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Vallate, foliate and fungiform human papillae gustatory cells. An immunocytochemical and ultrastructural study.

We studied the classifications, topographic distribution and cellular lines of taste bud components in vallate, foliate and fungiform papillae of young, mature and old men with light microscopy, SEM, and TEM. By identifying ultrastructural and immunocytochemical characteristics, three distinct sensorial cells were identified, along with a few basal cells: dark type I cells, light type II cells and light type III cells. These cells extend from the epithelial basal lamina to the gustatory canal, where their apical cytoplasm sends long microvillous expansions. Excluding those of the fungiform papillae-which never go beyond the lower third of the gustatory canal, and are always void of dense substance-the microvillous expansions continue to the external border of the taste pore. Dark type I cells are rich in free ribosomes, tubular RER and large dense granules. Light type II cells with scarce ribosomes and RER, do not have enough peculiar ultrastructural characteristics to be considered effector or phagocyte elements. Light type III cells are characterizes by dense core vesicles whose peculiar ultrastructural characteristics in the foliate and vallate papillae, should be considered a consequence of different functional phases. After comparative evaluation the authors hypothesized on the functional value of some ultrastructural aspects and on the dense core vesicles which are immunoreactive to 5-HT. They observed that all gustatory cells are involved in taste transduction based on behaviours caused by microvilli in the gustatory canal and gustatory cell relationships with nerve endings. Moreover the authors noted that age does not seem to influence taste perception.

Adult↗

Sperm ultrastructure of six Australian hylid frogs from two genera (Litoria and Cyclorana): phylogenetic implications.

The spermatozoa of four fossorial (Litoria alboguttata, Cyclorana brevipes, Cyclorana novaehollandiae and Cyclorana cryptotis) and two non-fossorial australian hylid frogs (Litoria aurea and Litoria moorei) together with previously examined Litoria (Hylidae: Anura) are compared. In spermatozoal ultrastructure (in particular the structure of the sperm tail) Cyclorana includes species which appear derived (apomorphic) relative to non-fossorial species of Litoria while the fossorial L. alboguttata groups with Cyclorana. All hylid species examined here are united by the bufonoid synapomorphy of a conical subacrosomal cone consisting of separate sheaves and the eubufonoid synapomorphy of a mitochondrial sheath or collar separated by a cytoplasmic canal from the centriolar region and tail. Spermatozoal symplesiomorphies for the Eubufonoidea, seen in Litoria (with the exception of L. alboguttata), are the well developed thin undulating membrane with juxta-axonemal and axial fibre. L. alboguttata, C. novaehollandiae and C. brevipes appear monophyletic in the apomorphic modification of the undulating membrane as a thick, dense structure. In L. alboguttata and C. novaehollandiae this structure retains a swelling, at the free edge, homologized with the axial rod. C. brevipes has a further apomorphy as the undulating membrane forms a parallel-sided dense structure with no separate differentiation of an axial fibre. C. cryptotis, however, retains the plesiomorphic sperm tail with a thin undulating membrane, juxta-axonemal and axial fibre. That these differences in spermatozoal ultrastructure have phylogenetic significance is endorsed by the similarity of the fertilization biology of the species examined. On the basis of sperm ultrastructure three separate lineages are discerned within Cyclorana s. lat.: 1) C. cryptotis; 2) L. alboguttata and C. novaehollandiae; and 3) C. brevipes. The evidence of sperm ultrastructure, supported by previously published molecular, morphological and karyological data, clearly places Litoria alboguttata within the genus Cyclorana.

Animals↗

Early ultrastructural lesions of apoptosis induced in vivo in two varieties of tissues (thymus and kidneys) after a single whole-body-gamma irradiation of adult mice.

Three groups of adult Swiss albino female mice with an average body weight of 20 grams were exposed to a whole-body irradiation by 137-cesium gamma rays at 2, 4 and 6 Gy (1.4 Gy/min.). For all groups, samples of thymus and kidney were taken 15 min., 2, 6 and 24 hrs. after irradiation and immediately prepared for observation by electron microscopy. In the thymus the earliest significant ultrastructural lesions are observed in the nuclei from the fifteenth minute. These lesions are characterized by condensation of chromatin, convolution and fragmentation of nuclei. Alteration of mitochondria and clarification of cytoplasm in a number of thymocytes are also observed depending on the dose of radiation. These lesions increased according to the length of the post-irradiation period and after six hrs. a great number of macrophages are observed in the thymus. Most often a single macrophage, contain several apoptotic thymocytes, with a maximum number up to eight. In the kidney, serious lesions were observed, affecting both, the nucleus and the cytoplasm of the proximal convoluted tubule cells (PCT). However in these cells, the most significant and earliest lesions consist of wide cytoplasmic clarifications, severe mitochondrial damages, associated with architectural modifications of the brush border and the beta-cytomembranes. These damaged cells have been observed to occur close to normal ultrastructural cells. Less significant ultrastructural alterations are also noticed in different varieties of glomerular cells. In contrast to the serious lesions of the PCT and glomerular cells, no ultrastructural alteration were observed in the distal tubule.

