Growth inhibition of murine melanoma by butyric acid and dimethylsulfoxide

Treatment of B16-F10 melanoma cells with dimethylsulfoxide (DMSO) or butyric acid (BA) inhibits cell growth and delays tumor appearance in syngeneic mice. Both agents induce morphological changes in these cells. Treatment of melanoma cells with DMSO results in a marked increase in tyrosinase activity and melanin content. BA, on the other hand, does not increase melanin content and decreases tyrosinase activity. The data show that there are marked differences in the effect of DMSO and BA on melanin biosynthesis, whereas both agents inhibit cell growth and cause a delay in tumor appearance. These findings indicate that decreased proliferation of melanoma cells and induction of melanin biosynthesis are not necessarily associated phenomena.     

Characterization of pancreatic-type tissue in the liver of rat induced by polychlorinated biphenyls

Pancreatic-type tissue induced in the livers of rats treated with polychlorinated biphenyls was characterized by transmission electron microscopy and high-resolution immunocytochemistry. The cells of pancreatic-type tissue were arranged as acini and in small groups. By electron microscopy the pancreatic-type tissue showed features very similar to normal pancreatic acinar tissue, such as well developed rough endoplasmic reticulum (RER), large numbers of mature zymogen granules, and a basally located nucleus. Protein A-gold imunocytochemical technique showed localization of amylase and trypsinogen over the zymogen granules and RER. These findings confirm that this tissue in the liver is morphologically and functionally identical to pancreatic acinar tissue.     

SOME ULTRASTRUCTURAL EFFECTS OF INSULIN, HYDROCORTISONE, AND PROLACTIN ON MAMMARY GLAND EXPLANTS

The effects of insulin, hydrocortisone, and prolactin on the morphology of explants from midpregnant mouse mammary glands were studied. Insulin promotes the formation of daughter cells within the alveolar epithelium which are ultrastructurally indistinguishable from the parent cells. The addition of hydrocortisone to the medium containing insulin brings the daughter cells to a new, intermediate level of ultrastructural development by effecting an extensive increase of the rough endoplasmic reticulum (RER) throughout the cytoplasm and an increase in the lateral paranuclear Golgi apparatus. When prolactin is added to the insulin-hydrocortisone medium, the daughter cells complete their ultrastructural differentiation. There is a translocation of the RER, Golgi apparatus, and nucleus and the appearance of secretory protein granules within the cytoplasm. There is excellent correlation between the ultrastructural appearance of the alveoli and their capacity to synthesize casein.     

Self-organization of endoplasmic reticulum aggregates in cells with overexpression of nucleoporin POM121

The nuclear pore complexes are complex protein structures located in the nuclear envelope, where they control the nuclear-cytoplasmic transport, and inside the stacks of endoplasmic reticulum cisternae, annulate lamellae. After overexpression of some nucleoporins, numerous granules are visible in the cytoplasm. According to the published data, these granules are the annulate lamellae. In the current paper, the structural organization of POM121-containing granules was analyzed using correlative light and electron microscopy. The ultrastructural study demonstrates that POM121-containing granules are not annulate lamellae but aggregates of endoplasmic reticulum membranes. Thus, overexpressed POM121 is not able to induce the annulate lamella formation. The mechanisms of self-organization of non-functional structures (such as the aggregates of endoplasmic reticulum membranes described here) and possible involvement of these mechanisms in the formation of cellular structures are discussed.     

Ultrastructural aspects of the goiter in cog/cog mice

Thyroids of congenitally goitrous (cog/cog) mice were studied with light and electron microscopy. The principal alteration in follicular cells was their largely overdistended rough endoplasmic reticulum (RER). Our findings resemble the ultrastructural features of human hypothyroid goiter caused by a thyroglobulin (TG) defect and thus support the previously suggested abnormalities of TG synthesis and/or processing in cog/cog mice.     

Morphometric analysis of hepatocytes from rats subjected to compound 48/80-induced anaphylactic shock

Morphometric and biochemical techniques were used to analyze hepatic glycogen, endoplasmic reticulum, and mitochondrial matrix granules in rats treated with compound 48/80 to induce an anaphylactic-like state of shock. Thirty minutes after insult there was a significant decrease in glycogen and mitochondrial matrix granules, an increase in rough endoplasmic reticulum (RER), and no change in smooth endoplasmic reticulum (SER). Less glycogen in experimental rats substantiated a previously described glycogenolytic response to compound 48/80. The decrease in matrix granules implies a loss and/or shift in intramitochondrial calcium as occurs in epinephrine-induced glycogenolysis in the rat. Since other glycogenolytic agents, e.g. glucagon, and starvation stimulate an increase in SER presumably from RER, the present morphological data suggest the increase in RER may precede proliferation of SER from RER.     

