Localization of apoVLDL-II,a major apoprotein in very low density lipoproteins,in the estrogen-treated cockerel liver by immunoelectron microscopy

Summary We have studied the sites of synthesis, assembly, and secretion of apoVLDL-II, a major apoprotein in very low density lipoproteins (VLDL), in the cockerel liver by immunoelectron microscopy. In the liver of the estrogen-treated cockerel, apoVLDL-II reaction products were localized in the cisternae of the nuclear envelope and the rough endoplasmic reticulum (RER). Such products were not observed in the smooth endoplasmic reticulum (SER). ApoVLDL-II reaction products were also located on the surface of lipid particles in the Golgi apparatus and secretory vesicles. Such lipid particles were not detected in the RER or SER. Some secretory vesicles containing the reaction products were seen during the process of fusion with the plasma membrane. Such fusion took place against the plasma membrane lining the space of Disse as well as the intercellular spaces. Reaction products also occurred in the sinusoids. These observations are compatible with the following sequence of events in the synthesis, assembly and secretion of apoproteins in VLDL in the cockerel liver: ApoVLDL-II is synthesized on bound ribosomes attached to the nuclear envelope and RER, and is discharged into their cisternae. The protein is probably transported to the Golgi apparatus where the assembly of this protein and its lipid components probably takes place. Secretory vesicles derived from the Golgi apparatus carry the VLDL particles to the plasma membrane where secretion of these particles takes place by exocytosis, and the VLDL are discharged into the sinusoid via both the space of Disse and intercellular spaces.This work was supported by Grants 78-1102 from the American Heart Association, and HL-16512 from the NIH     

Virus-neuron interactions in the mouse brain infected with Japanese encephalitis virus

The virus-host interactions between Japanese encephalitis (JE) virus and mouse brain neurons were analyzed by electron microscopy. JE virus replicated exclusively in the rough endoplasmic reticulum (RER) of neurons. In the early phase of infection, the perikaryon of infected neurons had relatively normal-looking lamellar RER whose cisternae showed focal dilations containing progeny virions and characteristic endoplasmic reticulum (ER) vesicles. The reticular RER, consisted of rows of ribosomes surrounding irregular-shaped, membrane-unbounded cisternae and resembled that observed in JE-virus-infected PC12 cells, were also seen adjacent to the lamellar RER. The appearance of the reticular RER indicated that RER morphogenesis occurred in infected neurons in association with the viral replication. The fine network of Golgi apparatus was extensively obliterated by fragmentation and dissolution of the Golgi membranes and their replacement by the electron-lucent material. As the infection progressed, the lamellar RER was increasingly replaced by the hypertrophic RER which had diffusely dilated cisternae containing multiple progeny virions and ER vesicles. The Golgi apparatus, at this stage, was seen as coarse, localized Golgi complexes near the hypertrophic RER. In the later phase of infection, RER of infected neurons showed a degenerative change, with the cystically dilated cisternae being filled with ER vesicles and virions. Small, localized Golgi complexes frequently showed vesiculation, vacuolation, and dispersion. The present study, therefore, indicated that during the viral replication the normal lamellar RER which synthesized neuronal secretory and membrane proteins was replaced by the hypertrophic RER which synthesized the viral proteins. The hypertrophic RER eventually degenerated into cystic RER whose cisternae were filled with viral products. The constant degenerative change which occurred in the Golgi apparatus during the viral replication suggested that some of the viral proteins transported from RER to the Golgi apparatus were harmful to the Golgi apparatus and that increasing damage to the Golgi apparatus during the viral replication played the principal role in the pathogenesis of JE-virus-infected neurons in the central nervous system.     

