Further enzyme histochemical observations on the segmentation of the proximal tubules in the kidney of the male rat

Summary The segmentation of the proximal tubules of the male rat kidney was studied by means of enzyme histochemical reactions. Soluble oxidoreductases (glucose 6-phosphate dehydrogenase, -glycerophosphate dehydrogenase, 3-hydroxysteroid dehydrogenase, NAD- and NADP-dependent isocitrate dehydrogenases, NAD-dependent malate dehydrogenase, NADP-dependent, decarboxylating malate dehydrogenase, uridine diphosphate glucose dehydrogenase) were demonstrated using methods which reduce enzyme diffusion (incubating in presence of polyvinyl alcohol) and eliminate interference from tissue tetrazolium reductases. Less soluble or insoluble enzymes (glucose 6-phosphatase, -hydroxybutyrate dehydrogenase, succinate dehydrogenase and tetrazolium reductases) were demonstrated by incubation in conventional watery media.Segmental differences were observed in respect to all enzymes studied, and most reactions clearly visualized the three segments known to exist from ultrastructural as well as previous histochemical studies: The pars convoluta includes the first (P₁) and most of the second (P₂) segment. The transition to the third segment (P₃) is in the beginning of the pars recta. Also these reactions revealed a difference between the first part of the P₃, which runs through the cortex in the medullary rays, and the terminal part transversing the outer stripe of the medulla. In most instances intensity of reaction decreased in the last portion of the P₃.A number of the enzymes studied were mainly or solely localized to the P₃ (glucose 6-phosphate dehydrogenase, -glycerophosphate dehydrogenases, -hydroxybutyrate dehydrogenase, 3-hydroxysteroid dehydrogenase, decarboxylating malate dehydrogenase and uridine diphosphate glucose dehydrogenase). Some possible functional implications of the findings are discussed.Supported by grants from Fonden til Lægevidenskabens Fremme and the Danish Medical Research Council. — Mr. Kaj L. Pedersen is thanked for valuable photographic assistance.     

Histological and histochemical observations on the testis of Gobius paganellus

Summary Histological and histochemical observations on the testis of Gobius paganellus during all seasons of the year are described. In the yearly reproductive cycle, spawning in the Gulf of Naples occured from June through August, testicular recovery and relative inactivity from September through December, and active spermatogenesis from January to May.Germ cells develop as clones from single primary spermatogonia, each clone in a follicle enveloped by cells which are interpreted as Sertoli cell homologues.Glandular tissue is present in large amounts both in the form of interstitial islets and as a large mass along the length of the mesorchium. Cholesterolpositive lipids and ⁵-3-hydroxysteroid dehydrogenase, presumptive evidence for steroid production, are present exclusively in this tissue.Lipids are present in the glandular cells in acidic and neutral forms. The acid fats are the more abundant, but neutral lipids increase in amount in the period April-June. There is, furthermore, a cyclic variation in lipid droplet size, small droplets being present in the spring (during active spermatogenesis) and fewer, larger droplets during the fall (post-spawning period). Phospholipids are lacking.Lactic dehydrogenase activity was weak in the relatively inactive postspawning period, but was much more intense during active spermatogenesis.The glandular tissue in the testis of Gobius is interpreted as homologous with the interstitial (Leydig) tissue found in the testes of higher vertebrates.This investigation was supported by research grant RG-6455 from the Division of General Medical Sciences, U.S. Public Health Service.Postdoctoral Fellow from the Division of General Medical Sciences, U.S. Public Health Service. Supported in part by U.S. Public Health Service Training Grant 5 Tl GM-136, Department of Biological Structure, University of Washington, School of Medicine, Seattle, Washington, U.S.A.     

Histological and histochemical observations on the forestomach of goat during pre-natal life.

Non-keratinized, stratified squamous epithelium lined the mucosa of the fore-stomach. The mucosal appendages in the form of ruminal papillae did not develop in the rumen during this study, however, omasal laminae and reticular ribs were evident in groups I and II, respectively. Cytoplasmic accumulations were evident in the middle layer of the epithelium, which is characteristic of this region. The thickness of the epithelium increased constantly in the rumen and reticulum, but in the omasum it remained constant with the increase in the size of the fetus. The corial papillae in the rumen, reticulum and omasum appeared at the 12.7-, 11.5-and 14.6-cm CVR stage, respectively. The muscularis mucosae appeared in the omasum at the 14.6-cm CVR stage in the first-order laminae. The tunica muscularis consisted of spirally arranged muscle fibre bundles. Cross-cut muscle fibre bundles below the tip of the omasal laminae descended downward with the increase in the size of the fetus and continued with the inner circular layer. Acid and alkaline phosphatases and fat could not be observed. The concentration of glycogen in the epithelium was maximum in group III and minimum in group II.     

