Cellular differentiation in the kidneys of newborn mice studies with the electron microscope

The structure of the kidney of the Swiss albino mouse changes progressively during the first 2 weeks after birth. Cells proliferate to form new nephrons, cells differentiate by acquiring specialized membranous components, and certain cytological features which are present at birth diminish in abundance or disappear. The differentiation of the cells of the cortical tubules has been studied using the light and electron microscopes. The tubules are partially and variably differentiated at birth. During the first 2 weeks after birth the brush border develops in the proximal tubules by the accumulation of numerous microvilli on the apical cell margins. Basal striations develop in proximal and distal tubules as an alignment of mitochondria, the result of what appears to be progressive interlocking of adjacent fluted cells. The mitochondria increase in number and size, accumulate homogeneous matrix, and acquire small, very dense granules. The collecting ducts develop tight pleating of the basal cell membranes, and dark cells containing numerous small cytoplasmic vesicles and microvilli appear. At birth there are dense irregular cytoplasmic inclusions presumed to be lipide in renal cells, the cytoplasmic granules of Palade are abundant, and there are large round bodies in the cells of the proximal tubules. The lipide inclusions disappear a few days after birth, and the cytoplasmic granules of Palade diminish in abundance as the cells differentiate. The large round bodies in the proximal tubules consist of an amorphous material and contain concentrically lamellar structures and mitochondria. They resemble the cytoplasmic droplets produced in the proximal tubules of adult rats and mice by the administration of proteins. The large round bodies disappear from the proximal tubules of infant mice during the first week after birth, but the concentric lamellar structures may be found in adult mice.     

Anatomy and fine structure of the alimentary canal of the spittlebug Lepyronia coleopterata (L.) (Hemiptera: Cercopoidea)

The alimentary canal of the spittlebug Lepyronia coleopterata (L.) differentiates into esophagus, filter chamber, midgut (conical segment, tubular midgut), and hindgut (ileum, rectum). The filter chamber is composed of the anterior extremity of the midgut, posterior extremity of the midgut, proximal Malpighian tubules, and proximal ileum; it is externally enveloped by a thin cellular sheath and thick muscle layers. The sac-like anterior extremity of the midgut is coiled around by the posterior extremity of the midgut and proximal Malpighian tubules. The tubular midgut is subdivided into an anterior tubular midgut, mid-midgut, posterior tubular midgut, and distal tubular midgut. Four Malpighian tubules run alongside the ileum, and each terminates in a rod closely attached to the rectum. Ultrastructurally, the esophagus is lined with a cuticle and enveloped by circular muscles; its cytoplasm contains virus-like fine granules of high electron-density. The anterior extremity of the midgut consists of two cellular types: (1) thin epithelia with well-developed and regularly arranged microvilli, and (2) large cuboidal cells with short and sparse microvilli. Cells of the posterior extremity of the midgut have regularly arranged microvilli and shallow basal infoldings devoid of mitochondria. Cells of the proximal Malpighian tubule possess concentric granules of different electron-density. The internal proximal ileum lined with a cuticle facing the lumen and contains secretory vesicles in its cytoplasm. Dense and long microvilli at the apical border of the conical segment cells are coated with abundant electron-dense fine granules. Cells of the anterior tubular midgut contain spherical secretory granules, oval secretory vesicles of different size, and autophagic vacuoles. Ferritin-like granules exist in the mid-midgut cells. The posterior tubular midgut consists of two cellular types: 1) cells with shallow and bulb-shaped basal infoldings containing numerous mitochondria, homocentric secretory granules, and fine electron-dense granules, and 2) cells with well-developed basal infoldings and regularly-arranged apical microvilli containing vesicles filled with fine granular materials. Cells of the distal tubular midgut are similar to those of the conical segment, but lack electron-dense fine granules coating the microvilli apex. Filamentous materials coat the microvilli of the conical segment, anterior and posterior extremities of the midgut, which are possibly the perimicrovillar membrane closely related to the nutrient absorption. The lumen of the hindgut is lined with a cuticle, beneath which are cells with poorly-developed infoldings possessing numerous mitochondria. Single-membraned or double-membraned microorganisms exist in the anterior and posterior extremities of the midgut, proximal Malpighian tubule and ileum; these are probably symbiotic.     

