The ammocoetes endostyle: its oxidative enzymes as an evidence of its homology with the thyroid of higher chordates.

Histochemical methods were used for the demonstration of activity of the following intracellular oxidative enzymes, unstudied hitherto, in the epithelial cells of the endostyle of the river lampre (Lampertr aluviatilis L.) ammocoetes: reduced NAD dehydrogenase (NADD), lactate dehydrogenase (LD), cytochrome oxidase (CO), succinate dehydrogenase (SD), alpha-glycerophosphate dehydrogenase (alphaGPD) and glucose-6-phosphate dehydrogenase (G6PD). The activities of NADD and LD in the iodophil and throidogenic cells of type 3, then of subtype 2c and partly types 4 and 5 of the endostylar epithelium and the hypobranchial duct-lining epithelium were particularly ithe larva proves the possibility of their participation in the formation of the thyroid gland in the period of metamorphosis. In type 1 cells of the ammocoetes, despite their fairly strong enzymatic reactivity, the oxidative activity does not change significantly during the ontogenetic stages examined. The data obtained make it possible to modify the present views on the genesis of the thyroid gland of the adult lamprey, namely, they indicate the participation of the type 6 cells of the hypobranchial duct-lining epithelium in the process of thyroidogenesis.     

Enzymic markers of thyroid C cells in some rodents.

The paper provides comparative data of the localization of histochemical reactions demonstrating the activities of alpha-glycerophosphate and succinate dehydrogenases, acid phosphatase, non-specific esterases and non-specific acetylcholinesterase in the C cells of thyroids of 26 animals belonging to 5 rodent species. The family Muridae is represented by the Wistar albino rat and albino mouse, the family Microtidae by the bank vole Clethrionomys glareolus (Schreber 1780), the field vole Microtus agrestis L. 1761, and the pine vole Pitymys subterraneus De Selys-Longchamps 1825. The observed enzyme activity differences were most conspicuous on comparing the rat and mouse thyroids and in a much less degree the Microtidae thyroids. Among the histochemical reactions tested that for succinate dehydrogenase proved to be least effective as a C cell marker, alpha-glycerophosphate dehydrogenase being better, and acid phosphatase and non-specific esterases the best (not in the rat thyroid). The reaction for non-specific cholinesterase (with some limitations) gave satisfactory results in the C cells of all animal's thyroids. The present paper continues earlier studies [19] on the morphology of the C cells in thyroid glands of the rodents of the families Muridae and Microtidae and aims at supplementing them with histochemical data of enzymic activities. It deals with enzyme reactions that are employed as C cell markers in Mammals other than Rodents.     

Morphological and histochemical studies on the prostate gland of developing and adult Paramphistomum cervi (Digenea: Paramphistomatidae)

Morphological and histochemical studies have been made on the development of the prostate gland of the digenetic trematode, Paramphistomum cervi during the course of its infection in sheep; histochemical characterization of prostate gland secretion in adult worms and its functional relationship with the transport of spermatozoa have also been studied. In 6-wk-old worms, the terminal portion of the male genital duct is formed of what appears to be a syncytial epithelium containing a relatively large number of nuclei compared to the rest of the male duct. Cellular organization of the prostate gland becomes conspicuous in 8-wk-old worms and the prostate gland is fully developed by 16 wk. Two types of prostate gland cells, characteristic of the adult, become distinct in 16-wk-old worms which contain spermatozoa in the lumen of their vas deferens and pars prostatica. Adult worms show two types of prostate gland cells: type I containing mainly glycoprotein and type II mainly phospholipid granules. Weak-to-strong activities of acid phosphatase, alkaline phosphatase, esterase, lipase, adenosine triphosphatase, tetrazolium reductase, NAD-diaphorase, isocitrate dehydrogenase, glyceraldehyde dehydrogenase, α-glycerophosphate dehydrogenase and glucose-6-phosphate dehydrogenase have been observed in the prostate gland. The release of prostate gland secretion and the passage of spermatozoa through the lumen of pars prostatica appear to be synchronized events.     

