Memoirs: The Vitellogenesis of Gasterosteus aculeatus (the stickleback) investigated by the Ultra-centrifuge

1938 ◽  
Vol s2-81 (321) ◽  
pp. 81-104
Author(s):  
B. N. SINGH ◽  
W. BOYLE

1. In the young oocytes the Golgi apparatus has the form of a solid body situated at one side of the nucleus. This body fragments and the pieces spread around the nucleus where they elongate and join up to form a network. This network moves out to the periphery of the cell where it fragments into pieces. No chromophobe part could be seen at any stage. 2. The mitochondria are seen in the earliest stages in the archoplasmic area. Here they occur as a number of scattered granules. In the later stages they are seen distributed in clumps throughout the cytoplasm. 3. Fat arises in the area which has been vacated by the juxta-nuclear Golgi apparatus (archoplasmic area). Prom here it spreads around the nucleus and out through the cytoplasm. There is no connexion between the Golgi apparatus and the secretion of fat. 4. The passage of material from the nucleoli through the nuclear membrane as fine granules has been described. These granules pass to the periphery of the egg where they swell up, become surrounded by a vacuole, and give rise to yolk. Only the nucleolar extrusions and the yolk stained with neutral red. 5. After centrifuging, the contents of the egg become separated into five distinct layers as follows, beginning at the centripetal pole: fat, cytoplasm, Golgi material, yolk, and nuclear material with mitochondria.

1930 ◽  
Vol s2-73 (291) ◽  
pp. 477-506
Author(s):  
VISHWA NATH

1. Observations on the living ovary. The earthworm ovary, as also that of the medicinal leech, is surprisingly favourable material for the study of the Golgi apparatus and the mitochondria in the living condition. The Golgi elements stand out very prominently in all stages of oogenesis as highly refractile spherules of a dark-greyish colour, performing a dancing movement in the cell. In the earliest oogonia situated near the septal insertion of the ovary there is a single Golgi spherule lying near the nuclear membrane. It probably divides at first into two and then into four, till in advanced oocytes there is a large number of Golgi elements distributed uniformly in the cytoplasm. The mitochondria in the earliest oogonia cannot be detected. Soon, however, they arise in the form of either a horseshoe closely fitting the nuclear membrane or a roundish mass, consisting of whitish granules, much less refractile than the Golgi elements. Gradually they spread out in the cytoplasm and perform a dancing movement. The Golgi elements and the mitochondria remain unaltered for a long time after the death of the cell. Attention is drawn to the excellent work of Foot and Strobell (1901), who described in the fresh egg of Allolobophora only two types of granules, namely, the ‘deutoplasmic’ or ‘osmiophile’ granules (Golgi elements) and the ‘archoplasmic’ or ‘yolk-nucleus’ granules (mitochondria). They have also shown only one osmiophile granule in their photographs of the earliest oogonia. 2. Observations on the living stained ovary. Neutral red and janus green B do not in any way improve the visibility of the inclusions, if indeed any improvement were desired. The Golgi elements do not at all stain with neutral red. The mitochondria may appear slightly blue with janus green. 3. Observations on fresh ovaries treated with osmic acid. The importance of this technique is greatly emphasized. After five to ten minutes' osmication the Golgi elements become copper-coloured, but they still appear solid. After half an hour's osmication they become slightly black and each element now shows very clearly a dark peripheral rim and a clear central area. The element is therefore not a solid or a semi-solid body, but a vesicle with a definite osmiophilic rim and a hollow interior. After two hours' osmication the vesicles become still blacker. 4. Experiments with the Centrifuge. The centrifuge very clearly reveals the existence of only two types of inclusions, namely, the Golgi elements and the mitochondria. There is neither yolk nor any other type of inclusion. 5. Observations on Fixed Preparations. If a Champy-fixed ovary is mounted whole, the Golgi elements appear as black granules. Within a month or so, however, they are decolorized by xylol. This proves the existence of fat inside the Golgi vesicle. In Champy-fixed sections, however, the vesicles are decolorized immediately after immersion in xylol. Kolatschev preparations demonstrate very satisfactorily the vesicular shape of the Golgi element. 6. The morphology of the Golgi apparatus in general is discussed in detail in the light of the recent work of Gatenby, Hirschler, Bowen, and others.