A RADIOAUTOGRAPHIC STUDY OF GLYCERIDE SYNTHESIS IN VIVO DURING INTESTINAL ABSORPTION OF FATS AND LABELED GLUCOSE

Radioautography was used to detect the synthesis of labeled glycerides in intestinal absorptive cells following injections of fatty chyme and glucose-6-H³ into ligated segments of upper jejunum of fasting rats. Absorption intervals ranged from 2 to 20 min. Labeling is evident throughout the cells in as short a time as 2 min. Most grains are present over droplets of absorbed fat beginning with those in the endoplasmic reticulum immediately subjacent to the terminal web. With longer absorption periods, frequent grains are present over accumulations of fat droplets in the Golgi cisternae and intercellular spaces. A similar pattern of grains is seen following absorption of either linoleic acid or safflower oil. By comparison, considerably less label is present in the cells when the fat is extracted with alcohol prior to radioautographic procedures, or when labeled glucose alone is absorbed. A significant incorporation of glucose label into newly synthesized glycerides is indicated and confirmed by scintillation counts on saponified lipid extracts. The grain distribution implies an involvement of the extreme apical endoplasmic reticulum in this synthesis.     

Cytochemical Localization of Pectinase Activity in Pollen Mother Cells of Tobacco During Meiotic ProphaseⅠand Its Relation to the Formation of Secondary Plasmodesmata and Cytoplasmic Channels

Pectinase activity was localized at the ultrastructural level in pollen mother cells of tobacco (Nicotiana tabacum L.) during meiotic prophaseⅠto elucidate its role in the biogenesis of secondary plasmodesma (sPD) and cytoplasmic channel (CC). At the leptotene stage the enzyme was mainly present in the cisternae of smooth endoplasmic reticulum (SER) and their derived vesicles, but absent in the Golgi body and Golgi vesicles. Later at the zygotene stage, when sPDs and CCs were actively formed, strong pectinase activity was observed not only in the SER cisternae and their derived vesicles but also in the cell wall, especially in the vicinity of or within both simple and branched plasmodesmata, notably along the middle lamellae, which also characterized the sites of CCs being formed. The presence of exocytotic vesicles containing reaction products suggests that pectinase shares the same excretive pathway as that used by cellulase for its delivery into the wall, i.e. in active form via smooth endoplasmic reticulum (ER) and its derived vesicles by exocytosis. In combination with cellulase, pectinase also promotes the secondary formation of plasmodesmata and CCs by specifically digesting the pectin in middle lamella.     

Localization of calmodulin in rat cerebellum by immunoelectron microscopy

Calmodulin, a multifunctional Ca(++)-binding protein, is present in all eucaryotic cells. We have investigated the distribution of this protein in the rat cerebellum by immunoelectron microscopy using a Fab-peroxidase conjugate technique. In Purkinje and granular cell bodies, calmodulin reaction product was found localized both on free ribosomes and on those attached to rough endoplasmic reticulum (RER) and the nuclear envelope. No calmoduline was observed in the cisternae of RER or the Golgi apparactus. Calmodulin did not appear to be concentrated in the soluble fraction of the cell under the conditions used. Rather, peroxidase reaction product could be seen associated with membranes of the Golgi apparatus the smooth endoplasmic reticulum (SER), and the plasma membrane of both cell bodies and neuronal processes. In the neuronal dendrites, calmodulin appeared to be concentrated on membranes of the SER, small vesicles, and mitochondria. Also, granular calmodulin was observed in the amorphous material. In the synaptic junction, a large amount of calmodulin was seen attached to the inner surface of the postsynaptic membrane, whereas very little was observed in the presynaptic membrane or vesicles. These observations suggest that calmodulin is synthesized on ribosomes and discharged into the cytosol, and that it then becomes associated with a variety of intracellular membranes. Calmodulin also seems to be transported via neuronal processes to the postsynaptic membrane. Calmodulin localization at the postsynaptic membrane suggests that this protein may mediate calcium effects at the synaptic junction and, thus, may play a role in the regulation of neurotransmission.     

