Electron microscopical observations on the brush border of proximal tubule cells of mammalian kidney

Summary The ultrastructure of the plasma membrane and the core of microvilli of proximal tubule cells has been investigated by electron microscopy using sectioned and negatively stained material. By the technique of negative staining, a particulated coat is disclosed on the outside of the plasma membrane of microvilli of brush borders isolated from rat, rabbit and ox. This coat is composed of 30 to 60 Å particles and is 150 to 300 Å thick and appears to be a distinguishing feature for the luminal plasma membrane (brush border) of proximal tubule cells. The plasma membrane of the basal part of tubule cells is found to be smooth. By thin sectioning, an axial bundle of 50 to 70 Å diameter filaments regularly arranged in an 1+6 configuration, one axially located filament being surrounded by a ring of six, is disclosed. The distance from the ring of filaments to the inner surface of the plasma membrane is 250–300 Å, the diameter of the ring 300 Å and the center-to-center distance between filaments 120 Å. Negative staining also discloses 60 Å filaments in microvilli of isolated brush borders. Broken off, single microvilli (fingerstalls) are observed with thin filaments projecting from their broken ends. Filaments up to 1 in length are seen. At high magnification, the filaments appear beaded and show strong resemblance with actin filaments isolated from skeletal muscle. Based on present evidence, it is postulated that microvilli constituting renal brush borders possess contractile properties, which may play a role in the absorption process operating at the luminal part of the cells.The authors are indebted to Miss Kirsten Sjöberg for skilled technical assistance, and to the Danish State Research Foundation and the Tuborg Foundation for financial support.     

Establishment of regional differences in brush border enzymatic activities during the development of fetal mouse small intestine

Summary In order to study the establishment of regional differences in brush border enzymic activities during the development of fetal mouse small intestine we have followed (1) the differentiation of microvilli by morphometry, and (2) the developmental pattern of three brush border enzymes (lactase, glucoamylase and alkaline phosphatase). From day 16 to day 19 of gestation, the height of duodenal microvilli increases 2.4 times on the absorptive cells located near the tip of the villi. During the same period in the upper half of the duodenal villi, the number of microvilli per square m rises by a factor of 2.4 and the microvillous surface area increases by a factor of 5.2. The differentiation of ileal microvilli follows a similar pattern but they are always shorter and less numerous than those of the duodenum. Lactase activity appears at 18 days of gestation; the other two brush border enzymes are first detected at 16 days of gestation. Afterwards all three enzyme activities increase rapidly and a decreasing gradient of activity is established from the proximal to the distal segment of the small intestine. Hence, the structural development of the microvilli and the appearance of brush border enzyme activities occur simultaneously and a proximo-distal gradient is already established at 16 days of gestation.Supported by MRC of Canada research grant MA-6069Mr. D. Malka was supported by a studentship from the F.C.A.C.Dr. D. Ménard is a chercheur boursier du Conseil de la Recherche en Santé du Québec     

Rapid synthesis of photoreceptor membrane and assembly of new microvilli in a crab at dusk

Summary Large areas of photoreceptor membrane are synthesized in the retinula cells of the crab Leptograpsus variegatus at dusk. Initially, new membrane differentiates from rough endoplasmic reticulum (ER) as large tubules of smooth ER. These tubules transform to concentric ellipsoids of closely apposed pairs of membranes (doublet ER), sometimes passing through an intervening crenate form. The new membrane is transported through bridges of cytoplasm that cross the palisade to the rhabdom region, from which the remains of the rhabdomeres that were built during the previous dusk have been dissolved. The degradation of the old microvilli of one rhabdomere is accomplished without affecting neighbouring rhabdomeres of other cells. New microvilli are assembled in situ from sheets of doublet ER, which are converted to tubules oriented in the same direction as the future microvilli. The cytoplasmic face of the ER remains the cytoplasmic face of the tubules, which become progressively narrower, partly by further longitudinal division, until the final diameter of the microvillus is reached. A central core is often seen in transverse sections of mature microvilli. It may be involved in the final consolidation, but rhabdomeric microvilli are not formed in the same manner as those of intestinal brush border cells. There is no evidence that new membrane passes through the Golgi compartment before incorporation into the rhabdom, as is the case for rod outer segment membrane in vertebrate photoreceptors.     

