Presence of pulmonary intravascular macrophages in the equine lung: Some structuro-functional properties

The pulmonary intravascular macrophages (PIMs) have been described in several species of animals. This study demonstrates for the first time that the equine lung has PIMs as resident phagocytes in its microvasculature. Their salient features such as globular surface coat, structures of the endocytic pathway, and related cell organelles closely resemble those of the calf, goat, and sheep. The exquisite organization of the coat globules in the form of a linear chain was structurally similar to the lipolytic lipase and the heparin-sensitive globular coat from PIMs of calf, goat, and sheep. Monastral blue (MB) when employed as a tracer to assess the phagocytic properties of equine PIMs induced similar modification of the globules of the coat into lipid droplets, reminiscent of neutral lipids. Lipid droplets (modified coat globules) were delivered into acid phosphatase-positive endosomes and lysosomes. Concurrently, the unaltered globules of the coat, probably internalized via fluid-phase constitutive pinocytoses, followed a different endocytic pathway. Large-scale platelet uptake by the PIMs was observed with thrombocytopenia in MB-treated ponies. The possible significance of hypothetical LDL-coat and the endocytic organelles as equivalents of synthetic apparatus of vasoactive lipids in the PIMs of horse needs to be assessed in future studies.© Willey-Liss, Inc.     

Presence of pulmonary intravascular macrophages in the equine lung: some structuro-functional properties.

The pulmonary intravascular macrophages (PIMs) have been described in several species of animals. This study demonstrates for the first time that the equine lung has PIMs as resident phagocytes in its microvasculature. Their salient features such as globular surface coat, structures of the endocytic pathway, and related cell organelles closely resemble those of the calf, goat, and sheep. The exquisite organization of the coat globules in the form of a linear chain was structurally similar to the lipolytic lipase and the heparin-sensitive globular coat from PIMs of calf, goat, and sheep. Monastral blue (MB) when employed as a tracer to assess the phagocytic properties of equine PIMs induced similar modification of the globules of the coat into lipid droplets, reminiscent of neutral lipids. Lipids droplets (modified coat globules) were delivered into acid phosphatase-positive endosomes and lysosomes. Concurrently, the unaltered globules of the coat, probably internalized via fluid-phase constitutive pinocytoses, followed a different endocytic pathway. Large-scale platelet uptake by the PIMs was observed with thrombocytopenia in MB-treated ponies. The possible significance of hypothetical LDL-coat and the endocytic organelles as equivalents of synthetic apparatus of vasoactive lipids in the PIMs of horse needs to be assessed in future studies.     

Morphology of pulmonary intravascular macrophages (PIMs) in ruminants: Ultrastructural and cytochemical behavior of dense surface coat

Recent studies have indicated that pulmonary intravascular macrophages (PIMs) are a resident cell population which in structure and function resemble mature macrophages of the mononuclear phagocyte system (MPS) in various domestic species, particularly the ruminants. The ultrastructural features of PIMs of the goat and calf lungs were studied by using vascular perfusion and direct airway instillation of fixatives. Staining with tannic acid as a component of paraformaldehyde-glutaraldehyde-based fixative revealed the presence of an electron-dense coat on the surface of the cell membrane of the PIMs. The surface coat disappeared after heparin infusion and after enzymatic digestion with lipolytic lipase, suggesting that the surface coat was predominantly lipoprotein in nature. The lipoprotein coat was organized in the form of a linear chain of spherical globules with a consistent periodicity created by the intervening translucent space between individual globules. The surface coat was separated from the outer-leaflet of the cell membrane by an empty space measuring 35–39 nm in width. PIMs possessed a significant number of coated pits and coated vesicles, the cell organelles of receptor-mediated endocytosis of lipoproteins. In concurrence with the coated pits and vesicles, microtubules, multivesicular bodies, and lipoprotein-positive vesicles were also observed. It is conceivable that PIMs are involved in lipid metabolism and are the major source of vasoactive substances, which significantly influence both the dynamics of pulmonary circulation and the surfactant turnover of the ruminant lung.     

