Viruses and annulate lamellae in Friend erythroleukemia cells

Virus formation in a clone of murine undifferentiated Friend erythroleukemia cells was examined by electron microscopy. Budding C-type particles were present at the cell surface. The principal site of intracisternal particle production was in elements of rough-surfaced endoplasmic reticulum disposed about the periphery of stacks of annulate lamellae. Serial sections demonstrated that these virus-laden cisternae were in direct continuity with the annulate lamellae. In addition, intracisternal particles occurred in membranous honeycomb structures present in the cytoplasm of many cells. Viral elaboration also was associated with stacks of cisternae of endoplasmic reticulum that were devoid of ribosomes, but that were coated with an extensive and continuous layer of dense material. In some instances, the outer nuclear membrane was coated with the same dense substance. It appears that in Friend erythroleukemia cells, a very substantial portion of their cytomembranes is devoted to synthesis of intracisternal particles.     

Ultrastructure of the absorptive cells in the small intestine of the rat during starvation

Summary The ultrastructure of the absorptive cells in the duodenum, jejunum and ileum after 7, 14 and 21 days of starvation was investigated using rats aged from 12 to 18 months weighing about 500 g. In the basal cytoplasm of the absorptive cells (in the duodenum and ileum of 21-day-starved rats and the jejunum of 14- and 21-day-starved rats), the following changes were found: atrophied mitochondrion-like bodies, small vesicles, a short and sparse rough-surfaced endoplasmic reticulum and a lack of density in a portion of the cytoplasm. Moreover, many autolysosomes of various sizes and shapes were encountered in the basal cytoplasm; occasionally these elements accumulated and appeared to fuse to one another. In contrast, in the apical cytoplasm of absorptive cells in the intestine of starved rats, the ultrastructure was similar to that of control rats. It was considered that the apical cytoplasm of the absorptive cells in the starved rat intestine might be preserved as long as possible during starvation in order to absord nutrients when they become available again.     

Ultrastructural localization of thiamine pyrophosphatase activity in the intestinal epithelial cells of adult rats

Ultrastructural localization of thiamine pyrophosphatase (TPPase) activity was studied in the small and large intestinal epithelial cells of adult rats. In the duodenal and jejunal absorptive cells, TPPase activity was localized on the microvilli, on the lateral cell membrane, and on the lysosome-like bodies. In the ileal epithelial cells, TPPase activity was detected on the lysosome-like bodies, and weak phosphatase activity was observed on the microvilli and on the cisternae of the rough-surfaced endoplasmic reticulum (rER). In the principal cells of the caecum, the ascending colon, and the descending colon, the lysosome-like bodies and the cisternae of the rER showed TPPase activity. Almost all Golgi lamellae, vesicles and GERL elements (NOVIKOFF 1964) in the duodenal absorptive cells were strongly TPPase positive. The 1st to 3rd of the inner Golgi lamellae and the vesicles of the jejunal, ileal, and large intestinal epithelial cells usually showed positive TPPase activity, while the GERL elements were totally negative.     

Ultrastructural localization of acid phosphatase activity in the jejunal absorptive cells of fasted rats

Ultrastructural localization of acid phosphatase activity was investigated in the jejunal absorptive cells of 21 d fasted rats (about 500 g). The enzyme activity was localized on the membrane of microvilli, the lateral cell membrane, the Golgi complex, the lysosomes and the GERL of Novikoff (a part of the smooth surfaced endoplasmic reticulum located in close proximity to the Golgi saccules) of jejunal absorptive cells. Moreover, the lysosomes of various sizes and shapes with acid phosphatase activity was characteristically encountered in the infranuclear cytoplasm. The lysosomes appeared to be autolysosomes .     

Annulate lamellae in human malignant tumors: report of three cases

During the course of applying electron microscopy to diagnostic surgical pathological specimens, three malignant tumors (malignant melanoma, fibrous mesothelioma, lymphoblastic lymphoma) were found to contain annulate lamellae, distinctive intracytoplasmic organelles composed of membrane stacks interrupted by constrictions or pores. In one case both annuli and lamellae were present, a combination rarely described in human tissue and in animal models. In this material, the annuli of the annulate lamellae were structurally similar to nuclear pores. It is postulated that the abundant fibrils are probably related to the unusual configuration of the annulate lamellae. A morphologic relationship of the annulate lamellae to both the endoplasmic reticulum (cases 1 and 2) and the nuclear membrane (case 3) supports the theory that annulate lamellae may be related to both of these structures.     