THE COLUMNAR CELLS OCCURRING IN THE PARIETAL LAYER OF BOWMAN''S CAPSULE : Cellular Fine Structure and Protein Transport

The renal corpuscles of adult, C3H Swiss, male mice contain testosterone-sensitive, columnar cells in the parietal layer of Bowman's capsule. A study of the normal fine structure of these cells reveals several distinctive characteristics: a microvillous brush border; apical tubular invaginations and apical tubules; an elaborate infolding of the basal surface membrane forming cellular compartments, which contain numerous mitochondria; and a complex group of membrane-limited cytoplasmic inclusions. This appearance is remarkably similar to the fine structure of cells in the proximal convoluted tubule. 1 hr after an in vivo injection of horseradish peroxidase, numerous protein-absorption droplets occur in the columnar cell cytoplasm. The speed and cytomorphology of protein transport by these capsular cells closely resemble the handling of peroxidase by the proximal convoluted tubule. Origins for these testosterone-sensitive cells are discussed briefly. Morphological evidence is presented for the differentiation of squamous cells in Bowman's parietal capsule into columnar cells, which appear structurally and functionally identical with proximal convoluted tubular epithelium.     

Ultrastructure and osmoregulatory function of the kidney in larvae of the Persian sturgeon Acipenser persicus

The localization of Na(+) , K(+) -ATPase (NKA) and the ultrastructural features of kidney were examined in larvae of the Persian sturgeon Acipenser persicus (L 31-41 mm total length and 182·3-417·3 mg). Investigations were conducted through light and electron microscopy and through immunofluorescence for NKA detection. The kidney nephrons consisted of a large glomerulus and tubules (neck, proximal, distal and collecting), which connected to the ureters. Posteriorly, ureters extended and joined together into a thin-walled ureter terminal sac. Ultrastructurally, the glomerular cells (podocytes) possessed distinctive pedicels that extended to the basal membrane. The proximal tubule (PT) showed two different cells. The cells lining the anterior part of PT possessed apical tall microvilli (c. 2·7 μm), a sub-apical tubular system, a basal nucleus and dense granules. Posteriorly in the cells, the sub-apical tubular system and granules were absent and round mitochondria associated with basolateral infoldings were found; the apical microvilli were reduced. Distal and collecting tubular cells showed the typical features of osmoregulatory cells, i.e. well-developed basolateral infoldings associated with numerous mitochondria. No immunofluorescence of NKA was detected in the glomeruli. A weak immunostaining was observed at the basolateral side of the cells lining the neck and PT. A strong immunostaining of NKA was observed in the entire cells of the distal tubules, collecting tubules and in some isolated cells of the ureters. In all immunostained cells, the basolateral region showed a much higher fluorescence and nuclei were immunonegative. In conclusion, the epithelial cells of kidney tubules had morphological and enzymatic features of ionocytes, particularly in the distal and collecting tubules. Thus, the kidney of A. persicus larvae possesses active ion exchange capabilities and, beside its implication in excretion, participates in osmoregulation.     

STUDIES ON THE DEPLETION AND ACCUMULATION OF MICROVILLI AND CHANGES IN THE TUBULOVESICULAR COMPARTMENT OF MOUSE PARIETAL CELLS IN RELATION TO GASTRIC ACID SECRETION

Gastric parietal cells in mice present a spectrum of microscopic appearances due mainly to variations in the abundance of the tubular and vesicular component of the cytoplasm and in the size and number of microvilli lining the intracellular canaliculi. Differences in the range of forms among parietal cells of fasting versus fed mice were not especially striking, but cells with very numerous tubules and vesicles were more common after fasting. However, in mice treated with drugs or hormones that induce acid secretion, parietal cells were more uniform in appearance. There was a marked reduction of these cytoplasmic membranes and a concomitant increase in both the number and size of microvilli. Measurements of acid secretion in control animals and in animals treated with acid secretagogues indicated hydrogen ion secretion contemporaneous with depletion of the cytoplasmic tubulovesicular membranes and with increase of the microvilli. In mice with inhibited acid secretion, parietal cells showed an accumulation of cytoplasmic tubules and vesicles and reduction in the numbers of microvilli. Stereological methods were used to quantitate 10 different parietal cell compartments. Tracer studies with lanthanum did not reveal continuity between the tubules and the plasma membrane. However, there were regions of close apposition between the tubulovesicular membranes and the cell membrane of the canaliculus, and instances where cytoplasmic tubules extended from the cell into the core of enlarged microvilli.     