On the fine structure of the aglomerular renal tubule in Lophius piscatorius

Summary The fine structure of the secretory tubules in the kidney of the aglomerular goose-fish (Lophius piscatorius) is described. The cells have a pyramidal shape, are joined together by multiple desmosomes, and share as main characteristics: abundant and deep inflections of the basal and lateral cell membranes; coated luminal plasma membranes forming multiple microvilli or a genuine brush border; moderate numbers of comparatively small mitochondria, usually unassociated with the basal and lateral plasma membrane specializations; numerous multivesicular bodies occuring in the apical cytoplasm; abundant large lysosome-like bodies in the intermediate regions of the cytoplasm; and comparatively poor development of endoplasmic reticulum and Golgi apparatus.The observations suggest that the cells perform both absorptive and secretory functions and are metabolically unusually active in autolytic and heterolytic work. Comparisons with other aglomerular species indicate that the ability for active secretory function is not necessarily dependent on a close association between plasma membrane and mitochondria; however, this ability does appear to require a markedly increased basal and/or lateral cell surface created by multiple invaginations of the plasma membrane. The abundance of desmosomes and associated structures appears to represent a unique structural specialization of the goosefish tubule, and indicates that the cells must be firmly anchored to one another to supply a rigid and mechanically continuous lining of the tubule. The multivesicular bodies probably represent endocytic vacuoles which fuse with apical vesicles and invaginate their outer membrane to form the internal vesicles; they appear to transform to ambilysosomes via a function as heterophagosomes and — later — combined hetero- and autophagosomes.Supported by grants from Karolinska Institutet, Fonden til Videnskabens Fremme and Konsul Johannes Fogh-Nielsen og fru Ella Fogh-Nielsens Légat. Part of the study was performed at the Zoological Station at Naples, Italy. The assistance of Mrs. Britt-Marie Karlsson is gratefully acknowledged.     

The fine structure of the retina of the honey bee drone

Summary The eye of the honey bee drone is composed of approximately 8,000 photoreceptive units or ommatidia, each topped by a crystalline cone and a corneal facet. An ommatidium contains 9 visual or retinula cells whose processes or axons pierce a basement membrane and enter the optic lobe underlying the sensory retina. The visual cells of the ommatidium are of unequal size: six are large and three, small. In the center of the ommatidium, the visual cells bear a brush of microvilli called rhabdomere. The rhabdome is a closed-type one and formed mainly by the rhabdomeres of the six large retinula cells. The rhabdomeric microvilli probably contain the photopigment (rhodopsin), whose modification by light lead to the receptor potential in the retinula cells. The cytoplasm of the retinula cells contains various organelles including pigment granules (ommochromes), and peculiar structures called the subrhabdomeric cisternae. The cisternae, probably composed of agranular endoplasmic reticulum undergo swelling during dark adaptation and appear in frequent connection with Golgi cisternae. Three types of pigment cells are associated with each ommatidium. The crystalline cone is entirely surrounded by two corneal pigment cells. The ommatidium, including its dioptric apparatus and corneal pigment cells, is surrounded by a sleeve of about 30 elongated cells called the outer pigment cells. These extend from the base of the corneal facet to the basement membrane. Near the basement membrane the center of the ommatidium is occupied by a basal pigment cell. Open extracellular channels are present between pigment cells as well as between retinula cells. Tight junctions within the ommatidium are restricted to the contact points between the rhabdomeric microvilli. These results are discussed in view of their functional implications in the drone vision, as well as in view of the data of comparative morphology.This work was supported by a grant from the Fonds National Suisse de la Recherche Scientifique.     