1965 ◽  
Vol s3-106 (73) ◽  
pp. 15-21
Author(s):  
JOHN R. BAKER

The exocrine cells of the mouse pancreas were fixed in potassium dichromate solution, embedded in araldite or other suitable medium, and examined by electron microscopy. Almost every part of these cells is seriously distorted or destroyed by this fixative. The ergastoplasm is generally unrecognizable, the mitochondria and zymogen granules are seldom visible, and no sign of the plasma membrane, microvilli, or Golgi apparatus is seen. The contents of the nucleus are profoundly rearranged. It is seen to contain a large, dark, irregularly shaped, finely granular object; the evidence suggests that this consists of coagulated histone. The sole constituent of the cell that is well fixed is the inner nuclear membrane. The destructive properties of potassium dichromate are much mitigated when it is mixed in suitable proportions with osmium tetroxide or formaldehyde.


1925 ◽  
Vol s2-69 (274) ◽  
pp. 291-316
Author(s):  
LESLIE A. HARVEY

1. The yolk-nucleus is merely a mass of mitochondria. 2. The mitochondria arise as a cap of threads over the nucleus, and this cap grows in size and density, migrates away from the nuclear membrane and breaks up into its component mitochondrial threads. These threads become evenly spread throughout the cytoplasm of the cell. 3. The mitochondria are not clearly defined in the very young oogonia. 4. The Golgi apparatus consists of numbers of Golgi elements lying separate in the cytoplasm. There is never any attempt at concentration of these elements round one central mass. 5. The Golgi elements are probably little platelets or spheroids somewhat resembling blood corpuscles in shape. They are not rods. As fixed by Da Fano technique, each element is a little plate with a very lightly impregnating centre and a very heavily impregnating rim. 6. The Golgi elements may probably arise from the cytoplasm. 7. The nucleus contains two nucleoli; an early arising karyosome, homogeneous and solid in structure, and a plasmo some arising later This plasmosome is liquid in consistency and contains an argentophil core. The karyosome disappears before the oocyte is half grown, but the plasmosome remains in the nucleus while the egg remains in the ovary. 8. No visible nucleolar extrusions into the cytoplasm were observed. 9. Yolk probably arises from the cytoplasm; no direct metamorphosis of either mitochondria, Golgi apparatus, or nucleolus into yolk was observed.


1962 ◽  
Vol s3-103 (62) ◽  
pp. 141-145
Author(s):  
R.A. R. GRESSON ◽  
L. T. THREADGOLD

That nucleolar material is extruded to the cytoplasm of the young oocyte of Blatta orientalis is confirmed by means of electron microscopy. The nucleolus and nucleolar extrusions are shown to contain RNA. In addition to the nucleolar extrusions, vesicle-like structures originate in the nuclear membrane and from there pass into the cytoplasm where they become indistinguishable from elements of the endoplasmic reticulum. When the nucleolar extrusions reach the cytoplasm they increase in size, come into close association with a few mitochondria, and migrate towards the periphery of the cell. It is concluded that the emission of material from the nucleolus and the passage of vesicles from the nuclear membrane to the cytoplasm are necessary prerequisites for the process of vitellogenesis.


The formation of mucus in goblet cells and its relation to the Golgi apparatus has been studied by various workers. Nassanow (1923) showed clearly that the mucin granules in the goblet cells of Triton originated in the Golgi apparatus, and so brought secretion in these cells into line with his theory of the bound secretion. More recently Clara (1926) has shown in the goblet cells of birds that the mucin first appears in the region next to the nucleus, between it and the gland lumen. Florey (1932, a, b ) has considered this more extensively in two recent papers, and for a number of mammals has shown that the mucin granules of goblet cells first form in the meshes of the Golgi network. In epithelial cells of the mouse vagina, undergoing conversion into mucous cells, he has found that the same process occurs. In a recent investigation of secretory formation in the salivary glands and pancreas it was found by the present author that in every cell type examined the young secretory granules first appeared in the basal region of the cell in relation to the mitochondria. Subsequent emigration occurred into the Golgi zone, where they underwent conversion into mature secretory granules. In the mucous cells of the salivary glands it was shown that these newly formed granules might be stained intravitam by Janus green or neutral red, and that in fixed preparations they stained selectively with acid fuchsin as described by Noll (1902), In the light of this work it appeared probable that while mucin formation might occur in the Golgi zone of the goblet cells as described by these authors, the origin of the granules might lie in the basal region of the cell.


