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PubMed2000

Morphologic characterization of the ice worm Mesenchytraeus solifugus.

Ice worms occupy a unique position in metazoan phylogeny in that they are the only known annelid that completes its life cycle in ice. The mechanism(s) associated with this adaptation are likely to occur at different levels, ranging from modification of their metabolism to changes in morphology. In this study, we examined specimens of Mesenchytraeus solifugus by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) in an effort to identify morphologic structures that may aid in its glacial habitation. We report that M. solifugus contains an elongated head pore at the tip of its prostomium, numerous sensory structures, and differentially oriented setae that curve abruptly at their distal end.

Alaska↗

PubMed1986

Quantitation and morphological characterization of rapid axon and dendritic growth from single cerebral hemispheric neurons in hydrated collagen lattice culture.

Quantitative and qualitative data are reported for single cerebral hemispheric neurons in a 3-dimensional hydrated collagen lattice (HCL) culture system. Individual neurons not in contact with other cells or cell processes, including synapses, rapidly displayed two morphologically distinct classes of processes that could be traced from origin to termination: long thin processes interpreted as being axons, and shorter tapering and sometimes branched processes interpreted as being dendrites. Axons and dendrites of single neurons that had at least one process longer than the cell body were measured on each of 3 days after plating using an image analysis system coupled to a phase-contrast microscope and a microcomputer. Mean lengths of axons and dendrites alone or combined as total new growth per neuron, increased 3- to 5-fold and were as high as 745, 694 and 1226 microns respectively after 3 days in HCL, although some individual axons measured over 1500 microns. Other indices of neuron growth and differentiation increased 1- to 5-fold including the number of primary processes, branch points, segments and growth cones. Phase-contrast microscopy, staining with Nissl and silver, and scanning and transmission electron microscopy demonstrated many single multipolar and other neurons with axons, dendrites and well-differentiated properties. The data show that individual central nervous system neurons have an inherent capacity to quickly express characteristic differentiated features and also to grow rapidly in HCL.

Animals↗

PubMed2002

Life span and renal morphological characterization of the SAMP1//Ka mouse.

The senescence-accelerated-mouse prone 1 (SAMP1) is considered to be a model of accelerated senility and it also develops severe kidney damage. The SAMP1//Ka mouse is a specific pathogen free (SPF) subline of SAMP1. The present study examined the life span of the SAMP1//Ka mouse and morphologically investigated the kidneys of this animal at 3, 4, 5, 9, 12, 15, 18 and 24 months of age. Males survived for an average of 25 months and females for 28 months. The median lifespan was 18 months for males and 20 for females. Focal cell infiltration and thickening of the basement membrane in the glomerular capsules or tubules appeared from 4 months of age. At 12 months old, glomerular lesions with expansion of the mesangial matrix and thickening of the basement membrane as well as scar lesions in the outer cortex appeared, and amyloid was deposited in the interstitium or glomeruli from 18 months of age. Morphometrically, although the area of the kidney sections was increased at 24 months of age, the diameter of the renal corpuscles, the number of nuclei of the proximal convoluted tubules and the percentage of renal corpuscles with a cuboidal glomerular capsule did not change with age. The results of the present study indicate that the life span of the SAMP1//Ka is increased and that their age-related renal changes differ from those of the original SAMP1.

Aging↗

PubMed1977

Spermatogenic cells of the prepuberal mouse. Isolation and morphological characterization.

A procedure is described which permits the isolation from the prepuberal mouse testis of highly purified populations of primitive type A spermatogonia, type A spermatogonia, type B spermatogonia, preleptotene primary spermatocytes, leptotene and zygotene primary spermatocytes, pachytene primary spermatocytes and Sertoli cells. The successful isolation of these prepuberal cell types was accomplished by: (a) defining distinctive morphological characteristics of the cells, (b) determining the temporal appearance of spermatogenic cells during prepuberal development, (c) isolating purified seminiferous cords, after dissociation of the testis with collagenase, (d) separating the trypsin-dispersed seminiferous cells by sedimentation velocity at unit gravity, and (e) assessing the identity and purity of the isolated cell types by microscopy. The seminiferous epithelium from day 6 animals contains only primitive type A spermatogonia and Sertoli cells. Type A and type B spermatogonia are present by day 8. At day 10, meiotic prophase is initiated, with the germ cells reaching the early and late pachytene stages by 14 and 18, respectively. Secondary spermatocytes and haploid spermatids appear throughout this developmental period. The purity and optimum day for the recovery of specific cell types are as follows: day 6, Sertoli cells (purity>99 percent) and primitive type A spermatogonia (90 percent); day 8, type A spermatogonia (91 percent) and type B spermatogonia (76 percent); day 18, preleptotene spermatocytes (93 percent), leptotene/zygotene spermatocytes (52 percent), and pachytene spermatocytes (89 percent), leptotene/zygotene spermatocytes (52 percent), and pachytene spermatocytes (89 percent).