Nuclear and cytoplasmic annulate lamellae in trophoblast giant cells of rat placenta

Single stranded profiles of nuclear annulate lamellae were identified in giant cells of rat trophoblast from the day when the chorioallantoic placenta first becomes vascularized, viz., day 12 post coitum, until the day before term, viz., day 22. Cytoplasmic annulate lamellae were observed only in giant cells from placentas at day 12. Occasionally cytoplasmic annulate lamellae were found in parallel array. Often the lamellar membranes were continuous with both granular and agranular membranes of endoplasmic reticulum; they closely resembled doubled outer nuclear membrane. Nuclear annulate lamellae resembled doubled inner nuclear membrane; and often the two were found in continuity. In addition, at later gestational ages (17 and 22 days), nuclear lamellae often were related anatomically to the variety of nuclear inclusions which characterize giant trophoblast cells during late pregnancy. A possible relationship of annulate lamellae to the synthesis of DNA, RNA and protein is considered.     

The primary human oocyte: Some observations on the fine structure of Balbiani's vitelline body and the origin of the annulate lamellae

The ultrastructural details of human oocytes from four primordial follicles and one early primary follicle are presented. A fifth primordial follicle is represented by a paraffin section stained by hematoxylin and eosin. The paranuclear Balbiani vitelline body, contsisting of a centrosome surrounded by endoplasmic reticulum, Golgi complexes, compound aggregates, annulate lamellae, and mitochondria is described. The annulate lamellae arise as an evagination from the outer leaflet of the nuclear envelope and interdigitate with folds of the endoplasmic reticulum which also is continuous with the outer leaflet of the nuclear envelope. Structural aspects of annulate lamellae are discussed in relationship to current ideas of nuclear membrane ultrastructure and to their possible role in nucleo-cytoplasmic transfer. A biographical note on the life of Edouard Gérard Balbiani is presented.     

The Coexistence of Ribosome-Lamella Complex and Annulate Lamellae in Chronic Lymphocytic Leukemia

An electron microscopic study of peripheral blood and lymph node from a patient with chronic lymphocytic leukemia revealed ribosome-lamella complexes in 11-16% of the peripheral lymphocytes and in 14% of the nodal lymphocytes. Annulate lamellae were seen in 2-8% of the peripheral lymphocytes and in 8% of the nodal lymphocytes. One peripheral blood specimen revealed coexistent annulate lamellae and ribosome-lamella complexes within the same cells in an additional 4% of the lymphocytes. The two structures were connected by continuous strands of apparent endoplasmic reticulum. Although annulate lamellae and ribosome-lamella complexes have each been described in chronic lymphocytic leukemia, albeit uncommonly, their unusual association with one another has not previously been documented and might offer a clue as to the origin and role of these interesting structures. The possible ramifications of these observations include relationship of the structures to endoplasmic reticulum and protein synthesis, to in vitro and possible clinical induction by drugs including “antitubulins”, and to induction in vitro by viral infection.     

FINE STRUCTURAL STUDY OF ANNULATE LAMELLAE COMPLEXES IN HUMAN TUMORS

Specific intracytoplasmic organelles, annulate lamellae and radial clsternae, have been studied In several human tumors. Annulate lamellae are observed In all cases of leiomyoma, leiomyosarcoma, rhabdomyosarcoma and malignant melanoma, whereas radial clsternae are only found in a case of leiomyosarcoma. Annulate lamellae are characterized by stacks of parallel arrayed long clsternae showing alternative arrangement of annuli and sacs. Some of these clsternae are connected directly with rough-surfaced endoplasmic reticulum and there is continuity with the lumen and membrane. Radial clsternae are mainly composed of two structures: numerous short clsternae, which are a variant of annulate lamellae, and numerous spherical particles derived from the clsternae. The clsternae are arranged parallel or radially around particles measuring up to 1100 A in diameter. These particles consisting' of an amorphous high electron dense material without distinct limiting membrane are organized in groups and vary In number. There is no evidence of a direct relationship between these structures and viral infection.     