Animals↗

Effect of beta-sitosterol on ultrastructure of liver cells in young and aged mice.

The effects of beta-sitosterol on liver ultrastructure were studied using young and aged male mice. Both groups of mice were fed a beta-sitosterol or a control diet. The beta-sitosterol diet contained 1% beta-sitosterol. Ultrastructural differences in the liver cells were evident between two groups fed the two diets. The number of peroxisomes was significantly greater in the liver cells of both young and aged mice fed the beta-sitosterol diet than in those fed the control diet. There were also significant differences in the maximum length of lipid droplets between the young and aged mice fed the beta-sitosterol diet. These results suggest that beta-sitosterol affects liver ultrastructure, and that some ultrastructural differences in liver cells exist between young and aged mice fed the beta-sitosterol diets.

Aging↗

The ultrastructure of ependymoma: personal experience and the review of the literature.

I report here the ultrastructure of 29 ependymal tumors. The ultrastructural pattern was florid and characteristic with a picture dominated by the presence of microlumina, cilia with basal bodies (blepharoplast), microvilli and long, interdigitating intercellular junctions of the zonulae adherentes (adhesive plaque junctions) type. Tumor cells themselves were not particularly peculiar but they formed typical patterns of rosettes (so called mini- or ultrastructural rosettes) cell gatherings around small, electron-lucent lumina which are filled with numerous microvilli. Empty microlumina were rare. The apical and lateral portions of the cells surrounding microlumina were sealed by intercellular junctions which are long, tortuous and clearly different from the zonulae occludentes (tight junctions) of epithelial tumors. Clusters of apparently "redundant" junctions were occasionally visible comprising segments of different lengths. Ependymoma cells contained myriads of 10 nm intermediate filaments (glial filaments), occasionally forming thick bundles, virtually identical to those encountered in astrocytic tumors and forming an ultrastructural correlate for the GFAP immunostaining. The glycogen granules were often remarkably numerous. Numerous cilia, with a typical 9+1 pattern or with a distorted pattern were frequently observed in longitudinal or cross-sections.

Brain Neoplasms↗

[Effect of thyrotropic hormone on the ultrastructure of the endothelium of thyroid gland perifolicular capillaries].

Under study was the alteration in the ultrastructure of the thyroid epithelium and endothelium of perifollicular capillaries influenced by exogenous thyrotropic hormone. The results obtained show that there is a direct relationship between the dosage of the injected thyrotopic hormone, the degree of the ultrastructural changes in the time of their appearance. It has been established that the alterations in the perifollicular capillary endothelium ultrastructure make their appearance and become maximum practically simulataneously with the alterations in the thyroid epithelium ultrastructure. The increased functional activity of the endothelium, thinner walls of capillaries and the appearnace of a greater amount of fenestrations against the background of the thyroid stimulation are likely to be factors contributing to penetration of non-hormonal iodine products (iodine tyrosines and products of incomplete hydrolysis of thyroglobulins) into the circulation, which can be observed under certain pathological conditions accompanied by increased thyrotropic stimulation--such as diffused toxic goiter and diffuse non-toxic goiter.

Animals↗

Ultrastructural in situ hybridization: a review of technical aspects.

Detection of nucleic acid sequence at the ultrastructural level has allowed us to better understand the expression of genes in some fields of application in cell biology. In situ hybridization at the ultrastructural level can be carried out using three different methods: on vibratome sections before embedding in epoxy resin, on ultrathin frozen section, or on ultrathin section of tissue embedded in hydrophilic resin such as Lowicryl. Before starting the detection of nucleic acid sequences at the electron microscope level, the experimenter has to choose various parameters; the type of tissue fixation, the probe and its label, and the in situ hybridization method, depending on the sensitivity, the resolution and the ultrastructural preservation required. This review of technical aspects, by describing the different methods of ultrastructural in situ hybridization, will help the experimenter to optimize each step of the hybridization procedure.

Animals↗

[Effect of vagotomy on the ultrastructural organization of the enterochromaffin cells of the rat duodenal mucosa].