Age Factors↗

PubMed2003

Morphological characterization of the spermatogonial subtypes in the neonatal mouse testis.

Spermatogenesis is the process of differentiation of diploid type A spermatogonia to haploid spermatozoa. Several subtypes of A spermatogonia have been characterized in the adult mouse testis. These include A-single (A(s)), A-paired (A(pr)), A-aligned (A(al)), and A1-A4. However, in the immature testis, very little information is available on subtypes and morphological features of type A spermatogonia. Six-day-old mouse testes, fixed either in Bouin solution or 5% glutaraldehyde, were embedded in paraffin and Epon, respectively. Thick sections (approximately 1 microm) of Epon-embedded tissue were stained with toluidine blue and revealed three subtypes of spermatogonia by light microscopy. The smallest spermatogonia (subtype I) appeared as single cells and exhibited a round or oval flattened nucleus with one or two prominent dense nucleoli and a characteristic unstained round and centrally located vacuole. These cells bound toluidine blue more avidly and appeared darker in comparison with the other cell types. Electron microscopy of thin sections (90 nm) revealed a finely granulated chromatin homogeneously distributed in the nucleus and sparse organelles in the cytoplasm. The second subtype of spermatogonia (subtype II) also displayed dark staining but was larger than subtype I; there was no central vacuole in the nucleus and heterochromatin clumps were observed. The largest subtype of spermatogonia (subtype III) showed large heterochromatin clumps and a pale staining nucleus. Intercellular bridges were noted between subtypes II and III. Based on the dye avidity, the three subtypes were classified as dark, transitional, and pale spermatogonia, respectively. Image analyses of 30 different cells of each subtype revealed a decline in gray-scale intensity from subtype I to III. Five-micrometer sections of paraffin-embedded tissue were immunoassayed with an antibody against the glial cell-derived neurotrophic factor family receptor alpha-1 (GFRalpha-1) receptor, a putative marker for undifferentiated spermatogonia, showing positive reaction only in germ cells. The pattern of GFRalpha-1 expression, coupled to the overall morphology of the cells, indicates that at this stage of development, mouse seminiferous tubules contain essentially A(s), A(pr), and possibly A(al) spermatogonia. Thus, the present study indicates the presence of subtypes of type A spermatogonia in the immature mouse testis similar to that described previously in adult monkey and man.

Animals↗

PubMed2006

Clinical features and morphological characterization of 10 patients with noninsulinoma pancreatogenous hypoglycaemia syndrome (NIPHS).