The ultrastructure of hemocytes inDactylopius confusus (Cockerell), and the role of granulocytes in the synthesis of cochineal dye

Summary The ultrastructural study of free circulating hemocytes in the adult cochineal scale,Dactylopius confusus (Cockerell), demonstrated five cell types: prohemocytes, typical granulocytes (T-granulocytes), oenocytoids, plasmatocytes, and granulocytes with modified sub-cellular structure to perform a special synthetic and secretory function, which we refer to as modified granulocytes (M-granulocytes). Prohemocytes showed undifferentiated sub-cellular structure of the basic stem cell type (i.e., high cytoplasmic density with numerous ribosomes, centrally located large nucleus with a distinct nucleolus, and poorly developed endoplasmic reticulum). The commonly observed typical granulocytes (T-granulocytes) had several smooth endoplasmic reticulum (SER) with dilated cisternae and many SER-derived membrane bounded granules of different sizes and electron density. Oenocytoids were identified by the presence of many crystals, RER-originated fine secretory granules, and an eccentric nucleus. Plasmatocytes were easily characterized by their variable shapes and irregular outline with pseudopodia-like cytoplasmic extensions, possession of an elongated lobed nucleus, multivesicular bodies, RER-derived membrane bounded, electron-dense, lysosomelike vacuoles, well-developed SER cisternae, and numerous pinocytic and SER-originated vesicles of different sizes along the peripheral region. M-granulocytes comprised the largest proportion of hemocytes in all samples observed. M-granulocytes were distinguished not only by the presence of membrane bounded granules of different sizes and electron density, but by the possession of large nuclei with distinct nucleoli, many mitochondria, and a highly developed network of rough endoplasmic reticulum (RER). M-granulocytes had abundant, rosette-shaped, RER-derived chains of fine secretory granules, which accumulated in the cytoplasm and vacuoles, and were ultimately deposited into the hemolymph by exocytosis. These fine granules gave a positive result with periodic acid-Schiff (PAS) test. Based on RER-synthesized fine secretory granules (M-granulocytes), their ultimate deposition into hemolymph, the red pigmentation of hemolymph, positive PAS histochemical test of these granules, and the high population of these hemocytes, no such cell type has been described in previous studies in insects. The sub-cellular structure of the granulocyte in this insect has been modified to perform a special synthetic and secretory function (i.e., possibly the synthesis of the red pigment found in hemolymph, which has been the source of commercially important cochineal dye).Abbreviations EM electron microscope - ER endoplasmic reticulum - LM light microscopy - MVB multivesicular body - PAS periodic acid-Schiff - RER rough endoplasmic reticulum - SER smooth endoplasmic reticulum - SG secretory granules - TEM transmission electron microscopy - UA uranyl acetate     

Tissue culture study on Mongolian gerbil's (Meriones unguiculatus) malignant melanoma

We tried to culture melanoma cells from a Mongolian gerbil's (Meriones unguiculatus) malignant tumor. In primary culture, most of cells have abundant melanin granules in their cytoplasm. Melanin granules decreased through 5 to 15 serial passages and disappeared after 15 th passage. The morphology of the cells varied from spindle to large polydendritic cells. Although typical melanin granules were not seen when the cells were stained by Masson-Fontana method, the cells were positive for DOPA reaction. Electron microscopically, most of the cells have well-developed Golgi apparatus and rough-surfaced endoplasmic reticulum with dilated cistern, and premelanosome-like granules were frequently observed in their cytoplasm.     

Increase of melanogenesis by retinoic acid: an ultrastructural and morphometric study

Melanin is a dark pigment protecting the skin against UV radiation in some organisms. Studies on invasion and metastasis using retinoic acid as inhibitor agent are well known, but its role in melanin production (melanogenesis), especially at ultrastructural level and using morphometry were not well studied. In the present study, we analyzed the effects of retinoic acid on the melanosomes in B16F10 melanoma cells. These organelles were identified and quantified using routine electron microscopy and the specific HMB45 antibody. Other approaches such as immunofluorescence, and flow cytometry were also used. Our results indicated that retinoic acid increased the melanogenesis process in B16F10 melanoma cells. Furthermore, this work also provided evidence that this substance interferes at the subcellular level altering the numerical density of melanosomes, as well as the relative volume of the nucleus and nucleolus. In addition, the cells displayed altered morphology and an increase in the percentage of the relative volume of melanosomes, mainly the stages II-III and IV, leading to melanin formation. Furthermore, a decrease in the cells number after retinoic acid treatment was also observed.     