Detailed investigation of the ultrastructure of the enterochromaffine cells of the duodenal mucous membrane was conducted by electron microscopy. These cells were studied 7, 14, 28, and 56 days after bilateral subphrenic vagotomy. Vagotomy resulted in the alteration of the ultrastructural orgainzation of the enterochromaffine cells. These changes were due to disturbances of 5-hydroxytryptamine (serotonine) secretion. The changes in the ultrastructure of the enterochromaffine cells were most expressed 7 and 56 days after the operation. Temporary and relative normalization of the ultrastructure was observed in 28 days.

Animals↗

Ultrastructure of collagen thermally denatured by microsecond domain pulsed carbon dioxide laser.

BACKGROUND: Clinical improvement in photodamaged skin after carbon dioxide (CO2) laser resurfacing is thought to result in part from thermal collagen shrinkage. The presence of such collagen has not been unequivocally demonstrated. To identify and characterize the morphological features of collagen after CO2 laser exposure, we irradiated ex vivo human facial skin and bovine calcaneus tendon with microsecond domain pulsed CO2 laser energy and examined specimens for histopathological and ultrastructural changes in collagen. OBSERVATIONS: In dermis and tendon, 3 zones of collagen structure were apparent on electron microscopy. The first, most superficial zone demonstrated loss of collagen structure. The second zone consisted of admixed normal collagen fibers and thickened collagen fibers. Zone 3 consisted of normal-appearing collagen fibers. CONCLUSIONS: Ultrastructural examination of irradiated collagen revealed distinct morphological zones of denatured collagen fibers. Partially denatured fibers had an increased diameter consistent with lineal shrinkage. Zonal distinction was undetectable by light microscopy. Ultrastructurally, the zones of denatured collagen located above the normal fibers correlated with the zone of altered material seen on light microscopy. These findings suggest that collagen fiber shrinkage does occur after pulsed CO2 laser irradiation and that this phenomenon contributed, at least in part, to the immediate tissue contraction observed clinically.

Animals↗

Lipofilling of the lips: ultrastructural evaluation by transmission electron microscopy of injected adipose tissue.

OBJECTIVE: To determine if facial autologous fat grafts preserve the morphologic features of adipocytes and guarantee a successful long-term outcome. METHODS: In a previous study, we performed lipofilling in 99 patients between January 1, 1999, and December 31, 2001. In all patients, we performed 3 fat injections at 28-day intervals. We injected 0.4 mL of adipose tissue in the parafrenal area of the upper lip for each treatment. After 4 months, we obtained a biopsy specimen from the same area. We performed an ultrastructural evaluation on freshly harvested fat at the time of harvesting, on stored fat (-30 degrees C) at 8-week and 12-month intervals, and on the biopsy specimens obtained 4 months after treatment. RESULTS: We observed good preservation of the ultrastructure in the harvested tissue. On histologic examination of the parafrenal area 4 months after grafting, some zones of the biopsy specimens showed putative adipocytes, fat cysts, and collagen fibers adequate for volume increase of the treated area. Ultrastructural images showed lipid droplets intermingled in the connective tissue, phagocytes with internal lipid droplets, and well-preserved adipocytes. CONCLUSIONS: This study demonstrates that by using a less traumatic surgical technique, it is possible to increase the cell survival rate of transplanted fat, thereby maintaining a certain number of viable cells and creating a volume increase in the grafted area. The multiple-stage injection technique seems to be a good method, especially when performed with fat stored at -30 degrees C.

Adipocytes↗

Congenital myopathy and cardiomyopathy with identical ultrastructural changes.

A 7-day-old girl with congenital hypotonia and unexplained episodes of bradycardia had a broad spectrum of similar skeletal muscle and myocardial degenerative ultrastructural abnormalities. Ultrastructural studies showed obliteration of cross striations, myofilament disorganization, streaming, smearing, clumping, and zigzag Z-band deformities. A decrease in glycogen, mitochondria, and T-tubular system occurred in the regions showing Z-band abnormalities of both skeletal muscle and myocardium. Concurrent structural cardiomyopathy should be considered in patients with congenital myopathies, particularly with unexplained cardiac conduction abnormalities or contractile insufficiency. Ultrastructural evaluation of skeletal and cardiac muscle may be necessary to define such disorders.

Biopsy↗

Ultrastructural analysis of extraocular muscle in chronic progressive external ophthalmoplegia.