Fine structure of the ordinary lateral line organ

Summary The lateral line organ of the spotted shark is characterized by its semi-cylindrical shape. Each organ (neuromast) is so closely apposed to the next that the individual neuromasts are almost continuous. The neuromast is composed of receptor cells, supporting cells and mantle cells. The receptor cells bear one kinocilium and up to 40 stereocilia. Bi-directional arrangement of the receptor cells as occurs in teleosts was demonstrated. Afferent and efferent nerve endings were found at the base of the receptor cells. The supporting cells extend from the basal lamina to the free surface. Long microvilli and a cilium-like ciliary rod project from the top of each supporting cell. The cell contains relatively few elements of the Golgi apparatus and little rough endoplasmic reticulum, but mitochondria and filaments are abundant. The mantle cell limits the lateral margin of the neuromast. It is distinguished from the supporting cell because of its long crescent-shaped nucleus and scarce, short microvilli. Myelinated nerve fibres are found in the subepithelial connective tissue but not in the epithelium.The fine structure of the shark lateral line organ suggests that this organ is in an intermediated step of evolution between that of lamprey and teleost.     

Immunocytochemical characterization of two types of microvillar cells in rodent olfactory epithelium

Microvillar cells (MCs) have been identified in the olfactory epithelium of various mammalian species from rodents to humans. Studies on properties and functions of MCs to date have yielded partially controversial results, supporting alternatively an epithelial or a neuronal nature of these cells. In the present study, single and double immunolabeling investigations were carried out using antibodies against cytoskeletal and integral membrane proteins in order to further characterize MCs in rat and mouse olfactory epithelium. Application of antibodies against ankyrin (ANK), a protein that links integral membrane proteins to the submembrane cytoskeleton, led to intense labeling of the basolateral membranes of numerous cells with characteristic MC morphology. ANK-immunoreactive (ir) cells bore an apical tuft of -actin-ir microvilli, were filled with cytokeratin 18 (CK18)-ir filamentous network, and extended a basal process that appeared to end above the basal membrane. Immunoreactions for villin, an actin-crosslinking protein particularly prominently expressed in brush cells in the gastrointestinal and respiratory tract epithelia, and for the -subunit of sodium-potassium ATPase (Na⁺, K⁺-ATPase), revealed that ANK-ir MCs fall into two subpopulations. The less frequent type I MCs displayed villin immunoreactivity in their apical microvilli and underneath the basolateral membranes; the more numerous type II MCs were negative for villin but possessed intense basolateral immunoreactivity for Na⁺, K⁺-ATPase. Strong reactivity for the epithelial-type integral membrane protein of adherens junctions, E-Cadherin, was localized in basolateral membranes of both types of MCs. Our results support an epithelial nature of ANK-ir MCs in rat and mouse olfactory epithelium. Type I MCs strongly resemble brush cells in their immunocytochemical characteristics, namely, their ANK reactivity, CK18 reactivity, and villin reactivity. The intense Na⁺, K⁺-ATPase reactivity of type II MCs implicates these cells in transport processes.     

Freeze-fracture study of taste bud pores in the foliate papillae of the rabbit

Summary In freeze-fractured specimens of taste buds from the foliate papillae of rabbits, the intercellular spaces are separated from the pore of the taste bud by zonulae occludentes of the tight-type. Below these tight junctions numerous desmosomes are found at irregular intervals. The epithelial cells adjacent to the pore are also joined by single strands of fusion. The microvilli arising from the neck of the type I cells have a high particle density. The microvilli of type II cells and especially the short microvilli of peripherally situated cells have a lower intramembranous particle density. The single microvillus of type III cells has a very large diameter and is longer than the other microvilli. It contains a few larger intramembranous particles and vesicle-like protrusions of the membrane facing the cytoplasm. Transverse fracturing reveals a filamentous fine structure in all microvilli. The physiological implications of these observations are discussed.Supported by grants from the Deutsche Forschungsgemeinschaft (Ja 205/5+6)     

