The Changes of the Vitality of Armillaria mellea and the Histochemicam Localization of Some Enzymes in the Hyphae Digested Period of Gastrodia elara

The changes of the vitality of Armillaria mellea of infecting corm of Gastrodia elata were observed by appling the live body staining method of acridine orange and by means of fluorescence microscopy. The green fluorescence of vitality was emitted by the first infected hyphae, the yellow one by the decrepit hyphae; but the orange to red fluorescence with the lost vitality were emitted by the fragmentary hyphae and clump form bodies. The large cells containing rich, RNA and protein had been confirmed by the method of the induced fluorescence which the acridine orange and by the method at pH 2.2 which the fast-green staining. The acid-phosphatase was mainly distributed within the cortical cells filled with the infected hyphae. There were few such deposits in the socalled large cells except their walls. The activity of the esterase was shown in the cortical cells filled with the infected hyphae. It were also shown in the clump form bodies and the collapsed nuclei of the large cells. The activities of adenosine triphosphatase (ATPase), peroxidase and polyphenol oxidase was notably shown in the cortical cells filled with the infected hyphae and the large cells.     

Ultrastructural cytology of the cyclic corpus luteum of the cow

Corpora lutea (CL) from cows on day 12 of the oestrous cycle were studied by electron microscopy to investigate whether, and how, different subpopulations of luteal cells can be identified in tissue sections. Tissues from 6 CL were examined, and nucleated profiles of luteal cells were classified as large, medium or small on the basis of their areas in electron micrographs. Cut-off points for area categories for large, medium and small-sized cells were based on diameters of greater than 25, 20-25 and less than 20 microns, respectively, if the measured areas were converted to a circular shape after correction for shrinkage. The only qualitative features which distinguished cells of large size from those of small size were the presence of clusters of secretory granules, and of exocytosis of these granules, in large cells only. However, these features were observed in only 59% of large cells, probably primarily due to sampling limitations in single sections. Other qualitative features which have been regarded as diagnostic of large or small luteal cells were observed in cells in all size categories. It was concluded that large and small luteal cells in the cyclic CL of the cow are distinguishable by their ultrastructural features. However, these data do not support the recent suggestion that the mid-cycle CL of the cow contain two subpopulations of large luteal cells in approximately equal numbers.     

Ultrastructural and histochemical study of previtellogenic oogenesis in the desert lizard Scincus mitranus (Squamata,Sauropsida)

The structure of the granulosa in reptilian sauropsids varies between groups. We investigated the follicle development in the desert lizard Scincus mitranus. In the germinal bed, oogonia, and primary oocytes were identified and found to be interspersed between the epithelial cells. Previtellogenesis was divided into three stages: early, transitional, and late previtellogenic stages. During the early previtellogenic stage (diplotene), the oocyte is invested by small epithelia cells that formed a complete single layer, which may be considered as a young follicle. The transitional previtellogenic stage was marked by proliferation and differentiation of the granulosa layer from a homogenous layer consisting of only small cells to a heterogeneous layer containing three cell types: small, intermediate, and large cells. The late previtellogenic stage was marked by high-synthetic activity of large cells and the initiation of cytoplasmic bridges between large granulosa cells and the oocyte. Small cells were the only type of granulosa cells that underwent division. Thus, these cells may be stem cells for the granulosa cell population and may develop into intermediate and subsequently large cells. The intermediate cells may be precursors of large cells, as suggested by their ultrastructure. The ultrastructure of the large granulosa was indicative of their high synthetic activity. Histochemical analysis indicated the presence of cholesterol and phospholipids in the cytoplasm of large cells, the zona pellucida, among the microvilli, in the bridges region, and in the cortical region of the oocyte cytoplasm. These materials may be transferred from large cells into the oocyte through cytoplasmic bridges and provide nutritive function to large cells rather than functioning in steroidogenesis or vitellogenesis.     

鹅掌楸油细胞的发育解剖学研究

鹅掌楸油细胞比相邻组织细胞分化,因而在叶肉细胞未完全分化的叶原基、幼叶以及未完全分化的幼茎中,都可看到已分化的油细胞。通常,在第二叶原基中可发现油细胞原始细胞,由于其染色深、细胞核大而易与周围组织的细胞相区别。以后,油细胞逐步液泡化,直至形成一个大的液泡,此时,细胞核呈扁平状,并与细胞质一起成为一薄层围绕着大液泡。当油细胞发育成熟后,细胞质及核开始解体,整个油细胞的腔由大液泡充满,成为油囊,在部分     

Organization of the teleostean nucleus rotundus.