Affinity cytochemical differentiation of glycoconjugates of small intestinal absorptive cells using Pisum sativum and Lens culinaris lectins

We studied the subcellular localization of glycoconjugates recognized by the garden pea and lentil lectins (Pisum sativum, PSA; Lens culinaris, LCA) in mature absorptive cells of duodenum and jejunum of fasted rats. PSA and LCA are mannose-, glucose-, and N-acetyl-glucosamine-recognizing lectins that bind with high affinity to fucosylated core regions of N-glycosidically linked glycans. The binding reactions were cytochemically demonstrated in a pre-embedment incubation system using peroxidase-labeled lectins. Both pea and lentil lectins bound with constituents of nuclear envelope and endoplasmic reticulum, cisternae of the Golgi apparatus, several Golgi-associated vesicles, lysosomes, and portions of the plasma membrane. PSA and LCA label was non-homogeneous in the endoplasmic reticulum; in the Golgi apparatus the reactions were most intense in the cis and medial cisternae of the stacks. For inhibition of the intense reactions apparent in the Golgi apparatus, in lysosomes, and at the plasma membrane, considerably higher concentrations of competitive sugars were necessary than for abolition of the endoplasmic reticulum label. This indicates that endoplasmic reticulum glycoconjugates bind at low affinities with pea and lentil lectins, and that high-affinity PSA/LCA-binding glycoconjugates, which may correspond to corefucosylated N-linked glycans, predominate in cis and medial Golgi cisternae, lysosomes, and at the plasma membrane.     

Immunolocalization of the oligosaccharide trimming enzyme glucosidase II

We used immunoelectron microscopy to localize glucosidase II in pig hepatocytes. The enzyme trims the two inner alpha 1,3-linked glucoses from N-linked oligosaccharide precursor chains of glycoproteins. Immunoreactive enzyme was concentrated in rough (RER) and smooth (SER) endoplasmic reticulum but not detectable in Golgi apparatus cisternae. Transitional elements of RER and smooth membraned structures close to Golgi apparatus cisternae contained labeling for glucosidase II. Specific labeling was also found in autophagosomes. These results indicate strongly that glucosidase II acts on glycoproteins before their transport to, and processing in Golgi apparatus cisternae, and suggest that an important transitional region for glucosidase II exists between RER and Golgi apparatus cisternae. Degradation in autophagolysosomes could form a normal catabolic pathway for glucosidase II.     

An electron microscopic study of hemoglobin synthesis in the marine annelid,Amphitrite ornata (Polychaeta: Terebellidae)

The heart-body of the marine worm Amphitrite, located within the supraesophageal dorsal vessel, is in the form of a cylinder the thin wall of which is deeply corrugated by luminal projections and folds along its entire length. It is anchored in places to the luminal surface of the dorsal vessel by an extracellular matrix containing collagen fibers. The luminal surfaces of both the heart-body and the dorsal vessel are covered by a basement membrane-like vascular lamina which in turn supports a discontinuous pseudoendothelium of littoral hemocytes. The cells of the heart-body constitute a pseudostratified, high columnar epithelium. They possess extensive rough endoplasmic reticulum (RER), a well developed Golgi zone, ferritin particles and granules, and several types of membrane-bound inclusions. Hemoglobin molecules identical to those in the circulation lie within cytoplasmic, membrane-bound vesicles. Analysis of our electron micrographs suggests the following sequence of hemoglobin production and secretion: Large quantities of a moderately dense flocculent material, probably globin, are synthesized in RER and move to the Golgi zone within partly rough- and partly smooth-surfaced transitional cisternae; small transport vesicles, formed from Golgi cisternae that have fused with transitional cisternae, convey the flocculent material from the convex to the concave face of the Golgi complex; a similar flocculent material and an amorphous, highly dense material are processed in the Golgi complex and are transferred to condensing vacuoles in which clearly identifiable hemoglobin molecules are first observed. Mature secretory vesicles containing only hemoglobin migrate to the cell periphery and discharge their contents by exocytosis. Hemoglobin molecules then cross the vascular lamina to reach the circulation.     

