Electron-lucent Lipid Droplets Research Articles

Changes in testicular histomorphometry and ultrastructure of Leydig cells in adult male Japanese quail exposed to di (n-butyl) phthalate (DBP) during the prepubertal period

Phthalate esters, such as di(n-butyl) phthalate, (DBP), are synthetic chemical pollutants commonly used as plasticizers in the manufacture of plastics. In the present study, we investigated the effects of DBP in the testes of adult male quails (Coturnix cortunix japonica) exposed by oral gavage to variable doses of DBP (0 [control], 1, 10, 50, 200, and 400 mg/kgbw−d), for 30 days during the prepubertal period, using histo-morphometric and ultrastructural techniques. Generally, significant decreases in seminiferous tubular diameter (STD) and epithelial height (SEH) were observed predominantly at the highest DBP doses (200 and 400 mg/kg), as compared to medium (50 mg/kg), and lowest doses (1 and 10 mg/kg) as well as the control group. Ultrastructurally, apparent dose-specific degenerative changes were observed in the Leydig cells. The lowest DBP doses (1 and 10 mg/kg) did not produce significant effects on Leydig cell ultrastructure, whereas, at the highest doses (200 and 400 mg/kg), the Leydig cells were remarkably conspicuous in the interstitium and appeared foamy. There was a preponderance of electron-lucent lipid droplets which crowded out the normal organelles of the cell, as well as increases in the number of dense bodies in the cytoplasm. The smooth endoplasmic reticulum (sER) was less obvious, compacted, and wedged between the abundant lipid droplets and mitochondria. Taken together, these findings indicate that pre-pubertal exposure of precocious quail birds to DBP, produced parameter-specific histometric tubular changes, as well as dose-dependent cyto-structural derangement of the Leydig cells; which consequently may lead to overt reproductive impairments in the adult bird in the environment.Graphical

The unusual cytoarchitecture of "vitelline follicles" in freshwater blood flukes of the genus Sanguinicola (Digenea, Aporocotylidae).

This is the first study assessing the cytoarchitecture of the vitellarium of members of the freshwater, teleost-infecting lineage of blood-flukes (Aporocotylidae). The vitelline cytoarchitecture of two innominate species of Sanguinicola from freshwater fishes in Russia showed that vitelline cells at different stages of maturation are widely distributed throughout much of the body and are mixed with other cell types. The latter feature indicates that use of the term “follicular vitellarium” is inappropriate for species of this genus. An additional characteristic of the vitelline cells in these Sanguinicola spp. is their ability to form long, pseudopodia-like extensions of the peripheral cytoplasm that contact neighbouring vitelline cells and sarcoplasmic extensions, forming both heterologous and homologous intercellular junctions. Within the vitelline duct lumen, the cytoplasm of mature vitelline cells is filled with regular clusters (0.5–1.0 μm in diameter), comprising 10–30 vitelline globules, which have heterogeneous contents and electron-lucent lipid droplets (1.1–1.7 μm in diameter), but no apparent modifications of vitelline globules occur within the vitelline duct. The flattened, ciliated, epithelial lining of the common vitelline duct contains intra-epithelial nuclei, its luminal surface bears shallow lamellae and adjacent cells are adjoined by apical septate junctions. All of these observations, when compared to the marine Aporocotyle simplex, likely represent additional characteristics supporting the divergent evolutionary lineages of marine and freshwater aporocotylids.

Ultrastructural investigations of vitellogenesis and oogenesis of Proctoeces sp. infesting Platycephalus indicus fishes caught from Alataka Harbor, Suez Gulf, Egypt.

