scholarly journals IN VIVO INCORPORATION OF 3H-GALACTOSE BY THYROID FOLLICULAR CELLS OF THE RAT, AS SHOWN BY ELECTRON MICROSCOPIC RADIOAUTOGRAPHY

1972 ◽  
Vol 20 (3) ◽  
pp. 220-224 ◽  
Author(s):  
A. HADDAD

Radioactive galactose was injected intravenously into rats and localized in thyroid follicular cells by electron microscopic radioautography at intervals ranging from 2.5 to 30 min after injection. The galactose label was mostly present in the Golgi apparatus at 2.5 min, with some of it in the adjacent rough endoplasmic reticulum. By 30 min, the label was found in apical vesicles and colloid. It was concluded that galactose is added to the carbohydrate side chains of incomplete thyroglobulin molecules during their travel through the cisternae of the endoplasmic reticulum into the Golgi apparatus; the uptake begins as this organelle is approached, but predominates within it. The thyroglobulin molecule which has thus been labeled is transported by the apical vesicles to the colloid.

1971 ◽  
Vol 49 (3) ◽  
pp. 856-882 ◽  
Author(s):  
A. Haddad ◽  
Meredith D. Smith ◽  
Annette Herscovics ◽  
N. J. Nadler ◽  
C. P. Leblond

The incorporation of fucose-3H in rat thyroid follicles was studied by radioautography in the light and electron microscopes to determine the site of fucose incorporation into the carbohydrate side chains of thyroglobulin, and to follow the migration of thyroglobulin once it had been labeled with fucose-3H. Radioautographs were examined quantitatively in vivo at several times after injection of fucose-3H into rats, and in vitro following pulse-labeling of thyroid lobes in medium containing fucose-3H. At 3–5 min following fucose-3H administration in vivo, 85% of the silver grains were localized over the Golgi apparatus of thyroid follicular cells. By 20 min, silver grains appeared over apical vesicles, and by 1 hr over the colloid. At 4 hr, nearly all of the silver grains had migrated out of the cells into the colloid. Analysis of the changes in concentration of label with time showed that radioactivity over the Golgi apparatus increased for about 20 min and then decreased, while that over apical vesicles increased to reach a maximum at 35 min. Later, the concentration of label over the apical vesicles decreased, while that over the colloid increased. Similar results were obtained in vitro. It is concluded that fucose, which is located at the end of some of the carbohydrate side chains, is incorporated into thyroglobulin within the Golgi apparatus of thyroid follicular cells, thereby indicating that some of these side chains are completed there. Furthermore, the kinetic analysis demonstrates that apical vesicles are the secretion granules which transport thyroglobulin from the Golgi apparatus to the apex of the cell and release it into the colloid.


1982 ◽  
Vol 58 (1) ◽  
pp. 79-93
Author(s):  
C.J. Flickinger

The synthesis, transport, and disposition of material labelled with [3H]mannose were studied by electron microscopic radioautography in normal amoebae and in cells that had internalized cell surface as a result of being induced to undergo pinocytosis. Control amoebae were injected with the precursor and placed in normal medium. The Golgi apparatus and rough endoplasmic reticulum were heavily labelled at the earliest intervals, while radioactivity of the cell surface peaked 12 h after injection of precursor. The experimental cells were injected, placed in bovine serum albumin solution from 15 to 60 min after injection, and then removed to normal medium until fixation. Incorporation of the precursor into the rough endoplasmic reticulum was near normal, but the proportions of grains associated with the Golgi apparatus and the cell surface were greatly reduced. The percentage of grains overlying vacuoles increased 12 h after injection, notably in the case of polymorphous vacuoles and dense vacuoles, both of which were identified as lysosomes with the acid phosphatase reaction. The results suggest that addition to the surface of components labelled with [3H]mannose was diminished following induction of pinocytosis. Incorporation of the precursor appeared to be shifted from cell surface material to lysosomal contents, possibly lysosomal enzymes. It is thought that this shift occurred in response to the need for the cell to digest unusually large amounts of endocytosed protein. Recycling of cell surface under these conditions is considered possible.


1969 ◽  
Vol 43 (2) ◽  
pp. 289-311 ◽  
Author(s):  
P. Whur ◽  
Annette Herscovics ◽  
C. P. Leblond

Rat thyroid lobes incubated with mannose-3H, galactose-3H, or leucine-3H, were studied by radioautography. With leucine-3H and mannose-3H, the grain reaction observed in the light microscope is distributed diffusely over the cells at 5 min, with no reaction over the colloid. Later, the grains are concentrated towards the apex, and colloid reactions begin to appear by 2 hr. With galactose-3H, the reaction at 5 min is again restricted to the cells but it consists of clumped grains next to the nucleus. Soon after, grains are concentrated at the cell apex and colloid reactions appear in some follicles as early as 30 min. Puromycin almost totally inhibits incorporation of leucine-3H and mannose-3H, but has no detectable effect on galactose-3H incorporation during the 1st hr. Quantitation of electron microscope radioautographs shows that mannose-3H label localizes initially in the rough endoplasmic reticulum, and by 1–2 hr much of this reaction is transferred to the Golgi apparatus. At 3 hr and subsequently, significant reactions are present over apical vesicles and colloid, while the Golgi reaction declines. Label associated with galactose-3H localizes initially in the Golgi apparatus and rapidly transfers to the apical vesicles, and then to the colloid. These findings indicate that mannose incorporation into thyroglobulin precursors occurs within the rough endoplasmic reticulum; these precursors then migrate to the Golgi apparatus, where galactose incorporation takes place. The glycoprotein thus formed migrates via the apical vesicles to the colloid.


