Development of the choroid plexus anlage and supraependymal structures in the fourth ventricular roof plate of human embryos: scanning electron microscopic observations

The developing anlage of the choroid plexus and supraependymal structures in the fourth ventricular roof plates of nine normal human embryos ranging from Carnegie stages 14 to 19 were investigated with scanning electron microscopy. In the human embryos at stage 18, the first semimacroscopic choroidal anlage developed in the form of bilateral evaginations that ran dorsomedially and caudally from the bilateral corners of the rhombencephalon. The anlage became evident with even smaller and parallel ridges in the embryo at stage 19. Embryos at earlier stages exhibited surface membrane modifications such as convexity, microvilli, cilia, and spherical protrusions at the middle one-third of the rhombencephalon, which corresponded to the future choroidal anlage region. Two morphologically different groups of supraependymal cells (SE cells) were elucidated throughout the stages examined. Type 1 SE cells has spindle or tear-drop-like bodies, frequently with one or more long cytoplasmic processes. Type 2 SE cells were globular, with numerous fine pseudopodial processes. Type 1 SE cells were distributed mainly at the future choroidal anlage regions or on the anlage itself and were less frequently located at the rostral end of the roof. We found no general pattern in the distribution of type 2 SE cells. Supraependymal fibers (SE fibers) were seen as fine processes that were distributed similarly to type 1 SE cells and extended transversely for a long distance.     

A comparative scanning electron microscopic analysis of the human cerebral ventricular system

Summary A scanning electron microscopic analysis of the adult human third ventricular wall revealed ultra-architectural differences between dorsal and ventral portions. In the brains of thirteen and sixteen week old human fetuses regional differences in the surface organization of lining ependyma were more sharply defined than those of the adult. Alterations in the luminal surfaces of ependyma may reflect differences in the functional capacity of various ventricular areas. The potential role of certain ependyma (tanycytes) and their putative participation in neuroendocrine events is discussed.Supported by U.S.P.H.S. Grant NS 08171.U.S.P.H.S. Career Development Awardee K 04 GM 70001.     

Surface defects in ventricular cells of brains of mouse embryos homozygous for the Loop-tail gene: scanning electron microscopic study.

Luminal surfaces in the mesencephalon and rhombencephalon in normal mouse embryos and those homozygous for Lopp-tail were studied by means of scanning electron microscopy. Ventricular cells in the ventrolateral regions of normal day-10 and -11 brains showed single apical cilia and microvilli, whereas those in ventromedial regions showed a dense network of microvilli and bulbous projections which tended to obscure the apical cilia and cellular outlines. Similar regional differences occurred in the Loop-tail brains, although there was a marked decrease in the number and density of microvilli and bulbous projections. At days 12-14 of gestation the latter brains also showed a flattening of cell surfaces, shallow depressions, and craterlike ruptures in the plasma membranes.     

Scanning electron microscopic observations of Thelazia callipaeda from human

Four females and a male nematode isolated from 2 patients who visited eye clinics in Seoul were identified as Thelazia callipaeda and their ultrastructures were observed by scanning electron microscopy (SEM). General features of the worms were slender and attenuated at both ends. Vaginal opening was located at 0.27 mm from the anterior end, and in front of the esophago-intestinal junction. In the body cuticle transverse striations varied characteristically through the body. The number of cuticular transverse striations was 400-650/mm at head portion, 250/mm at middle portion and 300-350/mm at tail portion. The SEM observation of the mouth part of the females showed 6 cord-like cuticular thickenings in hexagonal arrangement and an amphid was observed. A lateral line, a vaginal opening, a pair of phasmids, and an anus were identified in the body portion. A pair of papillae and 6 cord-like cuticular thickenings were on the mouth part of the male. It was difficult to observe structures at the tail of the male except wrinkle-like structures. Most of the larvae isolated from the uterus of a female worm were sheathed and thus cuticular striations were not seen. Others were un-sheathed and revealed cuticular striations. The oval membrane which encysted sheathed larvae was also observed. These are the 18th and 19th record of human thelaziasis in Korea as the literature are concerned.     