Heavy infestations with the intestinal parasite Proctoeces sp. were detected in the bar-tail flathead Platycephalus indicus collected from Alataka harbor, Suez Gulf, Egypt. Different developmental stages of vitellocyte and oocyte maturations of this digenean parasite were studied using transmission electron microscopy. The vitelline masses are covered with a delicate fibrous layer and enclosed with segregated circular muscle bundles. The first stage(S 1) of vitellocytes maturation is distinguished by a large centrally located oval nucleus and abundant mitochondria and free ribosomes fixed at a perinuclear reside. The second stage(S 2) of vitellocyte maturation has greater cytoplasmic volume, Golgi vesicles, granular endoplasmic reticulum and the vitelline globules became more electron-dense. The third stage (S 3) is characterized by an oval nucleus, shell globules with different sizes aggregated in clusters, electron-lucent lipid droplets and osmiophilic glycogen .The cytoplasm of mature vitellocytes (S 4) was gradually degraded, involving analogous sacs of granular endoplasmic reticulum situated at the peripheral and perinuclear regions, glycogen granules, lipid droplets and packed larger globule clusters. During vitellocyte developmental stages, the nucleo-cytoplasmic ratio decreased while the synthetic activities increased. Detailed ultrastructural characteristics of four different developmental stages of oocyte maturation were briefly described.

Susceptibility of chicken Kupffer cells to Chinese virulent infectious bursal disease virus

Infectious bursal disease (IBD) is an acute, highly contagious, and immunosuppressive avian disease caused by IBD virus (IBDV). Although the effects of IBDV on bursa of Fabricius in chickens have been well reported, the impacts of IBDV on liver after IBDV infection are still unclear. In the present study, specific pathogen free (SPF) chickens were experimentally inoculated with IBDV Chinese virulent strain BC6/85, and the cells in liver and bursa were examined by immunohistochemistry and transmission electron microscopy (TEM). The congestion of liver tissue and fatty degeneration of hepatocytes were characteristics of microscopical changes in chicken liver at 3 days post infection (d.p.i.), whereas there were follicular lymphoid necrosis, apoptosis, depletion, as well as edema and congestion in bursa. In addition, the number of IBDV-positive cells peaked at 4 d.p.i. in bursa and at 3 d.p.i. in liver, respectively. With respect to ultrastructural pathological changes of hepatocytes, mitochondria swelled and nucleus deformed into an irregular shape or its chromatin peripherally condensed which indicated that the hepatocyte was at the early stage of apoptosis, and the electron-lucent lipid droplets in a variety of sizes were observed within cytoplasm. Kupffer cells became "swollen-like" and the electron-density of their cytoplasm was lower than that of cells in uninfected group. Liver glycogen deposits significantly declined from 2 to 5 d.p.i. and recovered strongly at 6 d.p.i. More importantly, KLU01 (macrophage marker) positive (KUL01(+)) cells were infiltrated in bursa and liver in IBDV-exposed chickens by immunoperoxidase staining. To demonstrate the correlation between IBDV and macrophages in bursa and liver, we further investigated the colocalization of viral antigens and macrophages by double immunofluorescence labeling. At 4 d.p.i., the percentage of double positive cells (IBDV positive and KUL01(+) cells) accounted for 26.5 percent of the total IBDV positive cells or 57 percent of the total KUL01(+) cells in bursa. In comparison, the percentage of double positive cells in liver constituted 97 percent of the total IBDV positive cells or 99 percent of the total KUL01(+) cells. These results suggest that IBDV was susceptible to KUL01(+) cells in liver (mainly Kupffer cells) and replicated in the KUL01(+) cells. By comparison with the influence of IBDV on bursa, our findings were the first to elucidate the pathological changes in liver after IBDV infection on a microscopical and ultrastructural scale, and, especially, to gain the initial insight into the susceptibility of Kupffer cells to IBDV.