Morphometric and enzyme histochemical behaviour of the kidney of young rats before and after unilateral nephrectomy.

Four week old male Wistar rats were unilaterally nephrectomized, and one, two, four, fourteen, and twenty-eight days after the operation the remaining kidneys were removed for morphological examination--including measurement of diameters of the tubules--and for histochemical investigation of enzymatic activities in the nephrons. The following enzymes were examined: succinate DH, lactate DH, alpha-glycerophosphate DH, isocitrate DH, glucose-6-phosphate DH, cytochrome oxidase, adenosine triphosphatase, alkaline and acid phosphatases. In comparison with kidneys of control animals of the same age and sex, the hypertrophy of the remaining kidney was found to be, even in these young rats, entirely due to enlargement of preexisting nephrons. The measurements have indicated that the hypertrophy was most distinct between the second and fourth day after uninephrectomy. Four weeks after the operation the diameters of proximal and distal tubules were about 21% and 18% respectively bigger in the uninephrectomized animals than in the corresponding controls. This increase in size was accompanied by some changes in intensity of enzymatic staining reactions; after an initial diminishment, a graduate increase in activity of almost all the observed tubular enzymes was found; this seems to be the expression of the functional compensation by the remaining kidney.     

Enzyme histochemical observations on the segmentation of the proximal tubules in the kidney of the female rat.

The segmentation of the proximal tubules in the kidney of the female rat was studied by means of enzyme histochemical reactions and the results compared with those observed in male and recently described by Jacobsen and J0rgensen (1973 a). Reactions were performed for the following soluble, coezyme-dependent oxido-reductases: glucose 6-phosphate dehydrogenase, alpha-glycerophosphate dehydrogenase, 3 alpha-hydroxysteroid dehydrogenase, NAD-as well as NADP-dependent isocitrate dehydrogenases, NAD-dependent malate dehydrogenase, NADP-dependent, decarboxylating malate dehydrogenase, uridine diphosphate glucose dehydrogenase. Measures were taken to reduce enzyme diffusion and eliminate interference from tissue tetrazolium reductases. Furthermore, reactions were performed for a number of less soluble or insoluble enzymes: glucose 6-phosphatase, mitochondrial alpha-glycerophosphate dehydrogenase, beta-hydroxybutyrate dehydrogenase, succinate dehydrogenase and tetrazolium reductases. In the proximal tubules of the female rat all enzymes studied--except beta-hydroxybutyrate dehydrogenase--showed segmental differences, most of them clearly revealing three segments. Sex differences were found concerning all enzymes except uridine diphosphate glucose dehydrogenase and NADP-dependent isocitrate dehydrogenase. The most pronounced sex-related differences were seen in the third segment in which part the male rat showed highest activity in respect to tetrazolium reductases, NAD-dependent isocitrate dehydrogenase, succinate dehydrogenase, beta-hydroxybutyrate dehydrogenase, 3 alpha-hydroxysteroid dehydrogenase and glucose 6-phosphate dehydrogenase and the female in respect to glucose 6-phosphatase, alpha-glycerophosphate dehydrogenases, and NADP-dependent, decarboxylating malate dehydrogenase. A few of the enzymes exhibited minor sex differences in the first two segments.     

Histological and enzyme histochemical changes in the kidney of male bullhead (Cottus gobio) during the spawning period

Histological and enzyme histochemical studies were carried out on the excretory kidney of the male bullhead ( Cottus gobio ). During the spawning season striking morphofunctional changes were observed in the second proximal segment of the kidney tubule. The tubular epithelium was greatly hypertrophied, strongly basophilic and produced a PAS-positive secretion. The enzyme histochemical pattern also changed conspicuously during this time: the alkaline phosphatase activity in the brush border was greatly reduced; the acid phosphatase and non-specific esterase activity in the cytoplasm was distinctly elevated.     