In vivo monastral blue-induced lamellar-bodies in lysosomes of pulmonary intravascular macrophages (PIMs) of bovine lung: Implications of the surface coat

We previously reported that the pulmonary intravascular macrophages (PIMs) of sheep, goat, and calf lung contained a heparin and a lipolytic lipase sensitive surface coat by using tannic acid as a component of paraformaldehyde-glutaraldehyde-based fixative. The implication of this sensitivity was that the surface coat was predominantly comprised of lipoprotein-like substance. In this study we report that monastral blue (MB) used as a vascular tracer interacted with the coat globules and lost its original particulate appearance. Its precise localization in the PIMs was in combination with altered macromolecules of the surface coat in the form of lipid droplets, which conformed to the conventional view of neutral lipids. In contrast, pigment particles examined in their native state resembled metalic particles as electron-dense eliptical rods. The lipid droplets were subsequently internalized through endocytic route and found their access into the lysosomal compartments of PIMs at the electron microscopic level. Lamellar bodies (LLBs) arose from the lysosomal matrix after the entry of lipid droplets in the secondary lysosomes. Acid phosphatase activity was located in secondary lysosomes as well as in endosomes. These observations suggest that coat granules of the PIMs acted as a carrier of exogenous MB particles to deliver the complex to the lysosomal compartment where partial digestion lead to the formation of lamellar bodies. The implications of MB (cationic dye) as a vascular tracer for studying phagocytic index of PIMs in the light of their coat and the rapid development of LLBs are discussed. It is proposed that MB by initially combining with the surface coat provokes mobilization of intracellular lipid pools. In this way metabolism of vasoactive lipid in the PIMs is stimulated to influence the dynamics of pulmonary circulation in the calves.© Willey-Liss, Inc.     

Ultrastructural response of pulmonary intravascular macrophages to exogenous oestrogen in the bovine lung: translocation of the surface‐coat and enhanced cell membrane plasticity and angiogenesis

The pulmonary intravascular macrophages (PIMs) of domestic ungulates are recognised by their specific surface coat, consisting of linearly arranged globules along the external leaf of the plasma membrane. The coat is sensitive to in vitro digestion with lipolytic lipase (LPL), intravenous heparin and clinical exposure to halothane anaesthesia. The sensitivity to these experimental manipulations suggests that the globules of the coat are predominantly composed of lipoproteins (LDL). The present administration of oestradiol proprionate in castrated male calves potentiated the translocation of the surface coat into the endocytotic pathway of the PIMs. Concurrently with mobilisation of the coat, the plasma membrane was thrown into prominent arrays of lamellipodial extensions. The sprawling macrophages made extensive adhesive contacts with the lining endothelium of the capillaries. Consequently, the endothelial cells were highly attenuated and precariously maintained the integrity of the vascular wall. At some focal points, the vascular wall was penetrated by the filopodial processes of PIMs, which protruded into the perivascular space. Furthermore, there were signs of neovascularisation in the form of overt mitotic changes, sprouting and precursor capillary formation. It is conceivable that the evolving profile of angiogenesis is due to the vascular endothelial growth factor (VEGF) paracrine function of PIMs. Endothelial cell specificity has been considered an important advantage of VEGF for neovascularisation. It allows pleotrophic response of endothelial cells to proliferate and to assemble into endothelial tubes.     

Ultrastructural response of pulmonary intravascular macrophages to exogenous oestrogen in the bovine lung: translocation of the surface-coat and enhanced cell membrane plasticity and angiogenesis

The pulmonary intravascular macrophages (PIMs) of domestic ungulates are recognised by their specific surface coat, consisting of linearly arranged globules along the external leaf of the plasma membrane. The coat is sensitive to in vitro digestion with lipolytic lipase (LPL), intravenous heparin and clinical exposure to halothane anaesthesia. The sensitivity to these experimental manipulations suggests that the globules of the coat are predominantly composed of lipoproteins (LDL). The present administration of oestradiol proprionate in castrated male calves potentiated the translocation of the surface coat into the endocytotic pathway of the PIMs. Concurrently with mobilisation of the coat, the plasma membrane was thrown into prominent arrays of lamellipodial extensions. The sprawling macrophages made extensive adhesive contacts with the lining endothelium of the capillaries. Consequently, the endothelial cells were highly attenuated and precariously maintained the integrity of the vascular wall. At some focal points, the vascular wall was penetrated by the filopodial processes of PIMs, which protruded into the perivascular space. Furthermore, there were signs of neovascularisation in the form of overt mitotic changes, sprouting and precursor capillary formation. It is conceivable that the evolving profile of angiogenesis is due to the vascular endothelial growth factor (VEGF) paracrine function of PIMs. Endothelial cell specificity has been considered an important advantage of VEGF for neovascularisation. It allows pleotrophic response of endothelial cells to proliferate and to assemble into endothelial tubes.     

