Endocytosis in the uterine luminal and glandular epithelial cells of mice during early pregnancy

We have localized horseradish peroxidase (HRP) in the mouse uterus after intravenous administration on days 1 and 5 of pregnancy in an effort to understand how serum proteins reach the uterine lumen. Direct movement of HRP into uterine and glandular lumina was blocked by the epithelial tight junctions on both days. In luminal and glandular epithelial cells at both times, HRP was localized in endocytic vesicles along the basolateral membranes, multivesicular bodies (mvb), elongated dense bodies below the nucleus (bdb), and many small vesicles near the apical surface of the cells. The uptake of HRP was most extensive in the luminal epithelium on day 1: the number of tracer-containing apical vesicles and bdb was largest, and there were also clusters of vesicles containing the tracer above the nucleus. Acid phosphatase was localized on day 1 in mvb and bdb in both cell types, indicating that these structures are lysosomes. It appeared that HRP followed two pathways after basolateral endocytosis by the epithelial cells: it was transported to the apical region of the cells, where it was present in small vesicles that may release their contents into the uterine or glandular lumina, or it was transported to lysosomes. To investigate whether macromolecules may be transported from the uterine lumen to the stroma, we also studied endocytosis at the apical pole of luminal epithelial cells after intraluminal injection of HRP. There was no detectable uptake of HRP from the lumen on day 1, and no tracer was detected in the intercellular spaces or basement membrane region. On day 5, a large amount of HRP was taken up from the lumen into apical endocytic vesicles, mvb, and dense bodies, but tracer was not present in the Golgi apparatus, lateral intercellular spaces, or the basement membrane region at the times studied. These observations indicate that there was no transport of luminal macromolecules to the uterine stroma on day 1, while the possibility of transport on day 5 requires further study.     

Endocytosis in the epithelium of the mouse oviduct

We studied the pathway of serum protein transport into the lumen of the mouse oviduct by localizing several tracer proteins in the oviduct after intravenous injection on days 1, 5, and 11 of pregnancy. Fluorescent proteins were observed in the lamina propria and in vesicles in the lumenal epithelial cells mainly in the preampulla segment on days 5 and 11 of pregnancy. In the isthmus, there was much less fluorescence in the lamina propria and no fluorescent vesicles in lumenal epithelial cells. This is similar to previous observations on day 1 and indicates that the uptake of serum proteins into lumenal epithelial cells in the preampulla is not limited to the time when embryos are present in the oviductal lumen. Horseradish peroxidase (HRP) was present in the lamina propria of the preampulla on days 1 and 5, but direct tracer movement into the oviductal lumen was blocked by the epithelial junctional complexes. Within the epithelial cells, HRP was localized in endocytic vesicles along the basolateral membrane, multivesicular bodies (mvb), elongated dense bodies below the nucleus (bdb), and many small vesicles near the apical surface of the cells. Ferritin was also used as a tracer and was observed in the same locations as HRP. Acid phosphatase in the epithelial cells of the preampulla on day 1 was localized in mvb and bdb, indicating that these structures are lysosomes. It appeared that HRP and ferritin followed two pathways after basolateral endocytosis by the epithelial cells in the preampulla: 1) they were transported to apical vesicles that may release their contents into the oviductal lumen, or 2) they were transported to lysosomes.(ABSTRACT TRUNCATED AT 250 WORDS)     

The fine structure and histochemistry of the caecal epithelium of Calicotyle kröyeri (Monogenea: Monopisthocotylea)

The caecal epithelium of Calicotyle kröyeri consists of a single cell type which functions in the uptake and intracellular digestion of host epidermis and associated mucus. Each cell is columnar with a small basal nucleus and prominent nucleolus. Perinuclear cytoplasm contains narrow profiles of GER and mitochondria with numerous cristae. Golgi complexes are small and indistinct. Most of the cell is filled with vacuoles of heterogeneous content, the largest occupying the cell apex. There is in each cell an apical endocytotic complex comprising cell surface lamellae, apical vesicles and numerous tubular invaginations of the plasmalemma. The limiting membrane of all these components is structurally modified and bears a highly organized array of peg-like structures on its luminal surface. The complex is capable of ingesting particulate food material from the gut lumen for transfer, via vesicles, to the vacuoles for digestion. Most of the vacuoles represent the digestive elements of the cell and, histochemically, are reactive for protein, mucus and carboxylic esterases. Indigestible residues and lipid droplets accumulate in the large apical vacuole and are periodically released to the lumen by exocytosis. Small, undifferentiated caecal cells were occasionally observed in the epithelium, but their development has not been recorded.     

