Characterization of VAMP‐2 in the lung: implication in lung surfactant secretion

Lung surfactant is crucial for reducing the surface tension of alveolar space, thus preventing the alveoli from collapse. Lung surfactant is synthesized in alveolar epithelial type II cells and stored in lamellar bodies before being released via the fusion of lamellar bodies with the apical plasma membrane. SNAREs (soluble N‐ethylmaleimide‐sensitive fusion protein‐attachment protein receptors) play an essential role in membrane fusion. We have previously demonstrated the requirement of t‐SNARE (target SNARE) proteins, syntaxin 2 and SNAP‐23 (N‐ethylmaleimide‐sensitive factor‐attachment protein 23), in regulated surfactant secretion. Here, we characterized the distribution of VAMPs (vesicle‐associated membrane proteins) in rat lung and alveolar type II cells. VAMP‐2, ?3 and ?8 are shown in type II cells at both mRNA and protein levels. VAMP‐2 and ?8 were enriched in LB (lamellar body) fraction. Immunochemistry studies indicated that VAMP‐2 was co‐localized with the LB marker protein, LB‐180. Functionally, the cytoplasmic domain of VAMP‐2, but not VAMP‐8 inhibited surfactant secretion in type II cells. We suggest that VAMP‐2 is the v‐SNARE (vesicle SNARE) involved in regulated surfactant secretion.     

Electron microscopic observations on a new cell type in the fundus mucosa of the mouse stomach

Summary In the course of electron microscopic investigations of the fundus mucosa of the mouse stomach, a few cells of an unknown type were found by chance in the deep portions of the glands. These cells are characterized by two different kinds of specific granules in their cytoplasm, one of which being large and less dense, and the other one being small and dense. The large less dense granules resemble zymogen granules of the chief cell, which are formed by the rough endoplasmic reticulum and Golgi system. The small dense granules are quite similar to the secretory granules of the basal granulated cell, and are considered to be formed in the Golgi complex. Release figures of the small dense granule were not observed, numerous granules, however, were observed in close contact with the basal cell membrane. The occurrence of these two kinds of granules in one cell suggests that the basal granulated cell and the zymogenic cell originate from the same entodermal stem element.The author cordially thanks Professor Dr. Hisao Fujita, Department of Anatomy, Hiroshima University School of Medicine, for his kind advices and criticisms.     

A morphological and histochemical study of the subcommissural organ of young and old sheep

Summary Cytologic alterations with age are evident in the subcommissural organ of sheep, but evidence of a secretory function has not been found. In young lambs the cytoplasm has numerous, irregular, interconnecting, endoplasmic sacs with electron dense material. Lamellar cytoplasmic lipid bodies (lipid surrounded by membranous whorls) are scattered throughout the deep layer.With age, the cytoplasm contains electron-lucent, flocculent material and lamellar cytoplasmic lipid bodies are less apparent. In adolescent and adult sheep tightly-coiled membranous whorls (fingerprints) enclosing cytoplasmic constituents are noted. Gliosis is progressive. These changes probably indicate involution.Histochemical studies demonstrate distinct Golgi patterns in superficial and deep layers and a rich vascular network in the deep layer. Acid phosphatase activity is present and defines lysosomes, a larger cholesterol ester-containing body (Type II) and macrophages. Type II bodies are less frequent with age.Bioassays and chemical analyses should be interpreted with reference to the age of the experimental animal.This investigation was supported in part by Public Health Service Research Grants NB 05469-02, 5R01-HE05609-06 and GRS #FR-5428 from the National Institutes of Health.This work was done while Dr. Barlow was a Fulbright Scholar.     