Formation of vascular cords in the chick embryo ; scanning electron microscopic observations]

The morphological changes of haemangioblasts during their sorting out from the pleural mesoderm have been studied as well as their later association within vascular cords, using scanning electron microscopy. These transformations involve only changing the shape and the relationship of cells.     

Structural aspects of the zona pellucida of in vitro-produced bovine embryos: a scanning electron and confocal laser scanning microscopic study

Structural aspects of the bovine zona pellucida (ZP) of in vitro-matured (IVM) oocytes and in vitro-produced (IVP) embryos were studied in two experiments to find a tentative explanation for the zona's barrier function against viral infection. In Experiment 1, the ultrastructure of the outer ZP surface was studied. The diameter (nm) and the number of the outer pores within an area of 5000 microm(2) of 10 IVM oocytes, 10 zygotes, 10 8-cell-stage embryos, and 10 morulae were evaluated by scanning electron microscopy. In oocytes and morulae, the ZP surface showed a rough and spongy appearance with numerous pores. In zygotes, the ZP surface was found to have a smooth, melted appearance with only a few pores. In 8-cell-stage embryos, both surface patterns were found. The mean number (per 5000 microm(2)) and the mean diameter of the outer pores were different between the four stages of development (P < 0.001): 1511 pores in oocytes, 1187 in zygotes, 1658 in 8-cell-stage embryos, and 3259 in morulae, with mean diameters of 182, 223, 203, and 155 nm, respectively. In Experiment 2, the continuity of the meshes (network of pores) towards the embryonic cells was examined by confocal laser scanning microscopy. Therefore, the passage through and the location in the ZP of fluorescent microspheres, with similar dimensions as bovine viral diarrhea virus (BVDV, 40-50 nm) and bovine herpesvirus-1 (BHV-1; 180-200 nm), were evaluated. For all stages, the smallest beads were detected halfway through the thickness of the ZP, whereas the beads with a size of 200 nm were found only within the outer-fourth part of the ZP. It can be concluded that the intact ZP of bovine IVM oocytes and IVP embryos are constructed in such a way that BVDV and BHV-1 should not be able to traverse the ZP and reach the embryonic cells. However, the risk exists that viral particles can be trapped in the outer layers of the ZP.     

A scanning electron microscopic study of IAA-induced tumors in bean (Phaseolus vulgaris L.) embryos

A scanning electron microscopic study of indole-3-acetic acid (IAA) induced tumour in the hypocotyl region of bean embryos shows a distinct morphological, structural and topographical change from the non-treated bean embryos. The IAA-induced tumour surface, in the hypocotyl region, shows distinct cell enlargement, some cellular proliferation in the parenchymatous tissue, total destruction of the epidermis and stomata and some variation in trichome structures. In dole-3-acetic acid inhibits the normal growth of the epicotyl, and, as age progresses, adventitious roots appear all over the surface. When IAA-depletion occurs, epicotyl growth resumes, which indicates that this tumour formation in bean embryos is an IAA-dependent tumour.     

Effect of tribendimidine on adult Echinostoma caproni harbored in mice, including scanning electron microscopic observations

Food-borne trematodiasis is an emerging public health problem with more than 10% of the world's population at risk of infection, yet there are only 2 drugs available for treatment and morbidity control. We assessed the effect of a promising broad-spectrum anthelmintic drug, i.e., tribendimidine, with an experimental focus on adult Echinostoma caproni. Female NMRI mice were infected with 30 E. caproni for 2 wk and then administered single oral doses of tribendimidine ranging between 25 and 500 mg/kg. Three days post-treatment, mice were necropsied, and adult worms were recovered from their intestines. Worm burden reductions were assessed against untreated control mice. In addition, scanning electron microscopic observations were done on adult E. caproni recovered from mice given a single dose of 150 mg/kg tribendimidine intragastrically 2, 4, and 8 hr post-treatment. Worm burden reductions of 100% were achieved at doses of 125 mg/kg and above. Severe damage of the tegument, including extensive peeling, formation of blebs, and structural loss of the definition of collar and tegumentary spines already occurred within 2 hr after drug administration. Our findings call for further investigations using tribendimidine in other trematode-animal models, because this compound shows promising trematocidal activity.     

