in albino rats. The mean thickness of certain septa range from 3,1 to 6,2 mu, the mean thickness of the septa in different individuals being factually the same. A part of the alveolar surface of the septa represents the air-haematic barrier which occupies 51-64% of the total alveolar surface in different rats. The mean values of all the parameters in the animals studied are very close. It shows that the interalveolar septa are standard structures in their organization and dimensions both in the same animal and in different animals of the same species.     

Macrophage activation in the immune response

In inbred white rats, immunized with sheep erythrocytes, contents and phagocytic activity of the spleen and pulmonary macrophages have been studied on the 3d, 4th, 5th, 6th, 7th, 11th, 14th and 20th days of the experiment in the light, scanning and transmissive electron microscope, as well as the effect of the cells mentioned on proliferation of lymphoid elements. Maximal phagocytic activity of the splenic and alveolar macrophages is observed on the 7th day of the experiment. At the same time, certain drop in the lymphoid cells proliferation takes place. The change in the macrophages contents also influences proliferation of the lymphoid cells.     

Development and morphology of rat synovial membrane

The development of the knee joint and synovial membrane in Wistar rats was studied from the 13th fetal day until adulthood by means of light microscopy, transmission electron microscopy and scanning electron microscopy. Using acid phosphatase and peroxidase methods, the different cell types of synovial membrane could be identified.     

Effects of neonatal monosodium-L-glutamate treatment on rat alveolar macrophages

Treatment of neonatal rats with repeated doses of monosodium-L-glutamate resulted in changes of morphology and function of alveolar macrophages recovered from adult female rats. Numerous cellular lipid vacuoles and lamellar structures were observed in these alveolar macrophages under transmission electron microscopy. There was approximately 50% increase of cellular total lipid content measured by Oil-Red-O staining and colorimetric method. The yeast phagocytosis and intracellular killing ability, as well as the inhibitory effect on the growth of tumor cells were reduced in alveolar macrophages from neonatal MSG-treated rats.     

Postnatal alveolar development of the rabbit.

Previous studies of alveolarization have used rats or lambs; however, neither closely reflects human alveolar development. We characterized alveolar development in rabbits (n = 3-7 /group) at 28 days gestation (dg) to 9 mo to determine whether they followed the human pattern more closely. The right lung was made up of 30% alveolar and 50% duct space at 28 dg to 3 days and of 50 and 30%, respectively, at 14 days to 9 mo. Tissue fraction and alveolar wall thickness decreased by 40% 28 dg to birth. At birth, approximately 4.5% of the number of alveoli seen at 9 mo were present, with alveolar number increasing progressively well into adulthood. The rate of alveolar formation was high around birth, decreasing progressively with age. Alveolar volume increased more than twofold (28 dg to birth) and continued to increase postnatally to 16 wk. Surface fraction decreased by 17% (28 dg to 3 days), after which it remained uniform. Our findings suggest that the timing of onset of alveolarization in humans and rabbits is similar and that rabbits may be used to model postnatal influences on alveolar development.     

Detection of Fc receptors for IgA on rat alveolar and peritoneal macrophages.

The presence of Fc receptors for IgA on alveolar macrophages was determined by rosette assay and immunogold labeling. IgA-mediated phagocytosis by alveolar macrophages was observed. Results of these assays were compared between rats receiving no treatment and those receiving long-term cortisone administration. Sheep erythrocytes coated with dextran and an IgA monoclonal antibody specific for the alpha 1,3 linkages of dextran bound to 16% of alveolar macrophages. However, peritoneal macrophages did not form rosettes with dextran-IgA-coated erythrocytes. Immunogold labeling by transmission electron microscopy revealed that most Fc receptors for IgA were found on the membrane of pseudopodia of activated alveolar macrophages. Long-term cortisone administration diminished the phagocytosis and phagocytic index of alveolar macrophages, thereby contributing to decreased host resistance to infection (e.g., Pneumocystis carinii pneumonia).     

Morphological research on the effect of vagotomy on the parietal microflora of the stomach and duodenum

The effect of vagotomy on parietal microflora was studied in intact rats and rats with chronic experimental gastric ulcer using stereometry, as well as light, transmission and scanning electron microscopy. Vagotomy was found to increase the relative amount of parietal microflora on day 30 following denervation, especially in the duodenum and pylorus. Chronic gastric ulcers are also associated with a rise in the relative amount of parietal microflora.     

Structural and Ultrastructural Characteristics of Bone-Tendon Junction of the Calcaneal Tendon of Adult and Elderly Wistar Rats

Tendons are transition tissues that transfer the contractile forces generated by the muscles to the bones, allowing movement. The region where the tendon attaches to the bone is called bone-tendon junction or enthesis and may be classified as fibrous or fibrocartilaginous. This study aims to analyze the collagen fibers and the cells present in the bone-tendon junction using light microscopy and ultrastructural techniques as scanning electron microscopy and transmission electron microscopy. Forty male Wistar rats were used in the experiment, being 20 adult rats at 4 months-old and 20 elderly rats at 20 months-old. The hind limbs of the rats were removed, dissected and prepared to light microscopy, transmission electron microscopy and scanning electron microscopy. The aging process showed changes in the collagen fibrils, with a predominance of type III fibers in the elderly group, in addition to a decrease in the amount of the fibrocartilage cells, fewer and shorter cytoplasmic processes and a decreased synthetic capacity due to degradation of the organelles involved in synthesis.     