SPORE GERMINATION AND DEVELOPMENT OF THE YOUNG GAMETOPHYTE OF THE OSTRICH FERN (MATTEUCCIA STRUTHIOPTERIS)

Light is required for the germination of spores of Matteuccia struthiopteris. Histochemical studies show that dormant spores contain no starch, but have an abundance of storage protein granules. Starch accumulates in the numerous chloroplasts of the spore on exposure to light and becomes gradually more extensive. Protein granules disappear as germination progresses. Following this, the centrally located nucleus migrates toward the proximal spore face. Concomitant with the nuclear migration, an increase of cytoplasmic RNA surrounding the nucleus occurs. An equal nuclear division and unequal cell division give rise to a 2-celled gametophyte consisting of a large prothallial cell and smaller rhizoidal cell. A new peripheral wall forms around the entire protoplast at the time of nuclear migration, while a transverse wall forms after nuclear division. The rhizoid emerges through the split raphe along the proximal spore face; it is rich in cytoplasmic RNA but contains very few chloroplasts and little starch. Electron microscopy of the 2-celled stage revealed a greater concentration of mitochondria, Golgi bodies, and a more extensive endoplasmic reticulum in the rhizoid than was found in the prothallial cell, which, however, was far richer in chloroplasts and lipid bodies. As the rhizoid elongates and becomes more vacuolated, cytoplasmic RNA decreases as cytoplasmic protein increases. The rhizoid undergoes no cell divisions, while the prothallial cell retains the potential for further cell division. The possible significance of the distribution of storage products, cell organelles, and other cell components were considered in relation to the non-equational cell division and differentiation of the 2 cells.     

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.     

Ultrastructure of midgut endocrine cells in the adult mosquito, Aedes aegypti

The ultrastructure of endocrine cells in the midgut of the adult mosquito, Aedes aegypti, resembled that of endocrine cells in the vertebrate gastro-intestinal tract. Midgut endocrine cells, positioned basally in the epithelium as single cells, were cone-shaped and smaller than the columnar digestive cells. The most distinctive characteristic of endocrine cells was numerous round secretory granules along the lateral and basal plasma membranes where contents of the granules were released by exocytosis. Secretory granules in each individual cell were exclusively of one type, either solid or 'haloed', and for all cells observed, the range in granule diameter was 60-120 nm. The cytoplasm varied in density from clear to dark. Lamellar bodies were prominent in the apical and lateral cellular regions and did not exhibit acid phosphatase activity. The basal plasma membrane was smooth adjacent to the basal lamina, whereas in digestive cells the membrane formed a labyrinth. Some endocrine cells reached the midgut lumen and were capped by microvilli; a system of vesicles and tubules extended from beneath the microvilli to the cell body. An estimated 500 endocrine cells were distributed in both the thoracic and abdominal regions of the adult midgut. In one midgut, we classified a sample of endocrine cells according to cytoplasmic density and granule type and size; endocrine cells with certain types of granules had specific distributions within the midgut.     

Seasonal changes of the Leydig cells of viscacha (Lagostomus maximus maximus). A light and electron microscopy study

The Leydig cells of viscacha (seasonal rodent) show cytoplasmic hypertrophy and regional distribution during the breeding period (summer-autumn). The dominant organelles are smooth endoplasmic reticulum (SER) and mitochondria. A moderately well-developed Golgi, abundant lipid inclusions, dense bodies like lysosomes in different stages, and centrioles are observed. Extensive or focal desmosome and gap-like junctions between neighbouring Leydig cells are present. These cells exhibit an evident hypotrophy and an increase in the number of dense bodies during the gonadal regression in winter (July and August). Cells in different stages of involution are observed in this period. Their nuclei are irregular and heterochromatic. The cytoplasm contains few mitochondria. The vesicular SER is scarse. Irregular and large intercellular spaces with microvilli and amorphous material are present. The junctional complexes are absent. The nuclear and cytoplasmic volume and development of SER and mitochondria increase during the recovery period (spring). The lipid inclusions decrease. Dilatations of the intercellular space with microvilli and limited by focal desmosome-like junctions are observed. In conclusion, the Leydig cells of Lagostomus maximus maximus show deep changes alongside the reproductive cycle. The photoperiod variations, through pineal hypothalamus pituitary axis and the hormone melatonin, are probably responsible for them. Moreover, the fall of serum and tubular testosterone would be one of the factors responsible for gonadal regression.     