1984 ◽  
Vol 99 (6) ◽  
pp. 2011-2023 ◽  
Author(s):  
J W Wills ◽  
R V Srinivas ◽  
E Hunter

The envelope glycoproteins of Rous sarcoma virus (RSV), gp85 and gp37, are anchored in the membrane by a 27-amino acid, hydrophobic domain that lies adjacent to a 22-amino acid, cytoplasmic domain at the carboxy terminus of gp37. We have altered these cytoplasmic and transmembrane domains by introducing deletion mutations into the molecularly cloned sequences of a proviral env gene. The effects of the mutations on the transport and subcellular localization of the Rous sarcoma virus glycoproteins were examined in monkey (CV-1) cells using an SV40 expression vector. We found, on the one hand, that replacement of the nonconserved region of the cytoplasmic domain with a longer, unrelated sequence of amino acids (mutant C1) did not alter the rate of transport to the Golgi apparatus nor the appearance of the glycoprotein on the cell surface. Larger deletions, extending into the conserved region of the cytoplasmic domain (mutant C2), resulted in a slower rate of transport to the Golgi apparatus, but did not prevent transport to the cell surface. On the other hand, removal of the entire cytoplasmic and transmembrane domains (mutant C3) did block transport and therefore did not result in secretion of the truncated protein. Our results demonstrate that the C3 polypeptide was not transported to the Golgi apparatus, although it apparently remained in a soluble, nonanchored form in the lumen of the rough endoplasmic reticulum; therefore, it appears that this mutant protein lacks a functional sorting signal. Surprisingly, subcellular localization by internal immunofluorescence revealed that the C3 protein (unlike the wild type) did not accumulate on the nuclear membrane but rather in vesicles distributed throughout the cytoplasm. This observation suggests that the wild-type glycoproteins (and perhaps other membrane-bound or secreted proteins) are specifically transported to the nuclear membrane after their biosynthesis elsewhere in the rough endoplasmic reticulum.


1970 ◽  
Vol 48 (11) ◽  
pp. 1913-1922 ◽  
Author(s):  
H. W. J. Ragetli ◽  
M. Weintraub ◽  
Esther Lo

Mesophyll cells of Nicotiana glutinosa exposed to starvation in the light after leaf excision showed the following major changes: vacuolation of the cytoplast; rearrangement of chloroplast and nuclear material; accumulation of cytoplasmic crystals and of starch; evagination and increased perforation of the nuclear membrane; and lysis of the tonoplast. Rearrangement of chloroplast material resulted in extended, 3-dimensional arrays (pseudocrystalline bodies), composed of electron-dense globules 85–100 Å in diameter. The ultrastructural degeneration of cells within a leaf was unsynchronized, and the sequence of degenerative intracellular events was not fixed. These observations are compatible with an unequal distribution of "essential" metabolites among those cells. Cells suffering from starvation stress showed ultrastructural changes not observed in cells of aging attached leaves and vice versa. Thus, starvation stress does not merely accelerate the process of cell-aging, as it operates in attached leaves.


1931 ◽  
Vol s2-74 (296) ◽  
pp. 669-700
Author(s):  
DES RAJ BHATIA ◽  
VISHWA NATH

