Fine structure of the corpus Luteum of the raccoon during pregnancy

Summary Ultrastructure of the granulosa lutein cells of the raccoon from throughout pregnancy has been described. The lutein cells often from epithelial cords which are separated by the connective tissues, capillaries and lymphatics. Based on the arrangements and modifications of the cytoplasmic organelles and inclusions, three types of lutein cells have been recognized. The type I lutein cells predominantly contain tubular, agranular endoplasmic reticulum, juxtanuclear Golgi complexes, a few round to rod-shaped mitochondria, some free ribosomes, and occasional lipid droplets. Occasionally the tubular cristae of mitochondria and tubular smooth endoplasmic reticulum appear contiguous. The type II cells contain abundant lace-like and/or stacked fenestrated endoplasmic reticulum cisternae that frequently form membranous whorls, some tubular, agranular endoplasmic reticulum, mitochondria, and lipid droplets. Mitochondria are usually small, but unusual large ones also occur. The small, rod-to round-shaped mitochondria usually have tubular cristae; but the large, oval, elongate, and cup shaped mitochondria possess tubular, lamellar, plate like, and whorl-like cristae. The plasma membranes of the cells are complexly elaborated and folded, especially when apposing each other. In favorable sections, strands of fenestrated cisternae appose the folds of the plasma membranes. In general, the amount of cytoplasmic organelles and inclusions vary greatly in the cells. The type III cells predominantly contain lipid droplets and sparse cytoplasmic organelles. The type I and II cells are found throughout pregnancy, but the type III cells are observed from mid gestation to term. The cytological features of type I and II cells suggest that they probably secrete most of the steroids, whereas the type III cells primarily store lipids.This research was supported by UPSHS grant AM-11376 and NIH contract 69-2136.     

Pollenkitt formation in Ilex paraguariensis A.St.Hil. (Aquifoliaceae)

 We investigated the cellular and organelle transformations during the formation of the pollenkitt in the secretory tapetum of Ilex paraguariensis. After the dissolution of the callose surrounding the young microspores, the elaioplasts of the tapetum produce many globules of saturated and unsaturated lipids (plastoglobules). Further on, oleosomes with unsaturated lipids, synthesized in the endoplasmic reticulum, accumulate in the tapetal cytoplasm. In contrast to other species, the plastoglobule production seems to precede the oleosome synthesis. The tapetum shows signs of cellular maturation in the late vacuolated microspore stage, when the plastoglobules and oleosomes coalesce and form the pollenkitt mass. In mature stages of the tapetum the pollenkitt is released into the loculus. Finally, it is mainly deposited on the exine, according to the entomophilous character of this species. The mode of pollenkitt formation in Ilex para guariensis and its transfer to the pollen surface is slightly dissimilar to other Angiosperms. Received October 24, 2002; accepted December 2, 2002 Published online: March 20, 2003     

Cytological differentiation in the uropygial gland.

Ultrastructural changes were studied in the cells undergoing secretory differentiation in zone I of the tubules of the uropygial gland of White Plymouth Rock chickens. A layer of basal cells and four secretory stages are recognized as the cells migrate from the periphery to the lumen of tubules and progressively elaborate a secretion product. Basal cells, containing rough endoplasmic reticulum and free ribosomes, rest on the basement membrane and are the source from which secretory cells arise. Dilated perinuclear cisternae and the proliferation of smooth endoplasmic reticulum in the form of vesicles, invaginated sacs and cusp-shaped cisternae indicate the onset of lipgenesis in stage I cells. The perinuclear cisternae are more dilated and the endoplasmic reticulum is composed on saccules and cisternae in stage II cells. Stage III cells are characterized by concentric lamellae of endoplasmic reticulum surrounding secretory droplets. Dilated cisternae of endoplasmic reticulum and secretory droplets both contain a reticular substance. The perinuclear cisternae of stage III cells have returned to normal dimensions. Large mature lucent secretory droplets, lined with electron-dense material, fill the cytoplasm ostage IV cells which degenerate and release their secretory product into the tubule lumen. Spherical membrane-bound compartments containing a mottled substance of moderate electron density occur in basal cells and all subsequent secretory stages. These mottled bodies are surrounded by saccules of endoplasmic reticulum in stage II cells and are intimately associated with secretory droplets in stage III cells, but there is no evidence that they give rise to secretory droplets and their role in secretory differentiation is unknown.     