The digestive cells of the hepatopancreas in Aplysia depilans (Mollusca, Opisthobranchia): ultrastructural and cytochemical study

Digestive cells are the most abundant cell type in the digestive diverticula of Aplysia depilans. These are tall columnar or club shaped cells, covered with microvilli on their apical surface. A large number of endocytic vesicles containing electron-dense substances can be found in the apical zone, but the presence of many heterolysosomes of large diameter is the main feature of these cells. Glycogen particles and some lipid droplets were also observed. Peroxisomes with a circular or oval profile were common, but crystalline nucleoids were not detected in them, although a dense spot in the matrix was observed in a few cases. These organelles were strongly stained after cytochemical detection of catalase activity. The Golgi stacks are formed by 4 or 5 cisternae, with dilated zones containing electron dense material. Arylsulphatase activity was detected in the Golgi stacks and also in lysosomes. Cells almost entirely occupied by a very large vacuole containing a residual dense mass seem to be digestive cells in advanced stages of maturation. The observation of semithin and ultrathin sections indicates that these very large vacuoles are the result of a fusion among the smaller lysosomes. Some images suggest that the content of these large vacuoles is extruded into the lumen of the digestive diverticula.     

The ultrastructural investigation of the midgut in the quill mite Syringophilopsis fringilla (Acari,Trombidiformes: Syringophilidae)

The midgut of the females of Syringophilopsis fringilla (Fritsch) composed of anterior midgut and excretory organ (=posterior midgut) was investigated by means of light and transmission electron microscopy. The anterior midgut includes the ventriculus and two pairs of midgut caeca. These organs are lined by a similar epithelium except for the region adjacent to the coxal glands. Four cell subtypes were distinguished in the epithelium of the anterior midgut. All of them evidently represent physiological states of a single cell type. The digestive cells are most abundant. These cells are rich in rough endoplasmic reticulum and participate both in secretion and intracellular digestion. They form macropinocytotic vesicles in the apical region and a lot of secondary lysosomes in the central cytoplasm. After accumulating various residual bodies and spherites, the digestive cells transform into the excretory cells. The latter can be either extruded into the gut lumen or bud off their apical region and enter a new digestive cycle. The secretory cells were not found in all specimens examined. They are characterized by the presence of dense membrane-bounded granules, 2–4 μm in diameter, as well as by an extensive rough endoplasmic reticulum and Golgi bodies. The ventricular wall adjacent to the coxal glands demonstrates features of transporting epithelia. The cells are characterized by irregularly branched apical processes and a high concentration of mitochondria. The main function of the excretory organ (posterior midgut) is the elimination of nitrogenous waste. Formation of guanine-containing granules in the cytoplasm of the epithelial cells was shown to be associated with Golgi activity. The excretory granules are released into the gut lumen by means of eccrine or apocrine secretion. Evacuation of the fecal masses occurs periodically. Mitotic figures have been observed occasionally in the epithelial cells of the anterior midgut.     

Ultrastructural, histochemical and cytochemical characterization of intestinal epithelial cells in Aplysia depilans (Gastropoda, Opisthobranchia)

To improve the current knowledge about the digestive system in opisthobranchs, light and electron microscopy methods were used to characterize the epithelial cells in the mid‐intestine of Aplysia depilans. This epithelium is mainly formed by columnar cells intermingled with two types of secretory cells, named mucous cells and granular cells. Columnar cells bear microvilli on their apical surface and most of them are ciliated. Mitochondria, multivesicular bodies, lysosomes and lipid droplets are the main components of the cytoplasm in the region above the nucleus of these cells. Peroxisomes are mainly found in middle and basal regions, usually close to mitochondria. Mucous cells are filled with large secretory vesicles containing thin electron‐dense filaments surrounded by electron‐lucent material in which acidic mucopolysaccharides were detected. The basal region includes the nucleus, several Golgi stacks and many dilated rough endoplasmic reticulum cisternae containing tubular structures. The granular cells are characterized by very high amounts of flat rough endoplasmic reticulum cisternae and electron‐dense spherical secretory granules containing glycoproteins. Enteroendocrine cells containing small electron‐dense granules are occasionally present in the basal region of the epithelium. Intraepithelial nerve fibres are abundant and seem to establish contacts with secretory and enteroendocrine cells.     