Immunocytochemical localization of cathepsin D in lysosomes of cortical collecting tubule cells of the rat kidney

Immunocytochemical localization of cathepsin D in rat renal tubules was investigated by means of indirect immunoenzyme and protein A--gold techniques. By light microscopy, fine granular staining was seen in the mesangial cells of glomeruli. Heavy reaction deposits were present in the cortical tubular segments and some of the medullary collecting tubules. The proximal tubules contained a few positive granules. Other segments were negative for cathepsin D. By electron microscopy, gold particles representing the antigenic sites for cathepsin D were present in cytoplasmic granules and multivesicular bodies of the segment of the cortical collecting tubule. These cytoplasmic granules were presumed to be digestive vacuoles (secondary lysosomes) from their morphological profile. The proximal tubule cells contained the very weakly labeled secondary lysosomes. No specific labeling was noted in other segments of the nephron. Control experiments confirmed the specificity of the immunostaining. Quantitative analysis of the labeling density in each subcellular compartment also confirmed that the main subcellular sites for cathepsin D are the secondary lysosomes and multivesicular bodies. The labeling density in these granules of the lysosomal system varied widely with the individual granules, suggesting that there is a considerable heterogeneity of enzyme content among the granules of the lysosomal system. The prominent presence of cathepsin D in the cortical collecting tubule suggests a certain segment-specific function of this proteinase.     

ULTRASTRUCTURAL STUDIES OF SPERMATOGENESIS IN THE ANTHOCEROTALES. I. THE BLEPHAROPLAST AND ANTERIOR MITOCHONDRION IN PHAEOCEROS LAEVIS: EARLY DEVELOPMENT

Electron microscopic examination of thin sections showed that the blepharoplast of a young spermatid of Phaeoceros consists of two side-by-side centrioles and an accumulation of osmiophilic, granular matrix at their proximal ends. Lying between these nearly parallel organelles is a dark-staining body that will later disappear at the onset of flagellogenesis. For a brief period the centrioles are oriented perpendicular to the nuclear surface so that the granular matrix at their proximal ends is confluent with the nuclear envelope; furthermore, the nucleoplasm immediately in front of the centrioles becomes densely staining. The multilayered structure (MLS) develops directly under the centrioles. It comprises a band of 12 microtubules (the S₁ stratum) and three lower strata (S_2–4) whose constitutent lamellae are oriented at an oblique angle to the S₁ axis. While the S₁ tubules grow rearward over the nucleus which forms a beak adjacent to the posterior end of the lamellar strata, the centrioles are transformed into basal bodies with the distal growth of the axonemes and the proximal growth of the central cartwheels and lowermost triplets. The proximal ends of the basal bodies and the S₁ tubules overlying the lamellar strata are invested with osmiophilic matrix that extends down to the S₂ layer and may temporarily occlude the lamellar plates. At the onset of nuclear elongation an anterior mitochondrion becomes situated close beneath the lamellar strata which extend laterally beyond the S₁ tubules.     

Comparative histological, histochemical and ultrastructural studies of the nephron of selected snakes from the Egyptian area

The present study was aimed to compare and contrast the histochemical, histological and ultrastructural variations (microanatomical differences) in the nephrons of selected snake species, Eryx jaculus (Boidae), Psammophis sibilans (Colubridae), Naja haje (Elapidae) and Echis pyramidum (Viperidae) from Egypt. The structural studies were carried out by conventional light and electron microscopy. The nephron, the renal unit of snakes, consists of renal corpuscle, proximal tubule, intermediate segment, distal tubule and collecting tubule. The renal corpuscles have large capillaries with clear and dark fenestrated endothelial cells. The proximal tubule showed long microvilli, cytoplasmic vacuoles, developed endoplasmic reticulum and abundant mitochondria. The intermediate segment was lined by ciliated cells. The lining cells of the distal tubules showed few microvilli, abundant dense mitochondria and clear vesicles of mucous appeared in the terminal portion. The collecting tubules consisted of mucous cells. In summary, the ultra-structure studies of nephrons revealed several interspecies similarities and also some intra-species differences in species of snakes.     

Ultrastructure of spermiogenesis, spermatozoa and the tegument in Atriaster sp. (Platyhelminthes, Monogenea, Polyopisthocotylea, Microcotylidae)

Atriaster (Atriaster) sp., a gill parasite in the fish Diplodus cervinus caught near Dakar, Senegal, was studied by transmission electron microscopy. Spermiogenesis is similar to the general scheme in the Cercomeridae and particularly the polyopisthocotylean monogeneans, but shows two remarkable characteristics: (a) the zone of differentiation for each spermatid is separated from the common spermatid cytoplasmic mass by a region without longitudinal microtubules but with many mitochondria, herein termed the pre-zone of differentiation; (b) transverse sections of nuclei in late spermatids are polygonal with concave sides. These two characteristics are provisionally interpreted as autapomorphies, since observations in other species are lacking. The outer layer of the tegument is similar to that of other polyopisthocotylean monogeneans, and has microvilli and three kinds of cytoplasmic inclusions: granules with fibrous contents, granules with homogeneous contents, and elongate electron-dense bodies.     

