The ultrastructure of oocytes during oogenesis and oocyte degeneration associated with follicle cells in female Sinonovacula constricta(Lamarck, 1818) were investigated by electron microscope observations. Ovarian follicles are surrounded by a matrix of vesicular connective tissue cells(VCT cells). VCT cells contain large quantities of glycogen particles and several lipid droplets in their cytoplasm. It is suggested that VCT cells act as a source of nutrients for vitellogenesis during oogenesis. In early vitellogenic oocytes, several coated vesicles, which appear at the basal region of the oocyte, lead to the formation of membrane-bound vesicles via endocytosis. The uptake of nutritive materials in coated vesicles formed by endocytosis appears through the formation of coated pits on the oolemma during vitellogenesis. During the late stage of oogenesis, yolk precursors(yolk granules), mitochondria and lipid droplets are present in the cytoplasm of late vitellogenic oocytes. In particular, proteinaceous yolk granules containing several different components are intermingles and form immature yolk granules. In the mature oocyte, small immature yolk granules are intermingled and form large mature yolk granules. Vitellogenesis occurs through a process of autosynthesis, involving combined activity of the Golgi complex, mitochondria and rough endoplasmic reticulum in the cytoplasm of vitellogenic oocytes. The process of heterosynthesis is where extraovarian precursors are incorporated into oocytes by endocytosis at the basal region of early vitellogenic oocytes before the formation of the vitelline coat. Follicle cells appear to play an important role in vitellogenesis and oocyte degeneration. The functions of attached follicle cells to the oocyte during oocyte degeneration are phagocytosis and digestion of phagosomes originating from oocyte degeneration. After digestion of phagosomes, it is assumed that the function of follicle cells can permit a transfer of yolk precursors necessary for vitellogenesis and allows for the accumulation of glycogen and lipid during oocyte degeneration, which can be employed by vitellogenic oocytes. Follicle cells of S. constricta may possess a lysosomal system for induction of oocyte breakdown and might resorb phagosomes in the cytoplasm for nutrient accumulation during oocyte degeneration.
The present study was undertaken to demonstrate the surface structure of Paragonimus westermani metacercaria in Korea with special reference to the distribution of sensory papillae. Metacercariae were isolated from crayfish, one of the second intermediate tost of P. westermani in Bogil island, Chollanam-do (Province), Korea, where has been known as an endemic area of human paragonimiasis. Isolated metacercariae were encysted and examined with light. scanning and transmission electron microscopes for morphological features. On the surface of iBetacercariae, three types of sensory FaFillae were identified. Large domed papillae ($3~5{\mu\textrm{m}}$), which were covered with wrinkled plasma n!embrane of the worm, were distributed on the oral and ventral suckers only. On the oral sucker, these large domed papillae were 12~13 in number. On the other hand large domed papillae on the ventral sucker were constantly 6 in number and hexagonal in distribution. Small domed papillae ($2~3{\mu\textrm{m}}$), of which surface was more smooth than those of large ones, were distributed symmetrically on the ventral (30~32 pairs) and dorsal surfaces (40~42 Pairs). Ciliated Papillae ($0.8~1.5{\mu\textrm{m}}$) were observed about 5~6 in number around the oral sucker and 3~5 pairs each on the ventral and dorsal surface of the body. Single Fcinted spines covered the entire surface of the body except around the excretory pore. Spines on the anterior fart of the body were 0.9~2.0${\mu\textrm{m}}$ in length and $45~55/100{\mu\textrm{m}}$2 in number, and were gradually reduced in length ($0.4~1.4{\mu\textrm{m}}$) and in nuns.her ($12~27/100{\mu\textrm{m}}$2) toward the posterior part. The body wall of p. westermoni metacercariae was consisted with anucleated syncytium layer, fibrous interstitial layer and musclar layer. In the anucleated syncytium, biconcave ($0.15~0.55{\mu\textrm{m}}$) and spherical ($0.08~0.16{\mu\textrm{m}}$) secretory granules, which were transferred from epidermal cells via protoplasmic tubules, mitochondria and rihoEorses, T-ere observed. Spines originated around the basement membrane protruded externally. Epidermal cells were consisted with a nucleus and a cytoplasm, and connected to syncytium with protoplasmic tubules. In the cytoplasm many secretory granules, mitochondria, Golgi complex, endoplasmic reticula, ribosomes and lipid droplets were observed.