Nascent high density lipoproteins from liver perfusates of orotic acid-fed rats

Uniformly fatty livers from orotic acid-fed rats secreted almost no very low density lipoproteins (VLDL) but normal amounts of nascent high density lipoproteins (HDL) accumulated in perfusates. When lecithin:cholesterol acyltransferase (LCAT) was inhibited, nascent HDL were uniformly discoidal and lacked cholesteryl esters. Lipid and apoprotein compositions of nascent HDL from normal and fatty livers were similar whether LCAT was inhibited or not. Apolipoprotein B-100 was not detected in perfusates of uniformly fatty livers, but small amounts of apolipoprotein B-48 were present in HDL2 fractions. Nascent lipoproteins were not seen in Golgi compartments, but lipid-rich particles were clearly evident in endoplasmic reticulum cisternae adjacent to the cis face of the Golgi complex, suggesting that orotic acid blocks VLDL secretion by preventing translocation of nascent particles from the endoplasmic reticulum to the cis Golgi compartment. The accumulation of normal amounts of discoidal HDL in liver perfusates despite virtual absence of triglyceride-rich lipoproteins in Golgi secretory compartments, the space of Disse, and the perfusate is inconsistent with the concept that nascent HDL are exclusively a product of surface remnants cast off during lipolysis of chylomicrons and VLDL.     

The pineal gland of the gerbil,Meriones unguiculatus

Summary Electron microscopy was employed in a study of the pineal gland of the Mongolian gerbil (Meriones unguiculatus). It was determined that the gerbil pineal gland contains pinealocytes and glial cells with the pinealocytes being the predominant cell type. The pinealocytes contain numerous organelles traditionally considered as being either synthetic or secretory in function such as an extensive Golgi region, smooth (SER) and rough (RER) endoplasmic reticulum, secretory vesicles and microtubules. Other cytoplasmic components are also present in the pinealocytes (synaptic ribbons, subsurface cisternae) for which no function has been assigned. Dense-cored vesicles are rare. Vacuolated pinealocytes are present and appear to be intimately associated with the formation of the pineal concertions. Evidence presented supports the proposal that the concretions form within the vacuoles. Once the concretions reach an enlarged state, the vacuolated pinealocytes break down and the concretions are thus extruded into the extracellular space where they apparently continue to increase in size. The morphology of the glial cells was interpreted as indicative of a high synthetic activity. The glial cells contain predominantly the rough variety of endoplasmic reticulum and form an expansion around the wide perivascular area.Supported by NSF grant PCM 77-05734     

Ultrastructure of the lateral organs in larvalArgas (Persicargas) arboreus (Ixodoidea: Argasidae)

The ultrastructure of lateral organs (LO) in the larval tickArgas (Persicargas) arboreus is described before and after feeding and up to the 1st day of moulting. Three pairs of LO are associated with three pedal nerves arising from the synganglion. In unfed ticks, each LO is ensheathed by a neural lamella and consists of 6–7 neuronal cell bodies; their cytoplasm is mostly occupied by cisternae of rough endoplasmic reticulm (RER). In fully engorged ticks, the enlarged neuronal cells contain vacuolar cisternae of smooth endoplasmic reticulum (SER), coated vesicles and mitochondria. Golgi bodies are involved in the formation of neurosecretory granules which dominate, with the SER vacuoles, the cell cytoplasm before moulting. The vacuoles, coated vesicles and neurosecretory granules are similar to those found in the vertebrate steroid-secreting cells. Condensing vacuoles may fuse with lysosome-like bodies to form larger ones; these are possibly responsible for the cell breakdown when secretory products are no longer required. Ultrastructural observations of LO suggest that they are neuroendocrine glands and that, in engorged larvae, they may secrete a hormone involved in the control of moulting.     