In situ heparin‐induced peroxisomal reticulum and biogenesis of peroxisomes in pulmonary intravascular macrophages (PIMs) of caprine lung: an ultrastructural and cytochemical study

Pulmonary intravascular macrophages (PIMs) contain a unique electron‐dense globular surface‐coat which is sensitive to heparin treatment, halothane anesthesia, and the digestive effect of lipolytic lipase (LPL), suggesting that the coat is predominantly composed of lipoproteins. In the present study, evidence is presented that heparin, when administered intravenously in goats, potentiated both the translocation of the surface‐coat into the vacuolar system and the expansion of the Golgi apparatus. Sequentially, these changes were followed by proliferation of peroxisomes in combination with peroxisomal reticulum (PR), a transient precursor of this organelle. The peroxisomes, as well as PR, reacted positively for catalase after aldehyde fixation and 3,3′‐diaminobenzidine (DAB) staining. In addition to their role as phagocytes, the ultrastructural and cytochemical detection of peroxisomes suggests a functional capacity of the PIMs, which may be adaptable to the circulating level of free fatty acids (FAAs). Anat Rec 266:69–80, 2002. © 2002 Wiley‐Liss, Inc.     

Structural responses of pulmonary intravascular macrophages in lentivirus-infected and/or recombinant ovine interferon-τ–treated lambs

Ovine lentivirus (OvLV), a retrovirus, infects and disseminates to various tissue organs via monocytes. The differentiation of infected monocytes into macrophages is a prerequisite for viral replication, and the presence of infected macrophages in tissue organs induces chronic immunopathology such as lymphoid interstitial pneumonia. The pulmonary intravascular macrophage (PIM) is a recently identified mononuclear phagocyte in domestic animal species, including sheep. Recombinant ovine interferon-tau (roIFN-τ), a type I IFN originally named as the ovine trophoblast protein, has potent antiviral activity against OvLV and human immunodeficiency virus and prevents the development of OvLV-associated lung pathology. We investigated and compared the structural features of PIMs in OvLV-infected and/or roIFN-τ–treated 1-month-old lambs using transmission electron microscopy. The PIMs' numerical counts were performed in toluidine blue–stained sections of Epoxy-embedded lung tissues. A reduction in the number of PIMs was observed with OvLV infection and/or roIFN-τ treatment of lambs as compared to the control group (P ≤ 0.05). The majority of the PIMs in OvLV-infected and/or roIFN-τ–treated groups were devoid of their surface coat. The PIMs of OvLV-infected lambs exhibited signs of biosynthetic activation such as expanded rough endoplasmic reticulum, prominent Golgi complexes, and accumulation of secretory vesicles. A few PIMs contained OvLV-like structures. In roIFN-τ–treated OvLV-infected lambs, the lymphocytes had ruffled plasma membranes and were in intimate contact with the PIMs, as is observed during cytotoxic cell-mediated killing of target cells. Most of the PIMs in roIFN-τ–treated OvLV-infected lambs appeared smaller in size. Ovine lentivirus and roIFN-τ, individually or in combination, alter the integrity of the surface coat of PIMs and cause their disappearance from the lungs. Ovine lentivirus infection induces morphological changes that correlate with cytotoxic cell behavior between lymphocytes and PIMs in roIFN-τ–treated or placebo-treated lambs. The loss of PIMs, probably infected with OvLV, either through direct killing by roIFN-τ or indirectly by roIFN-τ–activated cytotoxic T lymphocytes may represent different aspects of therapeutic actions of this cytokine. Anat. Rec. 251:472–485, 1998. © 1998 Wiley-Liss, Inc.     