Unusual configurations of endoplasmic reticulum in human pituitary adenomas

Electron microscopic study of 435 surgically-removed human pituitary adenomas and 43 nontumorous adenohypophyses revealed unusual configurations of endoplasmic reticulum in 102 adenohypophysial tumors (23%) and in 12 nontumorous adenohypophyses (28%). These configurations classified as paired reticulum, annulate lamellae and ribosome-lamellar complexes were noted in various adenomatous and nontumorous adenohypophysial cells, indicating that they could not be used as specific markers for pituitary adenomas or for a particular adenohypophysial cell type. Paired reticulum was a common finding, whereas annulate lamellae and ribosome-lamellar complexes were rarely encountered. Whether or not these endoplasmic reticulum configurations could be considered as normal constituents of adenohypophysial cells was difficult to assess, since nontumorous cells studied were from patients who had various diseases and who had been treated with different hormones. The presence of endoplasmic reticulum configurations was neither related to age, sex of the patients nor degree of differentiation or endocrine activity of pituitary adenomas.     

The Coexistence of Ribosome-Lamella Complex and Annulate Lamellae in Chronic Lymphocytic Leukemia

An electron microscopic study of peripheral blood and lymph node from a patient with chronic lymphocytic leukemia revealed ribosome-lamella complexes in 11–16% of the peripheral lymphocytes and in 14% of the nodal lymphocytes. Annulate lamellae were seen in 2–8% of the peripheral lymphocytes and in 8% of the nodal lymphocytes. One peripheral blood specimen revealed coexistent annulate lamellae and ribosome-lamella complexes within the same cells in an additional 4% of the lymphocytes. The two structures were connected by continuous strands of apparent endoplasmic reticulum. Although annulate lamellae and ribosome-lamella complexes have each been described in chronic lymphocytic leukemia, albeit uncommonly, their unusual association with one another has not previously been documented and might offer a clue as to the origin and role of these interesting structures. The possible ramifications of these observations include relationship of the structures to endoplasmic reticulum and protein synthesis, to in vitro and possible clinical induction by drugs including “antitubulins”, and to induction in vitro by viral infection.     

Annulate lamellae in guinea pig Sertoli cells

Unilateral torsion of the spermatic cord was surgically induced in 24 Hartley strain guinea pigs. Groups of six animals were sacrificed 4, 8, 12, and 16 months after the surgery and testes were excised. Nine animals has severely damaged testes. Extensive study on the Sertoli cells of each of these nine animals was carried out after testicular tissues were processed for electron microscopy following the usual procedure. The majority of the affected seminiferous tubules contained a single layer of Sertoli cells without any differentiating germ cells. The Sertoli cells were vacuolated and contained highly lobulated nuclei. Each nucleus contained a nucleolus. The Sertoli cells contained an elaborate annulate lamellae system. The membranous portion of the annulate lamellae was associated with the rough and smooth endoplasmic reticulum. Close association of the annulate lamellae with the lysosomes and lipid droplets was found to be a consistent feature. Although it was not possible for us to predict the functional significance of these annulate lamellae, the present investigation clearly established for the first time, the presence of an annulate lamellae system in the Sertoli cell of a rodent species.     

Cadmium toxicity in the thyroid gland of pregnant rats

The toxic effects of cadmium on the thyroid gland of pregnant rats were studied with an electron microscope and an X-ray microanalyzer. Serum levels of thyroid hormones (T3 and T4) were also analyzed. Deterioration of the rough-surfaced endoplasmic reticulum occurred in the thyroid follicular epithelium on the fifth day of cadmium treatment. Large intracellular vacuoles, which arose from dilated cisternae of the rough-surfaced endoplasmic reticulum, were fused together, and marked swelling of the mitochondria was also noted. Thyroglobulin-secreting granules at the apical cytoplasm were decreased in number. By energy dispersive X-ray microanalysis, cadmium peaks were preferentially obtained from swollen mitochondria in the follicular epithelial cells. Serum levels of T3 and T4 were significantly decreased in cadmium-treated rats dams when compared to those of controls. In the present experiment, cycloheximide also caused degenerative changes in the rough-surfaced endoplasmic reticulum and the disappearance of thyroglobulin-secreting granules. Cycloheximide is a known inhibitor of protein synthesis on cytosolic ribosomes. These results indicated that accumulated cadmium in the mitochondria of thyroid follicular epithelial cells might disturb the oxidative phosphorylation of this organelle and the loss of energy supply possibly caused the inhibition of the synthesis and release of thyroid hormones.     