Ultrastructure of langur monkey epididymidis prior to and following vasectomy and vasovasostomy.

The ultrastructural changes in langur monkey epididymis prior to and following vasectomy or vasovasostomy were studied. The epididymal epithelium of the intact langur monkey was found to consist mainly of principal cells and basal cells and frequently apical or mitochondria rich cells were found. Besides these cells intraepithelial lymphocytes were also a consistent feature of the epididymal epithelium. Principal cells identified by means of the tuft of the stereocilia on their apical surface, bear well developed Golgi bodies, endoplasmic reticulum, vesicles, vacuoles and multivesicular bodies. This suggests their active involvement in absorption and secretion. Basal cells present at the base of the lamina bear a few cellular organelles and strong interdigitations with the adjacent cells. Apical or mitochondria rich cells were characterized by clusters of mitochondria in the apical region of the cell and few microvilli on their apical surface. Lymphocytes with a large nucleus and a pale rim of cytoplasm were also found at the base of the epithelium. Secretory and absorptive functions of principal cells of the epididymal epithelium were found to be increased after vasectomy, as indicated by bulging of the apical portion of the principal cells and membrane bound structure in the lumen. An extensive increase in the number of lysosomes, vesicles and vacuoles was also observed. An increase in the number of macrophages with spermatozoa remnants in the lumen of epididymis suggests that the principal mechanism for spermatozoa disposal following vasectomy is intraluminal endocytosis by macrophages. Changes following vasectomy persisted in vasovasostomized animals even after 12 months of recanalization, which may contribute to the failure of functional reanastomosis.     

Choanocyte-like cells in the digestive system of the starfish Marthasterias glacialis (Echinodermata)

Two types of choanocyte-like cells have been found in the digestive tract of the starfish. Type I choanocytes are in the lining epithelium of all organs of the digestive system. These are narrow, columnar cells strongly anchored basally and expanded apically into a protuberance projecting into the lumen. A prominent flagellum surrounded by microvilli projects from the center of this protuberance. Apical cytoplasm contains numerous mitochondria, secondary lysosomes, and multivesicular bodies. A distinctive characteristic of these cells is a filament bundle that traverses the length of the cell from its region of attachment on the rootlet of the flagellar basal body to its terminus on the basal plasma membrane. Between the attenuated basal ends of type I cells are the nerve fibers of an intraepithelial nerve plexus. Thickness of the plexus is correlated with the quantity of type I cells in the epithelium. Type II choanocytes are in the cuboidal coelomic epithelium that forms the outer layer of digestive tract organs. These cells are smaller than those of type I, and they have an apical collar surmounted by a ring of 13 microvilli. Within the collar is a cup-shaped depression with a central flagellum. Coated vesicles, secondary lysosomes, and phagocytic infoldings are observed in and near the collar cytoplasm. Filament bundles similar to those in type I choanocytes are also observed in coelomic epithelial cells that are sufficiently tall. Injection of peroxidase into the stomach and ferritin into the coelom results in phagocytic uptake of these macromolecules by type I and type II choanocytes, respectively.     

Ultrastructure and functional features of midgut of an adult water mite Teutonia cometes (Koch, 1837) (Hydrachnidia: Teutoniidae)

Shatrov, A. B. 2010. Ultrastructure and functional features of midgut of an adult water mite Teutonia cometes (Koch 1837) (Hydrachnidia: Teutoniidae). —Acta Zoologica (Stockholm) 91 : 222–232 The midgut of the adult water mite Teutonia cometes (Koch 1837) (Hydrachnidia: Teutoniidae) was investigated by means of transmission electron microscopy and on semi‐thin sections. The midgut is represented by a blind sac composed of the narrow ventriculus, two proventricular lateral diverticula and three pairs of postventricular caeca. A single‐layered epithelium consists of one type of endodermal digestive cells of quite different shape and size, which may form protrusions into the midgut lumen. The large nuclei are frequently lobed and contain one to three nucleoli. The apical cell membrane forms short scarce microvilli, between their bases the pinocytotic vesicles of unspecific macropinocytosis as well as the narrow pinocytotic canals are formed and immersed into the cell. The intracellular digestion of the food ingested into the midgut after extraintestinal digestion is predominant. The pinocytotic vesicles fuse with small clear vesicles of proposed Golgi origin to form secondary lysosomes. The digestive cells also contain small amounts of rough endoplasmic reticulum, variously structured heterolysosomes, residual materials in the form of both the small electron‐dense bodies and the large variously granulated substances, reserve nutritive materials such as lipid and glycogen, as well as clear vacuoles. Residual materials are obviously extruded from the cells into the gut lumen.     