Histopathology of the thyroid in amiodarone‐induced hypothyroidism

Amiodarone is well recognized as an anti-arrhythmic drug containing a high dose of iodine with considerable potential to cause thyroid dysfunction. The present patient was a 66-year-old Japanese woman who developed a cardiac arrhythmia and was given amiodarone as an anti-arrhythmic agent for approximately 3 months, until the day before her death. However, 19 days after starting amiodarone, serum testing indicated a hypothyroid status that was not recognized clinically. At autopsy, microscopy showed that most of the thyroid follicles were enlarged with dense colloid substance and lined by flattened follicular cells (involuted follicles). There were a small number of damaged follicles infiltrated by macrophages, which were immunopositive for HAM56. Sudan IV staining indicated many lipid droplets in follicular cells. Ultrastructurally the follicular cells contained large residual bodies composed of abundant electron-lucent lipid droplets of variable size. Although it is difficult to be certain of the direct link of amiodarone on the basis of a single case, it is reasonable to presume that this histopathology is associated with amiodarone-induced hypothyroidism and that involution changes represent the hypofunctional status of this drug-induced disorder. This is the first report on the histopathological findings of thyroid tissue from a patient with amiodarone-induced hypothyroidism.

Vitellocyte ultrastructure in the cestode Didymobothrium rudolphii (Monticelli, 1890): possible evidence for the recognition of divergent taxa within the Spathebothriidea

AbstractIn the spathebothriidean tapeworm Didymobothrium rudolphii (Monticelli, 1890) the fine structure of the vitellocytes at different stages of their development within the vitelline follicles, vitelline ducts and uterus was studied for the first time using transmission electron microscopy. The vitellocyte inclusions of D. rudolphii are shell globule clusters containing tightly packed shell globules associated with a matrix of moderate electron density, glycogen granules, large electron-lucent lipid droplets (up to 3 μm in diameter), and, occasionally, a lipid droplet may occur in the nucleus of the vitellocytes. The diameter of the clusters ranges from 0.4 to 2.5 μm, the number of shell globules in the clusters varies from 8 to 45, and the size of the globules ranges from 0.12 to 0.25 μm and they are of approximately homogeneous sizes within a cluster. Most vitellocyte lipid droplets have a heterogeneous configuration with a ‘cavity’ inside them when they are within vitelline ducts and intrauterine eggs. Vitellocytes of the eggs contain dark concentric bodies and lipid droplets. The interstitial tissue has a syncytial structure. The morphological parameters of the diameter and shape of shell globule clusters, arrangement of shell globules in clusters, number and diameter of globules within clusters, types of lipid droplets and presence of dark concentric bodies are compared with those of two other spathebothriidean genera, Cyathocephalus and Diplocotyle. The comparative data demonstrate that vitelline material morphology has unique features in three spathenothriidean genera and may be used as evidence for the recognition of separate taxa.

Xanthogranulomatous Appendicitis - An Incidental Finding of Localized Pathology

The clinical, histopathological, and electron microscopic features of an unusual case of xanthogranulomatous appendicitis are reported. The patient, a 37-year-old female, presented with typical signs of acute appendicitis and the appendix appeared slightly dilated at laparatomy. The histopathological sections showed numerous xanthoma cells mixed with inspissated fecaliths. Electron microscopy disclosed the presence of xanthoma cells filled with electron-lucent lipid droplets of variable size. The ultrastructural characteristics of these cells enabled the distinction of two types of lipid-laden histiocytes, in relationship to the size of the lipid droplets. Since the lipid droplets were seen also in cells other than histiocytes, it appears that these changes are secondary to a common mechanism,comprising factors such as obstruction, hemorrhage, inflammation,and local hypoxia.

Morphological characteristics of lipid accumulation in liver-constituting cells of acid lipase deficiency rats (Wolman's disease model rats).