Identification of brush cells in the alimentary and respiratory system by antibodies to villin and fimbrin

Summary Brush cells represent a population of epithelial cells with unknown function, which are scattered throughout the epithelial lining of both the respiratory system and the alimentary system. These cells are reliably distinguished from other epithelial cells only at the ultrastructural level by the presence of an apical tuft of stiff microvilli and extremely long microvillar rootlets that may project down to the perinuclear space. In the present study we show that brush cells can be identified in tissue sections even at the light microscopic level by immunostaining with antibodies against villin and fimbrin, two proteins that crosslink actin filaments to form bundles. In brush cells, villin and fimbrin are not only present in the actin filament core bundles of apical microvilli and their long rootlets but, in addition, both proteins are also associated with microvilli extending from the basolateral cell surface of the brush cells. Basolateral immunostaining specific for villin and fimbrin does not occur in any other epithelial cell type of the respiratory and alimentary tract. Thus immunostaining with antibodies against both proteins allows unequivocal identification of individual brush cells even in sectional planes that do not contain the brightly stained apical tuft of microvilli and their long rootlets.     

Elektronenmikroskopische Beobachtungen an den Sehzellen des Blutegels,Hirudo medicinalis L.

Summary The visual cell of the leech (Hirudo medicinalis) contains a big vacuole filled with a moderate dense substance (vitreous body). The wall of the vacuole consists of a system of microvilli (brush border) merging into the substance of the vitreous body. The cytoplasmic zone underneath the brush border has a special structure consisting of flattened sacs and vesicles giving a radial striation to this cell region.The cytoplasm is filled with a great number of mitochondria and small vesicles. Bundles of fine filaments are striking features of the cytoplasm; in the periphery of the cell they are in close contact with the cell membrane (half desmosomes). Elements of the endoplasmic reticulum and free ribosomes may also be present. The cells regularly contain membrane-bounded inclusions having a dense granular or cristalline content.The visual fibers (processes of the visual cells) show a certain similarity to the unmyelinated nerve fibers of higher animals. They are ensheathed into sheath cells possibly of glial nature. Processes of these cells often surround the visual cells and are sometimes embedded in their cytoplasm. Both visual and sheath cells are covered with a homogeneous basement membrane.The possible role of the structures and the problems of conduction from the brush border onto the cell surface are discussed.     

Cells with microvillous borders in the cerebral ganglion of Leptodora kindtii focke (Crustacea)

Summary Groups of large cells in the cerebral ganglion of Leptodora kindtii join in intricate patterns to enclose lacunar spaces. The cell surfaces bordering on these lacunae are covered by long, densely packed microvilli that all but fill the spaces. Near their brush borders the cells are joined by adhesion plates; for the rest they are separated from each other by glial septa. The possible significance of these structures is discussed.Dedicated to an inspiring teacher and loyal friend, Prof. F. Wassermann on the occasion of his 80th birthday, August 13, 1964.Supported by Grant No. NB-02145 from the United States Public Health Service. The expert assistance of Mrs. Cynthia Jones, Mrs. Sarah Wurzelmann, and Mr. Stanley Brown is gratefully acknowledged.     

Gap junctions between the follicle cells and the oocyte during oogenesis in an insect,Tribolium destructor/Coleoptera

Summary Oocyte-follicle cell gap junctions inTribolium occur in all oogenetic stages studied. During early previtellogenesis the junctions are found exclusively between lateral membranes of oocyte microvilli and the membrane of prefollicle cells. In late previtellogenesis and vitellogenesis the junctions are located between the tips of oocyte microvilli and the flat membranes of the follicle cells. During previtellogenesis gap junctions are infrequent, whereas in the phase of yolk accumulation their number increases considerably, exceeding 17 junctions/m² of the follicle cell membrane. It could be shown by microinjection of a fluorescent dye that gap junctions are in a functional state during vitellogenesis. Possible roles of heterologous gap junctions in oogenesis are discussed.     