Histochemical phenotypes of von Ebner's gland of ferret and their functional implications

Von Ebner's gland of ferret was examined by means of light microscopy, protein, mucosubstance and enzyme histochemistry, and neurohistology. Acinar cells were replete with granules containing neutral mucosubstances and disulphides, and showed strong diffuse acid phosphatase activity and weak granular staining for peroxidase. Staining for cytochrome oxidase, succinate dehydrogenase, and NADH and NAD(P)H dehydrogenases was also seen. Basolateral plasmalemma of acinar cells showed weak, ouabain-sensitive Na⁺,K⁺-ATPase activity. Ductal cells were of a simple appearance, contained thiols and showed variable staining for acid phosphatase, dehydrogenases and cytochrome oxidase. Variable amounts of -glucuronidase reaction product were localized in the glandular parenchyma, being marked in atrophic areas. Prominent stellate myoepithelial cells embracing acini and also basal ductal cells were demonstrated by alkaline phosphatase. Thiamine pyrophosphatase reaction product was concentrated in blood vessels around parenchyma, with little Golgi-like staining in acinar cells. Acetylcholinesterase activity was associated with an extensive network of nerve fibres embracing parenchyma, whereas catecholamine fluorescence was not seen. The results suggest that the acini of von Ebner's gland of ferret synthesise neutral secretory glycoproteins and peroxidase. Water mobilization is inconspicuous. Lysosomal activities feature in the parenchyma, possibly a consequence of processing secretory products in acini, absorption in ducts and/or adaptation atrophy. The gland receives a rich cholinergic-type innervation, and has extensive myoepithelial and microvascularbreak networks.     

Fine structural aspects of the black thyroid induced by minocycline, and the effects of a low iodine diet, propylthiouracil, thyroxine tablet and TSH, on the black discoloration of the rat thyroid

Fine structural aspects of the effect of minocycline, an antibiotic of the tetracycline group, on the rat thyroid were studied. In all the rats administered minocycline (100 mg/kg/day) for 21 days, diffuse black discoloration of the thyroid gland occurred. However, when the rats were fed on a low iodine diet, given propylthiouracil (PTU) or thyroxine tablet with minocycline the black pigmentation of the thyroid gland did not take place. On the other hand, black discoloration of the thyroid was accelerated in the rats administered TSH and minocycline simultaneously. Ultrastructurally, numerous dense bodies containing highly electron-dense deposits were seen in the supranuclear region of the follicular epithelial cells of the black thyroid. These dense bodies, which showed positive acid phosphatase activity, are considered to be lysosomes containing minocycline or its derivatives. It is speculated that minocycline is taken up into follicular epithelial cells with iodine, and that the black discoloration of the thyroid gland is intimately related to iodine metabolism.     

Morphological changes in the thyroid gland of rats during various phases of the estral cycle]

The functional state of the thyroid gland and the concentration of thyroid hormones in the peripheral blood were studied in 20 mature female albino rats during their estral cycle. Evaluation of the thyroid functional state was made according to data of histological, morphological (the diameter of folliculi, the height of the thyroid epithelium) and histochemical analysis (determination of NAD and NADP-dehydrogenase, succinatedehydrogenase, lactate dehydrogenase, peroxydase, acid and alkaline phosphatase) as well as biochemical determination of iodine bound with protein (IBP) in the blood plasma and investigation of the ratio of the parameters in question under conditions of the sex cycle. The cyclic changes of the morphological state of the thyroid gland attended by the phases of the estral cycle were revealed. The activation of the organ was observed in proestrus and estrus which was evidenced by high levels of activity of the enzymes under study, high concentration of IBP in the blood and increased height of thyreocytes. A decreased function of the thyroid parenchyma was observed at the period of metaestrus-diestrus.     