Pulmonary intravascular macrophages (PIMs) and cationised ferritin-induced phagocytosis of platelets: A morphological study

The pulmonary intravascular macrophages of goat contain a unique electron-dense coat consisting of globular units arranged in a linear fashion along the cell surface and closely conforming to the contours of the cell. The coat is sensitive to heparin treatment and to the digestive effect of lipolytic lipase, suggesting that the coat is predominantly composed of lipoproteins. During the present study, the globules of the coat showed a high affinity for cationised ferritin within 2–5 min of intravenous injection. The adherence of cationised ferritin molecules to the globular units of the surface-coat was accompanied by platelet aggregation followed by their rapid phagocytosis by the pulmonary intravascular macrophages. Furthermore, large vesicular-structures were also seen in the phagosomes of the pulmonary intravascular macrophages, sometimes lying side by side with the engulfed platelets. It is suggested that cationised ferritin as a multivalent substance acted as a cross-link between negatively charged membrane components of the platelets and cationised globular units of the surface coat of the PIMs. The electrostatic attraction thus created between the two cells led to adhesion and subsequent phagocytosis of platelets as well as of anionic plasma proteins by the pulmonary intravascular macrophages.     

In situ heparin-induced peroxisomal reticulum and biogenesis of peroxisomes in pulmonary intravascular macrophages (PIMs) of caprine lung: an ultrastructural and cytochemical study.

Pulmonary intravascular macrophages (PIMs) contain a unique electron-dense globular surface-coat which is sensitive to heparin treatment, halothane anesthesia, and the digestive effect of lipolytic lipase (LPL), suggesting that the coat is predominantly composed of lipoproteins. In the present study, evidence is presented that heparin, when administered intravenously in goats, potentiated both the translocation of the surface-coat into the vacuolar system and the expansion of the Golgi apparatus. Sequentially, these changes were followed by proliferation of peroxisomes in combination with peroxisomal reticulum (PR), a transient precursor of this organelle. The peroxisomes, as well as PR, reacted positively for catalase after aldehyde fixation and 3,3'-diaminobenzidine (DAB) staining. In addition to their role as phagocytes, the ultrastructural and cytochemical detection of peroxisomes suggests a functional capacity of the PIMs, which may be adaptable to the circulating level of free fatty acids (FAAs).     

In situ response of lung circulating leukocytes to volatile and injectable anesthetics in ponies. Enigma of stimulated pulmonary intravascular macrophages and lymphoproliferation: a comparative ultrastructural, cytochemical and morphometric study

The lipoprotein-rich surface coat of pulmonary intravascular macrophages (PIMs) shows acute sensitivity to halothane, isoflurane, as well as to intravenous anesthetics in horse and ponies. A single and/or multiple exposures lead to mobilization of the coat into the endocytic channels of the PIMs. Coupled with this translocation, the Golgi apparatus undergoes expansion and is enriched with acid phosphatase during multiple exposures to halothane as compared to isoflurane and barbiturate-treated animals. In concert with these changes, the PIMs are shaped into dendritic form to interdigitate with large aggregates of activated platelets inside the pulmonary capillaries. These changes are considered relevant to pathogenetic sequelae of an immunological lung injury. The present study was designed to determine the acute response of the surface coat of the PIMs to a separate single exposure of halothane, isoflurane, and xylazine in ponies. Ultrastructural and cytochemical changes in the PIMs were observed in parallel with morphometric evaluation of lymphoproliferation. The surface coat was initially translocated into the vacuolar system of the PIMs followed by exclusive hyperplasia of the Golgi complex and its enrichment with acid phosphatase in the halothane group. Lymphoproliferation was more vigorous in halothane group, which included frequent presence of B cells, plasma cells, agranular T cell blasts, and plasmacytoid-like T cells classified at EM level. Given an active lymphoproliferation and the presence of immune cells in halothane group, it is possible that PIMs triggered an immunological hypersensitivity response, similar to that of Kupffer cells mediating an immunological hepatic injury as a result of biotransformation of halothane into bioactive intermediates.     

Ultrastructure studies of Proteocephalus longicollis (Cestoda, Proteocephalidea): transmission electron microscopy of scolex glands

The ultrastructure of two types of secretory glands in the scolex of preadults of Proteocephalus longicollis is described for the first time in the present report. The gland cells contain extensive cisternae of granular endoplasmic reticulum and Golgi complexes, which participate in the production of secretory globules. Type I scolex glands produce electron-dense globules of various size. The secretory globules enter the secretory canal, openings of which were not observed in the preadults. The secretory product of type I was found at the inner sucker surface and in the tegument of the sucker edges. In addition, electron-dense globules in adult worms are secreted via an eccrine mechanism. Type II scolex glands are characterized by secretory globules of lower electron density and occur mainly in preadults. The electron-lucent, membrane-bound secretory globules are transported via microtubule-lined ducts opening to the exterior at the tegumental surface. Secretory globules of type II are released by an eccrine process. Received: 7 February 2000 / Accepted: 2 March 2000     