Adenocarcinoma of the fallopian tube

Summary A case of fallopian tube adenocarcinoma was studied by light and transmission electron microscopy. The neoplastic cells contained abundant mitochondria, moderate to large amounts of rough endoplasmic reticulum (RER) arranged in parallel arrays and often containing amorphous material, annulate lamellae, possible secretory vesicles, and glycogen. The presence of stacked RER and annulate lamellae together is unusual in papillary serous cystadenocarcinoma of the ovary, and has not been described in Fallopian tube adenocarcinoma. Golgi complexes were are. Small acini with projecting microvilli as well as junctional complexes were present, but cilia were not found. The electron microscopic findings suggest secretory activity, and are remarkably similar to those found in papillary serous cystadenocarcinomas of the ovary. The findings also support the hypothesis that ovarian serous tumors and adenocarcinomas of the Fallopian tube are derived from coelomic epithelium.     

Lipoprotein particles in the jejunal mucosa of postnatal developing rats

The jejunal mucosa of neonatal rats contains lipid particles of the same size, electron density and intracellular and extracellular distribution as particles identified by others in adult jejunum as lipoprotein particles. As in fetal jejunum obtained during the last three days of gestation, the jejunal mucosa of unsuckled newborn rats contains exclusively lipoprotein particles the size of very low density lipoproteins (VLDL). Within one day after initiation of suckling, there is in the mucosa a spectrum of lipoprotein particles ranging widely in size from those of VLDL particles to those of chylomicrons. These particles are seen in the endoplasmic reticulum and Golgi material of absorptive cells and within interepithelial cell spaces, the extracellular spaces of the lamina propria and lymphatic lacteals. VLDL-sized and chylomicron-sized particles are also seen, although in decreasing number, in the jejunal mucosa of 18-day-old suckling rats. However, in rats of comparable age, fasted for 48 or 72 hours, only VLDL-sized particles are seen in the jejunal mucosa. Ligation and transection of bile duct followed by fasting in rats of this age results in a marked decrease in the number of lipoprotein particles in absorptive cells. The results indicate that endogenous lipid contributes to the formation of VLDL particles whereas dietary triglycerides are needed for formation of chylomicrons.     

Lipoprotein particles in the jejunal mucosa of postnatal developing rats.

The jejunal mucosa of neonatal rats contains lipid particles of the same size, electron density and intracellular and extracellular distribution as particles identified by others in adult jejunum as lipoprotein particles. As in fetal jejunum obtained during the last three days of gestation, the jejunal mucosa of unsuckled newborn rats contains exclusively lipoprotein particles the size of very low density lipoproteins (VLDL). Within one day after initiation of suckling, there is in the mucosa a spectrum of lipoprotein particles ranging widely in size from those of VLDL particles to those of chylomicrons. These particles are seen in the endoplasmic reticulum and Golgi material of absorptive cells and within interepithelial cell spaces, the extracellular spaces of the lamina propria and lymphatic lacteals. VLDL-sized and chylomicron-sized particles are also seen, although in decreasing number, in the jejunal mucosa of 18-day-old suckling rats. However, in rats of comparable age, fasted for 48 or 72 hours, only VLDL-sized particles are seen in the jejunal mucosa. Ligation and transection of bile duct followed by fasting in rats of this age results in a marked decrease in the number of lipoprotein particles in absorptive cells. The results indicate that endogenous lipid contributes to the formation of VLDL particles whereas dietary triglycerides are needed for formation of chylomicrons.     