Changes in retinal fine structure induced in the crab Libinia by light and dark adaptation

Summary The cytological influence of light and dark adaptation (LA and DA) on the retinular cells of the spider crab Libinia emarginata has been studied by light and electron microscopy in four adaptive states: 17 hours darkness, 5 hours darkness, 5 hours diffuse light and 17 hours diffuse light. The rhabdom's fine structure is typical of decapods but its dual overall form and position mingle certain features of both apposition and superposition compound eye types. Distal and proximal retinal pigments both showed adaptive migration, but the distal pigment cells moved over a restricted range, and DA separated the retinular cell pigment granules into two groups, perinuclear and basilar.In the rhabdom no changes in its position, dimensions or microvillus fine structure were observed with LA or DA. But at the base of the rhabdom microvilli the rate of pinocytosis was strongly affected by the eye's adaptive state, being lowest after 17 hours DA and greatest after 17 hours LA; the wall of the 0.1 microvesicles so formed, looked like the membrane of the rhabdom microvillus and they were the same size as the vesicles in multivesicular bodies and in vesicular lamellar bodies.Three categories of complex cytoplasmic particles about 1 in diameter (multivesicular bodies, vesicular lamellar bodies and purely lamellar bodies) were all increased in number by decreased DA and by increased LA; similar quantitative effects occurred in the endoplasmic reticulum and in the ribosomes.The pinocytotic vesicles and the complex cytoplasmic bodies may represent part of an intracellular system to dispose of rhabdom metabolites whose production was initiated or increased by light absorption.Cytoplasmic and perirhabdomal vacuoles mainly distal in location, were also affected by light, but inversely; their maximal extent occurred after 17 hours DA; less DA or any LA significantly decreased their presence and aggregation.The data reported are of interest not only because they correlate retinal fine structure with the metabolism of vision but also because they provide a new and specific tool for distinguishing active from inactive neurosensory cells in the optic pathway.This research was initiated with the aid of U.S. Public Health Service Grant NB-03076 and has been continued with the support of U.S. Air Force Grant AFOSR-1064. The authors wish to thank Dr. Joseph G. Gall and Dr. William R. Adams for generously sharing their electron microscopic facilities; they are also grateful to Mrs. Mabelita Campbell for her collaboration on the light microscopy.     

The fine structure of photoreceptors in a marine flatworm

Summary The ocelli or eyes of the marine polyclad turbellarian Notoplana acticola are clustered on the paired dorsal nuchal tentacles and in two longitudinal bands lateral to the cerebral ganglion. The ocelli, studied by electron microscopy, were characterized as rhabdomeric and non-ciliary in origin. There are 60 to 80 ocelli per animal each enclosed in a fibrous capsule to which muscle fibers may attach. An ocellus consists of a pigmented eyecup into which 30 to 50 photoreceptor cells send dendritic processes through interruptions in or among pigment cell projections across the eyecup opening. The dendritic processes terminate in numerous long intertwined microvilli which fill the eyecup. The nucleated cell body of each photoreceptor cell lies outside the eyecup and projects an axonal process to the cerebral mass. Within the dendritic processes are observed mitochondria, ribosomes, neurotubules, multivesicular bodies, vesicles and vacuoles. The cell body contains smaller mitochondria, endoplasmic reticulum, ribosomes, vesicles and prominent Golgi complexes.After dark adaptation, there are some structural alterations in terms of swelling of microvilli, increased numbers of vacuoles associated with the microvilli and dendritic processes, and changes in the pigment cell projections.This work was supported by Grant No. GM 10292 from the U.S. Public Health Service to Professor Richard M. Eakin, Department of Zoology at the University of California, Berkeley, U.S.A., where this investigation was conducted during the author's sabbatical leave of absence from the University of Illinois, and by Grant No. 1 SO 1 FR 5369 from the U.S. Public Health Service to the University of Illinois at the Medical Center.I express appreciation to Professor Eakin for interesting discussions and generous hospitality to me as a guest in his laboratory, and to the John Simon Guggenheim Memorial Foundation for the Fellowship which I held during 1964–65. I thank Dr. John P. Marbarger, Director of the Aeromedical Laboratory for the electron microscope facilities used at the University of Illinois.     