Ultrastructure of the tubular nephron of Testudo graeca (Chelonia). A comparison between hibernating and non-hibernating animals

The tubular nephron of hibernating and non-hibernating specimens of Testudo graeca (Chelonia) was studied by means of conventional light and electron microscopy and histochemistry. The tubular nephron was composed of proximal, intermediate, distal and collecting tubules in both hibernating and non-hibernating animals. The cells of the proximal tubule showed long microvilli, cytoplasmic vacuoles, a developed endoplasmic reticulum and abundant mitochondria. Fat droplets were also observed. The intermediate segment was lined by ciliated and non-ciliated cells. The lining cells of the distal tubule presented few microvilli, abundant dense mitochondria and clear vesicles of mucous appearance in the terminal portion. Collecting ducts are composed of mucous and non-mucous cells. Mucous cells presented strong reaction to the histochemical techniques detecting sialo- and sulpho-mucins. During hibernation, a progressive vacuolar degeneration of the endoplasmic reticulum was observed in all the segments of tubular nephron, which may be caused by a massive intake of extracellular water into the cell.     

Fine structure of the Malpighian tubules in the housefly, Musca domestica

The epithelium of the Malpighian tubules in the housefly is comprised of four distinct cellular types. Type I cells are characterized by the presence of intimate associations between infoldings of basal plasma membrane and mitochondria. On the luminal surface, cytoplasm is extended into microvilli which contain mitochondria. Membrane-bound vacuoles in the cytoplasm seem to progressively accumulate granular material. Type II cells have dilated canaliculi. Microvilli lack mitochondria. The Type III cell has not been reported previously in Malpighian tubules. It has very well-developed granular endoplasmic reticulum which contains intracisternal bundles of tubules. Cytoplasm contains numerous electron dense bodies. Type IV cells occur in the common duct region of the Malpighian tubules. Mitochondria do not extend into the microvilli.     

Cytochemical correlates of structural sexual dimorphism in glandular tissues of the mouse. I. Studies of the renal glomerular capsule.

The parietal epithelium of Bowman's capsule has been analyzed by enzyme cytochemistry in kidneys of mice (C57BL/6J) from birth to 50 days of age. There is a greater tendency for cells in the central portions of the capsular crescent to be cuboidal in post-pubertal males than in pre-pubertal mice of either sex or in post-pubertal females where they are generally squamous; moreover, these heightened capsular cells have a distinct microvillous border. Cytochemical procedures were selected which might confirm the morphological suggestion that the cuboidal parietal epithelium possesses an absorptive capacity. The oxidoreductase activity of the mitochondria of the cuboidal cells of this layer is comparable to that of the columnar cells of the proximal convoluted tubule. The cytochrome oxidase activity of the mitochondria in both of these segments of the nephron is intense. This is in sharp contrast to the unreactive mitochondria in the squamous cells of the parietal epithelium. Furthermore, a striking heterogeneity in the degree of cytochrome oxidase activity is evident in the mitochondria of the cuboidal parietal cells as well as in the cells of the proximal tubules. In the former cells, active mitochondria were generally found near microvilli at the apical ends and in the areas of the basal infoldings whereas those in a central position were more frequently unreactive. The brush border of the cuboidal capsular epithelium had prominent alkaline phosphatase and aminopeptidase activities as has previously been observed in other brush borders. Functional capacity corresponding to the morphological and cytochemical specialization of the cuboidal capsular cells was demonstrated by their uptake of horseradish peroxidase. This exogenous protein tracer could be seen in apical vacuoles and phagosomes in the cuboidal parietal epithelium. The cytochemical resemblance of the cells of this epithelium to those of the proximal convoluted tubules suggests a similar involvement in resorption and perhaps in active transport. A possible relationship of this differentiation of the capsular epithelium to the proteinuria normal for adult male mice is discussed.     