The Golgi apparatus in the acinar cells of the developing embryonic pancreas: I. Morphology and enzyme cytochemistry

The Golgi apparatus apparatus of pancreatic acinar cells of rat embryos was studied during development from day 13 through day 20 of gestation. The morphological and enzyme cytochemical patterns varied characteristically in the course of cell differentiation. (1) A pronounced system of “rigid lamellae” characterized the area near the trans face of the Golgi stacks in the protodifferentiated and early phases of the differentiated states; by contrast, “rigid lamellae” were sparse in the terminal period of gestation. (2) Reaction product of acid phosphatase labeled the “rigid lamellae” in the protodifferentiated state, was extended across the majority of the stacked cisternae in the early differentiated state, but was restricted to the trans side again in the later periods of cell differentiation. (3) The early phase of the differentiated state was characterized by the tight association of the endoplasmic reticulum and Golgi cisternae on the trans side; the close spatial relationship of the two compartments was lessened after production of secretion granules had started. The findings are in line with those of recent studies on the Golgi organization in some other types of cells in different functional states, and they present the embryonic pancreatic tissue as another model for demonstrating the high flexibility of the Golgi complex. In agreement with the patterns previously found in the absorptive cells of the small intestine, the present results show that the close associations of the endoplasmic reticulum and cisternae of the trans Golgi side predominate in the early stages of cell differentiation.     

Unusual features of the nuclear envelope in human spermatogenic cells

Different types of human germ cells show unusual features of the nuclear envelope. Spermatogonial nuclei demonstrate two kinds of modifications. The first one is a series of intranuclear flattened cisterns, parallel to each other and to the inner aspect of the nuclear envelope. The second one is a nuclear envelope protrusion into the cytoplasm occupied by a double membrane-limited vesicle. Pores are found on the membrane of the vesicle facing the interior of the nucleus. In spermatocytes the nuclear pores are concentrated over certain areas and completely absent from others. In the regions where they are absent a single cytoplasmic cistern of rough endoplasmic reticulum is closely apposed to the outer membrane of the nuclear envelope. Early modifications of the nuclear surface appear in spermatids before the attachment of the acrosomic vesicle and may indicate an active role of the nuclear envelope in the morphogenesis of the acrosome. In round spermatids nuclear pores are absent from the area which is first related to the Golgi and later covered by the acrosomal cap. Single or multiple layers of cytoplasmic annulate lamellae are closely associated with the nuclear envelope over the pore rich areas. Frequently there are intranuclear accumulations of dense material adjacent to the annulate lamellae-nuclear pore complex. The chromatoid body is usually present on the cytoplasmic side of this complex. In the elongating spermatids most annulate lamellae are free in the cytoplasm, often in relation with Golgi and chromatoid body remnants near the axial filament. Few stacks of annulate la-mellae are noted adjacent to the pore rich nuclear regions. It is suggested that the described modifications are related to an active nuclear-cytoplasmic interaction.     

Ultrastructural morphology of light beta cells in pancreatic tissue implanted into the anterior eye-chamber of rats

The ultrastructure of light beta cells in pancreatic tissue implanted into the anterior eye-chamber of homologous rats was described using electron microscopical method. The light beta cells contain more mitochondria and rough-surfaced endoplasmic reticulum cisternae and dilatated smooth endoplasmic reticuli than the dark beta cells. Their cytoplasm are poorly stained and contain secretory beta granules of similar morphological form to those in the dark cells. The secretory granules are also more heterogenous. The light cells seem to represent the early secretory stages of the beta cells.     

Ultrastructural analysis of hepatitis B surface antigen production in vitro

Recently several continuous cell lines (among these PLC/PRF/5 cells) producing hepatitis B surface antigen (HBsAg) were established from human hepatocellular carcinomas. The cultured cells provide the first opportunity to study HBsAg synthesis and secretion in vitro. HBsAg, but not HBcAg, was localized by the fluorescent antibody technique in the cytoplasm and on the surface of the cultured cells. Under the electron microscope, th PLC/PRF/5 cells displayed morphologic characteristics of both hepatocytes and hepatocellular carcinoma cells. However, 22-nm. spherical or filamentous HBsAg particles were not seen in the cells, although spherical HBsAg particles were observed in the supernatant culture media. Therefore, the indirect immunoperoxidase technique was used to demonstrate HBsAg at the ultrastructural level. Electron-dense reaction product was detected along the nuclear envelope, on rough-surfaced endoplasmic reticulum, and in cisternae of endoplasmic reticulum. These findings suggest that HBsAg is synthesized on rough-surfaced endoplasmic reticulum and transferred into endoplasmic cisternae for processing and secretion. This mode of HBsAg production is identical with that observed in hepatocytes of patients infected with hepatitis B virus. The absence of detectable intracellular HBsAg particles suggests that the cultured cells secrete the particles very rapidly or that they may have a defect in intracisternal packaging of HBsAg into particles.