The nucleus rotundus of 21 species of teleosts was studied by a modified Bodian and the Golgi method to clarify the histological organization, with special reference to the cell lamination and the glomerular formation. The common components of the nucleus in all species are as follows: a thick fiber bundle which comes from the commissura horizontalis and enters the nucleus from the dorsal surface, many small cells, large cells, glomeruli, and a surrounding fibrous capsule. The nuclei of all species studied are classified into three types mainly on the distribution of the small cells, and to a lesser degree on the location of the large cells and the glomeruli. The first type of nucleus has small cells, large cells and glomeruli throughout its extent. In the second type of nucleus, many small cells form a peripheral cell layer, while the large cells and glomeruli are found all over the nucleus. The third type of nucleus is clearly laminated. It is composed of four layers arranged concentrically around a central fiber net in the following order: a glomerular layer, a fibrous layer, a small-cell layer, and a peripheral fibrous capsule. In some species, the large cells are located in the fibrous capsule, and all glomeruli contain a star-like structure, which corresponds to the tips of the large cell dendrites.     

Functional Organization of the Cardiac Ganglion of the Lobster, Homarus americanus

External recording and stimulation, techniques were used to determine which neurons and interactions are essential for production of the periodic burst discharge in the lobster cardiac ganglion. Burst activity can be modulated by brief single shocks applied to the four small cells, but not by similar stimulation of the five large cells, suggesting that normally one or more small cells primarily determine burst rate and duration. Repetitive electrical stimulation of large cells initiates spike activity in small cells, probably via excitatory synaptic and/or electrotonic connections which may normally act to prolong bursts and decrease burst rate. Transection of the ganglion can result in burst activity in small cells in the partial or complete absence of large cell spike activity, but large cells isolated from small cell excitatory synaptic input by transection or by application of dinitrophenol do not burst. Generally, transections which decrease excitatory feedback to small cells are accompanied by an increase in burst rate, but mean spike frequency over an entire burst cycle stabilizes at the original level within 10–30 min for various groups of cells whose spike-initiating sites are still intact. These and previous results suggest that the system is two layered: one or more small cells generate the burst pattern and impose it on the large cells which are the system's motorneurons.     

Observations on the Leydig cells in the male East African spring hare (Pedetes surdaster larvalis).

The morphology of Leydig cells of the testis of sexually mature and sexually immature spring hares was studied. The cytoplasm of the Leydig of cells the sexually immature spring hares was packed with large lipid droplets leaving little space for the other organelles. Smooth endoplasmic reticulum was poorly developed and occasionally formed concentric layers of fenestrated cisterns around the large lipid droplets. The Leydig of cells the sexually mature spring hares were almost devoid of lipid droplets and their cytoplasm was occupied by abundant tubular smooth endoplasmic reticulum. Cells which shared characteristics with both immature Leydig cells and undifferentiated mesenchymal cells were observed in the limiting membrane of the seminiferous tubulus. These Leydig-like cells may play a role in the differentiation of Leydig cells in the spring hare.     

Synthesis of the major oligosaccharide components of murine haemangioendothelioma cells

Murine haemangioendothelioma cells in culture synthesize lactosaminoglycan-type glycoproteins which are found both associated with cells and secreted into the culture medium. Pronase-derived glycopeptides, prepared from the [3H]glucosamine-labeled glycoproteins that were secreted into the culture medium were found to contain about 10% of the labeled products as large size (Mr greater than 5000) 3H-labeled glycopeptides. In contrast, 40% of the cellular 3H-glycopeptides were found to be of this large size class of glycopeptides. These large size glycopeptides did not bind to Con A-Sepharose but did bind to Datura stramonium-agarose, from which they were eluted with chitobiose. The glycopeptides which did not bind to Datura-lectin were sulfated complex-type oligosaccharides which were not degraded by endo-beta-galactosidase. The glycopeptides which bound to Datura-lectin were degraded by endo-beta-galactosidase (or keratanase) to yield Gal----GlcNAc----Gal and glycopeptides, which were resistant to further endo-beta-galactosidase digestion and which no longer bound to Datura lectin-agarose. A major [3H]glucosamine-labelled glycoprotein (Mr approx. 75000) was found to be susceptible to endo-beta-galactosidase degradation and is probably the major cellular constituent having lactosaminoglycan-type side chains in these cells. An in vitro assay to measure leucocyte-haemangioendothelioma interactions indicated that treatment of haemangioendothelioma cells with endo-beta-galactosidase reduced leucocyte binding to these cells by 80%.     

