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The application of scanning electron microscopy to the study of cell surfaces is limited in intact tissues, because extracellular material may often obscure the details of nonluminal surfaces. To remove connective tissue elements we have treated human skin and both kidney, and an autonomic ganglion of the rat with hydrochloric acid and collagenase. Regional variations in the basal surface of the nephron are noted following removal of the basement membrane. The basilar interdigitations of the cells of the proximal tubule appeared as parallel ridges encircling the tubule. Ridges on the parietal epithelium of Bowman's capsule were randomly arranged and alternated with smooth surfaces. The dermal surface of the human epidermis has an alveolar or honeycomb appearance due to the elevation of the epidermal ridges and numerous pits for the dermal pegs. At higher magnifications the basal surface of cells of the stratum germinativum possessed numerous and irregular projections. Neurons with their processes are evident in the autonomic ganglion. The soma of the neurons are enclosed by flattened satellite cells. Irregular spaces between opposed satellite cells are interpreted as regions for the passage of processes related to the ganglion cells. Nodes of Ranvier were clearly seen along nerve fibers. Some pitting of the nerve fibers was also noted. The HCl-collagenase method has the advantage of the removal of collagen and basement membrane while preserving the structural integrity of the cell surface.
Anat Rec 1976 Aug
PMID:Scanning electron microscopy of cell surfaces following removal of extracellular material. 18 20

Dogs are frequent subjects in experimental studies of renal physiology and pathology in spite of the paucity of information on their normal renal morphology. In this study, gross morphology, light microscopy, and scanning and transmission electron microscopy were used to describe dog renal anatomy. The dog has a multilobed kidney with the medulla fused into an elongate crest and a renal pelvis of elaborate shape. The outer zone of the medulla lacks a definitive outer stripe. The proximal tubule consists of four distinct anatomical segments. Dark cells are abundant in the collecting duct of the inner medulla. The majority of the nephron segments demonstrate remarkable similarities to those of the human kidney and less to those of the kidney of the laboratory rat.
Anat Rec 1979 May
PMID:Survey of the morphology of the dog kidney. 44 63

This study investigates the development of the vascular system of the puppy kidney (1-21 days after birth) after preparing casts of the renal vessels. At two days, the intrarenal vascular system distal to the afferent arteriole is strikingly different than that of the adult. The glomeruli of the outer cortex consist of a single dilated vessel while those of the mid and inner cortex posses an increasingly larger number of capillary loops. The efferent arterioles vary greatly in appearance from outer to inner cortex. Those in the nephrogenic zone are characteristically short and narrow and join a larger venous vessel termined a sinusoidal capillary. An efferent system somewhat similar to that of the adult is seen in the mid and inner cortex. One of the most obvious differences noted between the puppy and adult kidney is the relative lack of peritubular capillary networks throughout the cortex of the puppy kidney. The puppy possesses large, irregular vessels termed sinusoidal capillaries. The most rudimentary sinusoids are found in the outer cortex with more mature vessels in the inner cortex. The vascular arrangement of the efferent arteriole and sinusoidal capillary appears as a post-glomerular shunt. Functionally, the shunt would direct blood flow away from the proximal tubule and thus could result in a low extraction ratio and Tm for secreted solutes.
Anat Rec 1979 Jun
PMID:Development of the intrarenal vascular system of the puppy kidney. 46 22