Loss of Rab27 function results in abnormal lung epithelium structure in mice

Rab27 small GTPases regulate secretion and movement of lysosome-related organelles such as T cell cytolytic granules and platelet-dense granules. Previous studies indicated that Rab27a and Rab27b are expressed in the murine lung suggesting that they regulate secretory processes in the lung. Consistent with those studies, we found that Rab27a and Rab27b are expressed in cell types that contain secretory granules: alveolar epithelial type II (AEII) and Clara cells. We then used Rab27a/Rab27b double knockout (DKO) mice to examine the functional consequence of loss of Rab27 proteins in the murine lung. Light and electron microscopy revealed a number of morphological changes in lungs from DKO mice when compared with those in control animals. In aged DKO mice we observed atrophy of the bronchiolar and alveolar epithelium with reduction of cells numbers, thinning of the bronchiolar epithelium and alveolar walls, and enlargement of alveolar airspaces. In these samples we also observed increased numbers of activated foamy alveolar macrophages and granulocyte containing infiltrates together with reduction in the numbers of Clara cells and AEII cells compared with control. At the ultrastructural level we observed accumulation of cytoplasmic membranes and vesicles in Clara cells. Meanwhile, AEII cells in DKO accumulated large mature lamellar bodies and lacked immature/precursor lamellar bodies. We hypothesize that the morphological changes observed at the ultrastructural level in DKO samples result from secretory defects in AEII and Clara cells and that over time these defects lead to atrophy of the epithelium.     

A role for diacylglycerol in annexin A7-mediated fusion of lung lamellar bodies

Lung surfactant secretion in alveolar type II cells occurs following lamellar body fusion with plasma membrane. Annexin A7 is a Ca2+-dependent membrane-binding protein that is postulated to promote membrane fusion during exocytosis in some cell types including type II cells. Since annexin A7 preferably binds to lamellar body membranes, we postulated that specific lipids could modify the mode of annexin A7 interaction with membranes and its membrane fusion activity. Initial studies with phospholipid vesicles containing phosphatidylserine and other lipids showed that certain lipids affected protein interaction with vesicle membranes as determined by change in protein tryptophan fluorescence, protein interaction with trans membranes, and by protein sensitivity to limited proteolysis. The presence of signaling lipids, diacylglycerol or phosphatidylinositol-4,5-bisphosphate, as minor components also modified the lipid vesicle effect on these characteristics and membrane fusion activity of annexin A7. In vitro incubation of lamellar bodies with diacylglycerol or phosphatidylinositol-4,5-bisphosphate caused their enrichment with either lipid, and increased the annexin A7 and Ca2+-mediated fusion of lamellar bodies. Treatment of isolated lung lamellar bodies with phosphatidylinositol- or phosphatidylcholine phospholipase C to increase diacylglycerol, without or with preincubation with phosphatidylinositol-4,5-bisphosphate, augmented the fusion activity of annexin A7. Thus, increased diacylglycerol in lamellar bodies following cell stimulation with secretagogues may enhance membrane fusion activity of annexin A7.     

Nuclear membranous whorls

Summary Membranous whorls have been seen in the nuclei of peritoneal and testicular cells which had been subjected to various experimental manoeuvres. It seems likely that this is an early manifestation of cell degeneration which is demonstrated readily only by glutaraldehyde fixation, and to that extent can be regarded as a glutaraldehyde artifact. Acknowledgements. This work was supported by grants from the Medical Research Council, and the University of Sheffield Tuberculosis Research Fund, and by a grant to the Department from Unilever Ltd.I am grateful to Professor R. Barer for his advice and criticism, to Dr. G. A. Meek for guidance on electron microscopy, to Dr. E. J. Clegg for permission to use material from joint experiments. Technical and photographic assistance was provided by Messrs. P. GarLick and L. Murgatroyd and by Miss M. Tune.     