Ultrastructure and histochemistry of Citrus limon (L.) stigma

Citrus limon has a wet stigma which can be divided in two zones: a glandular superficial one formed by papillae, and a non-glandular one formed by parenchymatic cells. The stigmatic exudate is produced by the papillae after the latter have reached their ultimate size. The papillae of the mature pistil are of varying size and composition. Both the unicellular and multicellular ones are present. The cells at the base of the papillae are rich in cytoplasm, whereas the tip cells are vacuolated. Histochemical analysis has shown that the exudate of Citrus is composed of lipids, polysaccharides, and proteins. Our results indicate that the lipidic component is produced and secreted first, followed by production and secretion of the polysaccharidic component. The lipidic component of the exudate is produced in the basal papillae cells and accumulates as droplets in dilated parts of the smooth endoplasmic reticulum (SER). Subsequently the lipid droplets are transported to the plasma membrane, and transferred by the latter into the cell walls. Then the exudate component is accumulated in the intercellular spaces and in the middle lamellar regions of the walls. Subsequently, the polysaccharidic component of the exudate is produced and secreted by the tip cells of the papillae.Abbreviations RER rough endoplasmic reticulum - SER smooth endoplasmic reticulum     

Cytochemical and fine structural analysis of oogenesis in the gastropod, Ilyanassa obsoleta

An analysis of differentiating oocytes of the gastropod, Ilyanassa obsoleta, has been made by techniques of light and electron microscopy. Early previtellogenic oocytes are limited by a smooth surfaced oolemma and are associated with each other by maculae adhaerentes. Previtellogenic oocytes are also distinguished by a large nucleus containing randomly dispersed aggregates of chromatin. Within the ooplasm are Golgi complexes, mitochondria and a few cisternae of the rough endoplasmic reticulum. When vitellogenesis begins, the oolemma becomes morphologically specialized by the formation of microvilli. One also notices an increase in the number of organelles and inclusions such as lipid droplets. During vitellogenesis there is a dilation of the saccules of the Golgi complexes and cisternae of the endoplasmic reticulum. Associated with the Golgi complexes are small protein-carbohydrate yolk precursors encompassed by a membrane. These increase in size by fusing with each other. The “mature” yolk body is a membrane-bounded structure with a central striated core and a granular periphery. At maturity a major portion of the ooplasmic constituents such as as mitochondria and lipid droplets occupy the animal region while the bulk of the population of yolk bodies are situated in the vegetal hemisphere. The follicle cells incompletely encompass the developing oocyte. In addition to the regularly occurring organelles, follicle cells are characterized by the presence of large quantities of rough endoplasmic reticulum and Golgi complexes whose saccules are filled with a dense substance. Associated with the Golgi saccules are secretory droplets of varied size. Amongst the differentiating oocytes and follicle cells are Leydig cells. These cells are characterized by a large vacuole containing glycogen. A possible function for the follicle and Leydig cells is discussed.     

Juvenile hormone-controlled vitellogenin synthesis in Locusta migratoria fat body: Cytological development