Cytochemical characterization of glycoconjugates in the apocrine glands of the equine scrotal skin

Cytochemistry of glycoconjugates in the apocrine glands in the scrotal skin of the horse was studied using cytochemical methods for electron microscopy, particularly lectin cytochemistry. The secretory cells possessed a variable number of secretory vesicles, a well-developed Golgi apparatus, and abundant cisternae of the rough endoplasmic reticulum. Additionally, the basolateral plasma membrane formed numerous interdigitating folds. Glycoconjugates with vicinal diol groupings were present predominantly in the secretory vesicles, the Golgi apparatus, the surface coat of the plasma membrane, and the majority of the intracellular membranes. With lectin cytochemistry, the secretory vesicles of the glandular cells exhibited glycoproteins with different terminal sugars (alpha-D-mannose, beta-D-galactose beta-N-acetyl-D-glucosamine, and sialic acid). Several sugars were distinctly prominent in the surface coat of the plasma membrane of the secretory cells. The cytochemical properties of the complex glycoconjugates found are discussed in relation to the specific functions of the glandular secretions. These glands may have an important role in not only thermoregulation but protection of the scrotal skin, a specific body region.     

The sheep analogue of human CD59: purification and characterization of its complement inhibitory activity.

An inhibitor of the membrane attack complex of complement was isolated from the membranes of sheep erythrocytes. Fast protein liquid chromatography (FPLC) and affinity purification procedures for this sheep complement-inhibiting protein (SCIP) both yielded a pure protein with an apparent M(r) of 19,000 under reducing and non-reducing conditions. Incubation of the denatured protein with neuraminidase and Endo-F reduced the apparent M(r) to 18,000 and 15,000 respectively, while treatment with O-deglycosidase or phosphatidylinositol-specific phospholipase C (PIPLC) did not affect the apparent M(r). SCIP was detectable on erythrocytes and lymphocytes but not on platelets and could partially be removed by PIPLC treatment. Deglycosylation of the pure protein markedly reduced and PIPLC treatment abolished its activity. A monoclonal antibody (mAb) raised against sheep complement-inhibiting protein (SCIP) enhanced the susceptibility of sheep erythrocytes to lysis by homologous complement. SCIP inhibited complement after the stage of C5b-7 formation. Amino-terminal protein sequence was obtained and was shown to be similar to that of human CD59. All these features suggest that SCIP is the sheep equivalent of human CD59. Human CD59 has been reported to be species selective in that it inhibits complement from relatively few species. However, SCIP efficiently inhibited lysis of guinea-pig erythrocytes by complement from a wide range of species tested indicating that it is a potent and non-selective inhibitor of the membrane attack complex of complement (MAC).     

Pulmonary intravascular macrophages in domestic animal species: Review of structural and functional properties

In dogs, laboratory animals, and man, the clearance of bacteria and participates from blood occurs predominantly in hepatic Kupffer cells and splenic macrophages. In contrast, removal of blood-borne particulates in calves, sheep, goats, cats, and pigs occurs predominantly in pulmonary intravascular macrophages (PIMs). Review of recent studies indicates that PIMs are a resident cell population, junctionally adherent to the capillary endothelium of lungs and morphologically similar to hepatic Kupffer cells. PIMs are a pulmonary constituent of the mononuclear phagocyte system with respect to secretory, endocytic, and functional properties. Differentiated PIMs are rare in newborn pigs, and the majority of cells closely apposed to capillary endothelium consists of monocytes, which are occasionally in mitosis. In 7-day-old and older pigs, most cells apposed to capillary endothelium have characteristics of differentiated PIMs. This suggests a monocytic origin of PIMs in pigs. Perinatal colonization of lung capillaries by monocytes and their subsequent differentation into PIMs represent a component of postnatal lung development. Estimates of relative PIM numbers in ovine and porcine lung parenchyma suggest cell densities similar to that of rat hepatic Kupffer cells. Apart from phagocytic properties, PIMs participate in the removal and disintegration of aged and impaired blood cells. After phagocytic stimulation, isolated PIMs secrete oxygen radicals, which are essential for microbicidal function. Similarly, by secreting bioactive lipids, stimulated PIMs may contribute to regulation of pulmonary hemodynamics. After receiving minute amounts of bacterial endotoxin, pulmonary injury is pronounced in sheep, calves, pigs, and cats, but not in laboratory animals and dogs. This presumably is related to the secretion of bioactive lipids by PIMs.     