An environmentally-relevant mixture of organochlorines and its vehicle control, dimethylsulfoxide, induce ultrastructural alterations in porcine oocytes

Organochlorine chemicals accumulate in the environment, particularly in the Arctic, and constitute potential developmental hazards to wildlife and human health. Although some of their harmful effects are recognized, their mechanisms of action within the target cells need to be better understood. This study was designed to test the hypothesis that an environmentally-relevant organochlorine mixture alters oocyte ultrastructure in the porcine model. Immature cumulus-oocyte complexes (COCs), partially cultured (18 hr) COCs without treatment or exposed to the organochlorine mixture or its vehicle (0.1% dimethysulfoxide; DMSO) during culture were processed for light and transmission electronic microscopy (TEM). The organochlorines induced major ultrastructural changes in the COCs: decreased density of the lipid droplets, increased smooth endoplasmic reticulum (SER) volume and increased interactions among SER, mitochondria, lipid droplets and vesicles. We suggest that these ultrastructural changes facilitate energy formation necessary to produce metabolizing enzymes. Other ultrastructural changes may reflect some degree of organochlorine toxicity: fewer gap junctions and decreased electron density of the cortical granules. Unexpectedly, the DMSO control treatment also induced similar ultrastructural changes, but to a lesser degree than the organochlorine mixture. This study is the first to demonstrate the effect of environmental contaminants on mammalian oocyte ultrastructure.     

Comparative ultrastructure of Clara cells in neonatal and older cattle

Calf lungs were fixed with glutaraldehyde and examined by scanning (SEM) and transmission (TEM) electron microscopy to compare the ultrastructure of Clara cells in terminal bronchioles of neonatal calves and older cattle. In the neonatal calf, SEM revealed numerous smooth-surfaced Clara cells protruding above a similar number of ciliated cells, whereas in older animals the surface of Clara cells was lobulated. Thin sections examined by TEM revealed numerous cuboidal to columnar Clara cells with indented nuclei and a pale cytoplasm filled with faintly granular glycogen in the neonatal calf. Some cells were characterized by apical dense and/or pale membrane-bound granules or secretory droplets. Many cells had an apical tubular network of cisternae that were partly smooth and partly decorated with ribosomes. Ultrastructural comparison of Clara cells in a 2-day-old calf with those of 14- and 19-day-old, 4- and 5. 5-month-old, and 3.5-year-old cattle revealed a striking reduction in the amount of glycogen per cell after 14 days. The number of cells with apical granules was small at all ages, and the density of the secretory granules varied greatly in different cells. A variable amount of smooth endoplasmic reticulum (SER) was present but was less prominent than cisternae of ribosomal endoplasmic reticulum (RER). In older cattle, the limited amount of SER compared to the RER and secretory granules suggests that bovine Clara cells are more likely to be secretory than detoxifying.     

Ultrastructural morphometric analysis of the uterine epithelium during early pregnancy in the sheep

Stereological techniques were used to quantify ultrastructural changes in the caruncular epithelium during the pre- (Day 13), peri- (Day 16) and post- (Days 19 and 22) attachment periods of placentation. Tissues from Day-13 non-pregnant ewes were used as controls. Uteri for stereological evaluation were perfused via the uterine artery with 3% glutaraldehyde and separated into proximal, middle and distal regions. Tissues from caruncular areas were processed for electron microscopy. Volume fractions (Vv) of nuclei, mitochondria, lipid and cytoplasmic granules were estimated by point-counting volumetry. Surface areas per unit tissue volume (Sv) of mitochondrial membranes and cristae, Golgi, plasmalemma, endoplasmic reticulum and nuclear membranes were estimated by line-intersection counting. The only significant difference between pregnant and non-pregnant uterine epithelium at Day 13, a time before attachment, was a lower Sv of smooth endoplasmic reticulum (SER) in tissue from pregnant ewes. This value returned to control (non-pregnant Day 13) levels at Day 16, and was again significantly reduced at Days 19 and 22. The Vv of lipid decreased significantly at Day 16 and remained at reduced levels thereafter. These changes may reflect the effects of conceptus products on lipid storage and mobilization. The Sv of rough endoplasmic reticulum (RER) significantly increased on Day 16 of gestation, and remained elevated on Day 19. These results may reflect increased synthesis of protein for export at these times. In general, several of the values measured which may be indicative of cellular metabolism were reduced at Day 22 of pregnancy, perhaps suggesting diminished metabolism by the uterine epithelium after attachment of the trophoblast.(ABSTRACT TRUNCATED AT 250 WORDS)     