The Pancreatic islet are the clusters of endocrine cells scattered through out the exocrine pancreas. Transplantation of a sufficient pancreatic islets can normalize blood glucose level so that may prevent devastating complications of type I diabetes(IDDM) and other side effects of the IDDM. Recently, there are several approaches to transplant sufficient pancreatic islet, and it was comprised in increase or regeneration of the endogenous $\beta$-cell mass from donor's pancreas, but relatively few studies have been devoted to the morphological characters of the isolated and 3 day cultured pancreatic islets. We investigated morphological pattern of intracellular structure of isolated and 3 day cultured pancreatic islets. The morphological characters of the pancreatic islets were observed by scanning electron microscope and transmission electron microscope, and insulin distribution of the each islets were observed by transmission electron microscope, and were labeled with insulin antibody. Intracellular structures including nuclei, mitochondria, RER, Golgi complex and many secretory granules were normally appeared in the isolated pancreatic islets which was extracted immediately dornor's pancreas, however, There is a significant morphological changes between the 3 day cultured pancreatic islets and isolated islets. 3 day cultured pancreatic islet's $\beta$-cells had normal nuclei but increased cytoplasm mass and RER and developed Golgi complex. Insulin secretory granules were decreased in numbers rather than isolated pancreatic islet. In this study, the pattern of intracellular structure variation was examined during pancreatic islet culture. Most distinct features are variation of the insulin secretory granules, and developed RER, and dilated golgi complex. Therefore, we suggested that the various change of the morphological characters on cultured pancreatic islets were responsible for the function(biosynthesis and secretion of insulin) and growth. These results were also cultured islets have greater ability to recover and maintain normoglycemia than isolated islet transplantation.
The androgenic gland secretes a hormone, androgenic gland hormone, which is believed to act on the differentiation of the primary, secondary, and behavioral sex characteristics in most malacostracan crustaceans. This report presents the ultrastructural morphology of the androgenic gland in the freshwater prawn Macrobrachium nipponense. This gland, located in the coxopodite of the last pair of walking legs, was attached to the subterminal region of the sperm duct. The gland was composed of simple cellular strands, encased by a fibrous sheath. Microvilli were situated in the fibrous sheath, especially at the edge of each cellular strand. The androgenic gland cells had the large and round nucleus and rough endoplasmic reticulum arranged either in spirals or in concentric circles throughout the cytoplasm of the cell. They also had the well-developed Golgi complex and long mitochondria with flat and transverse cristae. The Golgi complex was similar to microvesicular cluster, but usually in the shape of typical dictyosomes, These features of androgenic gland cells coincides well with the protein/peptide secretion in their function. However, despite the apparent ultastructual equipment for protein/peptide secretion, no accumulation of materials secreted were noticed in the cytoplasm. Therefore it is strongly suggested that the transient transportation of the materials into the hemocoel has occurred just after synthesis.