Histological and enzyme histochemical investigation of the hemal nodes of the hair goat

Abstract

We investigated the structure of the hemal node in six healthy hair goats using histological and enzyme histochemical methods. After processing, tissue sections were stained with Crossman's trichrome, Gordon-Sweet's silver and Pappenheim's panoptic stains. Alpha-naphthyl acetate esterase (ANAE) and acid phosphatase (ACP-ase) were demonstrated in frozen sections. Hemal nodes were encapsulated by connective tissue and few smooth muscle cells. Several trabeculae originated from the capsule and extended into the hemal node. A subcapsular sinus was present beneath the capsule and was continuous with the deeper sinuses. Subcapsular and deep sinuses were filled with erythrocytes. The parenchyma consisted of lymphoid follicles, diffuse interfollicular lymphocytes and irregular wide lymphoid cords. Cortical and medullary regions were not distinct. ANAE (+) and ACP-ase (+) cells were located mainly in the germinal centers of the lymphoid follicles and also were scattered equally in the interfollicular region and lymphoid cords. Monocytes, macrophages and reticular cells displayed a diffuse positive reaction, whereas localized granular positivity was observed in lymphocytes. We demonstrated that the general structure of the hair goat hemal nodes is similar to that of other ruminant species.     

Histological and enzyme histochemical investigation of the hemal nodes of the hair goat

We investigated the structure of the hemal node in six healthy hair goats using histological and enzyme histochemical methods. After processing, tissue sections were stained with Crossman's trichrome, Gordon-Sweet's silver and Pappenheim's panoptic stains. Alpha-naphthyl acetate esterase (ANAE) and acid phosphatase (ACP-ase) were demonstrated in frozen sections. Hemal nodes were encapsulated by connective tissue and few smooth muscle cells. Several trabeculae originated from the capsule and extended into the hemal node. A subcapsular sinus was present beneath the capsule and was continuous with the deeper sinuses. Subcapsular and deep sinuses were filled with erythrocytes. The parenchyma consisted of lymphoid follicles, diffuse interfollicular lymphocytes and irregular wide lymphoid cords. Cortical and medullary regions were not distinct. ANAE (+) and ACP-ase (+) cells were located mainly in the germinal centers of the lymphoid follicles and also were scattered equally in the interfollicular region and lymphoid cords. Monocytes, macrophages and reticular cells displayed a diffuse positive reaction, whereas localized granular positivity was observed in lymphocytes. We demonstrated that the general structure of the hair goat hemal nodes is similar to that of other ruminant species.     

Histological, histochemical, and fine structural observations on the spleen of seals

Spleens of three species of Antarctic seals with different diving habits (Weddell seal, crabeater seal, and fur seal) have been studied with histological, histochemical, and electron microscopic methods. The spleens can be classified as nonsinusoidal, with capsule and trabeculae rich in innervated smooth muscle cells. The trabecular system is particularly well developed in the deep- and long-diving Weddell seal. As in other mammals the pulp can be divided into white and red pulp. In the white pulp, periarteriolar lymphatic sheaths and secondary lymphatic nodules occur; both are surrounded by a marginal zone rich in macrophages and eosinophils. The nodules can be observed frequently, which is in accordance with abundance of plasma cells in the red pulp. Well-developed white pulp and numerous plasma cells and eosinophils obviously reflect a high load of nematodes, which have mainly been found in lung and stomach. Additionally, in the red pulp morphological evidence for the following functions has been found: destruction of erythrocytes, erythropoiesis, and thrombopoiesis. In respect to blood flow through the red pulp, we interpret our observations in the following way: terminal branches of arterioles open into the space between the fibroblastic reticulum cells; blood draining from here is collected into pulp veins, which are mainly found near the trabeculae. Thus, the seals have an open vascular compartment in their spleens, as also occurs in the cat. The red pulp is innervated by numerous nerve fibers that seem to include both cholinergic and adrenergic ones. The target cells of these fibers seem to be the fibroblastic reticulum cells, whose state of contraction may influence the direction of blood flow through the red pulp.     

Effects of an organosilicon compound on the tubular apparatus of rat kidney. A histological and enzyme histochemical report

Male adult Wistar rats were treated for 8 weeks with an organosilicone [2,2-dimethyl-4-(chloromethyl)-1,3-dioxa-2-silacyclopentane] employing two different dosages (25 and 50 mg/kg body weight i.p. daily) and the changes in the tubular apparatus of the kidney were investigated employing histological and enzyme-histochemical methods. The effects, more pronounced at the higher dosage, were the following: The brush borders of the proximal tubules were stuck together and disintegrated; only few epithelial cells remained intact showing a decreased activity of nonspecific esterases and the increase of beta-hydroxybutyric acid dehydrogenase. The nuclei of most of the epithelial cells in the distal tubules were dislocated towards the enlarged lumen and the cytoplasma showed a decrease of nonspecific esterases and an increase of beta-hydroxybutyric acid dehydrogenase. The collagen fibers in the walls of the collecting tubules were dislocated and disintegrated with a discontinuous border of Mg2+-ATPase; the nuclei of the epithelial cells were pyknotic, the cytoplasma showed an increase of beta-hydroxybutyric acid dehydrogenase. The induced changes were partially reversible after a recovery period of 8 weeks.     