Two- to four-day-old beagle puppy kidneys were preapred for transmission and scanning electron microscopy and compared to similarly prepared adult tissues. Proximal tubules of puppy kidneys which contained nephrons in various stages of differentiation were examined and maturational changes were described. Lateral surface contours of proximal tubular cells were initially smooth and relatively unfolded, but progressively acquired complex processes that may be recognized as lateral ridges and lateral-basal processes. Basal projections began as short, stubby processes and gradually took on either a narrow, plate-like or finger-like appearance. Mitochondria lysosomes and apical vacuoles increased in number as the tubules matured. Mitochondria lacked orientation in outer cortical tubules, but exhibited some vertical arrangement within basal processes in inner cortical tubules. Despite features indicating advanced maturation of tubules in the inner cortex, puppy kidneys lacked the lipid droplets characteristic of the adult. Thus, differentiation of this portion of the developing nephron into S1, S2 and S3 segments was not possible at day 4. Morphometric analyses of the lateral and basal membrane surface concentration of proximal convulted tubules from the puppy revealed all cells to have a significantly smaller membrane area than that of the adult. However, the inner cortical cells of the puppy had a greater surface concentration than those of the outer cortex. The reduced transport capacity of the puppy proximal tubule may be realted to the lack of segmentation and/or reduced lateral-basal surface area.
Anat Rec 1979 Oct
PMID:Maturation of the proximal tubule in the puppy kidney: a comparison to the adult. 50 92

Simple dissection techniques of samples to be examined in the scanning electron microscope allow one to visualize easily the three-dimensional shape of epithelial cells in situ. Such preparations reveal a complex system of ridges and folds on the lateral surface of the cells whose intricacy can best be appreciated with SEM. In many epithelia there is a smooth apical band which corresponds to the region occupied by the junctional complex previously identified with conventional EM techniques. The secretion of chylomicra that result from a fatty meal can be observed. It is possible to study the distribution of concanavalin A binding sites on the lateral surfaces of the cells utilizing hemocyanin as a marker. In the case of the proximal tubule epithelium, the apical cell surface has many more binding sites than the lateral cell surface and there is a sharp demarcation at the level of the apical band. After blunt dissection the relationship of the basal surface of the cells with the basement lamina and the basement membrane can be appreciated as well. Possible physiological meaning of the morphological features observed is briefly discussed.
Anat Rec 1975 Oct
PMID:Scanning electron microscopy of epithelia prepared by blunt dissection. 110 52

Diuresis and natriuresis follow volume expansion by intravenous infusion of either blood or saline. However, the mechanisms underlying the renal responses seem different for the two types of expansion. It has been suggested that the response to saline expansion is due to changes in physical factors with alterations in the morphology of the lateral intercellular space (LIS). The present study examines the width of the LIS of the proximal tubule after a Ringer-Locke (1%, 2.3%, or 7% body weight) or whole blood (2.3% body weight) infusion and in nonexpanded control. After Ringer-Locke infusion, a significant widening of the LIS of the proximal tubule is seen at all levels of expansion. There is a linear relationship between the level of the Ringer-Locke infusion and the degree of expansion of the LIS. In contrast, after blood volume expansion, the width of the LIS is the same as nonexpanded controls. These studies are consistent with the view that the renal response to saline expansion is mediated at least in part by physical factors affecting the proximal tubule.
Anat Rec 1975 Jun
PMID:The effects of Ringer-Locke or blood infusions on the lateral intercellular spaces of the rat proximal tubule. 115 97

Studies were carried out, using transmission electron microscopy, of the cilia of the nephrons of rat and human kidneys. Cilia were observed in the parietal layer of Bowman's capsule, in the proximal tubule, the distal tubule, including the macula densa, and the collecting duct. They had a number of characteristic features, including the presence of a centriole adjacent to the basal body, long, slender cross-striated rootlets, and a typically organized basal body. The shaft of the cilia differed from the typical 9 + 2 pattern of organization. Near the base of the cilia the pattern was 9 + 0. In the middle portion, one or more of the peripheral doublets had been displaced centrally to give an 8 + 1 or a 7 + 2 pattern, while towards the tip the pattern became more irregular and the doublets were reduced to single microtubules. We have hypothesized that these cilia may be vestigial. They may, if motile, have some minor stirring function, or they may play a sensory role, as has been postulated for similarly structured cilia in other sites.
Anat Rec 1975 Jul
PMID:Fine structure of mammalian renal cilia. 115 3