Cytological development in the fat body of adult female Locusta migratoria, related to vitellogenin synthesis, has been studied by light and electron microscopy. In the newly-emerged adult, the cells are filled with lipid droplets, which indent the nucleus, and with fields of glycogen, while ribosomes and endoplasmic reticulum are scarce. Correlated with the onset of vitellogenin synthesis, about day 8 of adult life, the nucleus enlarges, lipid droplets and glycogen decrease, and rough endoplasmic reticulum and Golgi complexes become the most abundant organelles. These changes reflect a conversion of the principal role of the fat body from nutrient storage to the synthesis and secretion of protein. They are prevented by allatectomy, and restored by subsequent treatment with the juvenile hormone analogue, ZR-515. Late in the first gonotrophic cycle, about day 20, dense bodies, vesicle-containing bodies and lysosomes are seen, indicating recycling of cellular materials. Five days after ZR-515 treatment, when protein synthesis has declined, the rough endoplasmic reticulum appears in arrays adjacent to lipid droplets, possibly awaiting reactivation. By the use of ferritin-labelled antivitellin immunoglobulin, vitellogenin has been localized intracellularly in the RER saccules and Golgi vesicles, and extracellularly in channels between the folded plasma membranes, showing sites of accumulation and secretion of this protein.     

A comparative study of the fine structure of coleorhiza and root cells during the early hours of germination of rye embryos

Summary The sequences of changes which occur in the fine structure of root and coleorhiza cells of the rye embryo during the first 9 hours of germination are described. Quiescent cells from both tissues characteristically possess no vacuole, a cytoplasm densely packed with ribosomes, lipid droplets largely confined to a peripheral position, a greatly reduced endomembrane system, mitochondria with few cristae and nuclei in which the heterochromatin is condensed. Following imbibition the structure of root cells is elaborated slowly. Microtubules and dictyosomes appear, followed by the development of mitochondrial cristae and endoplasmic reticulum and the dispersion of lipid droplets. A similar pattern of events occurs within coleorhiza cells but at a much enhanced rate. By 6 hours the endomembrane system is highly organized but by 9 hours it has largely disappeared. These observations are discussed in relation to the penetration of the root through the coleorhiza.     

Membrane deformation by protein coats

Protein coats deform lipid membranes into spherical buds, which undergo fission at the neck to become vesicles. To induce membrane curvature, protein coats use basic tools including amphipathic helices and concave protein surfaces, and take advantage of the bulk properties of cellular membranes, such as loose lipid packing in the endoplasmic reticulum and cis-Golgi and the abundance of anionic lipids in the cytosolic leaflet of the plasma membrane. Protein scaffolds, sensors of membrane curvature and finely tuned reactions such as GTP hydrolysis permit the spatial and temporal organization of these tools, making protein coats self-organized molecular machines. Because biological membranes generally adhere to a cytoskeleton, the functioning of protein coats is coupled to other large remodeling events at the membrane interface.     

麻疯树种子发育过程中脂类物质变化的显微和超微结构观察

用石蜡切片、半薄切片和超薄切片方法研究了麻疯树种子发育过程中脂类物质的变化。结果表明:脂类物质主要储存于胚乳当中,当种子发育成熟时胚乳中迅速积累了大量的油脂;种皮在发育过程中脂类物质含量较多,成熟时外种皮硬化,内种皮含有大量的油脂;种子成熟时胚中含有少量的脂类物质。细胞内脂类物质含量比较多时,内质网、线粒体、质体和高尔基体数量也会较多。种子完全成熟时进行采收加工是最为合适的。     

A new paradigm for membrane-organizing and -shaping scaffolds

The clathrin, COPI and COPII scaffolds are paradigm vesicle coats in membrane trafficking. Recent advances in our understanding of the caveolar coat have generated a new paradigm. It represents those membrane coats, where a considerable part of the protein component is lipid modified, and integrated into the cytosolic leaflet of the vesicle membrane by a hairpin-like hydrophobic structure. Such coat proteins are permanently associated with membranes, and form oligomers early after synthesis. These oligomers assemble into a coat that has high affinity for particular lipids, creating lipid microdomains within the membrane. The combined protein-lipid structure should be considered as the scaffold that entraps ligands, either through affinity with the protein or with the lipid component, and that has the ability to shape membranes. Besides scaffolds assembled by caveolins, scaffolds assembled by reticulons and PHB domain-containing proteins such as the reggie/flotillin proteins fit this paradigm.     

Structure-function relationships during preovulatory development of porcine follicles following equine chorionic gonadotropin stimulation.