Compartmentalization of steroidogen esis by the equine corpus luteum

The presence of cytochrome P450C17 within equine follicles and corpora lutea (CL) was detected by immunostaining. Two different antibodies were used which had previously been shown by immunoblotting to cross-react with equine P450C17. Strong positive immunostaining was present in the theca-derived cells of the CL during the estrous cycle and pregnancy. In the CL from mares after Day 40 of pregnancy there were also occasional bands of positively stained cells which resembled the polyhedral-shaped theca cells seen in preovulatory follicles. The pattern of immunostaining suggested compartmentalization of steroidogenesis within the equine CL with small cells possessing the potential to produce androgen which could then be aromatized to estrogen by the large luteal cells.     

Asymmetric cell division in human leukemic promyelocytic cells (HL-60).

Small cells accounted for 8-9% of the human leukemic promyelocytic cells (HL-60). The diameter of the small cells was 8.44 μm, whereas that of the large cells, which were heterogeneous in cell size, was 11.0 μm. The small cells were produced from the large cells through asymmetric cell division, which was demonstrated by cloning experiments and by microscopy.     

The regulation of steroidogenesis is different in the two types of ovine luteal cells

Ovine luteal tissue contains two distinct steroidogenic cell types, small (8-20 microns) and large (greater than 20 microns), which differ based on morphological and biochemical criteria. Unstimulated small cells secrete low levels of progesterone, respond to LH or dibutyryl cAMP (dbcAMP) with enhanced secretion of progesterone, and contain most of the receptors for LH. The unstimulated large cells, conversely, secrete high levels of progesterone, have few, if any, receptors for LH, and do not respond to LH or dbcAMP with increased progesterone secretion. The lack of response to dbcAMP by large cells was investigated. Large cells incubated in the presence of cholesterol, ram serum, or 25-hydroxycholesterol did not demonstrate substrate limitation. Hormone-independent stimulation of adenylate cyclase by cholera toxin or forskolin resulted in increased adenylate cyclase activities (P less than 0.01), cAMP accumulation (P less than 0.05), and the binding of endogenous cAMP (P less than 0.05) by type I cAMP-dependent protein kinase in both small and large cells. These treatments were accompanied by enhanced secretion of progesterone (P less than 0.05) in small cells. In contrast, large cells did not respond with an increase in progesterone secretion under these conditions. These observations suggest that the high rate of secretion of progesterone in unstimulated large cells is not regulated by cAMP.     

Fine structure of the small,granule-containing cells in the superior cervical ganglia of hydrocortisone-treated early postnatal and adult rats

Summary Hydrocortisone injections into rats on postnatal days 3–9 caused an increase in the number of small granulecontaining cells in the superior cervical ganglia. These cells, corresponding to the small, intensely fluorescent cells, showed an extensive rough endoplasmic reticulum, a large Golgi apparatus and a very large number of granular vesicles. In addition to the granular vesicles, 70–160 nm in diameter, in which the dense core filled most of the vesicle, most cells of the hydrocortisone-injected rats contained also larger granular vesicles, up to 350 nm in diameter, in which the dense core was eccentrically located. A minority of the cells contained only granular vesicles 70–100 nm in diameter, which was the only type seen in the saline-treated control rats.Thirty days after discontinuation of the hydrocortisone treatment, most of the cells with large granular vesicles had disappeared, and only two profiles of such cells were seen on day 40. The other small cells contained only granular vesicles 70–160 nm in diameter, and these cells could not be distinguished from the small granule-containing cells of 40-day-old control rats treated early postnatally with saline.Hydrocortisone treatment, first on days 3–9 and subsequently on days 40–46, caused reappearance of the small granule-containing cells with large granular vesicles up to 350 nm in diameter, the dense core of which was eccentrically located. Hydrocortisone treatment on days 40–46 only was not followed by appearance of such cells in rats treated with saline on days 3–9.Abbreviations used in the Text PNMT phenylethanolamine-N-methyltransferase - SIF cell small intensely fluorescent cell - SGC cell small granule-containing cell The author is grateful to Professor Olavi Eränkö and Dr. Seppo Soinila for constructive criticism. Expert technical assistance by Miss Hanna-Liisa Alanen, Mrs. Marja-Leena Piironen and Mrs. Anne Reijula is gratefully acknowledged. This study has been supported by a grant from the Finnish Medical Foundation.     

Simian virus 40 small t antigen activates the carboxyl-terminal transforming p53-binding domain of large T antigen.