STUDIES ON SMALL INTESTINAL CRYPT EPITHELIUM : I. The Fine Structure of the Crypt Epithelium of the Proximal Small Intestine of Fasting Humans

Small intestinal crypt epithelium obtained from normal fasting humans by peroral biopsy of the mucosa was studied with the electron microscope. Paneth cells were identified at the base of the crypts by their elaborate highly organized endoplasmic reticulum, large secretory granules, and small lysosome-like dense bodies within the cytoplasm. Undifferentiated cells were characterized by smaller cytoplasmic membrane-bounded granules which were presumed to be secretory in nature, a less elaborate endoplasmic reticulum, many unattached ribosomes and, in some cells, the presence of glycogen. Some undifferentiated cells at the base of the crypts contained lobulated nuclei and striking paranuclear accumulations of mitochondria. Membrane-bounded cytoplasmic fragments, probably originating from undifferentiated and Paneth cells, were frequently apparent within crypt lumina. Of the goblet cells, some were seen actively secreting mucus. In these, apical mucus appeared to exude into the crypt lumen between gaps in the microvilli. The membrane formerly surrounding the apical mucus appeared to fuse with and become part of the plasma membrane of the cell, suggesting a merocrine secretory mechanism. Enterochromaffin cells were identified by their location between the basal regions of other crypt cells and by their unique intracytoplasmic granules.     

FUNCTIONS OF COATED VESICLES DURING PROTEIN ABSORPTION IN THE RAT VAS DEFERENS

The role of coated vesicles during the absorption of horseradish peroxidase was investigated in the epithelium of the rat vas deferens by electron microscopy and cytochemistry. Peroxidase was introduced into the vas lumen in vivo. Tissue was excised at selected intervals, fixed in formaldehyde-glutaraldehyde, sectioned without freezing, incubated in Karnovsky's medium, postfixed in OsO₄, and processed for electron microscopy. Some controls and peroxidase-perfused specimens were incubated with TPP,¹ GP, and CMP. Attention was focused on the Golgi complex, apical multivesicular bodies, and two populations of coated vesicles; large (> 1000 A) ones concentrated in the apical cytoplasm and small (<750 A) ones found primarily in the Golgi region. 10 min after peroxidase injection, the tracer is found adhering to the surface plasmalemma, concentrated in bristle-coated invaginations, and within large coated vesicles. After 20–45 min, it is present in large smooth vesicles, apical multivesicular bodies, and dense bodies. Peroxidase is not seen in small coated vesicles at any interval. Counts of small coated vesicles reveal that during peroxidase absorption they first increase in number in the Golgi region and later, in the apical cytoplasm. In both control and peroxidase-perfused specimens incubated with TPP, reaction product is seen in several Golgi cisternae and in small coated vesicles in the Golgi region. With GP, reaction product is seen in one to two Golgi cisternae, multivesicular bodies, dense bodies, and small coated vesicles present in the Golgi region or near multivesicular bodies. The results demonstrate that (a) this epithelium functions in the absorption of protein from the duct lumen, (b) large coated vesicles serve as heterophagosomes to transport absorbed protein to lysosomes, and (c) some small coated vesicles serve as primary lysosomes to transport hydrolytic enzymes from the Golgi complex to multivesicular bodies.     