Extraocular muscles are primarily involved in many mitochondrial diseases, but no reports exist regarding the morphological appearance of the muscles in cases of long-standing ocular myopathies. For this reason, muscle samples obtained from surgery in a sporadic case of chronic progressive external ophthalmoplegia (CPEO) were used for ultrastructural investigation and molecular analysis of mitochondrial DNA. Genetic testing revealed a heteroplasmic macrodeletion of about 5.0 kilobases in length, localized between the 9570- and 14619-base pair regions. Electron microscopy revealed focal areas of both disruption and abnormality of mitochondria in only some of the muscle fibers, producing "selective vacuolization." This ultrastructural pattern was highly selective and limited to some extraocular muscle fibers, sparing all the others. The "selective damage" observed in this case of CPEO resembles that case occurring in another mitochondrial disease, Leber hereditary optic neuropathy, where damage occurs only in the papillomacular bundle of the retina, sparing peripheral axons. It is possible that some anatomical and physiological factors play a leading role in both Leber hereditary optic neuropathy and ocular myopathies. The ultrastructural aspect herein observed needs to be further investigated to better understand whether a particular muscle fiber type is the target of mitochondrial impairment in CPEO.

Base Sequence↗

Ocular ochronosis. Clinicopathological, histochemical, and ultrastructural studies.

Presented here are the clinical, histological, histochemical, and ultrastructural findings of an eye with notable ochronotic changes. Ochronotic pigment is seen by light microscopy as amber globules or fiber-like structures in the cornea, conjunctiva, and sclera combined with degenerated collagen. In the area of a previous injury the intense involvement of the cornea, which is usually not notably involved, indicates the role of preexisting degeneration for the development of clinically visible pigmentation. Although the ultrastructure of the ochronotic pigment is similar to melanin, the chemical behavior is different and, it seems, similar to elastin. Ultrastructurally, most of the pigment granules are extracellular, partly altering the collagen fibers and fibrocytes. The electron-microscopically observed four patterns of deposition are interpreted as stages in the development of ochronotic deposits, according to biochemical findings in the literature.

Aged↗

Ultrastructure of human cataract in retinitis pigmentosa.

A second ultrastructural study of retinitis pigmentosa cataract showed differences from the previous case with respect to sex, age of cataract onset, age at time of cataract extraction, and mode of inheritance of retinitis pigmentosa. Furthermore, the cataract in the present study was limited clinically and preponderantly ultrastructurally to the posterior subcapsular area, which is typical of the location of cataracts associated with retinitis pigmentosa. The cataract was characterized by severe lens fiber disorganization, resulting in intricately distorted membrane configurations and varying cytoplasmic densities; however, "figure eight" configurations as noted in the other report were not observed. Aberrantly migrated epithelial cells were distributed unevenly in the posterior subcapsular zone. New capsular basement membrane, not mentioned in the other report, was conspicuous in our case. Thus, retinitis pigmentosa cataract may have diverse ultrastructural alterations.

Adult↗

CART peptide-immunoreactive neurones in the nucleus accumbens in monkeys: ultrastructural analysis, colocalization studies, and synaptic interactions with dopaminergic afferents.

Cocaine- and amphetamine-regulated transcript (CART) is a novel mRNA whose level of expression was found to be increased in the striatum after acute administration of psychomotor stimulants in rats. To define better the potential role of CART peptides in behavioural and physiologic changes induced by psychomotor stimulants, we analyzed the distribution, ultrastructural features, synaptic connectivity, and transmitter content of CART peptide-immunoreactive neurones in the nucleus accumbens in monkeys. Medium-sized CART peptide-immunoreactive neurones within a rich plexus of labelled varicosities were found mostly in the medial division of the shell of the nucleus accumbens in monkeys. At the electron microscope level, CART peptide immunoreactivity was exclusively associated with neuronal structures that included perikarya, dendrites, spines as well as nerve terminals packed with electron-lucent and dense-core vesicles. Most CART peptide-containing somata displayed the ultrastructural features of striatal output neurones. The majority of labelled terminals formed symmetric axodendritic synapses and displayed gamma-aminobutyric acid (GABA) immunoreactivity. CART peptide-immunoreactive somata were not immunoreactive for parvalbumin and somatostatin, two markers of striatal interneurones, nor for calbindin D-28k, a marker of a subpopulation of projection neurones. In double-immunostained sections, CART peptide-immunoreactive dendrites were found to be contacted by tyrosine hydroxylase-positive terminals which displayed the ultrastructural features of dopamine-containing boutons. These findings strongly suggest that CART peptides may be a cotransmitter with GABA in a subpopulation of projection neurones in the monkey accumbens. Furthermore, the fact that CART peptide-immunoreactive neurones receive direct synaptic inputs from dopaminergic afferents and are particularly abundant in the caudomedial division of the shell of the nucleus accumbens suggest that CART peptides might be involved in neuronal and behavioural changes that underlie addiction to psychomotor stimulants and feeding in primates.