A scanning electron microscopic survey of the brain ventricular system of the female armadillo

Summary The scanning electron microscope was used to survey the brain ventricular system of the female armadillo (Dasypus novemcinctus) with emphasis on the third ventricle. The walls of the lateral ventricles, aqueduct, and fourth ventricle are covered by long cilia. In the lateral ventricle, the cilia are arranged in groups; but in the aqueduct and fourth ventricle, they are evenly placed over the cellular surfaces. The ependymal cells of the third ventricle are densely ciliated except for the organum vasculosum and infundibular recess. The non-ciliated luminal surface of these areas has a pebblestone appearance punctuated by numerous microvilli and two types of supraependymal cells.Supported by Edward G. Schlieder Foundation GrantThe authors would like to thank Jacqueline Skaggs for her secretarial assistance and Garbis Kerimian for his photographic work     

Melting of myosin and tropomyosin: electron microscopic observations

A method was devised to maintain a very low angle (2-3 degrees) during the metal casting of specimens for electron microscopy. With this modified rotary shadowing procedure the melting of myosin and tropomyosin (TM) was investigated. When protein solutions were sprayed on mica sheets and then heated to melt alpha-helices, myosin molecules did not show any sign of chain separation but appeared to have collapsed into loose clumps. A few molecules showed separation of the two chains at the light meromyosin-heavy meromyosin hinge region. Heating myosin in bulk solution at 65 degrees C before spraying caused extensive fusing of the myosin heads. In contrast, in the case of TM, separation of the chains appeared to occur at temperatures at which the unfolding of alpha-helices had been shown by circular dichroism. Dissolution of TM and myosin in 0.5% SDS followed by 150-fold dilution led to single chain species. This method capable of detecting single chain peptides of melting TM whose thickness is of the order of 1 nm may be applicable to the study of the structure of proteins previously not considered possible.     

Collagenous network in cartilage of human femoral condyles. A light microscopic and scanning electron microscopic study

To determine the spatial arrangement of collagen fibrils in articular cartilage of the human femoral head, three healthy femoral heads, obtained at necropsy, were examined by light microscopy and scanning electron microscopy. Light microscopic observations revealed no collagen fibril organization. Scanning electron microscopic observations showed a fine fibrillar texture throughout the articular cartilage. At the articular surface, smooth and fibrillated areas were detectable. Underneath the articular surface, the collagen network in the superficial zone showed a tighter appearance when compared with the homogeneous collagen network of the matrix in the deeper zones. The calcified cartilage zone was well demarcated from the uncalcified cartilage. The arcade model of Benninghoff [Z. Zellforsch. Mikrosk. Anat. 2: 783-862 (1925)] could not be confirmed. It was concluded that the organization of collagen fibrils in hyaline cartilage shows a three-dimensional network of randomly oriented fibrils.     

Osteoarthrosis in guinea pigs: histopathologic and scanning electron microscopic features