Histological and ultrastructural study of the digestive tract of rice field eel, Monopterus albus

The histological characteristics of the digestive tract and the ultrastructure of mucosal cells of the stomach and intestine of rice field eel, Monopterus albus, are described to provide a basis for future studies on its digestive physiology. The digestive tract of the rice field eel is a long and coiled tube composed of four layers: mucosa, lamina propria‐submucosa, muscularis and serosa. The pharynx and oesophagus mucosa is lined with a stratified epithelium. The stomach includes the cardiac and pyloric portions and the fundus. Many gastric pits are formed by invaginations of the mucosal layer and tubular gastric glands formed by the columnar cells in the fundus. The intestine is separated from the stomach by a loop valve and divided into a proximal portion and a distal portion. The proximal intestinal epithelium consists of columnar cells with microvilli towards the lumen and goblet cells. The enterocytes are joined at the apical surface by the junctional complex, including the evident desmosomas. Numerous lysosomes and some vesicles are evident in the upper cytoplasm of the cells, and a moderate amount of endoplasmic reticulum and lysosomes are scattered in the supranuclear cytoplasm. The epithelium becomes progressively thicker and the folds containing large numbers of goblet cells are fewer and shorter in the distal portion of the intestine. At the ultrastuctural level, the columnar cells of the tubular gastric glands have numerous clear vacuoles and channels. A moderate amount of pepsinogen granules are present in the stomach. The enterocytes of the intestinal mucosa display a moderate amount of endoplasmic reticulum and lysosomes, and long and regular microvilli.     

Uptake and fate of luminally administered horseradish peroxidase in resting and isoproterenol-stimulated rat parotid acinar cells

The formation and fate of apical endocytic vesicles in resting and isoproterenol-stimulated rat parotid acinar cells were studied using luminally administered horseradish peroxidase (HRP) to mark the vesicles. The tracer was taken up from the lumen by endocytosis in small, smooth-surfaces "c"- or ring-shaped vesicles. About 1 h after HRP administration the vesicles could be found adjacent to the Golgi apparatus. At later times HRP reaction product was localized in multivesicular bodies and lysosomes; in isoproterenol-stimulated cells it was also present in autophagic vacuoles. HRP reaction product was never localized in any structure associated with secretory granule formation. These results suggest that the apical endocytic vesicles play a role in membrane recovery, but that they are degraded and not reutilized directly in secretory granule formation. Additionally, it was found that when isoproterenol was injected before HRP administration, the apical junctional complexes became permeable to the tracer, allowing it to gain access to the lateral and basal intercellular spaces. This permeability may provide an additional route whereby substances in the extracellular fluid could reach the saliva.     

Comparative midgut ultrastructure of unfed larvae and adult mites of Platytrombidium fasciatum (C.L. Koch, 1836) and Camerotrombidium pexatum (C.L. Koch, 1837) (Acariformes: Microtrombidiidae)

The midgut of unfed larvae and adult mites of Platytrombidium fasciatum (C.L. Koch, 1836) and Camerotrombidium pexatum (C.L. Koch, 1937) (Acariformes: Microtrombidiidae) was investigated by electron microscopy. The sac-like midgut occupies the entire body volume, ends blindly and is not divided into functionally differentiated diverticula or caeca. The midgut walls are composed of one type of digestive cell that greatly varies in shape and size. In larvae, the lumen of the midgut is poorly recognizable and its epithelium is loosely organized, although yolk granules are already utilized. In adults, the midgut forms compartments as a result of deep folds of the midgut walls, and the lumen is well distinguished. The epithelium is composed of flat, prismatic or club-like cells, which may contain nutritional vacuoles and residual bodies in various proportions that depend on digestive stages. In both larvae and adult mites, parts of cells may detach from the epithelium and float within the lumen. The cells contain a system of tubules and vesicles of a trans-Golgi network, whereas the apical surface forms microvilli as well as pinocytotic pits and vesicles. Lysosome-like bodies, lipid inclusions and some amount of glycogen particles are also present in the digestive cells. Spherites (concretions) are not found to be a constant component of the digestive cells and in adult mites occur for the most parts in the midgut lumen.     

Morphology of the epithelium of the extratesticular rete testis, ductuli efferentes and ductus epididymidis of the adult male rabbit

