Calbindin-D immunolocalization in developing chick thyroid: a light and electron microscopic study

Antiserum to calbindin-D, a 28 KD vitamin D-dependent calcium binding protein, was used to localize the protein immunocytochemically in developing chick thyroid by both light and electron microscopy. The protein first appeared in future follicular cells of developing thyroid tissue from 8-day-old embryos. The number of calbindin-D-containing cells increased rapidly to a near-plateau level at day 10; this concentration was sustained until day 15, and then declined to an undetectable level just before hatching. The protein was distributed throughout organelle-free areas of the follicular cell cytoplasm and extended into the nucleus; it was not present in the follicular colloid. Comparison of the time course of changes in calbindin-D content with known differentiative changes taking place in follicular cells suggests that the protein may function in some yet to be determined mechanism related to normal development of the thyroid.     

Histomorphologic and histochemical investigations in mannosidosis. A light and electron microscopic study.

Morphologic and histochemical studies have been performed at light and electron microscopic level on needle-liver biopsy specimens, circulating blood lymphocytes and fibroblast cultures from patients with mannosidosis. The findings demonstrated generalized storage phenomena of varying degrees in the various tissues examined. Histochemical findings were in agreement with the biochemical nature of the stored material. Enzyme histochemical methods indicated storage in the lysosomes, at least in the hepatocytes. The ultrastructural appearance of mannosidosis in itself has but a limited diagnostic significance since the morphology and distribution of vacuoles have characteristics in common with other storage diseases. Repeated liver biopsy disclosed extensive storage in the hepatic tissue. However, the progression of the disease was not accompanied by severe mechanical destruction or microcirculatory disturbances.     

Inorganic trimetaphosphatase as a histochemical marker for lysosomes in light and electron microscopy.

A new cytochemical method is presented for the light and electron microscopic localization of lysosomes in mineralized and soft tissues. Inorganic trimetaphosphate is used as substrate in a lead chelate incubation medium at pH 3.9. Lysosomes in several tissues are strongly reactive, and reaction product is frequently present in Golgi saccules and GERL. The reaction can be differentiated from acid glycerophosphatase activity, is relatively insensitive to fixation and demineralization procedures, and the reaction is often complete after short incubation times.     

An electron microscopic histochemical and X-ray microprobe study of spherites in a mussel

Transmission electron microscopic, histochemical and X-ray analytical microprobe techniques were used to study the inorganic-organic relationship in the spherites (calcospherules) from the mantle, i.e. subadjacent to the outer mantle epithelium, of the fresh-water mussel Amblema. These structures were shown to contain calcium which could be chelated by the flotation of sections on solutions of either formic acid or ethylene glycol bis-(beta-amino ethyl ether)-N, N1-tetra-acetic acid (EGTA), Analysis of both non-chelated and sections revealed a significant sulfur peak. Chelated spherites were also intensely stained with acid phosphotungstic acid (PTA), Such data is indicative of the presence of an organic glycoprotein (proteoglycan) matrix which could serve to bind mineral ions, thus forming organic-inorganic aggregates for calcium transport and homeostasis. In this regard, the spherites are analogous to both calcium phosphate containing mitochondrial granules and the initial calcification sites in vertebrate mineralizing tissues.     

On the specificity of the histochemical technique for sarcoplasmic reticular adenosine triphosphatase: a light and electron microscopic study.

The specificity of the histochemical localization of the calcium activated adenosine triphosphatase (ATPase) activity of the sarcoplasmic reticulum (SR) at pH 7.4 was studied using a calcium-citro-phosphate technique. The latter involves the splitting of ATP by ATPase producing phosphate ions which then react with calcium and citrate to form an insoluble reaction product. This reaction product was detected by both light and electron microscopy. Light microscopic examination showed a darkly stained continuous reticular pattern of reaction product which surrounded individual myofibrils. This reticular pattern of reaction product was distinctly dissimilar to that found when the histochemical reactions for mitochondrial or myofibrillar ATPase were performed. Ultrastructural investigations demonstrated the presence of discrete foci of electron dense reaction product in close association with the membranes of the SR in striated muscle fibres. Only occasional flecks were seen in the vicinity of mitochondria or myofilaments. The possibility is considered that the reticular pattern of staining achieved by the calcium-citro-phosphate technique may reflect the distribution of the "extra ATPase" of the SR, an enzyme implicated in the process of calcium uptake and muscle relaxation.     