Ultrastructural studies on the formation of yolk granules in the statoblast of a fresh-water bryozoan,Pectinatella gelatinosa

The formation of protein-carbohydrate yolk in the statoblast of a fresh-water bryozoan, Pectinatella gelatinosa, was studied by electron microscopy. Two types (I and II) of yolk cells were distinguished. The type I yolk cells are mononucleate and comprise a large majority of the yolk cells. The type II yolk cells are small in number; they become multinucleate by fusion of cells at an early stage of vitellogenesis. In both types of yolk cells, electron-dense granules (dense bodies) are formed in Golgi or condensing vacuoles, which are then called yolk granules. For the formation of yolk granules, the following processes are considered: 1. Yolk protein is synthesized in the rough-surfaced endoplasmic reticulum (RER) of the yolk cells. 2. The synthesized protein condenses in the cisternal space of the RER and is packaged into small oval swellings, which are then released from the RER as small vesicles (Golgi vesicles, 300-600 A in diameter). 3. The small vesicles fuse with one another to form condensing vacuoles, or with pre-existing growing yolk granules. 4. In the matrix of the condensing vacuoles or growing yolk granules, electron-dense fibers are fabricated and then arranged in a paracrystalline pattern to form the dense body. 5. After the dense body reaches its full size, excess membrane is removed and eventually the yolk granules come to mature. Toward the end of vitellogenesis of the yolk cells, the cytoplasmic organelles are ingested by autophagosomes derived from multivesicular bodies and disappear.     

Oogenesls in the terrestrial hermit crab,coenobita clypeatus (decapoda,anomura): II. Vitellogenesis

Oocytes from the land hermit crab, Coenobita clypeatus, in various stages of vitellogenesis were examined by light and electron microscopy. Early vitellogenic oocytes are characterized by accumulations of discrete vesicles of endoplasmic reticulum in the perinuclear cytoplasm. As oocytes develop, the endoplasmic reticulum becomes abundant, and numerous Golgi complexes are seen. There is a well developed Golgi-endoplasmic reticulum interaction. Within the confines of the reticulum are discrete intracisternal granules, which can be seen coalescing into electron-dense yolk bodies. Lipid accumulation is seen throughout the cytoplasm. Coincident with the burst of intra-oocytic metabolism are oolemma modifications and micropinocytosis, which provide ultrastructural evidence for extra-oocytic yolk production. The mature oocyte contains numerous yolk and lipid vesicles of varying electron density that comprise both intra- and extra-oocytic substrates.     

Electron microscopic immunocytochemistry of prolactin secreting cells (PRL-cells) and growth hormone secreting cells (GH-cells) in the anterior pituitary gland of partially hepatectomized rats

After 60% hepatectomy in rats, prolactin secreting cells (PRL-cells) and growth hormone secreting cells (GH-cells) of the anterior pituitary gland were distinctly identified by the protein A-gold procedure combined with electron microscopy. The animals were sacrificed by the decapitation at midnight at intervals of about 28, 76 and 124 h after the operation. The principal changes can be summarized as follows; (1) Hypertrophy of the Golgi complex, (2) Dilation of the rough endoplasmic reticulum (RER), (3) Increased numbers of secretory granules (markedly in GH-cells), and occurrence of granule extrusion or exocytosis, (4) Increased numbers of lysosomes, which were mostly seen at 124 h after the operation. These ultrastructural changes were remarkably observed in both PRL and GH-cells. Especially, the noticeable changes in PRL-cells after partial hepatectomy in the rat were new findings in this study. The above results suggest that hepatectomy induced synthesis and release of GH and PRL in cells from pars distalis.     