OBJECTIVE: Noninsulinoma pancreatogenous hypoglycaemia syndrome (NIPHS), characterized by postprandial neuroglycopaenia, negative prolonged fasts and negative perioperative localization studies for insulinoma, but positive selective arterial calcium stimulation tests and nesidioblastosis in the gradient-guided resected pancreas, is a rare hypoglycaemic disorder of undetermined aetiology. We analysed the clinical, morphological and immunohistological features to further clarify the aetiology and pathogenesis of this rare disease. PATIENTS: Ten consecutive patients with NIPHS (nine men and one woman, aged 29-78 years) were included in the study. Six of the 10 received a gradient-guided subtotal (70%) or distal (50%) pancreatectomy. In the remaining four patients, diazoxide treatment was initiated and the precise mechanism of its action was assessed by meal tests. RESULTS: All of the patients showed a combination of postprandial neuroglycopaenia, negative prolonged fasts (except one patient) and negative localization studies for insulinoma, but positive calcium stimulation tests and nesidioblastosis in the gradient-guided resected pancreas. Immunohistological studies of the resected pancreatic tissues revealed neither an increased rate of proliferation of beta-cells nor an abnormal synthesis and/or processing of either proinsulin or amylin. Evidence of overexpression of the two pancreatic differentiation factors, PDX-1 and Nkx-6.1, as well as the calcium sensing receptor (CaSR) was absent. Nevertheless, abnormal expression of islet neogenesis-associated protein (INGAP), a human cytokine expressed only in the presence of islet neogenesis, in ducts and/or islets, was identified in three of the five patients studied. All of the six patients who received a surgical operation were relieved of further neuroglycopaenic attacks, but one patient who received a subtotal pancreatectomy developed diabetes. In the remaining four patients who received diazoxide treatment, hypoglycaemic episodes were satisfactorily controlled with an attenuated response of beta-cell peptides to meal stimulation. CONCLUSIONS: Our results strengthen the existence of this unique clinical hypoglycaemic syndrome from beta-cell hyperfunction as well as the value of the selective arterial calcium stimulation test in its correct diagnosis and localization. The mechanisms underlying beta-cell hyperfunction and release of insulin to calcium, however, remain poorly characterized. Nevertheless, in a subset of patients with NIPHS, there exists some, as yet undefined, pancreatic humoral/paracrine factor(s) other than proinsulin, amylin, PDX-1, Nkx-6.1 and possibly glucagon-like peptide-1 (GLP-1) that are capable of inducing the INGAP gene and, if activated, will initiate ductal proliferation and islet neogenesis. As for the treatment, we recommend that diazoxide be tried first in each patient and, should it fail, a gradient-guided subtotal or distal pancreatectomy be attempted.

Adult↗

PubMed1991

Morphologic characterization of conjunctiva-associated lymphoid tissue in chickens.

Conjunctiva-associated lymphoid tissue (CALT) in the eyelids of chickens was studied by gross, histologic, and electron microscopic techniques. Structural features were characterized at 1 day of age and at posthatching week (PHW) 1, 2, 3, 4, 6, 8, 12, and 16. Beginning at PHW 1, prominent lymphoid nodules containing a heterogenous population of lymphocytes, lymphoblasts, and macrophages were first observed within conjunctival folds and fissures of the lower eyelid. Nodules contained germinal centers by PHW 2 and plasma cells by PHW 4. The epithelium associated with these nodules was flat, had short, irregular microvilli, contained intraepithelial lymphocytes, and lacked goblet cells. High endothelial venules were located at the base of lymphoid nodules and contained lymphocytes within and below the cuboidal endothelium. In the upper eyelid, CALT was morphologically similar to lymphoid tissue in the lower eyelid, but nodules were smaller and more random, lacked association with epithelial folds and fissures, and were clustered around the opening of the nasolacrimal duct. By PHW 12, CALT was characterized by basal germinal centers outlined by collagenous stroma, suprafollicular plasma cells, columnar epithelium with goblet cells, and fewer intraepithelial lymphocytes. On the basis of these features, CALT in chickens has morphologic characteristics similar to other components of the mucosal immune system and, therefore, may have a role in mucosal immunity.

Animals↗

PubMed1973

Golgi fractions prepared from rat liver homogenates. I. Isolation procedure and morphological characterization.

In devising a new procedure for the isolation of Golgi fractions from rat liver homogenates, we have taken advantage of the overloading with very low density lipoprotein (VLDL) particles that occurs in the Golgi elements of hepatocytes approximately 90 min after ethanol is administered (0.6 g/100 g body weight) by stomach tube to the animals. The VLDLs act as morphological markers as well as density modifiers of these elements. The starting preparation is a total microsomal fraction prepared from liver homogenized (1:5) in 0.25 M sucrose. This fraction is resuspended in 1.15 M sucrose and loaded at the bottom of a discontinuous sucrose density gradient. Centrifugation at approximately 13 x 10(6)g.min yields by flotation three Golgi fractions of density >1.041 and <1.173. The light and intermediate fractions consist essentially of VLDL-loaded Golgi vacuoles and cisternae. Nearly empty, often collapsed, Golgi cisternae are the main component of the heavy fraction. A procedure which subjects the Golgi fractions to hypotonic shock and shearing in a French press at pH 8.5 allows the extraction of the content of the Golgi elements and the subsequent isolation of their membranes by differential centrifugation.