Reduction of mitochondrial pyruvate dehydrogenase phosphatase activity in lactating rat mammary gland following starvation or insulin deprivation.

The initial rates of activation of inactivated pyruvate dehydrogenase from lactating rat mammary gland and from pig heart were employed to assay pyruvate dehydrogenase phosphatase activity in mammary gland mitochondrial extracts. 24 h-starvation or 3 h-deprivation of insulin diminished phosphatase activity compared to fed controls. Refeeding and insulin treatment of 24 h starved animals restored in 1 h control levels of phosphatase activity.     

Histochemical study of enzyme patterns in the human submaxillary gland

Summary The histochemical distribution of various enzymes, such as alkaline phosphatase, acid phosphatase, esterase, -glycosidase, aminopeptidase, succinic dehydrogenese and TPN diaphorase, in human submaxillary glands has been determined.Acini and ducts of human submaxillary gland were devoid of alkaline phosphatase activity, but this enzyme was observed in capillaries and somewhat in myoepithelium.Activities of acid phosphatase, esterase, -glucuronidase and -galactosidase were generally observed in the entire cytoplasm of serous acini; but the cytoplasm of mucous acini was either negative or showed only trace amounts.Aminopeptidase reaction of both acini and ducts was generally negative.Succinic dehydrogenase and TPN diaphorase activities were strongly active in intralobuler ducts. Serous acini exhibited less activity with these enzymes; and mucous cells showed still less and were almost negative. In serous acini, there was much greater activity of TPN diaphorase than of succinic dehydrogenase.With 7 Figures in the Text     

Cytochemical contributions to differentiating GERL from the Golgi apparatus

Synopsis Recent studies from our laboratory are described which deal with endocrine cells (insulinoma, -cells of the pancreas, thyroid epithelial cells), pancreatic exocrine cells, and hepatocytes. These emphasize the importance of the hydrolase-rich specialized region of endoplasmic reticulum, known as GERL, in secretory cells. Also reviewed in this paper are the varied molecular transformations which apparently occur in GERL in different cell types, as reported from other laboratories as well as our own. Evidence of the continuity of GERL with rough endoplasmic reticulum is presented. Two hydrolytic enzyme activities in GERL, in addition to acid phosphatase activity, are recorded. Finally, the use of cytochemical staining procedures in the study of microperoxisomes is briefly described. The Histochemical Journal lecture 1976. Delivered to the Histochemistry and Cytochemistry Section of the Royal Microscopical Society on 14 September 1976     

Quantitative histology of the rat thyroid. Influence of histologic techniques on the morphometric data

The influence of various histologic techniques on the results obtained by morphometric analysis of the rat thyroid gland was studied. The limits of thyroid follicles were more clearly defined in both silver-impregnated paraffin-embedded sections and resin-embedded semithin sections than in routinely stained paraffin-embedded sections, thus enabling more accurate measurements of thyroid structures. Due to its simplicity, the silver impregnation method is clearly useful for histomorphometric studies when large numbers of measurements are involved. C cells were easily identified in paraffin-embedded sections by immunohistochemical staining. The measurement of interstitial tissue in sections without immunostaining of C cells led to an overestimation of the volume fraction of interstitial tissue.     

Autoradiographic localization of 3H-GABA uptake in the thyroid gland of the rat

Summary The uptake of -aminobutyric acid (GABA) in the thyroid gland of the rat was studied autoradiographically following in vitro incubation.High-affinity GABA uptake was localized in follicle cells, whereas C cells (parafollicular cells) in general did not accumulate GABA by high-affinity transport. The follicle cells were also the main sites of low-affinity GABA uptake. Additionally, some nerve fibres were found to accumulate GABA.The predominant localization of GABA uptake in follicle cells is discussed in view of a presumed role of GABA in thyroid function.     