Plasma membrane biogenesis in the avian salt gland: A biochemical and quantitative electron microscopic autoradiographic study

The avian salt gland provides an ideal system for the study of plasma membrane (PM) biogenesis. Feeding ducklings 1% sodium chloride (salt stress) induces the secretory cells of the gland to synthesize large amounts of PM, which forms an extensive basolateral PM domain after 7–9 days of treatment. In the present study, the initial biosynthetic events following salt stress were investigated. In vivo studies using ³H-uridine indicated that increased rates of RNA synthesis could be detected by 2 hr after the beginning of salt stress and continued through at least 12 hr. Under in vitro conditions, increased rates of protein and glycoprotein synthesis (as monitored by ³H-leucine and ³H-fucose incorporation, respectively) were also detected after 2 hr and continued through 7–9 days. Increased levels of Na, K-ATPase, a specific secretory cell PM marker, were detected after 8 hr of treatment as monitored by specific activity and ³H-ouabain binding. Sodium dodecyl sulfate-polyacryl-amide slab gel electrophoresis coupled with fluorography indicated that both ³H-leucine and ³H-fucose were incorporated into partially purified preparations of Na, K-ATPase isolated after 12 hr. Light microscopic autoradiographic analysis of pulse-chase experiments indicated that in secretory cells of 12-hr salt-stressed glands, ³H-leucine- and ³H-fucose-labelled products reached the cell periphery by 1–2 hr after the initial pulse. The incorporation of both tritiated precursors was predominately associated with the secretory cells. Quantitative electron microscopic autoradiography indicated that ³H-leucine is initially taken up by elements of the rough endoplasmic reticulum (RER) and cytoplasm (5 min postpulse), subsequently transported to and concentrated within components of the Golgi apparatus (10 min of chase), and ultimately incorporated into all domains of the plasma membrane of secretory cells by 1–2 hr of chase. The data is consistent with a flow of newly synthesized membrane components from RER to Golgi to plasma membrane and is analogous to the pattern previously found for the synthesis and processing of PM proteins in a wide variety of cell types.     

In vivo interaction of pulmonary intravascular macrophages with activated platelets in microvessels of equine lung after multiple exposures to halothane, isoflurane, and thiamylal: a comparative ultrastructural and cytochemical study

The pulmonary intravascular macrophages (PIMs) of equines contain a unique electron-dense surface coat that is predominantly composed of lipoproteins. A single exposure of inhalatory halothane causes mobilization of the surface coat into the endocytotic system of the PIMs, followed by expansion of the Golgi apparatus and its enrichment with acid phosphatase. Simultaneously, the cells of the lymphocytic series show hyperplasia in the form of mitotic changes inside the microvascular compartment of the lung. Halothane is known to cause acute and chronic hepatotoxicity because of its biotransformation into trifluoroacytelated polypeptides. The present study was designed to examine the comparative effects of reexposures of inhalatory doses of halothane, isoflurane, and the intravenous barbiturate thiamylal sodium in ponies to evoke a stronger response in the PIMs after four exposures at increasing intervals of 1, 2, and 6 weeks. Ultrastructural and cytochemical evidence is presented that halothane induced translocation of the surface coat into the vacuolar system of the PIMs, followed by expansion of the Golgi apparatus and its enrichment with acid phosphatase. The cell membrane was thrown into extraordinary lamellipodial extensions, which enabled the PIMs to interact with platelets within the narrow confines of the pulmonary capillaries. The relationship between PIMs and platelets developed into large platelet aggregates. Isoflurane and thiamylal sodium did not affect the circulating platelets, although the surface coat was translocated into the endolysosomes in both situations. Although isoflurane is a lipid-soluble inhalant anesthetic similar to halothane, it is subject to very little biotransformation after use and in the present model demonstrates no immune response.     