Development of the proximal tubule in the fetal kidney was studied following ligation of both ureters of pregnant rats. The ligation was performed on days 15, 17, 19, and 21 of gestation and autopsy followed 24 hours after each operation. On fetal days 20 and 22, the proximal tubular length per unit volume (1 mm3) of whole kidney of the fetuses from the ligated mothers was significantly increased, while the ratio of total glomerular volume to total proximal tubular volume was remarkably decreased when compared to that of the fetuses from the non-ligated mothers. Apical vacuoles in proximal tubular cells were increased by the ligation on fetal days 20 and 22. These findings suggest that maternal bilateral ureteral ligation accelerates the growth and differentiation of the proximal tubule in fetal kidney, when the fetal kidney is functional in urine production.
Anat Rec 1990 Dec
PMID:Effects of maternal bilateral ureteral ligation on the development of the proximal tubule of the kidney in fetal rats: morphometry and electron microscopic study. 228 62

Gene expression for calbindin-D28k, the 28,000 relative molecular mass vitamin D-dependent calcium-binding protein, was measured in cells of the murine nephron by in situ hybridization on tissue sections (hybridization cytochemistry). Radiolabeled (35S-UTP), single-stranded RNA complementary to calbindin-D28k-mRNA (probe RNA) was prepared from linearized cDNA template and used for the hybridizations. Autoradiography was carried out and cellular levels of hybridization signal (silver grains) were quantified. After correction for background the concentration of silver grains was more than 350% greater in the distal tubule than in either the proximal tubule or the glomerulus. The relative cellular level of mRNA in the cytoplasm, as reflected in silver grains/cell, of the distal tubules with probe RNA was 3.4 times greater than that with control RNA. Cells of the distal tubule were the only apparent sites of specific hybridization with probe RNA. The presence of calbindin-D28k-mRNA in the distal tubule corresponded to the localization of calbindin-D28k by immunocytochemistry.
Anat Rec 1990 Jun
PMID:Cellular gene expression for calbindin-D28k in mouse kidney. 235 3

Immunoelectron microscopic localizations of cathepsins B and H and injected horseradish peroxidase (HRP) in the lysosomal system of rat kidney proximal tubules were investigated by a protein A-gold technique. Kidneys were fixed by perfusion at fixed intervals after intravenous injection of HRP. At 5 to 15 minutes after the injection, the endocytic apparatus--including the apical vesicles, tubules, and vacuoles (endosomes)--were stained for HRP, but they were negative for cathepsins. Within 15 to 30 minutes after the HRP injection, HRP-containing endosomes were fusing with preexisting lysosomes. In the S1 segment, they accumulated in the apical cytoplasm and formed giant phagosomes, which increased markedly in number and size after 1 hour. These phagosomes were composed of a peripheral clear matrix and electron-dense inclusions. The clear matrix was stained heavily for cathepsins and HRP, whereas the electron-dense inclusions were consistently negative for cathepsins and HRP. The same results also were obtained after the double-labeling and serial sectioning techniques. The dense inclusions were fragmented gradually as the phagosomes decreased in size. After 3 hours, the size and number of phagosomes returned to their normal state (before the HRP injection). These results indicate that the endocytic apparatus of the proximal tubule cells does not contain cathepsins. Phagosomes are formed by the fusion of endosomes containing the internalized protein with the preexisting lysosomes. The degradation of HRP in giant phagosomes occurred rapidly. The coexistence of cathepsins B and H with the endocytosed HRP suggests that these cystein proteinases are involved in the degradation of protein in heterophagosomes of the proximal tubule cells.
Anat Rec 1988 Aug
PMID:Involvement of cathepsins B and H in lysosomal degradation of horseradish peroxidase endocytosed by the proximal tubule cells of the rat kidney: II. Immunocytochemical studies using protein A-gold technique applied to conventional and serial sections. 305 13


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