Role of the glycerophosphate shunt in regulating oxidation of cytoplasmic NADH and ATP synthesis by liver mitochondria during rat ontogeny

The activity of alpha-glycerophosphate dehydrogenase rate of alpha-glycerophosphate oxidation and ATP synthesis were determined in the liver mitochondria of 1, 3, 12 and 24-month old rats in norm and two days after thyroxin daily administration in a dose of 0.25 mg per 100 g of body mass. The value of the studied parameters decreases with the age, most considerably--from one to three months and then--from 3 to 24-month age. This evidence for the fact that the role of glycerophosphate shuttle in the energy supply of cells falls with ageing; thyroxin administration evoked almost a two-fold increase of the mentioned parameters in one-month rats and more than three-fold in 12 and 24-month animals. A decrease in the alpha-glycerophosphate dehydrogenase activity with ageing is supposed to be connected with the thyroid gland function.     

Ontogeny and ultrastructure of somatostatin and calcitonin cells in the thyroid gland of the rat

Summary Calcitonin cells are relatively numerous in the thyroid gland of the rat. In contrast, somatostatin cells are very scarce except at the time of birth and a few days thereafter, when they are conspicuously numerous. Somatostatin cells of the thyroid gland, which are ultrastructurally similar to somatostatin cells in gut and pancreas, also contain immunoreactive calcitonin. It is not clear whether somatostatin cells in the rat thyroid gland produce calcitonin or accumulate calcitonin from the environment.     

Localization of some enzymes in the main venom glands of viperid snakes

Several secretory and nonsecretory enzymes were localized histochemically in the main venom gland of 13 viperid snakes. All secretory cells show the intracellular oxidative enzymes succinate dehydrogenase and monoamine oxidase. The granular reactions obtained for both enzymes resemble mitochondria in distribution. Distinctive cells with a very high succinate dehydrogenase activity are dispersed among the secretory cells of all species except Atractaspis. Nonspecific acid phosphatase activity is found in the supranuclear region of the secretory cells in species that do not secrete this enzyme and throughout the cytoplasm in snakes that secrete the enzyme. Nonspecific alkaline phosphatase activity occurs in the secretory cells of those snakes whose venom shows this activity. Leucine amino peptidase (aryl amidase) activity is found in the venom and in the secretory cells of all the species. In Vipera palaestinae both the venom and the secretory cells of the main venom gland contain nonspecific esterase, L-amino acid oxidase and phosphodiesterase activities. The localization of phosphodiesterase and L-amino acid oxidase do not show major differences between glands at different intervals from an initial milking. Adenosine-monophosphate phosphatase activity is localized in the supranuclear region of the secretory cells in the glands of Vipera palaestinae and Aspis cerastes. Its activity is found in the venom of Aspis only.     

Presence of chromogranin A, B and C in bovine endocrine and nervous tissues: a comparative immunohistochemical study

Summary Antisera against chromogranin A, B and C were used to study the distribution of these acidic proteins in bovine endocrine and nervous tissues. The three chromogranins occur together in several endocrine organs (adrenal medulla, anterior pituitary, endocrine pancreas) and in sympathetic ganglion cells. In the posterior pituitary, only chromogranin C and in the intermediate lobe only A and C are found. The parathyroid gland contains only A, and enterochromaffin cells are immunoreactive for A and B. Cells of the thyroid gland and some cells of the anterior pituitary apparently do not contain any chromogranins. It is concluded that the three chromogranins are not always stored together and that they are not present in all endocrine cells. This distinct localization of the chromogranins indicates some special, although still undiscovered, function for these proteins.     