Transitional epithelium of urinary tract in normal and dehydrated rats

Summary This report is a light microscopic histochemical and fine structural study of transitional epithelium of the urinary tract of normal and dehydrated rats. Four types of cells were recognized: basal, intermediate, squamous or luminal and bundle cells. The transitional epithelium of normal rat ureter and bladder shows distinct cytoplasmic staining of the squamous cells layer by PAS. The luminal free border stains more intensely with PAS. With the electron microscope, abundant cytoplasmic tonofilaments, free ribosomes and the characteristic thick-walled fusiform and round vesicles are observed, which were in greater number in the squamous cells. Lysosomes are identified with PAS, and Toluidine Blue 0, by their content of acid phosphatase and non-specific carboxylic esterase, and by their ultrastructural appearance. The bundle cell (Hicks, 1965) is characterized by histochemical technics. These cells form about 2.5% of the total cell population of normal transitional epithelium. The bundle cell contains basophilic metachromatic granules, which indicates the presence of a weakly acid mucosubstance. It is suggested that bundle cell granules are released in the intercellular spaces of transitional epithelium and that the mucosubstance may regulate flow of ions and metabolites in the epithelial intercellular channels.Several ultrastructural changes occur in the transitional epithelium of dehydrated rats: marked increase in number of thick-walled vesicles, development of polysomes, relative increase of cytoplasmic filaments and greater number of enlarged lysosomes. Bundle cells decrease in number. These ultrastructural changes promptly regressed by allowing the animal to drink water.It is suggested that the rate of formation of the characteristic vesicles of transitional epithelium, a function of membrane synthesis, may be under the control of the antidiuretic hormone.This investigation was supported in part by the Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina, through a travel grant to Dr. Monis, who would like to thank Dr. E. de Robertis for the use of the electron microscope facilities of the Instituto de Anatomía General y Embriología, Facultad de Medicina, Universidad de Buenos Aires.     

Ultrastructure of transitional epithelium of man

Summary Blocks of human normal renal pelvis and ureter obtained at the time of surgery were fixed in glutaraldehyde and osmium with or without ruthenium red, for electron microscopic observations. The transitional epithelium is arranged in three cell layers: basal, intermediate and superficial. All epithelial cells show numerous microvilli and contain the characteristic vesicles of transitional epithelium, bundles of cytoplasmic filaments, microtubules and numerous free ribosomes. The epithelial extracellular compartment is notably large and appears as an intricate, tridimensional network of canaliculi and cisternae which are wider in the intermediate and superficial layers and in which microvilli and cytoplasmic folds of vicinal cells are often attached or interdigitated. At these sites there are desmosomes.The surface of all transitional epithelial cells is covered by a fibrillar mucous coat which is more developed at the plasmalemma of the free border of luminal cells in which microvilli are also seen. Ruthenium red stains selectively the plasmalemma and the mucous coat of the free surface of the epithelium, indicating the presence of an acid polysaccharide. With this technic (Luft, 1965), it is observed, radiating from the plasmalemma, branching filaments which measure 100 Å in diameter forming a zone of varying density which is about 400 m wide and which corresponds, at the light microscopic level, to the luminal border of the transitional epithelial cells in which a sialomucin has been identified. The slender filaments have a beaded appearance. At the free border, superficial cells are attached by functional complexes in which tight junctions seal the epithelial intercellular space, which is opened at the level of the basement membrane where only desmosomes are observed.The ultrastructure of human transitional epithelium of urinary tract resembles the duct cells of the salt gland of certain marine birds (Fawcett, 1962) and the amphibian epidermis (Farquhar and Palade, 1965) in which there are active processes of transport. The mucous surface coat, selectively stained by the ruthenium red, contains a sialomucin (Monis and Dorfman, 1965, 1967).The ultrastructure and histochemistry of the mucous fluffy coat of man transitional epithelium and the observations of Porter and Tamm (1955), on the ultrastructure of preparations of the Tamm and Horsfall mucoprotein (1952) are bases for suggesting that transitional epithelium of urinary tract of man is the site of biosynthesis of certain urinary mucoids. Present investigations are directed to obtain evidence to substantiate this hypothesis.General Abbreviations B basal cell - E exfoliating cell - I intermediate cell - L lumen - S superficial cell - SC surface coat - bm basement membrane - ci cell infolding - d desmosome (macula adhaerens) - f fibroblast - fi cytoplasmic filaments - is intercellular space - jc junctional complex - ly lysosome - lym lymphocyte - mt microtubules - m mitochondria - mv microvilli - n nucleus - r ribosomes - rv round vesicle - zo zonula occludens - za zonula adherens Dr. Monis wishes to thank Dr. E. De Robertis for the use of the electron microscope facilities of the Instituto de Anatomía General y Embriologia, Facultad de Medicina, Universidad de Buenos Aires. — Prof. E. Trabucco and Dr. R. J. Borzone (Cátedra de Clinica Genitourinaria de la Facultad de Medicina, Universidad de Buenos Aires) generously supplied the specimens which were the bases of this study. — Thanks are due to Mrs. A. M. Novara and Mrs. Defilippi-Novoa for efficient technical help and to Miss Rosa Gentile for secretarial assistance. Photomicrography by Mr. M. A. Saenz.Dr. Zambrano is investigator (CNICT).     