THE FINE STRUCTURE OF NEURONS

1. Thin sections of representative neurons from intramural, sympathetic and dorsal root ganglia, medulla oblongata, and cerebellar cortex were studied with the aid of the electron microscope. 2. The Nissl substance of these neurons consists of masses of endoplasmic reticulum showing various degrees of orientation; upon and between the cisternae, tubules, and vesicles of the reticulum lie clusters of punctate granules, 10 to 30 mµ in diameter. 3. A second system of membranes can be distinguished from the endoplasmic reticulum of the Nissl bodies by shallower and more tightly packed cisternae and by absence of granules. Intermediate forms between the two membranous systems have been found. 4. The cytoplasm between Nissl bodies contains numerous mitochondria, rounded lipid inclusions, and fine filaments.     

Das Epithel der Tuba uterina des Neugeborenen Elektronenmikroskopische Befunde

Summary An electron microscopical study of the epithelium of the uterine tube was carried out in the newborn. Among the epithelial cells at least two morphologically well defined types can be distinguished: ciliated and non-ciliated cells.The ultrastructure of the cilia and related structures corresponds to what has been described by other authors in ciliated cells of various organs and of different species. Near the basal bodies of the cilia there is a concentration of vesicular mitochondria, which is thought to be evidence of a high metabolic activity in this region of the cell. Large opaque granules in the supranuclear zone of the ciliated cells are, it is suggested, paraplasmatic inclusions, perhaps supporting material for the ciliokinetic processes. There was no evidence of a secretory function of the ciliated cells.Among the non-ciliated cells, which in general show a straight lined luminal border with few microvilli, there are some cells containing dense granules, which are distributed throughout the cytoplasm and concentrated in the luminar side of the cell. The apical parts of these cells are protruding and sometimes digitated or branched; they contain accumulated granular materials and are separated from the rest of the cell after the formation of an intracellular plasmalemma. A similar detachment was found in an other cell type, but here the protruded apical parts of the cells are edematous and do not contain any visible secretory materials. It is uncertain if the detached cytoplasmic substances form a part of a specific secretory product; there are no secretory granules within the cytoplasm. On the contrary, the detachment of cytoplasmic parts may only accompany the excessive proliferation of cells which takes place during this period of growth.     

Further evidence for synthesis of screening pigment granules involved in the photosensory membrane turnover of the crayfish photoreceptor

Photosensory membrane degradation in crayfish occurs at first in multi-vesicular bodies (MVBs) and then, with the aid of lysosomal enzymes, in lysosome related lamellar bodies. In organ culture experiments with the isolated crayfish retina (Orconectes limosus) small screening pigment-like granules became visible under the electron microscope in such lamellar bodies and suggested a possible relation of photosensory membrane degradation and screening pigment granule synthesis. Chloroquine, an inhibitor of lysosomal activity, when added to the culture medium reduced the appearance of screening pigment-like granules in lamellar bodies, but led to the appearance of these granules in mature MVB's, indicating the involvement of lysosomal enzymes in the formation of pigmented lamellar bodies. In a second set of experiments the effect of bright light on the screening pigment granule ultrastructure of crayfish phoreceptors was investigated. It was found that after bright light exposure large numbers of little screening pigment granules (0.15-0.3 microns) were located between or close to rhabdomeral microvilli that were not at these sites in crayfish kept under natural light. MVB's were also reduced in size, and among the little screening pigmentary organelles granules of different electron density and morphology appeared. Additionally, vesicle flux to little screening pigment granules was detected. The screening pigment granules of the little type did not seem to be transported close to or between the microvilli, but appeared to be synthesized at these sites within little MVBs.     

In vitro culture of the proximal tubule of the bovine nephron

By means of a previously described method, viable pure tubules of the nephron were isolated in high yield from the outer cortex of the near-term foetal bovine kidney. The tubular suspension obtained was constituted almost exclusively of proximal segments (about 95%), whose cells were dispersed and grown as confluent primary cultures. The cultured proximal cells were shown to maintain in vitro, on glass or plastic surfaces, the same orientation as on the tubular basement membrane in vivo, with interdigitations extending from the base of the cells and along their full height. Numerous mitochondria and the typical cytoplasmic bodies of the proximal cell were retained in cells grown in vitro. A flagellum was seen in every cultured cell and was shown to be present in the proximal cell in vivo. There is a progressive change, in vitro, of the microvilli of the brush border, from a close-packed to a sparse distribution and to a decrease in height and a reduction in number. This in vitro regression to an earlier embryonic state was correlated with the ability of the proximal cells to synthesize in vitro an alpha-fetoprotein and with the loss in vitro of histiospecific antigen synthesis, confined in vivo to the brush border area. The confluent proximal cells became filled with microfilaments and microtubules, the significance of which is discussed.