Palaemon lamarrei 1. In the oogonia there are no granules which can be assigned to the category of mitochondria. They appear for the first time in young oocytes in the form of a juxta-nuclear heap of granules or in the form of a horseshoe closely embracing the nuclear membrane. Soon they arrange themselves in the form of a circum-nuclear ring which gradually expands towards the periphery of the oocyte without breaking away from the nuclear membrane. At the same time the marginal mitochondria of the ring grow in size till ultimately they give rise to albuminous yolk, which therefore appears for the first time in the peripheral regions of the cytoplasm (cf. Oniscus, King, 1926, and Rana tigrina, Nath, 1931). 2. A yolk-forming mitochondrium first swells up; but it is still poorly fixed and stained with Bouin-haematoxylin, like the unchanged mitochondria. The process of growth continues and the swelling mitochondria now show an internal differentiation in the form of minute granules or very small vacuoles. Such mitochondria are only slightly better fixed and stained with Bouin-haematoxylin. Gradually they are completely shorn of their lipoidal constituents, condensing at the same time more and more of protein material. Ultimately they give rise to albuminous yolk, sensu stricto, which is fixed and stained excellently in Bouin-haematoxylin. 3. In the earliest oocytes the nucleolus throws out into the cytoplasm deeply basophil pieces which are more or less uniformly dispersed. Soon they disappear. Hereafter the nucleolar extrusions are very minute, but they remain restricted to the perinuclear region. They never wander into the general cytoplasm or at least into its peripheral regions where protein yolk appears for the first time. A direct origin of the yolk granule from the extrusion must, therefore, be ruled out. But the possibility of the extrusions going into solution and thus indirectly contributing towards yolk cannot be eliminated. 4. Although the mitochondria can be easily observed in the fresh cover-slip preparations of young oocytes, the Golgi elements cannot be demonstrated unless the material is osmicated for at least twenty-two hours. Chemically the Golgi elements are lipoidal (fat-like). They are not stainable with neutral red. 5. In the oogonia and the earliest oocytes the Golgi elements exist in the form of vesicles, each vesicle showing a thick osmiophilic cortex and a central osmiophobic area. 6. During oogenesis many vesicles grow enormously in size, store up neutral fats inside them, and give rise to the fatty yolk as in Lithobius, spider, Otostigmus, Luciola, cockroach, Dysdercus, and Ophiocephalus (Nath, and Nath and collaborators), in Oniscus (King), in saw-flies (Gresson), and in Helix (Brambell). 7. The vacuolar system is absent in the prawn and also in the crab. Paratalphusa spinigera. 8. The Golgi elements of the crab behave exactly like those of the prawn, but the mitochondria, on the other hand, remain inactive and have no visible relationship with yolk formation. 9. In the crab also there are well-marked nucleolar extrusions. As in the prawn a prominent circum-nuclear ring of these granules is established early in oogenesis. But, unlike the prawn, granules from this ring continue to wander into the cytoplasm at the periphery of which they directly grow into the albuminous yolk.


1964 ◽  
Vol s3-105 (70) ◽  
pp. 219-226
Author(s):  
JENNIFER M. BYRNE

The neutral red granule cycle in the mouse exocrine pancreas was studied with the electron microscope in order to discover what changes appear at an ultrastructural level in cells treated with neutral red. There are no changes in the endoplasmic reticulum, the nucleus, the Golgi apparatus, the zymogen granules, or the mitochondria of stained cells when compared with normal tissue. Osmiophil inclusions are found which in their size and distribution correspond to the neutral red granules seen under the light microscope. Such inclusions are not seen in normal tissue. They resemble morphologically the lysosomes of various tissues.


1936 ◽  
Vol s2-79 (313) ◽  
pp. 73-90
Author(s):  
R. H. J. Brown

1. The Golgi apparatus may appear as a network or incomplete reticulum; it is lighter than the other cytoplasmic inclusions but its form makes its displacement difficult. Its parts never approach the periphery of the cell. The neutral-red bodies have no part in its composition. 2. There exists a separate canalicular system which is connected with the surface of the cell, and otherwise is of similar dimensions to the Golgi apparatus. It is thought to represent the trophospongium of Holmgren. It is unaffected by the centrifuge. 3. The vacuome appears in the form of isolated granules which can be osmicated after staining in neutral red. They are lighter than the cytoplasm and are separate from the Golgi apparatus, though on account of their similar density they are thought to have some spatial connexion with it. 4. The mitochondria are in the form of rods and granules which are very slightly denser than the cytoplasm, and show no evidence of having any connexion with the Golgi apparatus. 5. The Nissl substance occurs as large irregular bodies in the fixed material. It is thought to be in a diffuse form in the living cell. It is much denser than the cytoplasm.


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