Apolipoprotein B100 exit from the endoplasmic reticulum (ER) is COPII-dependent,and its lipidation to very low density lipoprotein occurs post-ER

Hepatic apolipoprotein B100 (apoB100) associates with lipids to form dense lipoprotein particles in the endoplasmic reticulum (ER) and is further lipidated to very low density lipoproteins (VLDL). Because the VLDL diameter can exceed 200 nm, classical ER-derived vesicles may be unable to accommodate VLDLs. Using hepatic membranes and cytosol to reconstitute ER budding, apoB100-containing vesicles sedimented distinct from those harboring more typical cargo but contained Sec23. Moreover, ER exit of apoB was inhibited by dominant-negative Sar1. Budding required Sar1 regardless of whether oleic acid (OA) was added to stimulate apoB lipidation; therefore, either large apoB100-lipoproteins reside in secretory vesicles, or full lipidation occurs post-ER. Using membranes from cells incubated in the presence or absence of OA, we determined that apoB100-lipoproteins in ER vesicles had not become lipidated to VLDLs. VLDL particles resided in the Golgi, but not the ER, after fractionation of OA-treated cells. We conclude that apoB100-lipoproteins exit the ER in COPII vesicles, but under conditions favorable for VLDL formation final lipid loading occurs post-ER.     

The mystery of the unstained Golgi complex cisternae

The Champy-Maillet OsKI reaction has been used upon Golgi complexes to show two kinds of staining. It stains material being processed as it passes along the secretory pathway of the rough endoplasmic reticulum (RER) and Golgi cisternae (GC) up to crystallization in secretory vesicles. It also stains separately the environment within parts of the GC. This GC staining may occur in all compartments (transition vesicles, saccules, condensing vacuoles), but it is characteristically missing from any one of them. The unstained cisternae may be explained if outer saccules are made from either stained or unstained transition vesicles, both of which occur. The presence of empty, unstained transition vesicles is dictated by the surface to volume ratios of microvesicles in relation to saccules. Most transition vesicles must return their membrane to the endoplasmic reticulum, but from time to time it is presumed that they fuse to make a saccule. Saccules, stained and unstained, then mature through the stack. OsKI reactions with tissues and test molecules suggest that in the RER and GC the stain detects labile--S . S--bridges before they lock the tertiary configuration of proteins.     

Distribution of microsomal triglyceride transfer protein within sub-endoplasmic reticulum regions in human hepatoma cells

Very low-density lipoprotein (VLDL) particles are formed in the endoplasmic reticulum (ER) through the association of lipids with apolipoprotein B (apoB). Microsomal triglyceride transfer protein (MTP), which transfers lipid molecules to nascent apoB, is essential for VLDL formation in ER. However, little is known of the distribution and interaction of MTP with apoB within ER. In this study, distribution patterns of apoB and MTP large subunit (lMTP) within ER were examined. Microsomes prepared from HuH-7 cells, a human hepatoma cell line, were further fractionated into rough ER (RER)-enriched subfractions (ER-I fraction) and smooth ER (SER)-enriched subfractions (ER-II fraction) by iodixanol density-gradient ultracentrifugation. ApoB was evenly distributed in the ER-I and the ER-II fractions, while 1.5 times more lMTP molecules were present in the ER-I fraction than in the ER-II fraction. lMTP and apoB were coprecipitated both in the ER-I and in the ER-II fractions by immunoprecipitation whenever anti-apoB or an anti-lMTP antibodies were used. ApoB-containing lipoprotein particles showed a lower density in the ER-II fraction than those in the ER-I fraction. From these results, it is suggested that MTP can function in both rough and smooth regions of ER in human hepatoma cells.     

Periodate-acid treatment of sections permits on-grid immunogold localization of pea seed vicilin in ER and Golgi

Summary The legume seed reserve protein vicilin has been localized in developing pea cotyledons by immunogold labelling on sections of glutaraldehyde-osmium-fixed, resin-embedded tissue. By treating sections with periodate and acid before antibody labelling, a 20-fold increase in specific antibody binding was observed. The densest label occurred over vacuolar contents, previously identified as the site of protein storage, and over electron-dense cisternae occasionally seen to be continuous with the vacuolar contents. Vicilin was also associated with the rough endoplasmic reticulum, the implied site of synthesis, and in electron-dense Golgi vesicles which, we suggest, are a vehicle by which newly synthesized protein is relocated into the vacuole.     