Ultrastructure of germ cells, the cyst epithelial cells and interstitial cells during spermatogenesis of the stone flounder, Kareius bicoloratus (Pleuronectidae) sampled on the west coast of Korea were investigated by electron microscopic observations. In the primary spermatocyte, the synaptonemal complexes appear in the zygotene stage of the prophase during maturation division. In the growing testis, especially, the interstitial cells (Leydig cells) appear near the primary, secondary spermatocytes and spermatids. Well-developed interstitial cells (steroid hormone secreting cells) which are located in the interlobular space in growing testis have three morphological characteristics of a vesicular nucleus, mitochondria with tubular cristae and smooth endoplasmic reticulum. During spermatogenesis, the primary and secondary spermatocytes attach to the cyst epithelial cell (Sertoli cell) having an elongated ovoid or triangular nucleus and several mitochondria in the cytoplasm. In the growing testis, lipid droplets, the mitochondrial rosettes and glycogen particles appear in the cytoplasm of the cyst epithelial cells near the secondary spermatocytes and spermatids. Particularly, the mitochondria, endoplasmic reticulum, little lipid droplets and the large amount of glycogen particles are present in the cytoplasm of the cyst epithelial cell in the late growing testis. In the late stage of spermiogenesis, the proximal centriole is joined to the nuclear envelope, the distal centriole forms the basal body of the flagellum and gives rise to the axial filament of the flagellum. No acrosome of the sperm is formed as seen in other teleost fish. The head of the spermatozoon is approximately $3{\mu}m$ in length and its tail is about $30{\mu}m$ in length. The axoneme of the tail flagellum of the spermatozoon consists of nine outer doublet microtubules at the periphery and two centrial singlet microtubules at the center. The spermatozoon of this species has two axonemal lateral fins. Especially, the cyst epithelial cells which located near groups of gametes in the various stages, show three functions: nutrition, phagocytosis and steroidogenesis. Especially, the nuclei of cyst epithelial cells in the recovery stage of the testicular developmental stages appear to be irregular in shape after spermiation. Of three functions of the cyst epithelial cell, several characteristics of phagocytosis are showed in the cytoplasm of the cyst epithelial cells in the recovery stage of the testicular developmental stages. At this stage, therefore, it is assumed that the cyst epithelial cells are involved in degeneration and resorption of undischarged germ cells after spermiation.
Kim, Eun Ji;Kim, Guen Tae;Kim, Bo Min;Lim, Eun Gyeong;Kim, Sang-Yong;Kim, Young Min
Journal of the Korean Society of Food Science and Nutrition
/
v.45
no.9
/
pp.1257-1264
/
2016
Extracts from Artemisia annua $Linn\acute{e}$ (AAE) have various functions (anti-malaria, anti-virus, and anti-oxidant). However, the mechanism of the effects of AAE is not well known. Thus, we determined the apoptotic effects of AAE in AGS human gastric carcinoma cells. In this study, we suggested that AAE may exert cancer cell apoptosis through the Akt/mammalian target of rapamycin (mTOR)/glycogen synthase kinase (GSK)-$3{\beta}$ signal pathway and mitochondria-mediated apoptotic proteins. Activation by Akt phosphorylation resulted in cell proliferation through phosphorylation of tuberous sclerosis complex 2 (TSC2), mTOR, and GSK-$3{\beta}$. Thus, de-phosphorylation of Akt inhibited cell proliferation and induced apoptosis through inhibition of Akt, mTOR, phosphorylation of GSK-$3{\beta}$ at serine9, and control of Bcl-2 family members. Inhibition of GSK-$3{\beta}$ attenuated loss of mitochondrial membrane potential and release of cytochrome C. Bax and pro-apoptotic proteins were activated by their translocation into mitochondria from the cytosol. Translocation of Bax induced outer membrane transmission and generated apoptosis through cytochrome C release and caspase activity. We also measured 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, lactate dehydrogenase assay, Hoechst 33342 staining, Annexin V-PI staining, 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide staining, and Western blotting. Accordingly, our study showed that AAE treatment to AGS cells resulted in inhibition of Akt, TSC2, GSK-$3{\beta}$-phosphorylated, Bim, Bcl-2, and pro-caspase 3 as well as activation of Bax and Bak expression. These results indicate that AAE induced apoptosis via a mitochondrial event through regulation of the Akt/mTOR/GSK-$3{\beta}$ signaling pathways.