Expression of the simian virus 40 large T antigen (large T) in F111 rat fibroblasts generated only minimal transformants (e.g., F5 cells). Interestingly, F111-derived cells expressing only an amino-terminal fragment of large T spanning amino acids 1 to 147 (e.g., FR3 cells), revealed the same minimal transformed phenotype as F111 cells expressing full-length large T. This suggested that in F5 cells the transforming domain of large T contained within the C-terminal half of the large T molecule, and spanning the p53 binding domain, was not active. Progression to a more transformed phenotype by coexpression of small t antigen (small t) could be achieved in F5 cells but not in FR3 cells. Small-t-induced progression of F5 cells correlated with metabolic stabilization of p53 in complex with large T: whereas in F5 cells the half-life of p53 in complex with large T was only slightly elevated compared with that of (uncomplexed) p53 in parental F111 cells or that in FR3 cells, coexpression of small t in F5 cells led to metabolic stabilization and to high-level accumulation of p53 complexed to large T. In contrast, coexpression of small t had no effect on p53 stabilization or accumulation in FR3 cells. This finding strongly supports the assumption that the mere physical interaction of large T with p53, and thus p53 inactivation, in F5 cells expressing large T only does not reflect the main transforming activity of the C-terminal transforming domain of large T. In contrast, we assume that the transforming potential of this domain requires activation by a cellular function(s) which is mediated by small t and correlates with metabolic stabilization of p53.     

Embryonic development of the heart. I. A light and electron microscopic study of myocardial development in the early chick embryo

Embryonic chick myocardium (stages 8+ to 12?) was studied by light and electron microscopy. The myocardium, which is initially comprised of radially oriented cells with large intercellular spaces gradually becomes more tightly packed. Intercellular spaces decrease and the cells assume a circumferential orientation. Myocardial cells remain epithelial throughout formation of the functional tubular heart and specialized epithelial junctions (apical junctional complex or terminal bars) undergo modification to form intercalated discs. Embryonic myocardial cells contain large amounts of free ribosomes and particulate glycogen, the latter often associated with portions of granular reticulum. Unlike developing skeletal muscle. The amount of granular reticulum contained in the myocardial cell cytoplasm is large and, along with a hypertrophied Golgi apparatus, suggests that these cells may have a secretory function. These organelles persist during the initial period of fibril formation. Myofibrils apparently form from non filamentous precursor material and not by alignment of sequentially synthesized components.     

The Structure and Development of the Pineal Complex in the Lanternfish Triphoturus mexicanus (Family Myctophidae)

The pineal complex of the lanternfish Triphoturus mexicanus was studied by light and electron microscopy. Receptor, supportive and ganglion cells were identified in the pineal end-vesicle. The presence of receptor cells and the prominent pineal window strongly suggest a photoreceptive function. Numerous unmyelinated nerve fibers were also found in the end-vesicle. These converge to form part of the pineal stalk. Two kinds of cells (Types I and II) were distinguished in the dorsal sac, which is very well developed. Type I cells are non-ciliated ependymal cells and contain large parcels of glycogen. Type II cells contain large lipidlike inclusions and a distinct band of filaments around the entire periphery of the cell. Receptor cells appear very early in the development of T. mexicanus. They seem to originate from ependymal cells lining the roof of the third ventricle.     

Peroxidase activity in the epithelium of the digestive tract of the bullfrog, Rana catesbeiana

Peroxidase activity was examined cytochemically in the mucosal epithelium along the length of the digestive tract from the esophagus through the large intestine during the development of the bullfrog, Rana catesbeiana. In the tadpole of this species, cells with peroxidase activity were found abundantly in the esophagus, stomach, and large intestine; and the types of such cells differed according to the region: ciliated cells and mucous cells in the esophagus; ciliated cells in the stomach; and brush cells, absorptive cells, and goblet cells in the large intestine, respectively. After metamorphosis, however, peroxidase activity was observed exclusively in absorptive cells and goblet cells in the large intestine. Peroxidase activity was commonly demonstrated in apical vesicles or granules, to some degree in rough endoplasmic reticulum, and in some elements of the Golgi apparatus. Furthermore, reaction product was also found in mucus covering the luminal surface of such epithelial cells. These findings indicate that peroxidase-positive cells, which may have the ability to synthesize peroxidase as a secretory product, were distributed mainly in three regions of the digestive tract in tadpoles (esophagus, stomach, and large intestine), but were centered in one specific region, the large intestine, after metamorphosis. Concomitantly, the variety of types of peroxidase-positive cells decreased during metamorphosis. Our results indicate that some of the peroxidase in the digestive tract may have a secretory origin and may play a role in the defense against microorganisms.     

Epibolic extension of the presumptive ectodermal layer of embryos of the newt Cynops pyrrhogaster before and during gastrulation.