Animals↗

PubMedDate not supplied

Morphological characterization of marginal (lamina I) neurons immunoreactive for substance P, enkephalin, dynorphin and gamma-aminobutyric acid in the rat spinal cord.

Neurons of the rat spinal cord were immunostained for substance P, enkephalin and dynorphin in colchicine-treated animals, and for gamma-aminobutyric acid (GABA). Lamina I stained cells were classified in the four neuronal groups of our previous morphological classification of marginal cells (See Lima and Coimbra, 1986), according to their configuration in the three main anatomical planes. Most lamina I cells exhibiting substance P-immunoreactivity belonged in the group of flattened neurons. Most enkephalinergic cells were pyramidal neurons, while GABA-immunoreactive cells included all multipolar stained neurons and some fusiform neurons. Dynorphin-immunoreactive cells could be fusiform, pyramidal or flattened. The different neurochemical nature and supraspinal projection patterns are suggestive of functional specificity for each group. It is likely that each immunocytochemical subset in each cell group includes tract cells acting at their projection target and intrinsic neurons with local functional roles.

Animals↗

PubMed1989

Morphologic characterization of alveolar macrophages from subjects with occupational exposure to inorganic particles.

Alveolar macrophages recovered by bronchoalveolar lavage from 43 nonsmoking or greater than 5-yr ex-smoking subjects with occupational exposure to inorganic particles (asbestos, n 1/2 19; silica, n 1/2 10; coal, n 1/2 14) were evaluated by light microscopy and transmission and scanning electron microscopy to determine the morphologic changes resulting in these cells from chronic inorganic particulate inhalation. Alveolar macrophages from dust-exposed subjects, including those who had been free of exposure to particles for more than 1 yr, contained particles of higher proportion than did those of normal unexposed subjects. Most of these particles were located within phagolysosomes. The frequency of multinucleated alveolar macrophages was significantly higher in the dust-exposed groups. Ultrastructural studies showed alterations of the morphologic aspects of the surfaces of alveolar macrophages from the dust-exposed subjects, including increased numbers of rufflings, filopodia, pinocytotic vesicles, subplasmalemmal linear densities, and increased frequency of macrophage-macrophage and macrophage-lymphocyte interactions. Furthermore, the numbers of lysosomes were significantly increased in alveolar macrophages from the dust-exposed subjects. Together, these morphologic changes are consistent with the sequelae of phagocytosis, and they emphasize both the role of alveolar macrophages in eliminating inorganic particles from the alveolar spaces and the consequences this role has in alveolar macrophage activation.

Adult↗

PubMed2002

Electrophysiological and morphological characterization of the medulla bilateral neurons that connect bilateral optic lobes in the cricket, Gryllus bimaculatus.

The medulla bilateral neurons (MBNs) in the cricket brain directly connect two optic lobes and have been suggested to be involved in mutual coupling between the bilateral optic lobe circadian pacemakers. Single unit analysis with intracellular recording and staining with Lucifer Yellow was carried out to reveal morphology and physiology of the MBNs. Neurons having a receptive field in the rostral part of the compound eye showed greater response and a higher sensitivity to light than those having receptive fields in the ventro-caudal or dorsal portions. The MBN showed diurnal change in their responsiveness to light; the light-induced response in the night was about 1.3, 5 and 2 times of that in the day in MBN-1s, -3s and -4s, respectively. These results suggest that the MBNs mainly encode the temporal information by the magnitude of light-induced responses. The differences in magnitude of light-induced responses and of daily change in photo-responsiveness among MBNs may suggest that each group of MBNs plays different functional role in visual and/or circadian systems.

Journal Article↗

PubMed2003

Morphological characterization of elastin-mimetic block copolymers utilizing cryo- and cryoetch-HRSEM.