Animals↗

Ultrastructural and confocal laser scanning microscopic examination of TUNEL-positive cells.

TdT-mediated dUTP-biotin nick end labelling (TUNEL) has been widely used for detecting cells with DNA fragmentation or apoptotic cells. However, since the concept of apoptosis is based on cellular ultrastructure, it is important to identify the morphological features of TUNEL-positive cells. In this study, we performed TUNEL and electron microscopic observation on serial semithin and ultrathin sections of pancreas from bilaterally adrenalectomized rats with caerulein-induced pancreatitis. TUNEL-positive cells were identified with two different ultrastructural patterns. One was characteristic of apoptosis, with condensed nuclei, intact mitochondria, and zymogen granules. The other pattern was one of marked cellular degeneration, possibly representing the end stage of cell death. Cells which did not demonstrate these ultrastructural patterns were not labelled by the TUNEL method. The three-dimensional structure of TUNEL-positive cells was also investigated by confocal laser scanning microscopy (CLSM), which showed the apoptotic nuclei exhibited various three-dimensional structures. These results confirm the utility of the TUNEL method in detecting apoptosis; application of the technique reported in this study will contribute to the further characterization of individual TUNEL-positive cells.

Acute Disease↗

Ultrastructural characterization of human herpesvirus 8 (Kaposi's sarcoma-associated herpesvirus) in Kaposi's sarcoma lesions: electron microscopy permits distinction from cytomegalovirus (CMV).

Kaposi's sarcoma (KS) has been shown by molecular techniques to be associated with infection with human herpesvirus 8 (HHV8/KSHV), but specific ultrastructural characterization of the virus has been impaired by the frequent presence in these lesions of other herpesviruses, particularly cytomegalovirus (CMV). Since the ultrastructural appearance of HHV8/KSHV has been studied in the cell line KS-1 uninfected with other viruses including CMV, it was possible to undertake a comparative study of CMV and HHV8/KSHV in KS lesions. HHV8/KSHV was sparsely present and lytic infection was restricted to endothelial cells. The following specific ultrastructural features allowed distinction between HHV8/KSHV and CMV: the viral particles were more delicate and less numerous in cases of HHV8/KSHV infection; the viral tegument was more electron-dense in CMV than in HHV8/KSHV; dense bodies characteristic of CMV were absent in HHV/KSHV; complete CMV viral particles were more variable in size and generally larger (150-200 nm) than HHV8/KSHV (120-150 nm); and finally, the viral envelope was more pleomorphic in CMV than in KSHV/HHV8. Similarities between CMV and HHV8/KSHV included the basic structure of the nucleocapsids and the presence of capsids lacking central DNA cores (so-called non-infectious enveloped particles). These observations show that electron microscopy can be used to identify HHV8/KSHV and confirm the relationship between HHV8/KSHV and KS.

Cytomegalovirus↗

Quantitative and ultrastructural changes in glia and pericytes in the parietal cortex of the aging rat.

The frequency of astrocytes, microglia plus oligodendrocytes, and pericytes displaying nuclei was analyzed and quantified in 160-microm-wide strips of the parietal cortex (Par1 region) from young and aged Wistar rats. The study was performed on two groups of rats aged 3-4 and 32-36 months. Quantifications of the glial cell types and pericytes were made in 1-microm-thick sections stained with toluidine blue. Ultrathin sections were also made to analyze the ultrastructural features of these cells during aging. Astrocytes and pericytes increased in number by about 20% and 22%, respectively, with age. These increases were most significant in layers II-IV and V for both cellular types. Clusters of astrocytes were common in these layers of aging rats. The ultrastructural analysis also indicated changes in all cell types that stored inclusions and vacuoles with age, which were particularly abundant in microglial cells. End-feet astrocytes and pericytes surrounding the vascular wall also contained vacuoles and inclusions, and consequently the vascular wall increased in thickness. In conclusion, the aging process increased astrocyte and pericyte populations, but not microglia plus oligodendrocyte populations, in the rat parietal cortex. Although no significant change in nuclear size could be observed in any cell type, all glial cells as well as pericytes underwent morphological ultrastructural changes. These modifications may result from the need to correct possible homeostatic imbalances during aging.

Aging↗