The fine structural localization of acid phosphatase in the gut epithelial cells of the slug,Avion ater (L.)

Summary Acid phosphatase, generally thought of as a lysosomal enzyme and indeed widely employed as a lysosomal marker, has been found associated with the Golgi complex of all cell types from the crop, intestine and digestive gland ofArion ater. Reaction product was also detected within the multivesicular bodies and cytoplasm of columnar cells from the crop and the multivesicular bodies of mucous cells from the intestine. A vacuolar localization was obtained in the digestive cells of the intestine and digestive gland. Secretory protein granules in the calcium cells of the same gland and apical vacuoles in the so-called thin cells also showed a positive reaction.This work was undertaken as part of a slug research project under the direction and co-ordination of Dr. D. K.Roach, supported by A.R.C. Assistance was given by Mr. T. R.Mainwaring in the preparation of tissue for electron microscopy.I would like to thank Professor J.Brough and Professor D.Bellamy for providing facilities and encouragement.     

Electron microscopic observations of young rat liver

Summary Electron microscopic observations on normal liver tissue of four-day-old rats reveal the presence of numerous lamellar structures (lamellar bodies). These can be contained within parenchymal cells or in bile canaliculi, Disse's space, and in the lumen of blood sinusoids. Such bodies can also be found in Kupffer and endothelial cells.The lamellar bodies within hepatic cells are generally seen in very intimate relation to glycogen particles and lipide droplets, but in some to agranular endoplasmic reticulum and Golgi membranes as well.On the basis of this intimate relation to intracellular glycogen granules and lipide droplets, it is presumed that lamellar bodies represent a special intermediate stage in carbohydrate and lipide metabolism.Discontinuities in the endothelial layer of intrahepatic sinusoids are described.This work was supported in part by a N.A.T.O. research fellowship of the Consiglio Nazionale delle Ricerche, Roma.Assistant Professor in the Department of Veterinary Anatomy, Histology and Embryology (Dir.: Prof. A. de Girolamo), University of Naples, Naples, Italy.     

The fine structure of the mouse adrenal X zone

Summary Electron microscopically the adrenal X zone was examined in the fourteen SMA female mice aged 40 and 70 days. At these ages, the X zone showed no signs of degeneration. The X zone cell was somewhat smaller than the permanent cortical cell.The mitochondria in the X zone cell were quite bizarre in shape, provided with tubules or cristae. Many intramitochondrial bodies very similar to the cytoplasmic lipid droplets were found in the X zone. A few lipid droplets and globules were also noticed in this zone. The lipid droplets may possibly be formed within the mitochondria.The light and dark cells were differentiated. For the light cells, scant mitochondria and tubular granular endoplasmic reticulum were characteristic in contrast to the abundant mitochondria and multi-lamellated agranular endoplasmic reticulum in the dark cells. The cellular variety in density was discussed with regard to steroid synthesis.The author wishes to express his sincere appreciation to Prof. H. Tauchi, The 2nd Department of Pathology, Nagoya University School of Medicine, for kind advice, to Dr. M. Hoshino for helpful suggestion, and to Mr. J. Aoki for excellent technical assistance.     