Ultrastructure of the protonephridial system of regenerating Stenostomum sp. (Plathelminthes,Catenulida)

Summary The ultrastructure of the flame bulbs, protonephridial capillaries and duct of fully developed and regenerating Stenostomum sp. is described. Flame bulbs are formed by a single cell whose nucleus is located basally or laterally to the weir. The weir is formed by a single row of transverse ribs connected by a thin membrane, apparently of extracellular matrix. Internal leptotriches arise from the proximal cytoplasm and extend in a (usually) single row along the weir and into the lumen of the distal cytoplasmic tube. Many or all leptotriches do not fuse with the distal cytoplasm. Two cilia are each anchored in the proximal cytoplasm by a cross-striated vertical and lateral rootlet, the latter bent forward and extending for some distance into one of the two cytoplasmic cords along the weir. Each cord contains the lateral rootlet in its proximal part, as well as many microtubules. The distal cytoplasmic tube contains two (longitudinal) junctions, i.e. lines of contact between cell processes of the same, terminal cell. Occasionally, more than two junctions were seen, apparently due to branches of the terminal cell in contact with each other. Flame bulbs join capillaries lined by several canal cells type I, containing few or no microvilli but lateral flames. Such capillaries join a duct (or ducts?) lined by canal cells type II with many long microvilli. The large protonephridial duct is lined by numerous cells with lateral flames and many long microvilli. In regenerating tissue (10.5 hours after cutting) some flame bulbs were free, i.e. not connected to capillaries, and some capillaries openly communicated with the surrounding intercellular space. In the presence of a single row of ribs in the weir, of internal leptotriches, and of vertical and lateral ciliary rootlets, the flame bulb of Stenostomum sp. resembles that of other Plathelminthes much more closely than hitherto thought. The species differs from non-catenulid plathelminths mainly in the large number of glandular cells lining the large protonephridial ducts, in the transverse orientation of the ribs in the weir and in the presence of only two cilia in the flame.     

Selective release of inner core proteins from intestinal microvillus membrane by lithium diiodosalicylate

Summary Lithium diiodosalicylate (LIS) was used to selectively solubilize proteins from purified intestinal brush border membrane vesicles. Incubation of the vesicles with increasing concentrations of LIS resulted in the progressive release of proteins with total disruption of the membranes being obtained at 200 mM. Maximum selectivity was observed at 20–30 mM LIS which preferentially released actin and other non-glycosylated proteins while all the glycoproteins remained associated with the membrane. Electron micrographs showed that, after LIS treatment, brush border vesicles are partially disrupted and have lost their inner core of microfilaments. Sucrase, trehalase, leucylnaphthylamide hydrolase, -glutamyl transpeptidase and alkaline phosphatase all retained more than 70% of their activities and remained associated with the membrane fraction after LIS solubilization (30 mM). The results indicate that lithium diiodosalicylate treatment provides an efficient method for the separation of cytoskeletal proteins from intrinsic membrane glycoproteins and should be very useful for the purification of microvilli proteins and for the study of membrane-protein interactions.Abbreviations LIS Lithium 3,5-diiodosalicylate - LNAase leucylnaphthylamide hydrolase - Tris Tris (hydroxymethyl) aminomethane     

The sieve tube wall and its relation to translocation

Summary The sieve tube wall possesses a broad inner layer often with pronounced radial striations. The plasmalemma of the sieve tube appears to penetrate this wall in the form of a brush border of irregular microvilli, greatly increasing its surface area. It is suggested that this is the site of active transport of potassium, which circulates electroosmotically through the sieve plate pores and back through the thick wall. The function of the companion cells is the care and maintenance of the active brush border sites; in conjunction with their activity in supplying high-energy intermediates movement in the column acts regeneratively and fully polarises the plates. Many of the lamellar stacks and curvilinear membrane aggregates hitherto regarded as endoplasmic reticulum are, it is suggested, plasmalemma displaced from the wall. These findings have important consequences for the electroosmotic theory.     