Spontaneous cartilage degeneration of the femorotibial joint of male Hartley guinea pigs, 61 to 365 days old, was studied by light microscopy (LM) and scanning electron microscopy (SEM) to determine the incidence, age at onset, and to characterize the early changes. Knee joints of 61 day old animals were histologically and ultrastructurally normal. Focal minimal degeneration characterized by cell and proteoglycan loss with surface fibrillation was first observed by LM on the medial tibial plateau (MTP) in two of five 89 day old animals. Mild lesions characterized by focal surface disruption, primarily in the area of medial tibial plateau not covered by the meniscus, were observed in three of five 89 day old animals by SEM. Light microscopic alterations in knee joints of 4, 5, and 6 month old animals consisted of varying degrees of focal chondrocyte death, decreased toluidine blue matrix staining, and surface fibrillation. Small chondrocytic clones were first observed in medial tibial cartilage of 6 month old animals with moderate focal degeneration. Ultrastructurally, 4, 5, and 6 month old animals generally had moderate to severe fibrillation involving primarily the area of the medial tibial plateau not covered by the meniscus. Tibial osteophyte formation, mild synovial hyperplasia, medial femoral and meniscal cartilage degeneration, were first seen by LM in 9 month old animals. Lesions in 1 year old animals were similar, but more severe and included subchondral sclerosis of medial tibial and femoral bone. Bilateral fibrillation of greater than 50% of the medial tibial articular surface was observed in all 1 year old animals by SEM.(ABSTRACT TRUNCATED AT 250 WORDS)     

A scanning electron microscopic study of trichomes of IAA-induced tumours in bean embryos (Phaseolus vulgaris L.).

A scanning electron microscopic study of trichome morphology of bean embryo, growing in semi-solid synthetic medium, has been undertaken. The work shows some distinct differences in trichome morphology as well as variation in their development, due to the effect of indole-3-acetic acid (IAA).     

A combined scanning and transmission electron microscopic study and electron probe microanalysis of human pineal acervuli

Summary Untreated, decalcified and trypsinized acervuli from human pineal bodies were studied with the scanning and transmission electron microscope as well as by electron probe microanalysis. The mulberry-like acervuli are composed of a various number of spherical lobes (135–800 m) between which clustered groups of globuli (4–14 urn in diameter) are observed. The acervular lobes are very probably formed by an aggregation of these globuli. Small round particles 125–500 Å in diameter are observed on the surface of the pineal concretions. These are not influenced by either decalcification or trypsin treatment. The acervular mineral corresponds morphologically to hydroxyapatite. The electron probe microanalysis reveals the existence of calcium and phosphorus as main components of the acervuli. Small quantities of magnesium and strontium were also detected.Dedicated to Professor Berta Scharrer on the occasion of her 70th birthdayWith the technical assistance of Mr. P.A. MilliquetThe author wishes to thank Mr. Bauer and Mr. Fryder (Nestec SA, La Tour de Peilz) for the use of the Cambridge Stereoscan electron microscope and Dr. T. Jalanti (C.M.E., Lausanne) for his help with the use of the X-ray microanalyser     

Use of human organs fixed long ago for the scanning electron microscopic analysis

Normal human organs fixed 10-15 years ago in formol and Bouin as well as organs prepared from cadavers used for the anatomical dissections show a surprisingly well preserved structure in the scanning electron microscope. Such organs can be successfully used for the teaching of the 3-dimensional microanatomy.     

A scanning electron microscopic study of the dorsal surface of the human tongue

To study the dorsal surface of the human tongue using a scanning electron microscopy (SEM), tissue specimens were taken from the anterior part of the tongues of 15 individuals aged from 21- to 28-years-old. The formalin-fixed samples were processed routinely for SEM. With SEM the surface of the normal tongue mucosa was shown to be rather evenly covered by filiform papillae, with some fungiform papillae scattered among them. Filiform papillae consisted of two parts: the body and hairs. The mucosal surface of the body was smooth; the squamous epithelial cells were polygonal, and their boundaries were prominent. On the surface of the superficial epithelial cells were parallel or branching microplicae. Each filiform papilla had 6-10 hairs, which were scaled and covered by an extensive plaque of microorganism. The upper surface of the fungiform papillae was smooth; only a few desquamating cells were seen. The superficial cells had a pitted appearance and cell boundaries overlapped. Taste pores, up to 3 pores in a single papilla, were found on the upper surface. Desquamation was more pronounced on the base of the fungiform papillae than on the upper surface. In almost all fungiform papillae some hairs protruded from the base. Parallel microplicae were found on the surface of the superficial cells of the base. The structure and function of the human tongue, as well as the microplicae of its superficial cells, are compared to those of various species of animals.