Lysosomal acid lipase is a hydrolase essential for the intracellular degradation of cholesteryl esters and triglycerides. In the laboratory, rats with congenital deficiency of lysosomal acid lipase and marked accumulation of cholesteryl ester, cholesterol free and triglyceride in livers (Wolman's disease rat or Yoshida rat) that corresponded to human Wolman's disease were found and maintained. The morphological characteristics of accumulated lipids in the livers of affected rats were examined also. Many small lipid droplets and lipid crystals were found in the cytoplasms of hepatocytes and ED1-positive and ED2-positive foamy Kupffer's cells, respectively. Electron microscopically, many electron-lucent lipid droplets with limiting membrane were found in hepatocytes. Foamy Kupffer's cells had many multivesicular bodies with limiting membrane, which contained crivilinear bodies, lipid droplets and crystal clefts. At areas of aggregation of foamy Kupffer's cells forming islets, there were many desmin-positive Ito cells. Small lipid droplets with limiting membrane were also found in the cytoplasm of Ito cells and endothelial cells. These findings, which were obtained by morphological methods, indicated that triglyceride and both cholesteryl ester and free cholesterol accumulated in lipolysosomes mainly in hepatocytes and Kupffer's cells, respectively, and suggest that lysosomal acid lipase could participate in dissolution of the membrane.

Structural changes in the aorta of the hereditary hypertriglyceridemic rat.

Structural changes in the ascending thoracic aorta of hereditary hypertriglyceridemic (hHTG), insulin-resistant, and hypertensive rats were studied using transmission electron microscopy. Normotensive Wistar rats were used as controls. The most-pronounced morphological changes were observed in the tunica intima. Endothelial cells of hHTG rats formed a continual layer around the whole circumference. Subendothelial space was enlarged. Some endothelial cells were delaminated from the subendothelial space by big lipid droplets that were often present in the subendothelial space, and the endothelial cells bulged out towards the lumen. Big electron-lucent lipid droplets were present in the majority of the endothelial cells and occupied the main part of the cytoplasm. Degenerative microvesicular and membranous material was present in the cytoplasm. Increased numbers of vesicles of Golgi apparatus and cisternae of endoplasmic reticulum were found. Similar morphological alterations, but in less-extended form, were observed in smooth muscle cells. The organization and orientation of smooth muscle cells were essentially intact. In muscle cells, lipid droplets were localized in close relation to Golgi complex and in dilated cisternae of the sarcoplasmic reticulum. Lipid droplets, degenerative material, myelin figures, myelinoid membranes, and vesicular components were also sporadically found in the intercellular space among muscle cells. This pilot morphological investigation provides further arguments for a thorough and more-focused electron microscopy study of conductance arteries of the hHTG rats.

Ultrastructure of the cells of the ovarian interstitial gland in hypophysectomized rats. The effects of stimulation of the ovarian plexus and of denervation.

The fine structure of the interstitial gland of the ovary was studied in hypophysectomized rats and in hypophysectomized rats after denervation of the ovary or stimulation of the ovarian plexus. Hypophysectomized rats were used to eliminate gonadotropic influences on interstitial cells. In hypophysectomized rats, there was a large amount of intercellular space and cells had irregularly shaped nuclei and a large nuclear-cytoplasmic ratio. Prominent cytoplasmic features included small mitochondria with an electron-dense matrix, rough and smooth endoplasmic reticulum, polysomes and large osmiophilic lipid droplets. Interstitial cells from stimulated ovaries had reduced intercellular space and a reduced nuclear-cytoplasmic ratio. Mitochondria had tubular cristae; smooth endoplasmic reticulum-surrounded lipid droplets, and large polysomes were present. After section of the ovarian plexus, intercellular space was increased and filopodia were numerous. Cytoplasmic features included mitochondria with a dense matrix and indistinct cristae, large electronlucent lipid droplets, and variously sized multivesicular structures. These observations suggest that stimulation of the ovarian plexus in hypophysectomized rats causes regressed cells to assume the fine structural features of active steroidogenic cells. In contrast, interruption of the ovarian nerve supply causes a qualitative and quantitative increase in ultrastructural features characteristic of regressed steroidogenic cells. These responses of interstitial gland cells to denervation and stimulation provide morphological evidence for a functional role for the adrenergic nerves to this ovarian compartment.