Presence of cells combining features of two different cell types in the colonic crypts and pyloric glands of the mouse

A small number of epithelial cells which combine features of two cell types were observed in the descending colon and pyloric stomach of the mouse. In the descending colon, where the base of the crypts is mainly composed of poorly differentiated “vacuolated” cells, a few of these cells contain, besides the characteristic “vacuoles,” mucous globules identical to those in mucous cells or, less frequently, dense granules such as are found in entero-endocrine cells. Because there is evidence that the poorly differentiated vacuolated cells give rise to the other cells of the epithelium, those which also contain mucous globules or dense granules are likely to be differentiating into mucous cells or enteroendocrine cells respectively. In the pyloric stomach, where the glands are mainly composed of mucous cells, some of which are poorly differentiated, a few of the latter exhibit, besides the mucous globules, entero-endocrine type granules or features of caveolated cells. It is likely that the poorly differentiated mucous cells give rise to the other gland cells; and, therefore, those mucus-containing cells which also display dense granules or caveolated cell features are taken to be differentiating into entero-endocrine or caveolated cells respectively. Most of the cells containing two kinds of secretory materials are believed to be stem cells which initially contain a few vacuoles (colon) or mucous globules (pylorus) but are differentiating into a cell containing a different type of secretion. Rare observations of two kinds of secretory materials in a mature cell suggest that the transitional period may be prolonged, perhaps indefinitely.     

Ultrastructure and lectin cytochemistry of the cloacal pelvic glands in the male newt Triturus marmoratus marmoratus

The cloacal organ of Salamandridae species contains four glands: pelvic, dorsal, ventral, and Kingsbury's glands. Pelvic glands have been studied only by light microscopy with conventional methods, and consist of multiple tubular serous glands with a prismatic epithelium which contains numerous PAS positive secretory granules. The present report is an ultrastructural and lectin cytochemistry characterization of the pelvic glands of Triturus marmoratus marmoratus throughout the reproductive cycle. Our methods consisted of conventional electron microscopy, and colloidal‐gold lectin cytochemistry of the following lectins: WGA, ConA, LcA, UEA‐I, PNA, SBA, and HPA. In the prereproductive period, the glands showed a tall epithelium which consisted of two cell types, dark and clear cells, surrounded by elongated, myoepithelial cells. Both dark and clear cells showed the ultrastructural characteristics of secretory cells, and exhibited many secretory granules in the apical cytoplasm. Areas showing densely packed, degenerating cell organelles—which were not surrounded by membrane—were observed in the dark cells whereas the clear cells showed large heterolysosomes. In the postreproductive period the number of secretory granules decreased, the rough endoplasmic reticulum was less developed, and areas of degenerating organelles were absent. In addition, small basal cells appeared. The results of the lectin histochemistry study were similar in both reproductive periods. In the epithelial cells, the rough endoplasmic reticulum, the Golgi complex, and secretory granules exclusively labeled to ConA. In all cell types, the nuclei reacted to all lectins while the cytosol only reacted to LcA lectin. The ultrastructural and histochemical characteristics of the pelvic glands of T. marmoratus suggest that these glands could be homologous to the mammalian seminal vesicles and prostate. Anat Rec 254:196–204, 1999. © 1999 Wiley‐Liss, Inc.     

Prevalence of natural ovine fasciolosis shown by demonstrating the presence of serum circulating antigens

The prevalence of fasciolosis in sheep (Galicia, Northwest Spain) kept under field conditions was determined by using a sandwich-enzyme-linked immunosorbent assay (sELISA). Serum Fasciola hepatica circulating antigens were captured by means of a rabbit polyclonal IgG antibody to F. hepatica excretory/secretory products. Results were compared to those obtained by faecal sedimentation and an indirect ELISA (iELISA) and excretory/secretory antigens. Prevalences were 39.1% by sELISA, 30.4% by faecal sedimentation and 56% by iELISA; 83.3% of the sheep were positive to any one of the three tests. We observed that 59.5% of the sheep examined had active fasciolosis, 29.1% (117) had antigenaemia, 20.4% (82) passed eggs, and 40 (10%) were positive to both probes. We conclude that there is a high prevalence of fasciolosis in sheep from the studied region, and that the combination of sELISA and coprological sedimentation is extremely helpful for demonstrating current fasciolosis, so its application can be strongly recommended for epidemiological surveys.