1992 ◽  
Vol 40 (3) ◽  
pp. 257 ◽  
Author(s):  
RC Jones ◽  
M Lin

The genital ducts of Heterodontus portusjacksoni are lined by a ciliated epithelium. In the ductuli efferentes the epithelium is low and contains numerous intraepithelial leucocytes which often contain large dense bodies. All epithelial cells are ciliated and are characterised by apical vesicles, vacuoles and glycogen granules, some rough endoplasmic reticulum, dense bodies and lipid droplets, and a Golgi apparatus. The initial segment of the ductus epididymidis is lined by a very tall epithelium of ciliated and non-ciliated cells. The non-ciliated cells contain numerous apical vesicles, a large Golgi apparatus and numerous mitochondria and secretory granules in close association with an extensive endoplasmic reticulum. The terminal segment of the ductus epididymidis is lined by a low columnar epithelium. A proximal region, occupying part of the head of the epididymis, is similar to the epithelium in the ductuli efferentes. Distally, all the epithelial cells are ciliated. They are characterised by considerable dilated endoplasmic reticulum, a Golgi apparatus, apical vesicles, and numerous mitochondria and secretory granules. The secretory tubules of Leydig's glands are lined by a very tall epithelium with non-ciliated cells containing extensive, dilated, rough endoplasmic reticulum, a large Golgi apparatus, and numerous mitochondria and secretory granules. The significance of the structural differentiation of the duct is discussed in relation to the evolution of the mammalian epididymis.


1979 ◽  
Vol 149 (1) ◽  
pp. 17-26 ◽  
Author(s):  
JWM Van Der Meer ◽  
RHJ Beelen ◽  
DM Fluitsma ◽  
R Van Furth

Monoblasts, promonocytes, and macrophages in in vitro cultures of murine bone marrow were studied ultrastructurally, with special attention to peroxidatic activity. Monoblasts show peroxidatic activity in the rough endoplasmic reticulum and nuclear envelope as well as in the granules. The presence of peroxidatic activity in the Golgi apparatus could not be determined. Promonocytes have peroxidase-positive rough endoplasmic reticulum, Golgi apparatus, nuclear envelope, and granules, as previously reported. During culture, cells are formed with peroxidatic activity similar to that of monocytes or exudate macrophages (positive granules; negative Golgi apparatus, RER, and nuclear envelope); we call these cells early macrophages. In addition, transitional macrophages with both positive granules and positive RER, nuclear envelope, negative Golgi apparatus (as in exudate- resident macrophages in vivo), and mature macrophages with peroxidatic activity only in the RER and nuclear envelope (as in resident macrophages in vivo) were found. A considerable number of cells without detectable peroxidatic activity were also encountered. Our finding that macrophages with the peroxidatic pattern of monocytes (early macrophages), exudate-resident macrophages (transitional macrophages), and resident macrophages (mature macrophages), develop in vitro from proliferating precursor cells deriving from the bone marrow, demonstrates once again that resident macrophages in tissues originate from precursor cells in the bone marrow. Therefore, this conclusion can no longer be challenged on the basis of a cytochemical difference between monocytes and exudate macrophages on the one hand and resident macrophages on the other.


1977 ◽  
Vol 74 (3) ◽  
pp. 992-1015 ◽  
Author(s):  
J Paiement ◽  
CP Leblond

In the hope of localizing thyroglobulin within focullar cells of the thyroid gland, antibodies raised against rat thyroglobulin were labeled with the enzyme horseradish peroxidase or with (125)I-radioiodine. Sections of rat thyroids fixed in glutaraldehyde and embedded in glycol methacrylate or Araldite were placed in contact with the labeled antibodies. The sites of antibody binding were detected by diaminobenzidine staining in the case of peroxidase labeling, and radioautography in the case of 125(I) labeling. Peroxidase labeling revealed that the antibodies were bound by the luminal colloid of the thyroid follicles and, within focullar cells, by colloid droplets, condensing vacuoles, and apical vesicles. (125)I labeling confirmed these findings, and revealed some binding of antibodies within Golgi saccules and rough endoplasmic reticulum. This method provides a visually less distinct distribution than peroxidase labeling, but it allowed ready quantitation of the reactions by counts of silver grains in the radioautographs. The counts revealed that the concentration of label was similar in the luminal colloid of different follicles, but that it varied within the compartments of follicular cells. A moderate concentration was detected in rough endoplasmic reticulum and Golgi saccules, whereas a high concentration was found in condensing vacuoles, apical vesicles, and in the luminal colloid. Varying amounts of label were observed over the different types of colloid droplets, and this was attributed to various degrees of lysosomal degradation of thyroglobulin. It is concluded that the concentration of thyroglobulin antigenicity increases during transport from the ribosomal site of synthesis to the follicular colloid, and then decreases during the digestion of colloid droplets which leads to the release of the thyoid hormone.