Differentiation of the golden hamster oviduct epithelial cells during postnatal development: an electron microscopic study

The ultrastructural changes in the process of differentiation of the epithelial cells of the golden hamster oviduct during postnatal development were investigated by means of electron microscopy. In the epithelium of the ampulla of the neonatal oviducts, no differentiated ciliated cells or secretory cells were identified. In these undifferentiated cells, free ribosomes were well developed, but rough endoplasmic reticulum (RER) and the Golgi apparatus were undeveloped. Cells undergoing ciliogenesis were first identified at 3.5 days after birth, and some ciliated cells appeared at 4.5 days. In the nonciliated cells, marked changes in the organelles were observed at this time. Subsequently, some nonciliated cells containing well-developed RER and Golgi apparatus were observed at 9.5 to 10.5 days after birth, and a few mature secretory cells were observed at 10.5 days. An increase in secretory granules occurred in the secretory cells at 12.5-15.5 days after birth. Many fully mature ciliated and secretory cells were observed at 15.5 days after birth. After 20.5 days after birth, the epithelium was identical with that of the adult golden hamster. Quantitative data indicated that the differentiation of ciliated cells began earlier and took place over a more extended period of time than did that of the secretory cells in the golden hamster oviduct during postnatal development.     

Olfactory mucosa ultrastructure in the short-tailed shrews,Blarina brevicauda and Blarina carolinensis

The olfactory mucosae of the northern short-tailed shrew, Blarina brevicauda, and the southern short-tailed shrew, Blarina carolinensis, were examined by light and electron microscopy. A well-developed olfactory epithelium was observed that included all of the cells necessary to provide for a sensitive olfactory system, suggesting that olfaction plays a major role in the behavior of these animals. There were no significant differences between the olfactory mucosae of these two species. The general features of the olfactory epithelium in these shrews were similar to those reported for several other macrosmatic mammals. A new type of supporting cell, called the light supporting cell, was observed in these shrews. The light supporting cell cytoplasm exhibited very little staining by light microscopy and had low electron density by transmission electron microscopy compared to that of the more common dark supporting cell. The light supporting cell had a convex apical surface with microvilli and lacked the large amounts of smooth endoplasmic reticulum (SER) typical of the apical cytoplasm of the dark supporting cell. In the lamina propria of the mucosa, the Bowman's glands consisted of two cell types, one with electron-lucent, alcian blue-positive granules, and the other with electron-dense PAS-positive granules. The cell with electron-lucent granules contained large amounts of SER and small clumps of rough ER. The cells with electron-dense granules had large amounts of RER and little SER.     

Observations on the role of smooth endoplasmic reticulum in glucocorticoid-induced hepatic glycogen deposition

We have studied by quantitative electron microscopy the relationship of specific hepatic cellular organelles to glycogen synthesis using dexamethasone, a potent synthetic glucocorticoid, to induce glycogen deposition in livers of adrenalectomized rats. Chemical and ultrastructural glycogen determinations revealed that the livers of fasted adrenalectomized rats had very low glycogen levels. Dexamethasone caused a time-related increase in hepatic glycogen which was the result of increases in the number of hepatocytes depositing glycogen and the amount of glycogen in each cell. The surface density of smooth endoplasmic reticulum (SER) in centrilobular and periportal hepatocytes also increased after treatment with dexamethasone; this increase preceded glycogen deposition. The newly deposited glycogen was spatially associated with membranes of SER, and a continued increase in SER surface density was correlated temporally with the increasing glycogen volume density. In both centrilobular and periportal hepatocytes, the suface density of rough endoplasmic reticulum (RER) initially decreased after dexamethasone administration but later increased. These data support the hypothesis that dexamethasone-induced enhancement of SER is functionally associated with the increase in glycogen, and that although the initial increase in SER may occur through transformation of RER to SER, later increases in SER require synthesis of new membranes.     

The Ultrastructure of Circulating Hemolymph Cells of the Marine Snail Cerithidea californica (Gastropoda: Prosobranchiata)

Two morphologically distinct blood cell-types, the granulocyte and hyalinocyte, are found in the hemolymph circulation of the marine prosobranch Cerithidea californica. Granulocytes, measuring 12.7 µ (9.0–15.0 µ) in diameter, possess well-defined ectoplasmic and endoplasmic regions of the cytoplasm, granules of moderate to heavy electron density, tubular rough endoplasmic reticulum (RER), short vesicles of smooth endoplasmic reticulum (SER), and a large cytoplasm to nucleus ratio. Two morphological variants of this cell-type are distinguished depending upon the presence or absence of dense granules or RER. Hyalinocytes, measuring 5.3 µ (4.0–8.0 µ) in diameter, are distinguished from gran ulocytes by possessing a smaller cytoplasm to nucleus ratio and a general lack of dense cytoplasmic granules and SER.