Porcine follicular maturation begins by recruitment from a continually proliferating pool of small antral follicles; those receiving the appropriate stimulus differentiate rapidly through a series of structural and functional changes. Such ovarian activity can be induced in prepubertal gilts with a single injection of equine chorionic gonadotropin (eCG). Average follicular diameter in eCG treated females increased from approximately 2 mm before stimulation to 3.5 mm by 24 hr after injection, with subsequent growth to ovulatory size (8 or 9 mm) by 96 hr. Both theca and granulosa layers increased in thickness and complexity, and a prominent capillary bed evolved immediately outside the basement membrane separating the two layers. Cytoplasmic organelles associated with increased metabolic activity and steroidogenesis proliferated within the first 24 hr. Progressive changes included increasing amounts of lipid and rough and smooth endoplasmic reticulum, with the latter occurring in vesicular or lamellar forms and as lipid-associated whorls. Bizarre mitochondrial forms also appeared, often associated with lipids. The amount and proportion of rough and smooth endoplasmic reticulum shifted dramatically as follicles matured. By 24 hr, rough endoplasmic reticulum in thecal cells increased from 4.2 to 7% of cell volume, while the amount in granulosa cells increased from less than 3.5% to more than 10%; the quantity remained relatively constant in the theca but declined to prestimulation values in the granulosa layer. Rough endoplasmic reticulum predominated over smooth in the first 24 hr following stimulation but the proportions were then reversed, so that more than 10% of both layers was composed of smooth endoplasmic reticulum by the time ovulation was imminent. Some follicles had or were in the process of ovulating by 96 hr. Their walls were collapsed into prominent folds with the two cell types beginning to mix. Slight undulations and some regions of discontinuity were observed in basement membranes of large unovulated follicles at this time. In specimens collected at 96 hr poststimulation and processed for retention of lipid, lipid-like material was noticeable in the extracellular matrix surrounding cells that contained organelle configurations suggestive of steroidogenesis.     

Intracellular origin and secretion of milk fat globules

The cream or fat fraction of milk consists of fat droplets composed primarily of triacylglycerols that are surrounded by cellular membranes. In this review we discuss what is known about how these droplets are formed in and secreted by mammary epithelial cells during lactation. This secretion mechanism, which appears to be unique, is unlike the exocytotic mechanism used by other cell types to secrete lipids. Milk fat globules originate as small, triacylglycerol-rich, droplets that are formed on or in endoplasmic reticulum membranes. These droplets are released from endoplasmic reticulum into the cytosol as microlipid droplets coated by proteins and polar lipids. Microlipid droplets can fuse with each other to form larger cytoplasmic lipid droplets. Droplets of all sizes appear to be unidirectionally transported to apical cell regions by as yet unknown mechanisms that may involve cytoskeletal elements. These lipid droplets appear to be secreted from the cell in which they were formed by being progressively enveloped in differentiated regions of apical plasma membrane. While plasma membrane envelopment appears to be the primary mechanism by which lipid droplets are released from the cell, a mechanism involving exocytosis of lipid droplets from cytoplasmic vacuoles also has been described. As discussed herein, while we have a general overview of the steps leading to the fat globules of milk, virtually nothing is known about the molecular mechanisms involved in milk fat globule formation, intracellular transit, and secretion.     

STRUCTURE-FUNCTION RELATIONSHIPS IN THE ADIPOSE CELL : I. Ultrastructure of the Isolated Adipose Cell

A method is described for preparing isolated rat adipose cells for electron microscopy. The ultrastructure of such cells and their production of ¹⁴CO₂ from U-glucose-¹⁴C were studied simultaneously in the presence of insulin or epinephrine. Each adipose cell consists of a large lipid droplet surrounded by a thin rim of cytoplasm. In addition to typical subcellular organelles, a variety of small lipid droplets and an extensive system of membranes characterize the cell's cytoplasm. A fenestrated envelope surrounds the large, central lipid droplet. Similar envelopes surround cytoplasmic lipid droplets occurring individually or as aggregates of very small, amorphous droplets. Groups of individual droplets of smaller size also occur without envelopes. The system of membranes consists of invaginations of the cell membrane, vesicles possibly of pinocytic origin, simple and vesiculated vacuoles, vesicles deeper in the cytoplasm, flattened and vesicular smooth surfaced endoplasmic reticulum, and Golgi complexes. Neither insulin nor epinephrine produced detectable ultrastructural alterations even when cells were incubated under optimal conditions for the stimulation of ¹⁴CO₂ evolution. Structural responses of the isolated adipose cell to hormones, if such occur, must, therefore, be dynamic rather than qualitative in nature; the extensive system of smooth surfaced membranes is suggestive of compartmentalized transport and metabolism.     