A study on the isolation of intestinal brush borders in saline

Summary Attempts to remove brush borders from rat intestinal mucosal scrapings in a saline medium are described. Variation of the pH of the medium showed that at about pH 6.5, morphologically intact columnar epithelial cells could be isolated and a suitable method for preparing whole cells is suggested. When the pH was raised to 8.5, the brush borders were liberated. Studies by electron microscopy revealed that a considerable amount of granular cytoplasmic material remained attached to the terminal web. This attached material could be removed, however, by adding EDTA to a suspension of the brush borders.The supernatants and the brush borders sedimented by centrifugation were investigated for contaminants from other regions of the cell. In general, the levels of contamination were higher in the brush borders isolated in saline than when isolated in the presence of EDTA. Phospholipid: cholesterol ratios for brush borders isolated in saline varied from 1.121 to 1.211, whereas in whole cells the ratio was about 3.51.The results are discussed in relation to previous studies.     

Ultrastructure of the tubule of the aglomerular teleost Nerophis ophidion

Summary The tubules in the aglomerular kidney of Nerophis ophidion are composed of cells showing different types of specializations of their plasma membrane. All cells possess a luminal brush border composed of microvilli, and show presence of vesicles with 100 Å thick unit membranes — some containing electron dense material —, tubular elements, multivesicular bodies, and plasma membrane invaginations in their apical cytoplasm. These features suggest an absorptive function of the cells. The apical portions of the cells are supplied with typical cilia.Some cells have abundant basilar plasma membrane invaginations usually lacking cytoplasmic organelles. Other cells appear to form interdigitating basilar cytoplasmic processes containing mitochondria; still other cells have smooth basilar cell membranes. These findings are discussed with reference to the known secretory function of the tubules and are compared with tubular fine structure in other species. It was concluded that urine formation by tubular secretion may occur in cells with different types of basilar cell membrane specializations.The occurrence of coated vesicles associated with invaginated basilar plasma membranes indicates transport of proteins (from peritubular blood vessels ?) at these sites.The tubule cells have abundant smooth surfaced endoplasmic reticulum and large and numerous active Golgi zones.Supported by grants from Fonden til Videnskabens Fremme and Therese och Johan Anderssons Minne. Part of this study was done at the Zoologica Stazione, Naples.The assistance of Miss Britt-Marie Petterson and Mr. Magnus Norman is gratefully acknowledged.     

Molecular Model of the Microvillar Cytoskeleton and Organization of the Brush Border

Background

Brush border microvilli are ∼1-µm long finger-like projections emanating from the apical surfaces of certain, specialized absorptive epithelial cells. A highly symmetric hexagonal array of thousands of these uniformly sized structures form the brush border, which in addition to aiding in nutrient absorption also defends the large surface area against pathogens. Here, we present a molecular model of the protein cytoskeleton responsible for this dramatic cellular morphology.

Methodology/Principal Findings

The model is constructed from published crystallographic and microscopic structures reported by several groups over the last 30+ years. Our efforts resulted in a single, unique, self-consistent arrangement of actin, fimbrin, villin, brush border myosin (Myo1A), calmodulin, and brush border spectrin. The central actin core bundle that supports the microvillus is nearly saturated with fimbrin and villin cross-linkers and has a density similar to that found in protein crystals. The proposed model accounts for all major proteinaceous components, reproduces the experimentally determined stoichiometry, and is consistent with the size and morphology of the biological brush border membrane.