Epibolic extension of the presumptive ectodermal layer (PEL) was investigated in embryos of the newt Cynops pyrrhogaster before and during gastrulation. The PEL was composed of only one layer of columnar cells at all stages examined. The cells of the PEL became elongated from the blastula to the early gastrula stage. They were most elongated at the early gastrula stage and then shortened during gastrulation. Present observations suggest that changes in cell shape of the PEL play an important role in the control of the epibolic extension of the newt embryos. The morphology and movement of the isolated cells from the PEL were examined in an attempt to elucidate the role of cell movement in epibolic extension of the PEL. Blebbing and vermiform cells which showed active cell movement appeared at the early blastula stage. The blebbing cells, which formed large hyaline blebs that moved around the circumference of each cell, appeared in large numbers at the early blastula stage. The frequency of the blebbing cells decreased from the early blastula to the early gastrula stage and increased again during gastrulation. The vermiform cells, which had an elongated cell body and moved in a worm-like manner, increased in frequency from the early blastula to the early gastrula stage. The relative number of such vermiform cells was maximal at the early gastrula stage and decreased abruptly during gastrulation. These results suggest that the elongation of the cells of the PEL is controlled by the active cell movement which resembles that of a worm.     

Diagnosis of sexual cycle by means of vaginal smear method in the chinchilla (Chinchilla lanigera)

An investigation was made as to whether the sexual cycle and pregnancy can be determined by means of vaginal smear in chinchillas. This study represents the first attempt to record changes which occur in the pattern of exfoliated cells in chinchilla's vaginal smear during anoestrus, proestrus, oestrus, metoestrus and pregnancy. Fifteen female chinchillas aged from 8 months to 3 years and bred through harem breeding method were used. The major change during proestrus was an increase in the proportion of superficial cells, with a corresponding decrease in other cells. Goblet cells were observed in the smears prepared by strong aspiration during this cycle. Neutrophils, small and large intermediates and parabasal cells were not found in the smear during oestrus and the smear consisted of superficial cells only. In the proportion of neutrophils, small and large intermediates and parabasal cells increased during metoestrus. In addition, metoestrum and foam cells were found in this cycle. In anoestrus; superficial and parabasal cells were present in small numbers. Also small and large intermediate cells as well as neutrophils were present. Traces of foam and metoestrum cells were found. During pregnancy, neutrophils generally of medium density were present, parabasal; small and large intermediate cells were present at low or medium density, and superficial cells were only present in trace amounts.     

Some observations on the fine structure of the corpus striatum of the rat brain

Summary The rat corpus striatum was perfused vitally with glutaraldehyde, immersed in OsO₄ and then observed under an electron microscope.Numerous small cells in the neostriatum show a simple cytoplasmic structure, while the large cells possess a complicated fine structure. These are differentiated under the elctron microscope into two kinds, which seem to have functional differences. The large pallidal cells containing much pale cytoplasm are covered with many varied axonal boutons from the cell body to the dendritic terminal making numerous axo-somatic or axo-dendritic trunk synapses. Numerous axo-dendritic, or spine synapses are recognized in the neostriatal neuropil.These numerous axon terminals, which belong to striatal nerve cells or other nuclei of the brain, are classified morphologically into several types. At least five types of synaptic vesicles are distinguished by their size or by the presence of fine dense granules on their membranes, and seem to be specific to the neostriatum.Many myelin interruptions and several kinds of glial cells in the corpus striatum are observed. Moreover, the ventricular wall of the caudate nucleus, namely, the ependyma, and two kinds of subependymal cells are described and discussed with reference to the subependymal layer.     

Mean diameter of nucleolar bodies in cultured human leukemic myeloblasts is mainly related to the S and G2 phase of the cell cycle

Mean diameter of nucleolar bodies (nucleoli without the perinucleolar chromatin) per cell was studied in human leukemic myeloblasts represented by K 562 and Kasumi 1 cell lines which originated from chronic and acute myeloid leukaemia. The measurement of mean diameter of nucleolar bodies in specimens stained for RNA was very simple. Such approach eliminated the variability of the perinucleolar chromatin discontinuous shell which might influence the measured nucleolar size as suggested by earlier studies. Ageing of K 562 myeloblasts produced a significant decrease of cells in S+G2 phase of the cell cycle accompanied by a significant reduction of mean diameter of nucleolar bodies (MDNoBs) per cell. In contrast, treatment of Kasumi 1 myeloblasts with histone deacetylase inhibitor - Trichostatin A - produced a large incidence of resistant cells in S+G2 phase which were characterised by a large increase of MDNoBs. Thus, MDNoBs in leukemic myeloblasts might be a helpful tool to estimate the incidence of cells in the S+G2 phase at the single cell level in smear preparations when the number of cells is very small.