E600-resistant Esterase Research Articles

Organic secretory products, adaptive responses and innervation in the parotid gland of ferret: A histochemical study

To qualify cellular events of possible pathophysiological significance in the parotid of ferret, tissue obtained post-mortem from mature animals of either sex was examined by light microscopical histochemistry for calcium, protein, amino acids, mucosubstances and hydrolases, and by neurohistology. Calcium was localised in acinar cells replete with granules containing protein, disulphides and usually carboxylated mucosubstances. Acid phosphatase activity was basally concentrated in the acinar cells. The granular luminal region of striated ductal cells showed protein, tryptophan, disulphides, neutral mucosubstances, and E600-sensitive esterase and Naphthol AS-D chloroacetate esterase activities, whereas their basal region showed acid phosphatase activity. Strong periluminal activity of acid phosphatase and E600-resistant esterase characterised the collecting ducts. Cholinesterase activity was associated with an extensive network of nerve fibres embracing parenchyma. Catecholamine fluorescence was not seen. beta-glucuronidase reactive macrophages abounded in the interstices. The results suggest that while the acini in the parotid of ferret secrete polyionic glycoproteins, shielded by calcium, the striated ducts secrete tryptophan-rich products comprising neutral glycoproteins and showing proteolytic activity. Innervation is of the cholinergic type and parenchymal lysosomal activity, possibly related to autophagy of stored secretory products and heterophagy of luminal material, is brisk. Macrophages contribute to maintaining the glandular microenvironment, wherein secretory activity appears to be lethargic.

Morphological phenotypes and functional capabilities of submandibular parenchymal cells of the ferret investigated by protein, mucosubstance and enzyme histochemistry.

Submandibular glands obtained post-mortem from mature ferrets of both sexes were examined with the use of light microscopical histochemical methods for proteins, mucosubstances and enzymes associated with cell functions or organelles. Demilunar cells showed carboxylated mucosubstances that were mainly non-sulphated, and diffuse activity for peroxidase, E600-sensitive esterase and acid phosphatase. Thiol groups were also detected in these cells. Central acinar cells showed sulphated mucosubstances, disulphides and reticular staining for thiamine pyrophosphatase. Intercalary ducts showed diffuse activity for NADH and NAD(P)H dehydrogenases. Striated ducts contained protein, tryptophan, disulphides, neutral mucosubstances and E600-sensitive esterase periluminally. Basally, the striated ductal cells showed variable activity for peroxidase, cytochrome oxidase, succinate dehydrogenase and acid phosphatase. Basolateral plasma membranes of these cells exhibited ouabain-sensitive Na,K-ATPase activity. The collecting ducts were characterized by variable periluminal staining for acid phosphatase, beta-glucuronidase, acid beta-galactosidase and E600-resistant esterase. The results suggest that the histological appearances of the acini of the submandibular gland of the ferret are dependent on the synthesis of secretory acid glycoproteins, that the striated ducts are involved with the secretion of tryptophan-rich product comprising neutral glycoproteins and showing esterase activity and with marked transport of ions and that the collecting ducts are involved with absorption.

Histochemical findings in the rat gastric mucosa during starvation.

The influence of starving on the activity of enzymes of the rat gastric mucosa was investigated by selected histochemical methods. Beside the conventional methods of enzymatic histochemistry the technique of semipermeable membranes was used in the proof of lysosomal enzymes. Dehydrogenases were proved in aqueous and also in gel media with PMS. During the starvation in the parietal cells a marked increase took place in the activity of acid phosphatase, E-600 resistant esterase, less in beta-glucuronidase. High activity of the lysosomal enzymes in macrophages did not change during starvation. Nor did any changes took place in the activity of alkaline phosphatase in the endothelium of the capillaries. The chief cells in the control and starving animals, in contrast to the human gastric mucosa, did not contain any non-specific esterase. Concerning dehydrogenases, parietal cells with a different activity of these enzymes were observed both in starved and control animals. In the rat gastric mucosa starving induced changes in the activity of the enzymes which mark important organelles of the cells. Thus it is possible to consider the observed histochemical changes as a functional manifestation of morphological damage of cellular structures which are affected during starvation.