Conclusions/Significance

The model presented here will serve as a structural framework to explain many of the dynamic cellular processes occurring over several time scales, such as protein diffusion, association, and turnover, lipid raft sorting, membrane deformation, cytoskeletal-membrane interactions, and even effacement of the brush border by invading pathogens. In addition, this model provides a structural basis for evaluating the equilibrium processes that result in the uniform size and structure of the highly dynamic microvilli.     

Analytical isolation of plasma membranes of intestinal epithelial cells: Identification of Na,K-ATPase rich membranes and the distribution of enzyme activities

Summary A procedure was developed for the analytical isolation of brush border and basal lateral plasma membranes of intestinal epithelial cells. Brush border fragments were collected by low speed centrifugation, disrupted in hypertonic sorbitol, and subjected to density gradient centrifugation for separation of plasma membranes from nuclei and cole material. Sucrase specific activity in the purified brush border plasma membrane was increased fortyfold with respect to the initial homogenate. Basal lateral membrane were harvested from the low speed supernatant and resolved from other subcellular components by equilibrium density gradient centrifugation. Recovery of Na, K-ATPase activity was 94%, and 61% of the recovered activity was present in a single symmetrical peak. The specific activity of Na, K-ATPase was increased twelvefold, and it was purified with respect to sucrase, succinic dehydrogenase, NADPH-cytochromec reductase, nonspecific esterase, -glucoronidase, DNA, and RNA. The observed purification factors are comparable to results reported for other purification procedures, and the yield of Na, K-ATPase is greater by a factor of two than those reported for other procedures which produce no net increase in the Na, K-ATPase activity.Na, K-ATPase rich membranes are shown to originate from the basal lateral plasma membranes by the patterns of labeling that were produced when either isolated cells or everted gut sacs were incubated with the slowly permeating reagent³⁵S-p-(diazonium)-benzenesulfonic acid. In the former case subsequently purified Na, K-ATPase rich and sucrase rich membranes are labeled to the same extent, while in the latter there is a tenfold excess of label in the sucrase rich membranes. The plasma membrane fractions were in both cases more heavily labeled than intracellular protein.Alkaline phosphatase and calcium-stimulated ATPase were present at comparable levels on the two aspects of the epithelial cell plasma membrane, and 25% of the acid phosphatase activity was present on the basal lateral membrane, while it was absent from the brush border membrane. Less than 6% of the total Na, K-ATPase was present in brush border membranes.     

Cyclical changes in the fine structure of bovine endometrial gland cells

Summary Cyclical changes in bovine endometrial gland cells were investigated in six heifers, three at estrus and three at day twelve of the estrous cycle in the luteal phase. The epithelium is generally low at estrus but high in the luteal phase. There are ciliated and non-ciliated cells. The ciliated cells are fewer and lighter and show inconspicuous cyclical changes.The secretory cells show more prominent changes. At estrus, their free border is flat with short microvilli. The conspicuous rough-surfaced endoplasmic reticulum may synthesize protein for later secretion. The Golgi complex seems inactive. The high number of cytosegresomes and dense bodies might express cell regression caused by endocrine changes.In the luteal phase, the cells are lighter with long microvilli. The Golgi apparatus shows vacuoles and immature secretory droplets. Secretory vacuoles with light contents occur in the apical cytoplasm. Some of them appear to discharge their contents into the lumen. This is interpreted as evidence of merocrine secretion. Accumulations of tubular, smooth-surfaced endoplasmic reticulum, masses of glycogen granules, and several fat droplets are present.Some lymphocytes and degranulated granulocytes are seen near the basement membrane, more frequently at estrus.Financial support for this study was received from Anslaget för främjande av medicinsk forskning vid Veterinärhögskolan.The authors express their gratitude to Prof. A. Bane and Dr. J.-E. E. Ringmar, Department of Obstetrics and Gynecology, for their help with the selection and clinical control of the animals and for keeping them in good condition.Post doctoral fellow, No. 43-KO-52 (1968) from the Educational Ministry of Japan.