Vacuolar ATPase Regulates Surfactant Secretion in Rat Alveolar Type II Cells by Modulating Lamellar Body Calcium

Lung surfactant reduces surface tension and maintains the stability of alveoli. How surfactant is released from alveolar epithelial type II cells is not fully understood. Vacuolar ATPase (V-ATPase) is the enzyme responsible for pumping H⁺ into lamellar bodies and is required for the processing of surfactant proteins and the packaging of surfactant lipids. However, its role in lung surfactant secretion is unknown. Proteomic analysis revealed that vacuolar ATPase (V-ATPase) dominated the alveolar type II cell lipid raft proteome. Western blotting confirmed the association of V-ATPase a1 and B1/2 subunits with lipid rafts and their enrichment in lamellar bodies. The dissipation of lamellar body pH gradient by Bafilomycin A1 (Baf A1), an inhibitor of V-ATPase, increased surfactant secretion. Baf A1-stimulated secretion was blocked by the intracellular Ca²⁺ chelator, BAPTA-AM, the protein kinase C (PKC) inhibitor, staurosporine, and the Ca²⁺/calmodulin-dependent protein kinase II (CaMKII), KN-62. Baf A1 induced Ca²⁺ release from isolated lamellar bodies. Thapsigargin reduced the Baf A1-induced secretion, indicating cross-talk between lamellar body and endoplasmic reticulum Ca²⁺ pools. Stimulation of type II cells with surfactant secretagogues dissipated the pH gradient across lamellar bodies and disassembled the V-ATPase complex, indicating the physiological relevance of the V-ATPase-mediated surfactant secretion. Finally, silencing of V-ATPase a1 and B2 subunits decreased stimulated surfactant secretion, indicating that these subunits were crucial for surfactant secretion. We conclude that V-ATPase regulates surfactant secretion via an increased Ca²⁺ mobilization from lamellar bodies and endoplasmic reticulum, and the activation of PKC and CaMKII. Our finding revealed a previously unrealized role of V-ATPase in surfactant secretion.     

Entwicklung und Lage der Augenpigmente bei Verschiedenen Drosophilamutanten

Ohne ZusammenfassungMit II TextabbildungenHerrn Professor Dr. Dr.A. Reichensperger zum 80. Geburtstag gewidmet.     

Synaptic relationships in the plexiform layers of carp retina

Summary The synaptic contacts made by carp retinal neurons were studied with electron microscopic techniques. Three kinds of contacts are described: (1) a conventional synapse in which an accumulation of agranular vesicles is found on the presynaptic side along with membrane densification of both pre- and postsynaptic elements; (2) a ribbon synapse in which a presynaptic ribbon surrounded by a halo of agranular vesicles faces two postsynaptic elements; and (3) close apposition of plasma membranes without any vesicle accumulation or membrane densification.In the external plexiform layer, conventional synapses between horizontal cells are described. Horizontal cells possess dense-core vesicles about 1,000 Å in diameter. Membranes of adjacent horizontal cells of the same type (external, intermediate or internal) are found closely apposed over broad regions.In the inner plexiform layer ribbon synapses occur only in bipolar cell terminals. The postsynaptic elements opposite the ribbon may be two amacrine processes or one amacrine process and one ganglion cell dendrite. Amacrine processes make conventional synaptic contacts onto bipolar terminals, other amacrine processes, amacrine cell bodies, ganglion cell dendrites and bodies. Amacrine cells possess dense-core vesicles. Ganglion cells are never presynaptic elements. Serial synapses between amacrine processes and reciprocal synapses between amacrine processes and bipolar terminals are described. The inner plexiform layer contains a large number of myelinated fibers which terminate near the layer of amacrine cells.This work was supported by an N.I.H. grant NB 05404-05 and a Fight for Sight grant G-396 to P.W. and N.I.H. grant NB 05336 to J.E.D. The authors wish to thank Mrs. P. Sheppard and Miss B. Hecker for able technical assistance. P.W. is grateful to Dr. G. K. Smelser, Department of Ophthalmology, Columbia University, for the use of his electron microscope facilities.     

Macrophages containing langerhans cell granules in normal lymph nodes of the rabbit

Summary Light and electron microscopic studies on macrophages of normal rabbit lymph nodes showed two types, one with little phagocytic activity and many features similar to those of epidermal Langerhans cells. Among these are characteristic Langerhans cell granules. From these findings it is concluded that the Langerhans cells may be derived from lymph node macrophages.The helpful advice and criticism of Dr. Toshio Nagano, Department of Anatomy, are gratefully acknowledged. The discussion with Dr. Mitsumasa Itoh, Department of Dermatology, was also helpful.     