Histochemical localization of enzymes in the normal rat gastric mucosa using the technique of the semipermeable membranes and the other methods

In the paper we observed histochemically the distribution and activity of 16 enzymes in the normal rat gastric mucosa. The lysosomal enzymes were demonstrated by the method of semipermeable membranes (LOJDA 1972). At the proof of dehydrogenases aqueous and gel media were used. The parietal cells of the gastric mucosa contained a moderate activity of acid phosphatase, E-600 resistant esterase, and only a very slight activity of beta-glucuronidase and N-acetyl-beta-D-glucosaminidase. The macrophages of the interstice contained a high activity of beta-glucruonidase, acid phosphatase, E-600 resistant esterase and a low activity of N-acetyl-beta-D-glucosaminidase. The chief cells of the rat gastric mucosa, in contrast to the human, did not contain nonspecific esterase and also in them acid phosphatase was mostly lacking. The alkaline phosphatase was found only in the endothelium of the capillaries of the gastric mucosa. The parietal cells contained high activities of succinate dehydrogenase, alpha-glycerophosphate dehydrogenase, beta-hydroxybutyrate dehydrogenase, NADH tetrazolium reductase, a lower activity of NADPH tetrazolium reductase, as well as other soluable dehydrogenases. At the examination of dehydrogenases using aqueous as well as gel media with PMS during optimal short incubation periods, we found more and less active forms of parietal cells. The different oxidoreductase capacity of parietal cells in normal rat gastric mucosa can point to their unequal-functional load at the production of hydrochloric acid. The findings obtained are compared with the findings in older papers concerning different experimental animals and with the distribution of enzymes in the human gastric mucosa.

The effects of ductal ligation on the parenchyma of salivary glands of cat studied by enzyme histochemical methods.

Submandibular, sublingual and parotid glands of cat have been studied following periods of ductal ligation ranging from 1 day to 1 year. An increased prominence of granules of acid phosphatase, beta-glucuronidase and E600-resistant esterase reaction products was sometimes seen in acinar cells and atropic striated ductal cells, and probably represents increased lysosomal enzymic activity, which may be of importance in the adapation of the parenchyma to the altered environment. Other reaction products often appeared to be at normal levels in parenchymal structures that were not very atrophic, and at reduced levels in those that were very atrophic, suggesting a reduction of functional activity in these structures.

Minor salivary glands of man: enzyme and mucosubstance histochemical studies.

Lower labial, buccal and sublingual glands have been examined. The lower labial glands contained a greater proportion of seromucous cells than the buccal and sublingual glands. In the latter glands there was a greater proportion of mucous cells that are stainable with the Alcian Blue critical electrolyte concentration technique with 0.2m MgCl2 and above than in the lower labial glands. Granules of acid phosphatase, β-glucuronidase and E600-resistant esterase reaction product, presumbly, representing lysosomal enzyme activity, were usually more prominent in seromucous than in mucous cells. Thiamine pyrophosphatase and inosine diphosphatase activities were demonstrated in a Golgi-like distribution, which was more prominent in mucous cells. Diffuse non-specific esterase, NADH-and NADPH- diaphorases and peroxidase activities were more prominent in seromucous cells. However, diffuse acid phosphatase activity was more prominent in mucous cells. Granules of β-glucuronidase, acid phosphatase and E600-resistant esterase reaction product were seen in ductal cells, and were often concentrated in the supranuclear region. Arylamidase activity was present lining the lumen of some ducts and acini.

Salivary glands of the cat: a histochemical study.

The submandibular, sublingual and parotid glands of the cat have been studied. Mucosubstance histochemistry demonstrated acidic mucosubstances with varying properties in the acini. Thiamine pyrophosphatase and nucleoside diphosphatase reaction products were seen with a Golgi-like appearance in acinar cells. Granules of acid phosphatase, β-glucuronidase and E600-resistant esterase reaction products, presumably representing lysosomal enzyme activities, were seen in acinar and ductal cells. Diffuse acid phosphatase and β-glucuronidase reaction products were seen in central cells of the submandibular acini, and diffuse non-specific esterase reaction product was seen in acinar and ductal cells. Arylamidase reaction product was associated with some acinar cells. Reaction product from a peroxidase technique was seen in demilunar cells of the submandibular acini, in parts of the sublingual acini, in parotid acini, and in ductal cells. Cytochrome oxidase and succinate dehydrogenase reaction products were seen most strongly in striated ducts, whereas NADH- and NADPH-diaphorase reaction products were seen at a high level throughout the ducts.