The fine structure of the epithelium lining the extratesticular rete testis, ductuli efferentes and ductus epididymidis of the rabbit has been investigated. In the ductuli efferentes the epithelium is composed of two cell types, principal cells and ciliated cells. The latter type is distinguished from principal cells by the presence of cilia projecting into the lumen and the position of the nucleus in the apical half of the cell. Principal cells in this segment are characterized by micropinocytotic vesicles on the surface plasma membrane and a variety of small dense bodies scattered throughout the cytoplasm. In the ductus epididymidis basal cells replace ciliated cells as the second cell type, but differences between various segments of the epididymis are related to the fine structure of the principal cells. In the proximal caput epididymidis (Nicander's region 1) the principal cells are tall with long microvilli. They typically contain a small Golgi apparatus and a cluster of dense bodies adjacent to the nucleus. In the distal caput epididymidis (Nicander's regions 2-5) the apical cytoplasm of principal cells is filled with numerous micropinocytotic vesicles and large multivesicular bodies; these features are interpreted as signs of absorptive activity. The multivesicular bodies are absent from the cytoplasm of principal cells in the corpus epididymidis (Nicander's region 6) and, instead, numerous elements of smooth endoplasmic reticulum, a large Golgi apparatus, lipid droplets and dense bodies characterize principal cells in this segment. Towards the proximal cauda epididymidis (Nicander's region 7), the number of dense bodies (lysosomes) in the cytoplasm increases considerably. In the globose cauda (Nicander's region 8), the principal cells are reduced in height, and in addition to the features described in region 7, are characterized by a concentric array of rough endoplasmic reticulum in the basal cytoplasm. These observations are discussed in relation to the role of the epididymis in promoting the maturation and survival of spermatozoa.     

Ultrastructural changes in the oviductal epithelium of Merino ewes during the estrous cycle

Isthmic and ampullary oviductal epithelia sampled from Merino ewes at days -1, 1, 3, and 10 of the estrous cycle (estrus = day 0) were studied by scanning and transmission electron microscopy after fixation by vascular perfusion. Secretory cells, ciliated cells, and lymphocytelike basal cells were observed in both isthmic and ampullary epithelium at all stages of the estrous cycle studied and their ultrastructural features were analyzed. Synthesis of lamellated secretory granules occurred in the ampullary secretory cells during the follicular and early luteal phases, and their contents were released by exocytosis into the oviductal lumen during the luteal phase. Granule release was associated with nucleated apical protrusion of these cells into the oviductal lumen. No such secretory activity was displayed by isthmic secretory cells even though a few cells contained nonlamellated granules. Apocrine release of apical vesicles and accompanying cytoplasmic material from apical protrusions of ciliated cells occurred in the isthmus around estrus but not in the ampulla. This unexpected feature has not previously been reported in any other mammal. Dendritic basal cells were distinguished in the lower part of the epithelium by their heterochromatic nuclei, electron-lucent cytoplasm, and lack of attachment zones. No migration of basal cells was observed, and their ultrastructural features were similar in the ampulla and isthmus and at all stages of the estrous cycle examined. The function of these lymphocytelike cells in the epithelium is uncertain, but the presence of phagocytic bodies and lysosomes in 20% of them may indicate a phagocytic role.     

Internalization of cationized ferritin by isolated pancreatic acinar cells

The internalization of cationized ferritin (CF) was studied in isolated pancreatic acinar cells in vitro. Horseradish peroxidase (HRP) was used in conjunction with CF to compare internalization of soluble-phase and membrane-bound tracers. The mode of internalization of CF was dependent upon tracer concentration and origin of the plasma membrane (apical vs. lateral-basal). At the lower tracer concentrations (0.19 and 0.38 mg/ml), internalization from the apical cell surface occurred via small vesicles. The tracer then appeared in multivesicular bodies, in tubules, and in irregular membrane-bound structures. After 15 min, CF particles were seen in many small vesicles near the Golgi apparatus, but not in the Golgi saccules. In contrast, at the lateral-basal cell surface the CF particles tended to form clusters. These clusters were more pronounced at higher CF concentrations (0.76 and 1.5 mg/ml) and were associated with elongated cellular processes, which seemed to engulf CF accumulations in a phagocytic manner. Once internalized, CF was found primarily in large irregular structures which appeared to migrate slowly toward the nucleus, reaching a juxtanuclear position after approximately 30 min. CF was observed in lysosomes after 30-45 min and by 90 min most of the CF was confined to large vacuoles and to trimetaphosphatase-positive lysosomes. Similar routes were observed when cells were double-labeled with CF and HRP, where endocytic structures showed co-localization of both tracers. The results of this study indicate the importance of the Golgi region in the intracellular sorting of internalized apical membrane. Furthermore, this work confirms the presence of distinct endocytic pathways at the apical and lateral-basal cell surfaces.     