Microlipid droplets in milk secreting mammary epithelial cells: evidence that they originate from endoplasmic reticulum and are precursors of milk lipid globules

Microlipid droplets, structures with diameters less than 0.5 micron, resemble larger cytoplasmic lipid droplets of milk secreting mammary epithelial cells in triacylglycerol core and surface coat composition. Previously, evidence was obtained that microlipid droplets fuse with and support growth of cytoplasmic lipid droplets, which are immediate precursors of large milk lipid globules. Morphological observations suggested that microlipid droplets may also be secreted directly from mammary epithelial cells, yielding the very small lipid globules of milk. The secretion mechanism, which involves envelopment of triacylglycerol droplets in apical plasma membrane, appeared to be the same for microlipid droplets as for larger cytoplasmic lipid droplets. Microlipid droplets appeared to originate by blebbing from cisternae of endoplasmic reticulum. By immunogold cytochemical localization and by immunological identification of electrophoretically separated polypeptides, endoplasmic reticulum, micro- and cytoplasmic lipid droplets, and milk lipid globules had a number of common polypeptides. Kinetics of incorporation of radiolabeled palmitate or glycerol into triacylglycerols and phospholipids were consistent with a possible endoplasmic reticulum origin of microlipid droplets and with the view that microlipid droplets may be secreted directly from the cell or may fuse with cytoplasmic lipid droplets.     

THE FINE STRUCTURE OF TESTICULAR INTERSTITIAL CELLS IN GUINEA PIGS

In guinea pig testes perfused with either glutaraldehyde or osmium tetroxide fixative, the cytoplasm of the interstitial cells contains an exceptionally abundant agranular endoplasmic reticulum. The reticulum in central regions of the cell is a network of interconnected tubules, but in extensive peripheral areas the reticulum is commonly organized into closely packed, flattened cisternae which are fenestrated. Occasional small patches of the granular reticulum occur in the cytoplasm and connect freely with the agranular reticulum. The mitochondria have a dense matrix and contain cristae and some tubules. The Golgi complex is disperse and shows no evidence of secretory material. The cytoplasm also contains lipid droplets. Lipofuscin pigment granules are probably polymorphic residual bodies and contain three components: (1) a dense material which at high magnification shows a 75-A periodicity; (2) a medium-sized lipid droplet; and (3) a cap-like structure. In glutaraldehyde-perfused testis the interstitial cell cytoplasm appears to have the same density from cell to cell, and the agranular reticulum is tubular or cisternal but not in the form of empty vesicles. Thus the "dark" and "light" cells and the vesicular agranular reticulum sometimes encountered in other fixations may be artifacts. Biochemical results from other laboratories, correlated with the present findings, indicate that the membranes of the agranular endoplasmic reticulum in guinea pig interstitial cells are the site of at least two enzymes of androgen biosynthesis, the 17-hydroxylase and the 17-desmolase.     