Cold stress influence plant growth through a wide range of growth characters. Adverse effects of low temperature to plant growth come from results of colligative and complex physiological responses to cold stress. To evaluate more exactly cold tolerance of crop plant, it is needed to observe physiological changes induced by cold stress and to analyze relationships between intraspecific variations in physiological factors related to cold tolerance and the extent of cold tolerance in the field. Therefore, the composition and unsaturation ratio of fatty acids in phospholipid, a constituent of membrane, the transition-temperature in respiratory activity of mitochodria, the chlorophyll fluorescence as a factor related to photosynthesis were investigated in rice plant and data on these factors were compared with the degree of cold tolerance obtained in the field experiment. Also, effects of hardening and Mn++ treatment were evaluated as a method to reduce chilling injuries. The unsaturation ratio of fatty acids, whether rice plants were grown in a natural condition or under the chilling stress, was higher in the cold- tolerant varieties and was significantly correlated with the degree of cold tolerance (1-9) observed in the field experiment. And it was also increased by chilling treatment or hardening treatment, due to a reduction in palmitic acid content and an increase in linolenic acid content. The transition-temperature of respiratory activity of mitochodria isolated from etiolated rice seedlings ($25^{\circ}C$, two week-grown in the dark), was correlated with the degree of cold tolerance in the field, cold -tolerant varieties showing a lower transition-temperature. It was not influenced by growth stages. The intensity of chlorophyll fluorescence was highly correlated with the degree of cold tolerance, cold-tolerant varieties having a higher fluorescence intensity. By foliar application of Mn, the transition-temperature of respiratory activity was lowered as much as 0-2$^{\circ}C$ in all tested varieties. Soil application of Mn induced more significant effect in cold-susceptible varieties with a possibility of reducing chilling injuries. On the whole, there were high correlationships among the degree of cold tolerance, the unsaturation ratio of fatty acids in phospholipid, the transition- temperature of respiratory activity and chlorophyll fluorescence except for a few varieties. The transition- temperature of respiratory activity appeared to be negatively correlated with the unsaturation ratio of fatty acids. and the chlorophyll fluorescence to be positively correlated with the unsaturation ratio. This implies that these physical and physiological factors were very closely related to cold tolerance and can be used as an effective index of the evaluation of cold tolerance of crop plant. But other factors as well as three factors discussed above are needed to be considered colligatively and altogether with a systematic analysis for the more exact evaluation of cold tolerance. in rice cultivars. in rice cultivars.
Purpose: Mitochondria play a crucial role in preserving skeletal muscle mass, and damage to mitochondria leads to muscle mass loss. This study investigated the effects of oxypeucedanin hydrate, a furanocoumarin isolated from Angelica dahurica radix, on myogenesis and mitochondrial function in vitro and in zebrafish models. Methods: C2C12 myotubes cultured in media containing 0.1, 1, 10, or 100 ng/mL oxypeucedanin hydrate were immunostained with myosin heavy chain (MHC), and then multinucleated MHC-positive cells were counted. The expressions of markers related to muscle differentiation, muscle protein degradation, and mitochondrial function were determined by quantitative reverse transcription polymerase chain reaction. To investigate the effects of oxypeucedanin hydrate on mitochondrial dysfunction, Tg(Xla.Eef1a1:mito-EGFP) zebrafish embryos were treated with 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI) with or without oxypeucedanin hydrate and analyzed for mito-EGFP intensity and mitochondrial length. Results: Oxypeucedanin hydrate significantly increased MHC-positive multinucleated myotubes (≥ 3 nuclei) and increased the expression of the myogenic marker myosin heavy chain 4. However, it decreased the expressions of muscle-specific RING finger protein 1 and muscle atrophy f-box (markers of muscle protein degradation). Furthermore, oxypeucedanin hydrate enhanced the expressions of markers of mitochondrial biogenesis (peroxisome proliferator-activated receptor-gamma coactivator 1 alpha, transcription factor a mitochondrial, succinate dehydrogenase complex flavoprotein subunit A, and cytochrome c oxidase subunit 1) and mitochondrial fusion (optic atrophy 1). However, it reduced the expression of dynamin-related protein 1 (a mitochondrial fission regulator). Consistently, oxypeucedanin hydrate reduced FOLFIRI-induced mitochondrial dysfunction in the skeletal muscles of zebrafish embryos. Conclusion: The study indicates that oxypeucedanin hydrate promotes myogenesis by improving mitochondrial function, and thus, suggests oxypeucedanin hydrate has potential use as a nutritional supplement that improves muscle mass and function.