The fine structure of sensory receptor processes in the auricular epithelium of the planarian,Dugesia tigrina

Summary The marginal epithelium of the lateral auricles of the planarian, Dugesia tigrina, includes a cell type with surface cilia and microvilli, a basal nucleus, and dense cytoplasm containing secretory vacuoles, Golgi elements, mitochondria and ribosomes. Through channels within the epithelial cytoplasm, cellular processes, interpreted as extensions of neurosensory receptor cells located in the subepidermis, project to the surface. The receptor processes, containing microtubules, mitochondria, vesicles and an agranular tubular reticulum, project beyond the epithelial cell surface; one or two cilia each emerge from a basal body in the apex of the projection. Close to the point of emergence to the epithelial surface, each cylindrical receptor process is surrounded by a collar-like septate junction between adjacent plasma membranes. The cilia of the projections differ from those of the epithelial cells in diameter, density of matrix and in the banding patterns of the rootlets. A few projections appear with the apex and basal body retracted below the epithelial surface. The possible function of these ciliated processes in sensory reception is discussed.This work was supported by Grant No. SO 1 FR 5369 from the U.S. Public Health Service to the University of Illinois at the Medical Center.I thank Dr. J. P. Marbarger, Director of the Research Resources Laboratory, for use of the electron microscope facilities, Miss Irena Kairys for technical help, Miss Marie Jaeger for assistance with photography, and Mr. Robert Parshall for the drawing.To Professor Arthur Wagg Pollister, I respectfully dedicate this article on the occasion of his retirement from Columbia University.     

Alveolar type II cells: studies on the mode of release of lamellar bodies.

There is increasing evidence that type II alveolar cells are capable of synthesizing surface active material like that obtained from the airways. However a number of problems remain to be solved before it can be stated conclusively that type II cells synthesize the surface active material of the terminal airspace. Among these problems is that of secretion. A number of previous studies have given evidence of the release of lamellar bodies by merocrine secretion. In this study morphologic evidence is presented which supports the view that secretion of lamellar bodies is accomplished by exocytosis. At the apical surface of type II cells, sites can be found where the limiting membrane of the lamellar body is clearly fused with the type II cell plasma membrane and an open channel exists between the contents of the lamellar body and the alveolar space. At these sites the lamellar contents extrude into the airspace with consequent loss of the highly compact organization of intracellular lamellar bodies. The intactness and continuity of the membranes can be traced for the full extent of the exocytosis site. Freeze-etch replicas of the membranes of type II cells show depressions which may represent the sites of discharged lamellae. In addition, tongue-like folds are seen which could be explained as the extensions of cytoplasm which surround the releasing lamellar body and which may flap over the exocytosis pit after discharge. Micrographs of the alveolar space show disorganized lamellar whorls which appear to be unravelling to produce tubular myelin. In view of the unusually large size and lipid composition of lamellar bodies, a mechanism involving hydration of mucopolysaccharide contents as an aid to expulsion of lamellar contents is suggested.     

Scanning electron microscopic studies of cilia

Summary A simple method for the preparation of ciliated epithelia for study with the scanning electron microscope is described. Ciliary groups are well preserved and it is possible to discern individual cilia and work out their numbers and orientation. Following scanning electron microscopical study some of the material was prepared for transmission electron microscopy and the ultrastructure of the tissue was found to be surprisingly well preserved. The tracheal epithelium of the rabbit, the olfactory epithelia of the goldfish and the rabbit, and the sensory epithelia in the statocyst of a cephalopod mollusc were examined with the scanning electron microscope to demonstrate the possibilities of the method. Acknowledgements. We would like to thank Professor J. Z. Young for his continued interest and support. The scanning electron microscope was purchased with a grant provided by the Science Research Council to Dr. Boyde, Mr. R. Willis helped in the initial stages of the study, Mr. G. Savage provided help with the goldfish material, Mr. S. Waterman provided much photographic assistance, and Mrs. N. Finney the secretarial assistance.