The ovary of the Guppy, Poecilia reticulata

The problem of the endocrine significance of the atretic follicle in the ovary of fishes has been approached with histological and cytological methods in the Guppy, Poecilia reticulata. The atretic follicles are subdivided into four stages: α-, β-, γ-, and δ-stage. During the period of pregnancy, lasting approximately one month, the atretic follicles were irregularly present. In 50 out of 73 animals, they were entirely absent. This appeared to have no influence upon a normal pregnancy; consequently, endocrine activity of the atretic follicle is not probable. The results of the cytochemical test also pointed in this direction. Enzymes, essential for steroid synthesis could not be proved to be present. Acid phosphatase and E-600 resistant esterase, however, were found in considerable amounts, indicating the presence of lysosomal activity. These results clearly show that the atretio follicle of the Guppy does not produce steroid hormones. Consequently, it is preferable to use the name Corpus atreticum instead of Corpus luteum praeovulationis. Moreover, it appears from cytochemical results that the degeneration process of the follicle can be subdivided into two phases: primarily the resorption of the oocyte during the α-stage, and secondly the regression of the granulosa cells during the subsequent stages.

Lysosomal changes associated with hyperoxia in the isolated perfused rat liver

Changes in lysosomes associated with hyperoxia were studied in the isolated rat liver, perfused by the method of Mayes and Felts (1966). Increased PO2 in perfused blood resulted in the formation of enlarged lysosomes and autophagic vacuoles. Demonstration of acid phosphatase (both by light and electron microscopy), β-glucuronidase, aryl sulfatase, and E600-resistant esterase in these organelles identified them as lysosomes. Electron-microscopic observations show that except for changes in the lysosomes, none of the other cellular organelles are distinctly affected. The formation of autophagic vacuoles and the cell damage are prevented if perfusion is carried out with blood oxygenated with air or if the liver is derived from a rat treated with chloroquine. These findings are attributed to alteration in the permeability of the lysosomal membrane resulting from hyperoxia. In turn, this permeability change releases hydrolytic enzymes into the cytoplasm leading to cell death, particularly in the centrilobular areas of the liver in fed and fasted rats. A special role in this process is envisaged for the smaller pericanalicular lysosomes, as indicated by a reduction in their number and a change in their distribution.

Nonspecific esterases in rat prostatic epithelial cells.

Prostates from normal and recently castrated rats were examined for the localization of esterase activity using Holt's indoxyl acetate method. Tissue fixed in formalin mixtures containing sucrose showed preservation of abundant cytoplasmic esterase activity while tissue fixed in formalin mixtures lacking sucrose had little cytoplasmic esterase activity. Esterase activity in droplets was abundant after both fixations. The difference could not be explained by ferroferricyanide inhibition, pH or osmolality of the fixative, and is attributed to a protective effect of sucrose during fixation. E600 greatly reduced cytoplasmic esterase activity in normal and castrated rats. Esterase activity in droplets was resistant to E600 in both normals and castrates. Ribonuclease extraction reduced cytoplasmic esterase activity in both normal and castrated rats only slightly, indicating no association of cytoplasmic esterases with ribosomal ribonucleic acid. Cytoplasmic esterase activity is reduced in castrates in which the epithelial cells of the prostate also show a decrease in endoplasmic reticulum and ribosomes. Similarity of location of droplet esterase activity and of lysosomes marked by acid phosphatase and reduction in droplet esterase activity by saline extraction are cited as evidence for a lysosomal localization of E600-resistant esterase.

Lysosomal enzymes in human endometrium: A histochemical study of acid phosphatase, nonspecific esterase, and E-600 resistant esterase

1.1. Three lysosomal enzymes: Acid phosphatase, nonspecific esterase, and E-600 resistant esterase, have been investigated histochemically in the human endometrium through the menstrual cycle.2.2. Cyclic variations of these enzymes were observed in the glandular epithelium, endometrial stroma, and macrophages, in relation to the various stages of the menstrual cycle.3.3. The relationships of these enzymes to the lysosomes and their regulation by ovarian sex steroid hormones is briefly discussed.4.4. The nonspecific esterase activity of the endometrial epithelium appears to be due the action of a lysosomal cathepsin or cathepsins.5.5. A more active role than hitherto believed is postulated for the endometrial macrophages, especially in relation to spermatocyte survival in the uterus.