The G protein of vesicular stomatitis virus has free access into and egress from the smooth endoplasmic reticulum of UT-1 cells

We have investigated the role of the smooth endoplasmic reticulum (SER) of UT-1 cells in the biogenesis of the glycoprotein (G) of vesicular stomatitis virus (VSV). Using immunofluorescence microscopy, we observed the wild type G protein in the SER of infected cells. When these cells were infected with the mutant VSV strain ts045, the G protein was unable to reach the Golgi apparatus at 40 degrees C, but was able to exit the rough endoplasmic reticulum (RER) and accumulate in the SER. Ribophorin II, a RER marker, remained excluded from the SER during the viral infection, ruling out the possibility that the infection had destroyed the separate identities of these two organelles. Thus, the mechanism that results in the retention of this mutant glycoprotein in the ER at 39.9 degrees C does not limit its lateral mobility within the ER system. We have also localized GRP78/BiP to the SER of UT-1 cells indicating that other mutant proteins may also have access to this organelle. Upon incubation at 32 degrees C, the mutant G protein was able to leave the SER and move to the Golgi apparatus. To measure how rapidly this transfer occurs, we assayed the conversion of the G protein's N-linked oligosaccharides from endoglycosidase H-sensitive to endoglycosidase H-resistant forms. After a 5-min lag, transport of the G protein followed first order kinetics (t1/2 = 15 min). In contrast, no lag was seen in the transport of G protein that had accumulated in the RER of control UT-1 cells lacking extensive SER. In these cells, the transport of G protein also exhibited first order kinetics (t1/2 = 17 min). Possible implications of this lag are discussed.     

Synthesis,secretion and immunoelectron microscopic demonstration of apolipoprotein B-containing lipoprotein particles in the visceral rat yolk sac

Summary Electron microscopic investigations on the involvement of the fetal membranes of the rat (visceral yolk sac) in the lipid metabolism revealed the occurrence of lipoprotein-sized particles located in cisternal Golgi stacks, Golgi vesicles and secretory vesicles of the cells of the visceral yolk sac epithelium as well as in distended areas of the intercellular space between adjacent epithelial cells. Application of the protein A-gold technique with specific anti-apoB antiserum resulted in a specific location of immunogold both over the different compartments of the lipoprotein pathway (RER, Golgi complex, secretory vesicles) as well as over the distended intercellular spaces, thus confirming these particles to be lipoproteins in nature. Isolated visceral epithelial cells prepared by a tryptic digestion method exhibited some ultrastructural alterations, such as a loss of apical brush border, a change from columnar to spherical cell shape, a decrease in phagolysosomes, but an increase in autophagosomal structures after 6 h incubation at a vitality rate of at least 85%. Within this period the epithelial cells secreted measurable amounts of apoB-containing lipoproteins into the medium floating in the density classes d<1.006 g/ml, d=1.006–1.020 g/ml and d=1.020–1.064 g/ml. The production of the lipoproteins was partly inhibited by cycloheximide indicating the secretion of particles with preformed as well as newly synthesized apoB. Negative staining of the particles revealed an average diameter of 34 nm of VLDL, 31 nm of IDL and 24 nm of LDL. In summary, our studies demonstrate that in the feto-placental unit of the rat the fetal membranes are capable of synthesizing and secreting lipoproteins. The cells of the visceral yolk sac epithelium were shown to be the producers of apoB-containing particles.Abbreviations apo apolipoprotein - ER endoplasmic reticulum - IDL intermediate density-lipoprotein - LDL low density-lipoprotein - VLDL very low density-lipoprotein - PBS phosphate-buffered salt solution - RER rough endoplasmic reticulum - TEM transmission electron microscopy     