1991 ◽  
Vol 39 (8) ◽  
pp. 1077-1087 ◽  
Author(s):  
H Tamaki ◽  
S Yamashina

We studied the ultrastructure and cytochemistry of mitotic parotid acinar cells in vivo after induction of mitosis by isoproterenol injection. With entrance of the cells into the division cycle, the Golgi apparatus lost its characteristic stacked structure and internal polarity among the cisternae, appearing as fragments distributed throughout the cytoplasm. These fragments consisted of electron-lucent vesiculotubular structures and electron-dense 70-nm vesicles; neither component showed thiamine pyrophosphatase activity, a marker for trans cisternae of the Golgi apparatus, but the 70-nm vesicles showed a positive reaction for osmium impregnation, indicating retention of the cis nature. The rough endoplasmic reticulum was dilated and fragmented. Recovery of the structure of Golgi apparatus and rearrangement of rough endoplasmic reticulum occurred in daughter cells during telophase. These changes were the same as those observed after drug-induced inhibition of protein transport. The secretory granules were not dispersed but were divided into two groups with which centrioles were closely associated. Both groups migrated with the centrioles as far as the next interphase. The distribution of 5'-nucleotidase on the luminal plasma membrane showed no change during the process of division, thus demonstrating that surface polarity was maintained during mitosis. These changes in organelle structure and distribution may be due to the conversion of cell function from a secretory to a mitotic action.


1969 ◽  
Vol 43 (2) ◽  
pp. 237-249 ◽  
Author(s):  
Charles A. Ashley ◽  
Theodore Peters

The sites of synthesis of proteins and their subsequent migration in rat liver have been studied during a 75 min period after labeling of liver-slice proteins by exposure to leucine-H3 for 2 min. Incorporation of the label into protein began after 1 min and was maximal by 4 min. Electron microscopic radioautography showed that synthesis of proteins in hepatocytes occurs mainly on ribosomes, particularly those in rough endoplasmic reticulum and, to some extent, in nuclei and mitochondria. Most of the newly formed proteins leave the endoplasmic reticulum in the course of 40 min, and concurrently labeled proteins appear in Golgi bodies, smooth membranes, microbodies, and lysosomes. A likely pathway for the secretion of some or all plasma proteins is from typical rough endoplasmic reticulum to a zone of reticulum which is partially coated with ribosomes, to the Golgi apparatus, and thence to the cell periphery. The formation of protein by reticuloendothelial cells was measured and found to be about 5% of the total protein formed by the liver.


Development ◽  
1978 ◽  
Vol 43 (1) ◽  
pp. 107-121
Author(s):  
Tomohisa Hirobe ◽  
Takuji Takeuchi

Electron microscopic observations on normally differentiating and α-MSH (melanocytestimulating hormone)-treated epidermal melanocytes of newborn mouse skin were carried out. The process of melanocyte differentiation from premelanosome-containing melanoblasts was investigated in detail with respect to melanosomes as markers. Melanoblasts containing unmelanized premelanosomes gradually decreased in number after birth, while the number of melanocytes rapidly increased. The epidermis of α-MSH-treated 3-day-old mice and normal 6-day-old mice contained melanocytes with numerous fully melanized melanosomes, and with no or only a few melanoblasts. Changes in other organelles in differentiating melanocytes were also noticeable. Golgi apparatus and RER (rough endoplasmic reticulum) decreased in number during the normal or α-MSH-induced differentiation of the epidermal melanocytes, though the number of mitochondria showed no notable change. The number of SER (smooth endoplasmic reticulum) per cell did not change in the cells of newborn mice, while in α-MSH-treated cells the number increased significantly. These results led us to an assumption that Golgi apparatus or RER transforms into other forms of organelles including melanosomes and SER during the differentiation of melanocytes.


Author(s):  
Sant S. Sekhon

Although there have been numerous studies concerning the morphogenetic changes accompanying the maturation of insect sperm, only a few deal with the sperm differentiation in the dragonflies. In two recent electron microscopic studies Kessel, has comprehensively treated the erlationship of microtubules to the nucleus and mid-piece structures during spermiogenesis in the dragonfly. The purpose of this study is to follow the sequential nuclear and cytoplasmic changes which accompany the differentiation of spermatogonium into a mature sperm during spermatogenesis in the dragonfly (Aeschna sp.).The dragonfly spermatogonia are characterized by large round nuclei. Loosely organized chromatin is usually unevenly distributed within the spermatogonial nuclei. The scant cytoplasm surrounding the nucleus contains mitochondria, the Golgi apparatus, elements of endoplasmic reticulum and numerous ribosomes (Fig. 1).


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