An electron microscopic and histochemical study of the secretory cells inHydra viridis

Summary The fresh water coelenterateHydra viridis possesses a unique distribution of mucous and serous secretory cells in the gastrodermis. The mucous cells are found only in the hypostome, a region devoid of the serous zymogen cells. On the other hand, the zymogen cells are found extending from the tentacles to the peduncle. Histochemical stains indicated that the two hypostomal mucous cells, spumous and granular, secreted an acid mucopolysaccharide, and incorporated radiosulfate. The radiosulfate label was not sensitive to hyaluronidase digestion, but was removed by acid methanolysis. In contrast, the secretory product of the zymogen cell was rich in proteins and a PAS-positive moiety (unsulfated).The ultrastructure of these cells was correlated with their histochemical staining properties. It was demonstrated that glutaraldehyde preserved the ultrastructure of the secretory granules better than osmium, and also preserved more components within the granules. The mucous cell granules contained an electrolucent and an electron dense component. The cells were both PAS-positive and alcianophilic. After osmium fixation the dense component was lost and the cells were primarily alcianophilic. Osmium also failed to preserve the electron dense component in the zymogen cells.Observations of corresponding thick and thin sections showed a cell containing granules similar to the granules seen in mouse Paneth cells. The dense core was osmiophilic and the lighter halo was alcianophilic.These results lead us to conclude that the electrolucent filamentous component is an alcianophilic acid mucopolysaccharide and the dense granular component is probably a PAS-positive material.

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Zusammenfassung Der FrischwassercölenteratHydra viridis weist eine einzigartige Verteilung von mukösen und serösen sekretorischen Zellen in der Gasterodermis auf. Die mukösen Zellen finden sich nur im Hypostom, in welchem seröse Zymogenzellen fehlen. Die Zymogenzellen andereseits finden sich von den Tentaklen bis zum Pedunkulus. Histochemische Methoden zeigten, daß die zwei Typen hypostomaler muköser Zellen, d.h. spumöse und granuläre, ein saures Mukopolysaccharid ausscheiden und radioaktives Sulfat einbauen. Der Radiosulfat-Markierer war nicht sensitiv gegenüber Hyaluronidase, konnte aber entfernt werden mit saurem Methanol. Im Gegensatz dazu war das Produkt der Zymogenzellen reich an Proteinen und enthielt PAS-positives Material.Die Feinstruktur dieser Zellen war korreliert mit diesen histochemischen Befunden. Glutaraldehyd erhielt die Feinstruktur der Sekretgranula besser und fixierte mehr Komponenten als Osmium. Die Granula der mukösen Zellen enthielten elektronendichte und -durchsichtige Komponenten; diese Zellen färbten sich mit PAS und Alcyan. Nach Osmium-Fixierung war die elektronendichte Komponente abwesend und die Zellen waren hauptsächlich alcyanophil. Auch in den Zymogenzellen vermochte Osmium nicht, die elektronendichte Komponente zu erhalten. Beobachtungen an alternierenden dicken und dünnen Schnitten zeigten eine Zelle mit Körnern ähnlich den Granulen von Maus Paneth-Zellen. Das dichte Zentrum dieser Granula war osmiophil, der hellere Halo alcyanophil.Diese Resultate lassen uns schließen, daß die elektronen-durchsichtige filamentöse Komponente ein alcyanophiles Mukopolysaccharid ist; das dichte, zentrale Material ist wahrscheinlich PAS-positiv.

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This paper was prepared from a thesis submitted in partial fulfillment for the degree of Master of Arts.