The trans Golgi face in rat small intestinal absorptive cells

In the small intestine cell differentiation from immature crypt cells to mature absorptive cells localized along the villi is accompanied by alterations in the organization of the trans Golgi side. In immature crypt cells the transmost Golgi cisterna is usually located closely adjacent to the other cisternae thus being a component of the stack. Concomitantly with cell differentiation the transmost cisterna of an increasing number of Golgi stacks sets off from the other cisternae being then located at various distances to the stacks. This transmost cisterna has, as in several other cell types, been interpreted as "GERL" (Golgi associated endoplasmic reticulum lysosomes [20, 28]) and thus, has been postulated to represent a specialized region of the endoplasmic reticulum. Our results, however, have shown that the cytochemical staining pattern which has been used as a basis for the differentiation of GERL from Golgi components is not present in crypt cells nor in mature absorptive cells of the proximal small intestine: identical cisternae react for thiamine pyrophosphatase, inosine diphosphatase, and acid phosphatase. Thiamine pyrophosphatase and inosine diphosphatase--enzymes characteristic for Golgi cisternae--are apparent over transmost cisternae defined as GERL, too, and in addition, acid phosphatase--postulated as GERL-marker--is demonstrable over stacked Golgi cisternae. This overlapping cytochemical reaction, as well as the alterations during cell differentiation, indicate that those structures which have been described as GERL are to be interpreted as Golgi components rather than as endoplasmic reticulum. On the other hand, endoplasmic reticulum is a constant component of the trans Golgi face in undifferentiated crypt-base cells and in maturing cells of the crypt-top region. From its localization closely adjacent to trans Golgi cisternae it may be termed "Golgi-associated endoplasmic reticulum"; however, these cisternae of endoplasmic reticulum are constantly devoid of acid phosphatase. No indications exist for continuities with the thiamine pyrophosphatase-, inosine diphosphatase-, and acid phosphatase-positive transmost Golgi cisternae, and for an engagement in production of lysosomes.     

Ultrastructure of pinealocytes of the cotton rat,Sigmodon hispidus

Summary Fine structural features of pinealocytes of cotton rats (Sigmodon hispidus) were examined. Golgi complexes, mitochondria, endoplasmic reticulum and polysomes are usual organelles seen in the perikaryonal cytoplasm of pinealocytes. Many non-granulated vesicles (40 to 80 nm in diameter) and a few granulated vesicles (about 100 nm in diameter) are associated with the Golgi cisternae. Occasionally, the cisternae contain granular materials. The perikaryonal cytoplasm of pinealocytes is characterized by the presence of inclusion bodies. These bodies are usually round in shape, not bounded by a limiting membrane and composed of fine granular or filamentous materials of high electron-opacity, which are similar in appearance to the substance seen in the nucleolonema. Pinealocyte processes, filled with abundant non-granulated vesicles and some granulated vesicles, are mainly found within the parenchyma and occasionally in perivascular spaces.Supported in part by NSF grant no. PCM 77-05734 and NIH grant no. HD-10202 (Morphology Core)     

Intestinal lipoprotein assembly

PURPOSE OF REVIEW: The assembly of intestinal lipoproteins is critical for the transport of fat and fat-soluble vitamins. In this review we propose a nomenclature for these lipoproteins and have summarized recent data about their intracellular assembly and factors that modulate their secretion. RECENT FINDINGS: The assembly and secretion of intestinal lipoproteins increases with the augmented synthesis of apoB, apoAIV and lipids. Chylomicron assembly begins with the formation of primordial, phospholipid-rich particles in the membrane, and their conversion to large chylomicrons occurs in the lumen of the smooth endoplasmic reticulum. Chylomicrons are transported from the endoplasmic reticulum via specialized vesicles to the Golgi for secretion. The identification of genetic mutations in chylomicron retention disease indicates that Sar1b may play a critical role in this process. In addition to chylomicron assembly, intestinal cells have been shown to transport dietary cholesterol via apoB-independent pathways, such as efflux. SUMMARY: Understanding the mechanisms involved in the intracellular transport of chylomicrons and chylomicron-independent secretion pathways are expected to be the next frontiers in the field of intestinal lipoprotein assembly and secretion.