Elastin-mimetic block copolymers were produced by genetic engineering. Genetically driven synthesis permitted control of the final physiochemical characteristics of the block copolymers. We designed BB and BAB block copolymers in which the A-block was hydrophilic and the B-block was hydrophobic. By designing the copolymers in this manner, it was proposed that they would self-assemble into micellar aggregates that, at high concentration, would form thermoreversible hydrogels. To analyze the three-dimensional fine surface morphology of the copolymers, to the resolution level of a few nanometers, we employed cryo-HRSEM. This method provided vast expanses of the specimen in its frozen hydrated state for survey. In our initial cryo-HRSEM studies, we observed the protein filaments and micelles surrounded by lakes of vitreous ice. Upon examination at low and intermediate magnifications, there was an extensive honeycomb-like filamentous network. To delineate the fine morphology of the hydrogel network at high magnification and to greater depths, we cryoetched away unbound water from the sample surface, in high vacuum, prior to chromium deposition. By using this technique, we were able to visualize for characterization purposes the fine fibril networks formed from the micellar aggregates over the surface of the hydrogel.

Amino Acid Sequence↗

PubMed2001

Biochemical and morphological characterization of an intracellular membrane compartment containing AMPA receptors.

AMPA receptors cycle rapidly in and out of the postsynaptic membrane, while NMDA receptors are relatively immobile. Changing the distribution of AMPA receptors between intracellular and surface synaptic pools is an important means of controlling synaptic strength. However, little is known about the intracellular membrane compartments of neurons that contain AMPA receptors. Here we describe biochemical and morphological characteristics of an intracellular pool of AMPA receptors in rat brain. By velocity gradient centrifugation of microsomal light membranes from rat brain, we identified a membrane fraction enriched for AMPA receptor subunits GluR2/3 but lacking NMDA receptors. This membrane compartment sedimented more slowly than synaptosomes but faster than synaptic vesicles and cofractionated with GRIP, PICK-1 and syntaxin-13. Morphological examination of this fraction revealed round and tubular vesicles ranging from approximately 50 to 300 nm in diameter. Immunocytochemistry of cultured hippocampal neurons showed that a significant portion of AMPA receptors colocalized with syntaxin-13 (a SNARE protein associated with tubulovesicular recycling endosomes) and with transferrin receptors. Taken together, these results suggest that a pool of intracellular GluR2/3 resides in a syntaxin 13-positive tubulovesicular membrane compartment, which might serve as a reservoir for the dendritic recycling of AMPA receptors.

Animals↗

PubMed2005

Hybridization between Helicoverpa armigera and Helicoverpa assulta (Lepidoptera: Noctuidae): development and morphological characterization of F1 hybrids.

Reciprocal hybridizations between Helicoverpa armigera (Hübner) and Helicoverpa assulta (Guenée) were studied. The cross between females of H. armigera and males of H. assulta yielded only fertile males and sterile individuals lacking an aedeagus, valva or ostium bursae. A total of 492 larvae of the F1 generation were obtained and 374 of these completed larval development and pupated. Only 203 pupae were morphologically normal males, the remaining 171 pupae were malformed. Larvae and pupae that gave rise to morphologically abnormal adults exhibited longer development times. Sterility was not only associated with malformed external sex organs, but also a range of abnormalities of the internal reproductive system: (i) loss of internal reproductive organs, (ii) with one to three copies of an undeveloped bursa copulatrix; or (iii) with one or two undeveloped testes. Normal male hybrid adults showed higher flight activity in comparison with males of both species. In contrast, the cross between females of H. assulta and males of H. armigera yielded morphologically normal offspring (80 males and 83 females). The interaction of the Z-chromosome from H. assulta with autosomes from H. armigera might result in morphological abnormalities found in hybrids and backcrosses, and maternal-zygotic incompatibilities might contribute to sex bias attributed to hybrid inviability.

Animals↗

PubMed1989

Morphologic characterization of acute injury to vascular endothelium of skin after frostbite.