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This work was supported by the National Institutes of Health grant no. GM-11218.

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I wish to thank Dr.Marcus Singer for permission to use the E. M. facilities in the Dept. of Anatomy, Case Western Reserve University, and Dr.Joseph A.Grasso for instruction in the techniques of electron microscopy and the use of his facilities.     

Pigment cell development in rhesus monkey eyes: an electron microscopic and histochemical study

The ontogeny of pigment cells in the eyes of rhesus monkeys was studied by electron microscopy and histochemistry.In 60- to 80-day-old fetuses, the pigment epithelium of the iris and retina has already differentiated whereas stromal melanocytes of the uveal tract differentiate much later. The morphological and histochemical difference between melanocytes of the iris stroma and the choroid suggests that during embryonic development melanocytes migrate from the iris toward the ciliary body and choroid.Similarly, melanosomes of pigmented epithelial cells may have their origin in the epithelium of the anterior layer of the iris, which is metabolically more active than both the posterior layer and the pigment epithelium of the ciliary body and retina.     

A histochemical and electron microscopic study of a fast- and a slow-twitch muscle in genetically spastic mice

Fast and slow muscle fibers were studied in the flexor digitorum longus (FDL) and soleus (SOL) muscles, respectively, in control and spastic mice. HIstochemical and electron microscopic studies indicated an increased number of mitochondria, a decreased deposition of glycogen and a vesiculation and distension of the sarcoplasmic reticulum in many fast-twitch fibers of the spastic FDL. Similar findings were not evident in the slow-twitch fibers of the spastic SOL. Since the spastic condition causes increased muscular activity as a result of more rapid and prolonged nerve impulse firing, these findings reinforce the idea that a muscle fiber's oxidative capabilities are a function of its activity.     

Lectin-binding pattern in normal human gastric mucosa. A light and electron microscopic study

Normal human gastric mucosal cells were examined by light and electron microscopy using lectins as a probe. The ABC method was used with biotinylated lectins for light microscopy and HRP-labeled lectins for electron microscopy. The human gastric mucosal cells revealed specific binding patterns for each lectin by light microscopy. Among the lectins tested, in particular, DBA gave a characteristic pattern. It specifically stained the supranuclear region of surface epithelial cells and the perinuclear region of parietal cells. By electron microscopy, the stacked cisternae and the vesicles of the Golgi apparatus of the surface epithelial cells were positive for the DBA staining. These results show that the DBA-positive supranuclear region observed by light microscopy corresponds to the Golgi apparatus. In the parietal cells, DBA, RCA and ConA bound to the intracellular secretory canaliculi which are invaginations of the cell membrane running around the nucleus in the cytoplasm. Therefore, the tubular perinuclear positive region observed by light microscopy corresponds to the membranes of the intracellular secretory canaliculi. In addition, the ConA reagent stained the endoplasmic reticulum, Golgi apparatus, nuclear envelope, and cell membrane of the parietal cell, which explains the diffuse cytoplasmic staining observed at the light microscopic level with this lectin. Lectins have proved to be very useful for the evaluation of in situ cytochemical aspects of the glycoconjugates characteristic to human gastric mucosal cells.     

Elastase-1-secreting acinar cell carcinoma of the pancreas. A cytologic, electron microscopic and histochemical study

A case of acinar cell carcinoma of the pancreas was diagnosed by fine needle aspiration cytology during surgery. The cytologic characteristics of this neoplasm are described. Electron microscopy disclosed numerous zymogen granules in the tumor while histochemistry demonstrated the presence of elastase-1. Serum elastase-1 levels were markedly elevated. The cytologic differential diagnosis of pancreatic tumors is discussed.     

Three-dimensional scanning electron microscopic study of the normal hamster olfactory epithelium

Brain Cell Biology - The olfactory epithelium of the adult hamster (Mesocricetus auratus) was studied using the scanning electron microscope. A method that produced fractures in the epithelium...