Lipid droplet formation on opposing sides of the endoplasmic reticulum

In animal cells, the primary repositories of esterified fatty acids and alcohols (neutral lipids) are lipid droplets that form on the lumenal and/or cytoplasmic side of the endoplasmic reticulum (ER) membrane. A monolayer of amphipathic lipids, intermeshed with key proteins, serves to solubilize neutral lipids as they are synthesized and desorbed. In specialized cells, mobilization of the lipid cargo for delivery to other tissues occurs by secretion of lipoproteins into the plasma compartment. Serum lipoprotein assembly requires an obligate structural protein anchor (apolipoprotein B) and a dedicated chaperone, microsomal triglyceride transfer protein. By contrast, lipid droplets that form on the cytoplasmic face of the ER lack an obligate protein scaffold or any required chaperone/lipid transfer protein. Mobilization of neutral lipids from the cytosol requires regulated hydrolysis followed by transfer of the products to different organelles or export from cells. Several proteins play a key role in controlling droplet number, stability, and catabolism; however, it is our premise that their formation initiates spontaneously, solely as a consequence of neutral lipid synthesis. This default pathway directs droplets into the cytoplasm where they accumulate in many lipid disorders.     

Oleosomes (Spherosomes) from Daucus carota suspension culture cells

Isolated oleosomes from Daucus carota L. cells are lipid droplets consisting mainly of triacylglycerols (>97%) and very little protein (1–2%). The boundary between the lipid phase and the cytosol, which is visible on electron micrographs, is not built up by a true phospholipid-containing unit or half unit membrane. Enzymatic activities of lipid metabolism were not found to be associated with oleosomes with the exception of very low (contaminating) acyl-CoA:1,2-diacylglycerol acyltransferase (EC 2.3.1.20) and relatively high acyl-CoA hydrolase (EC 3.1.2.2) activities. The triacylglycerols exhibited a half life time of about 70 h, which is below the generation time of the cells (80–90 h). The fatty acid pattern of triacylglycerols was very similar to that of polar cellular membrane lipids.     

Lipid inclusions in cells and their changes during the desiccation and reactivation of yeast organisms

The formation of lipid granules in Saccharomyces cerevisiae cells was shown to begin in the exponential phase of the culture growth and to be associated with the activity of the endoplasmic reticulum. Lipid granules can be formed (1) when the endoplasmic reticulum cisterns extend and (2) when the endoplasmic reticulum membranes separate relatively small regions of the cell cytoplasm. Yeast cells contain spherosomes which differ in their structure. Lipid granules merge together upon dehydration of yeast cells. The authors discuss possible participation of these granules in the reparation of damages of the intracellular membranes.     

Ultrastructure of endopolyploid antipodals inAconitum vulparia Rchb.

Summary The ultrastructure of the antipodals ofAconitum vulparia Rchb. was studied in mature embryo sacs. Antipodal cell wall thickness varies in different parts of the cells. The antipodals resemble transfer cells with distinctly marked wall ingrowths which are particularly well developed in the chalazal part and between the antipodals. A few plasmodesmata occur in the cell wall between the antipodals and the central cell. The cytoplasm is rich in ribosomes which occur free or bound to the membranes of the well developed endoplasmic reticulum. Only in the micropylar region of the cells are some larger vacuoles found. The antipodals contain numerous mitochondria, plastids and apparently active dictyosomes. Vesicles with electron dense contents, microbodies, multivesicular bodies as well as lipid droplets and small multiple concentric cisternae are also present in the cytoplasm. The giant endopolyploid nuclei have lobed outlines, especially at the chalazal side of the nuclei.Ultrastructural features, especially the occurrence of numerous free ribosomes and the development of extensive rough endoplasmic reticulum, suggest high metabolic activity in the growing and differentiating antipodals of this species.     

Structure and function of lipid droplet assembly complexes

Cells store lipids as a reservoir of metabolic energy and membrane component precursors in organelles called lipid droplets (LDs). LD formation occurs in the endoplasmic reticulum (ER) at LD assembly complexes (LDAC), consisting of an oligomeric core of seipin and accessory proteins. LDACs determine the sites of LD formation and are required for this process to occur normally. Seipin oligomers form a cage-like structure in the membrane that may serve to facilitate the phase transition of neutral lipids in the membrane to form an oil droplet within the LDAC. Modeling suggests that, as the LD grows, seipin anchors it to the ER bilayer and conformational shifts of seipin transmembrane segments open the LDAC dome toward the cytoplasm, enabling the emerging LD to egress from the ER.