High extracellular glucose concentration was reported to suppress intracellular $Ca^{2+}$ clearing through altered sarcoplasmic reticulum (SR) function. In the present study, we attempted to elucidate the effects of pyruvate and fatty acid on SR function and reveal the mechanistic link with glucose-induced SR dysfunction. For this purpose, SR $Ca^{2+}$-uptake rate was measured in digitonin-permeabilized H9c2 cardiomyocytes cultured in various conditions. Exposure of these cells to 5 mM pyruvate for 2 days induced a significant suppression of SR $Ca^{2+}$-uptake, which was comparable to the effects of high glucose. These effects were accompanied with decreased glucose utilization. However, pyruvate could not further suppress SR $Ca^{2+}$-uptake in cells cultured in high glucose condition. Enhanced entry of pyruvate into mitochondria by dichloroacetate, an activator of pyruvate dehydrogenase complex, also induced suppression of SR $Ca^{2+}$-uptake, indicating that mitochondrial uptake of pyruvate is required in the SR dysfunction induced by pyruvate or glucose. On the other hand, augmentation of fatty acid supply by adding 0.2 to 0.8 mM oleic acid resulted in a dose-dependent suppression of SR $Ca^{2+}$-uptake. However, these effects were attenuated in high glucose-cultured cells, with no significant changes by oleic acid concentrations lower than 0.4 mM. These results demonstrate that (1) increased pyruvate oxidation is the key mechanism in the SR dysfunction observed in high glucose-cultured cardiomyocytes; (2) exogenous fatty acid also suppresses SR $Ca^{2+}$-uptake, presumably through a mechanism shared by glucose.
The development of notochordal cells of nucleus pulposus was studied with electron microscope in human fetuses ranging from 30 mm to 260 mm crown-rump length. At 30 mm fetus, primitive notochordal cells were large with central nucleus, few organelles, and their cytoplasm usually contained dense glycogen and fine filaments. Notochordal cells at all ages contained bundles of fine filaments of indeterminate nature. One unusual feature of fetal notochordal cells was the consistent presense of rough endoplasmic reticulum surrounding poorly developed mitochondria. At 50 mm fetus, notochordal cells formed dense masses with interdigitating cell membranes connected by a variety of cell to cell junctions. With increasing age, the cell connections became slender threaded cytoplamic extending from cell and enclosed large extracellular space. Chondrocyte-like cells appeared to be separated by large volumes of extracellular matrix. Viable notochordal and condrocyte-like cells existed in specimen from all age. The extracellular spaces were filled with fibrillar and granular material by 90 mm fetus. Necrotic cells were distinguished by loss of their membrane integrity, vacuolization of their organelles, and the presence of dense osmiophilic masses. In adult tissue, notochordal cells became rounded or irregular in shape and developed a pericellular matrix consisting of collagen fibrile, and dense particle. The structure of notochordal cells and their persistance in the nucleus pulposus after fetal life suggested that they may have a significant role in the formation and maintenance of the nucleus pulposus. The presence of Golgi complex and well-developed endoplasmic reticulum in chondrocyte-like cells suggested that they are capable of producing and maintaining the extracellular matrix.
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