Histological and histochemical observations in the stomach of the Senegal sole, Solea senegalensis

An histological and histochemical study was conducted on the stomach of adult Senegal sole, Solea senegalensis specimens. The stomach was made up of four distinct layers: mucosa, lamina propria-submucosa-, muscularis and serosa. Surface epithelial, glandular and rodlet cells were present in the mucosa. Cells of the columnar epithelium contained a basal nucleus. Numerous mitochondria, granular endoplasmic reticulum and Golgi apparatus consisting of several parallel cisternae and vesicles were observed in the cytoplasm of these cells. The lysosomes were small, round and dense. The gastric glands were numerous in the pyloric and fundic regions but absent in the cardiac stomach. These glands were formed by two cell-types: light and dark cells. The light cells were characterised by numerous mitochondria, while dark cells had slightly fewer mitochondria and a tubulo-vesicular system. Rodlet cells similar to those observed in other teleostean fish were present among the epithelial cells. Although the epithelial cells of the mucosa contained a weak presence of neutral and acid mucopolysaccharides/mucosubstances, these substances were abundant in the lamina propria-submucosa. Proteins rich in arginine, lysine, cysteine and cystine were rarely present in the mucosa and lamina propria-submucosa of stomach, while proteins rich in tyrosine were abundant in these layers. Acid phosphatase, and ATP-ase (pH 7.2 and 9.4) activities were detected in the mucosa and lamina propria-submucosa. Alkaline phosphatase activity was not detected.     

Histological and histochemical observations of the hemolymph cells in the crayfish, Orconectes virilis

Hemolymph cells of Orconectes virilis were stained during the months of August to November by a variety of histological and histochemical techniques. Cells were classified as hyaline cells, small granulocytes, and large granulocytes. Presence of mitochondrial enzymes was indicated by tests for succinic dehydrogenase and cytochrome oxidase. Reaction to test for the hydrolytic enzyme, leucine acylnaphthylamidase was intense in the granules of the large granulocytes. The PAS reaction indicated a mucopolysaccharide at the cell membrane. Lipid was found in all three hemolymph cell types of Orconectes virilis at the time of this study.     

Morphometric and enzyme histochemical behaviour of the kidney of young rats before and after unilateral nephrectomy

Summary Four week old male Wistar rats were unilaterally nephrectomized, and one, two, four, fourteen, and twenty-eight days after the operation the remaining kidneys were removed for morphological examination—including measurement of diameters of the tubules—and for histochemical investigation of enzymatic activities in the nephrons. The following enzymes were examined: succinate DH, lactate DH, α-glycerophosphate DH, isocitrate DH, glucose-6-phosphate DH, cytochrome oxidase, adenosine triphosphatase, alkaline and acid phosphatases. In comparison with kidneys of control animals of the same age and sex, the hypertrophy of the remaining kidney was found to be, even in these young rats, entirely due to enlargement of preexisting nephrons. The measurements have indicated that the hypertrophy was most distinct between the second and fourth day after uninephrectomy. Four weeks after the operation the diameters of proximal and distal tubules were about 21% and 18% respectively bigger in the uninephrectomized animals than in the corresponding controls. This increase in size was accompanied by some changes in intensity of enzymatic staining reactions; after an intial diminishment, a graduate increase in activity of almost all the observed tubular enzymes was found; this seems to be the expression of the functional compensation by the remaining kidney. Paper dedicated to Professor Dr. Drs. h. c. W. Bargmann (Kiel) in honour of the 70th anniversary of his birthday.     

A histochemical study about the influence of lytic enzymes on plasma membrane enzyme activities in rat liver and kidney.

1. The effect of lipolytic, glycolytic and proteolytic enzymes on the activities of plasma membrane enzyme activities in rat liver and kidney has been investigated by a pretreatment of tissue sections with the lytic enzymes. 2. The action of the proteolytic enzymes causes a very strong decrease of leucyl-beta-naphthylamidase activity, whereas the activities of ATP-ase, 5'-nucleotidase and alkaline phosphatase show a lesser decrease. This indicates a different membrane anchorage of leucyl-beta-naphthylamidase as compared to that of the phosphatases. 3. Treatment with glycolytic enzymes results in a decrease of 5'-nucleotidase and ATP-ase activity, whereas liver alkaline phosphatase and leucyl-beta-naphthylamidase show an increase in activity. 4. Treatment with phospholipase C gives about the same results. The very strong decrease of 5'-nucleotidase activity indicates a great dependence on phospholipids.