Cyclo-oxygenase inhibitors and free-radical scavengers protect the skin against necrosis induced by frostbite. However, the tissue component(s) that determine the evolution of skin necrosis and the mechanism of this pharmacologic protection are not precisely defined. We have studied freezing injury to rabbit ears by serial biopsies examined by light and electron microscopy. The morphologic evidence of skin injury due to freezing was localized exclusively in the endothelial cells, particularly in the arterioles. Within 1 hour, the entire microvasculature demonstrated endothelial damage. Intravascular platelet aggregation occurred just after thawing and closely paralleled the endothelial cell injury. Very few neutrophils were seen initially (at 10 minutes). By 1 hour, leukocyte aggregates were present, and they further increased at 6 hours. Swelling of the interstitium started 10 minutes after thawing, while extravasation of erythrocytes began to appear by 6 hours. Parenchymal elements of skin were relatively free of damage. In the ear cartilage, the chondrocytes showed evidence of damage immediately after freezing. The administration of superoxide dismutase (SOD) during thawing (reperfusion) did not qualitatively alter any of the initial morphologic changes induced by freezing. We conclude that the endothelial cell is the initial target of injury induced by freezing, an initial injury that is mediated by a non-free-radical-mediated mechanism. It is likely that this acute injury ultimately compromises blood flow and leads to skin necrosis.

Animals↗

PubMed1987

Isolation and morphologic characterization of human ovarian carcinoma cell clusters present in effusions.

Serous, mucinous, endometrioid and clear cell human ovarian carcinoma cells were isolated as multicellular aggregates from patient effusions by filtration on nylon mesh of defined porosity and examined by light microscopy. The cell clusters ranged from compact to loosely adherent groups of cells to spheroids with a central lumen surrounded by a cell monolayer. There was considerable variation in cluster morphology between effusions from different patients as well as within effusion from the same patient. Apparent budding of clusters was observed as well as different stages of cluster growth and development. This was observed for all histologic types studied. Electron microscopy of serous, mucinous and clear cell types showed that cells forming clusters were attached to each other by desmosomes, demonstrating that cluster formation did not result from a nonspecific stickiness of cells. Irregular microvilli were present on the external periphery of the various carcinoma cells and a prominent glycocalyx was present on the surface of mucinous carcinoma cells. Extensive interdigitation of cytoplasmic extensions and extended villi was present in mucinous and serous clusters which appeared to strengthen cluster cohesiveness. Nuclei were irregular with prominent nucleoli frequently present. The cell clusters usually remained intact and viable in culture but generally did not attach to glass or plastic substrata, whereas mesothelial cells and nonactivated histiocytes rapidly attached. When carcinoma cell clusters did attach, they were resistant to detachment by trypsin-EDTA treatment, in contrast to the nonmalignant cells.

Adenocarcinoma↗

PubMed1991

Morphological characterization of human juxtaglomerular apparatus.

The recent demonstration of the endothelial fenestration at the juxtaglomerular part of the afferent arteriole facing mesangial and granular cells and finger-like protrusions of urinary space into the region of the lacis in experimental animals like the rat, the mouse, and Tupaia belangeri led us to propose the hypothesis of a "short loop" feedback mechanism. To establish whether a further species possesses these morphological features, the JGA region was examined in the human kidney. Tissue was obtained by needle biopsy from patients and conventional electron-microscopical procedures including serial sections were utilized to reevaluate the morphology of JGA. Both the endothelial fenestration in the wall of the afferent arteriole and protrusions with filtration slits of urinary space into the lacis, were observed with remarkable heterogeneity. The occurrence of these features of JGA in such diverse mammalian species as rat, mouse, Tupaia, and man suggests that these structures may be involved in regulatory process and the proposed "short loop" feedback mechanism may be a general phenomenon.

Animals↗

PubMed2005

Morphological characterization of molecular complexes present in the synaptic cleft.

We obtained tomograms of isolated mammalian excitatory synapses by cryo-electron tomography. This method allows the investigation of biological material in the frozen-hydrated state, without staining, and can therefore provide reliable structural information at the molecular level. We developed an automated procedure for the segmentation of molecular complexes present in the synaptic cleft based on thresholding and connectivity, and calculated several morphological characteristics of these complexes. Extensive lateral connections along the synaptic cleft are shown to form a highly connected structure with a complex topology. Our results are essentially parameter-free, i.e., they do not depend on the choice of certain parameter values (such as threshold). In addition, the results are not sensitive to noise; the same conclusions can be drawn from the analysis of both nondenoised and denoised tomograms.

Animals↗