Ultrastructural Differences between the Propagative and Dispersal Forms in Pine Wood Nematode, Bursaphelenchus lignicolus, with Reference to the Survival

The ultrastructure of cuticle and intestine was compared between the dispersal and propagative forms of Bursaphelenchus lignicolus. With growth of the propagative form, the thickness of cuticle increased gradually, while the ratio of cuticle to body diameter decreased. The cuticle of dispersal 3rd-stage larvae (LXIIIs) was the thickest of all stages in the two forms. In the dispersal 4th-stage larvae (LXIVs), the ratio of basal layer was the largest (67% of cuticle). The external cortical layer was thicker in the dispersal form than in the propagative one. The large electron-lucent lipid droplets deposited deeply in the intestinal and hypodermal cells of LXIIIs. These lipid droplets were partly mobilized to glycogen granules with a development of LXIIIs to LXIVs.

Electron microscopic observations on postnatal development of the X zone in mouse adrenal cortex.

The postnatal development and involution of the X zone in the mouse adrenal cortex of both sexes were examined using the light and electron microscopes. At 0–5 days of age, no special cell group could be distinguished for the developing X zone in the inner cortex. The inner cortical cells contained spherical or ellipsoidal mitochondria with vesiculotubular cristae, vesiculotubular smooth endoplasmic reticulum (sER) and electron-lucent lipid droplets. The first sign of the developing X zone was the appearance of small groups of cells in juxtamedullary region differing from the cells in other part of inner cortex at 8 days. The electron microscopy showed that such cells contained nuclei of somewhat irregular outline and some parallel stacks of flattened sER. At 10–11 days, a thin layer of small eosinophilic cells were clearly identified as the developing X zone light microscopically in both sexes. Electron microscopically, the X zone cells showed a much dense cytoplas, which contained abundant sER, many mitochondria and numerous ribosomes. The typical X zone cells were characterized by the formation of peculiar mitochondrial complexes and whorled pattern of the sER. Mitoses were often found in the X zone, where mitotic cells even contained the whorled sER and bizarre mitochondria characteristic of the typical X zone cells. In the male the X zone rapidly involuted and might disappear by 30 days of age, whereas in the female X zone persisted as a thicker layer with the earlist sign of fatty degeneration. The origin of the X zone cell and the process of formation of its characteristic organelles are discussed.

On the fine structure of alteration of the adrenal cortex in hypophysectomized rats

The adrenal cortex of normal and hypophysectomized rats was observed by electron microscopy 5–90 days after the operation and, chiefly, the degeneration mechanism of the cortical tissue was discussed. In normal rats mitochondrial inner structures are generally tubular in shape in glomerulosa cells, vesicular (and tubulo-vesicular) in fasciculate cells and tubulo-vesicular (and vesicular) in reticularis cells. Some mitochondria in a few glomerulosa cells show long straight tubular crests in normal as well as hypophysectomized rats. Usually lipid droplets in normal glomerulosa, fasciculate and reticularis cells are surrounded by elements of smooth endoplasmic reticulum, which are very closely associated with mitochondria. About 10–25 days after hypophysectomy, most mitochondria in fasciculate and reticularis cells show tubular crests, and numerous electron lucent lipid droplets are accumulated in these cells. These droplets occupy most parts of the cytoplasm and are sometimes in contact with one another or with mitochondria. The elements of smooth endoplasmic reticulum and Golgi apparatus are markedly reduced in number and size. About 10–90 days after the hypophysectomy, the degenerative cortical cells containing irregularly shaped nuclei, heterogeneously dense bodies in various sizes, and a large number of electron lucent lipid droplets are seen. The author speculates that most fasciculate and reticularis cells degenerate because of a disorder in the lipid metabolism, since ACTH release, which is necessary for synthesis of corticosteroid hormones from cholesterol, is absent in hypophysectomy. The fatty degeneration of the fasciculate and reticularis cells is considered to be a main cause for attenuation of the cortical tissue. Sometimes atrophied cortical cells without lipid droplets are also found 60–90 days after hypophysectomy.