An electron microscopical study of absorbing cells in the posterior caput epididymidis of rabbits

Summary The columnar cells in regions 3 and 4 of the ductus epididymidis in rabbits display ultrastructural features characteristic of absorbing cells. The stereocilia show basal anastomoses and often a fibrillar core continuous with a fibrillar web in the apical cytoplasm. Numerous invaginations of the slightly downy apical cell membrane and many thick-walled apical vesicles and vacuoles contain an opaque substance similar to that seen in the lumen. The vacuoles often contain small vesicles or bodies, probably formed from the vacuolar wall by budding. Numerous bodies or vacuoles with moderately dense contents are seen in the Golgi area and in the supranuclear and intranuclear cytoplasm in region 3. In region 4 they are denser and mainly seen above the nucleus. A high acid phosphatase activity was demonstrated in most dense and some light bodies. India ink introduced by way of the rete testis was taken up from the lumen into apical invaginations, vesicles and vacuoles and slowly transferred to denser bodies below the Golgi apparatus.These observations are interpreted as evidence for a resorption of substances from the lumen by a pinocytotic process, and for their storage and perhaps digestion in the dense bodies, which appear to have a lysosomal character. The Golgi apparatus is large with many vesicles of two types and empty cisternae but few typical Golgi vacuoles. The partly granular endoplasmic reticulum is very well developed and has opaque contents. Microtubules run from the terminal bar region into the Golgi area. Thick-walled vesicles occur throughout the cytoplasm, sometimes in continuity with the cell membrane. The basal parts of the cell borders often interdigitate.Supported by a grant from the Swedish State Medical Research Council.     

Intracellular pathways of endocytosed transferrin and non-specific tracers in epithelial cells lining the rete testis of the rat

Summary The uptake and pathway of different markers and ligands for fluid-phase, adsorptive and receptor mediated endocytosis were analyzed in the epithelial cells lining the rete testis after their infusion into the lumen of these anastomotic channels. At 2 min after injection, diferric transferrin bound to colloidal gold was seen attached to the apical plasma membrane and to the membrane of endocytic coated and uncoated pits and vesicles. The injection of transferrin-gold in the presence of a 100-fold excess of unconjugated diferric transferrin revealed no binding or internalization of transferrin-gold. Similarly, apotransferrin-gold was neither bound to the apical plasma membrane nor internalized by these cells. These results thus indicate the presence of specific binding sites for diferric transferrin. At 5 min, internalized diferric transferrin-gold reached endosomes. At 15 and 30 min, the endosomes were still labeled but at these time intervals the transferrin-gold also appeared in tubular elements connected to or associated with these bodies or seen in close proximity to the apical plasma membrane. At 60 and 90 min, most of the transferrin-gold was no longer present in these organelles and was seen only exceptionally in secondary lysosomes. These results thus suggest that the tubular elements may be involved in the recycling of transferrin back to the lumen of the rete testis. The coinjection of transferrin-gold and the fluid-phase marker native ferritin revealed that both proteins were often internalized in the same endocytic pit and vesicle and shared the same endosome. However, unlike transferrin, native ferritin at the late time intervals appeared in dense multivesicular bodies and secondary lysosomes. When the adsorptive marker cationic ferritin and the fluid-phase marker albumin-gold were coinjected, again both proteins often shared the same endocytic pit and vesicle, endosome, pale and dense multivesicular body and secondary lysosomes. However, several endocytic vesicles labeled only with cationic ferritin appeared to bypass the endosomal and lysosomal compartments and to reach the lateral intercellular space and areas of the basement membrane. The rete epithelial cells, therefore, appear to be internalizing proteins and ligands by receptor-mediated and non-specific endocytosis which, after having shared the same endocytic vesicle and endosome, appear to be capable of being segregated and routed to different destinations.     

The digestive gland of Viviparus ater (Mollusca, Gastropoda, Prosobranchia): an ultrastructural and histochemical study

The digestive gland of Viviparus ater was studied using histochemical and ultrastructural methods. Only one cell type was observed in the tubule epithelium of the gland. The cells are involved in an endocytotic process mediated by clathrin-coated vesicles and in the intracellular digestion of food materials (thus they can be regarded as digestive cells). The different stages of digestion and exocytotic extrusion of residual bodies into the tubule lumen were shown by electron microscopy. Very few, small mucocytes are scattered among the digestive cells. Calcium concretions, glycogen-containing cells and endocrine cells are scattered in the area of connective tissue present among the digestive tubules.