Immunocytochemical study on the C cells in pig thyroid glands

The distribution of pig thyroid C cells was demonstrated by a immunoperoxidase method using antiserum to porcine calcitonin. The average number of C cells per 4 mm2 counted in the deep and superficial regions was 127 and 55, respectively. The number in the cranial, middle and caudal regions was 60, 128 and 83, respectively. The number of C cells per follicular and C cells (C cell concentration) in deep and superficial regions was 4.9 and 2.6%. It was 3.2, 4.9 and 3.3% in the cranial, middle and caudal regions, respectively. These results indicate that the number and concentration of C cells were not only larger in deep than superficial regions but also in the middle region than in cranial and caudal in pig thyroid gland. These results also indicate that the C cell distribution in pig thyroid gland is the same as that of dog, human, cat, rat, guinea pig, and mouse.     

Electron microscopic and ultracytochemical investigations of thyroid C cells in primary hyperparathyroidism

The subject of study were C Cells of the human thyroid in course of primary hyperparathyroidism caused by the presence of a single adenoma. C cells were estimated by routine electron microscopic technique and by cytochemical cholinesterase (ChE) reaction according to Iwayama in electron microscopic technique. The presence of single C cells and of a small agglomeration of these cells, parafollicularily localized was found in the thyroid gland. An electron dense reaction product was noticed on membraneous structures of C cells of the thyroid.     

Enzyme histochemical studies on rat lungs 2. Normal enzyme distribution in the alveolar tissue of the mature lung

Synopsis The activity and distribution of the following eighteen oxidative and hydrolytic enzyme systems have been investigated in the lung of the adult rat: reduced NAD dehydrogenase, reduced NADP dehydrogenase, succinate dehydrogenase, malate dehydrogenase, isocitrate dehydrogenase, lactate dehydrogenase, glucose-6-phosphate dehydrogenase, glucose dehydrogenase, glutamate dehydrogenase, -hydroxybutyrate dehydrogenase, acid phosphatase, alkaline phosphatase, glucose-6-phosphatase, adenosine triphosphatase, 5-nucleotidase, non-specific esterase, cytochrome oxidase and -glucuronidase.The low concentration of cells in sections of inflated lung may have made histochemical demonstration of some enzymes impossible because the enzyme concentration was below that detectable by the method employed.The carboxylic acid cycle and the hexose monophosphate shunt were potentially active but fatty acid metabolism was not indicated.The granular reaction sometimes encountered in alveolar cell cytoplasm may be useful for differentiating alveolar cell types, but further cytochemical studies are required to resolve the possible metabolic differences of alveolar cells.     

Solid cell nests of the thyroid gland. Optic and immunohistochemical study at autopsies.

In order to establish the prevalence of solid cell nests (SCN) in adult thyroids, we studied 100 consecutive glands at necropsy. These were serially sectioned and stained with routine and immunoperoxidase techniques in order to detect calcitonin, carcinoembryonic antigen, thyroglobulin and keratin. SCN may be considered as normal thyroid gland components, and they share with C cells a common origin in the ultimobranchial body.     

Change in the adrenal cortex of rats in stress after hypophysectomy, thyroidectomy and castration

To elucidate factors producing destructive changes in the adrenal cortex under severe stress, the following experiments were performed: 1) severe combined stress (KC) including unilateral adrenalectomy and fixation in supin position for 24 h (control group); 2)hypophysectomy KC; 3) thyroidectomy KC; 4) bilateral orchiectomy KC; 5) injection of vitamine "E" for 14 days + KC. The adrenals were studied by a complex of morphometrical and histochemical methods. Hypophysectomy prevented estructive changes in the adrenal cortex under KC. This effect was not connected with hypofunction of the thyroid gland and the testes, as their excision (in rats without hypophysectomy) did not prevent the development of necrobiotic changes in the adrenal cortex. A suggestion is made on a "corticolytic" effect of ACTH which reveals itself under severe stress. Destruction of adrenocorticocytes is supposed to result from activation of peroxide oxidation of lipids. This is indirectly supported by a large number of lipofuscin containing cells, by decrease in the activity of alkaline phosphatase and ATPh-ase, by activation of acid phosphatase and also by decreasing alterative phenomena at the injection of vitamine "E".