Polyfunctional role of the alveolar brush cells in the rat lung

By means of scanning and transmission electron microscopy it was demonstrated that the number of vacuoles located in the apical part of cytoplasm in alveolar brush cells of the regenerating rat lung increases, hyperplasia of Golgi-complex takes place and the activation of the protein-synthetising apparatus is evident. The immature surfactant material (osmiophilic lamellar bodies) and secretory dense core vesicles were found in the cytoplasm of alveolar brush cells. Intramuscular injections of colchicin to rats (0.1 mg/100 g body weight) 6 times during 24 hours before decapitation does not influence the number, topography and structure of microfibrilla bundles contained in a sufficient amount by alveolar brush cells. At the same time a part of microvilli of alveolar brush cells undergoes destruction and resorption under the action of colchicin. The data on ultrastructural organization of alveolar brush cells show that they are able to fulfill several functions: absorptive, contractile and secretory.     

Characterization of rat calvarial nonunion defects

This study examined the healing of nonunions by describing the histology and ultrastructural appearance of craniotomy defects as a model. Bone defects (3, 4 and 8 mm) were created in the calvaria of adult rats. Central and peripheral specimens of 8-mm defects were retrieved at 1, 3, 7, 10, 14, 21, 28 and 42 days and examined using both light and transmission electron microscopy. Specimens from the 3- and 4-mm defects were retrieved at 28 days and examined using light microscopy. In all sizes of defects, bony repair was consistently localized to the dural side of the defect. The 3- and 4 mm defects demonstrated the greatest degree of osseous bridging and evidence of normal osseous repair throughout the defect. The 8-mm defects repaired in general with the formation of nonunions which contained a small amount of bone at the periphery and fibrous connective tissue. Bone formation was evident at 10 days in the peripheral regions of the 8-mm defects and exhibited bony peninsulas with normal primary calcification fronts. Matrix vesicles containing hydroxyapatite-like crystals were present. In contrast, the central regions of the 8-mm defects were characterized by several islands of cartilage-like cells which stained metachromatically with toluidine blue. Transmission electron microscopy of this region at 14 days demonstrated a dense, collagenous extracellular matrix with matrix vesicles infiltrating the collagen bundles. There was no evidence of crystal formation in the matrix vesicles nor of calcification in the collagenous matrix. At 21 days, both the central and peripheral regions of the 8-mm calvarial nonunions were characterized by dense fibrous connective tissue repair and inactive fibroblasts.(ABSTRACT TRUNCATED AT 250 WORDS)     

Apoptotic bone cells may be engulfed by osteoclasts during alveolar bone resorption in young rats

The alveolar bone is a suitable in vivo physiological model for the study of apoptosis and interactions of bone cells because it undergoes continuous, rapid and intense resorption/remodelling, during a long period of time, to accommodate the growing tooth germs. The intensity of alveolar bone resorption greatly enhances the chances of observing images of the extremely rapid events of apoptosis of bone cells and also of images of interactions between osteoclasts and osteocytes/osteoblasts/bone lining cells. To find such images, we have therefore examined the alveolar bone of young rats using light microscopy, the TUNEL method for apoptosis, and electron microscopy. Fragments of alveolar bone from young rats were fixed in Bouin and formaldehyde for morphology and for the TUNEL method. Glutaraldehyde-formaldehyde fixed specimens were processed for transmission electron microscopy. Results showed TUNEL positive round/ovoid structures on the bone surface and inside osteocytic lacunae. These structures--also stained by hematoxylin--were therefore interpreted, respectively, as osteoblasts/lining cells and osteocytes undergoing apoptosis. Osteoclasts also exhibited TUNEL positive apoptotic bodies inside large vacuoles; the nuclei of osteoclasts, however, were always TUNEL negative. Ultrathin sections revealed typical apoptotic images--round/ ovoid bodies with dense crescent-like chromatin--on the bone surface, corresponding therefore to apoptotic osteoblasts/lining cells. Osteocytes also showed images compatible with apoptosis. Large osteoclast vacuoles often contained fragmented cellular material. Our results provide further support for the idea that osteoclasts internalize dying bone cells; we were however, unable to find images of osteoclasts in apoptosis.     

Prolonged immobilization-induced stress delays alveolar bone healing. A histometric study in rats

The purpose of the present study was to investigate the effect of prolonged immobilization-induced stress on reparative bone formation, using the rat alveolar healing as an experimental protocol. Stress was attained by immobilization for 2 hours a day, beginning three days before extraction of the upper right incisors and continuing until sacrifice. The stress condition was assayed on the basis of plasma corticosterone concentration (measured by double-antibody radioimmunoassay), which increased by 2.5 to 4 times in rats submitted to immobilization. The volume density of neoformed bone filling the alveolar socket was quantified by a histometric differential point-counting method 7 to 21 days following tooth extraction. Stress caused a significant delay in reparative bone increment, somewhat related to impairment of coagulum remission and/or organization.