On the Identity of the ‘Neurofibrils’, ‘Nissl complex’, ‘Golgi Apparatus’, and ‘Trophospongium’ in the Neurones of Vertebrates

1961 ◽  
Vol s3-102 (60) ◽  
pp. 481-493
Author(s):  
G. B. DAVID ◽  
A. W. BROWN ◽  
K. B. MALLION

1. Classical histological methods demonstrate 4 cytoplasmic networks in fixed vertebrate neurones: ‘neurofibrils’, ‘Nissl complex’, ‘Golgi apparatus’, and ‘trophospongium’. The work described in this paper was undertaken to find out whether the 4 networks of classical histology correspond to 4 structures recognizable as such in the living neurone, or to only one structure, which may be coloured in characteristic ways by the classical methods. 2. A single continuous network, comprising features traditionally associated with the four classical networks of the fixed cell, can be isolated by micro-dissection and detected by interference microscopy in living vertebrate neurones. 3. When living neurones are centrifuged at a moderate angular velocity, a single continuous network remains visible under the interference microscope. There does not appear to be enough clear space left for 3 other voluminous structures. 4. When living neurones are centrifuged at a high angular velocity, a single continuous network is pushed to the centrifugal pole of the cell. The remainder of the cell then contains only small separate objects. 5. A single continuous network can be detected by interference microscopy in the cytoplasm of fixed, unstained neurones. When the same cells are dyed by a Nissl method, the cytoplasmic network seen in the unstained cells becomes deeply coloured. When the same cells are bleached and then silvered by a Golgi method, the objects that before had bound the dye now are blackened by the reduced silver. The same effect is obtained when the Nissl method is used after silvering. 6. It is inferred from the results of these experiments that there is only one cytoplasmic network in living normal adult vertebrate neurones. This network is demonstrated in fixed neurones of the same type with varying degrees of faithfulness by the classical methods. It is suggested that the terms ‘neurofibrils’, ‘Nissl complex’, ‘Golgi apparatus’, and ‘trophospongium’ be abandoned.

1960 ◽  
Vol s3-101 (54) ◽  
pp. 207-221
Author(s):  
G. B. DAVID ◽  
K. B. MALLION ◽  
A. W. BROWN

1. It was accidentally found that methods of silvering synaptic end-feet sometimes blackened Golgi's ‘internal reticular apparatus’ in neurones of the central nervous system of the cat. 2. A method of achieving this consistently was worked out: (a) paraffin sections are coated with a collodion membrane; (b) the collodion membrane is soaked in silver nitrate; (c) the silver nitrate is reduced to metallic silver with a buffered formaldehyde solution; (d) steps (b) and (c) are repeated until the sections appear quite black; (e) the silver attached to structures other than the Golgi apparatus is removed with a ferricyanide/thiosulphate bleach; (f) the section is ‘toned’ with gold chloride, fixed in thiosulphate, and washed thoroughly; (g) the section is dehydrated, cleared, and finally mounted in Canada balsam, DPX, or similar media. Results: Golgi-apparatus, black; connective-tissue fibres, black; axons, grey to black; everything else is light grey or colourless. 3. A tentative hypothesis is advanced to explain the results obtained. 4. The following advantages are claimed for the new method: the cytoplasmic reticulum thus blackened resembles that seen in living neurones with the interference microscope; special methods of fixation are not required; the cytoplasmic reticulum of given cells can be studied before and after silvering; and serial sections of the same piece of tissue can be used for histochemical purposes.


1969 ◽  
Vol 43 (3) ◽  
pp. 456-479 ◽  
Author(s):  
J. Metuzals ◽  
C. S. Izzard

The giant nerve fiber of the squid (Loligo pealii L.) has been investigated in situ, and in fresh and fixed preparations, by differential interference microscopy and electron microscopy. A continuous, three-dimensional network, composed of threadlike elements, was disclosed in the axoplasm. The threadlike elements in the axoplasm are twisted as a whole into a steep, right-handed helix. In a peripheral ectoplasmic region, the elements are more parallel to one another and more densely packed than in a central endoplasmic core. The threadlike elements can be resolved into a hierarchy of decreasing order of size. Successive levels of the hierarchy are formed by the association of smaller elements into larger ones. The following levels in the hierarchy of network elements have been distinguished: 1–3-µ-wide threads, 0.1–0.35-µ-wide strands, and 70–250-A-wide unit-filament strands. The differential interference microscope selects, from the network, threads oriented at a specific angle to the long axis of the axon. The specific angle depends upon the orientation of the long axis of the axon relative to the direction of shear. It is postulated that the network configuration is expressed in the solid-state properties of the axoplasm essential for the normal functioning of the nerve fiber.


1954 ◽  
Vol s3-95 (31) ◽  
pp. 271-304
Author(s):  
H. G. DAVIES ◽  
M. H. F. WILKINS ◽  
J. CHAYEN ◽  
L. F. LA COUR

1. The total mass M of substances other than water (the dry mass) in the living cell can be obtained from the expression M = φA/χ, where φ is the optical path difference (o.p.d.) due to the cell and A its projected area. The method makes use of the fact that the refractive increments α(χ = ioocα) of most substances in cells are approximately the same, and independent of concentration. Values for χ have been tabulated. Inaccuracies in the measurement of dry mass due to variations in χ (using λ average = 0.18) will be less than ± 10 per cent, in cells containing nucleic acids, proteins, and lipoproteins. When appreciable quantities of other substances are present the inaccuracy may be somewhat greater. When the total dry mass of living cells is determined in a medium other than water (e.g. isotonic solution), a correction term involving the thickness must be determined; this correction is often small. 3. The total dry masses and, in some cases, the concentrations of dry substance in a variety of biological objects including Amoebae, pollen grains at various stages of development, nuclei of cells in tissue culture, and sperm heads have been determined. In Tradescantia bracteata, during development from the microspore to the mature pollen grains, the dry mass increases by about tenfold. The dry masses of mature pollen grains were measured before and after successive digestion with ribonuclease, which removed about 4 to 14 per cent, of the dry mass, and with trypsin, after which about 40 per cent, of the original dry mass remained. In living ram sperm heads the ratio of deoxyribose nucleic acid to total dry mass determined by ultra-violet and interference microscopy respectively is 40 per cent. This is in good agreement with the value 45 per cent, obtained by bulk biochemical methods. 4. The interference microscope has been used to measure the refractive index of cells and, hence, the concentrations of dry substances in them, by immersing them in media of different known refractive indices. The application of this method to fixed cells is discussed theoretically. In experiments on fixed ram sperm heads the expected linear relationship between o.p.d. and refractive index of the immersion medium was obtained. Data on the average concentration of dry substance in ram sperm heads, the localized refractive index, and concentration in the denatured submicroscopic particles in the head, the percentage of the head volume occupied by them, and the geometrical thickness of the head were obtained. 5. Factors affecting the accuracy of the measurements of o.p.d., such as glare in the microscope, light scatter or absorption by the object, &c, are outlined.


1959 ◽  
Vol s3-100 (51) ◽  
pp. 339-368
Author(s):  
S. K. MALHOTRA

A reticulum can be seen by interference microscopy in the cytoplasm of the living neurone of vertebrates. The reticulum consists of irregular, massive bodies and thin strands. There are also well-defined spaces in the cytoplasm, in contact with the reticulum; they are usually crescentic. The massive bodies are the objects commonly called Nissl bodies. The thin strands are the basiphil threads clearly recognized by Nissl himself as constituting a part of his basiphil material. The classical ‘Golgi apparatus’ of the cell-body of the neurone of vertebrates consists of a deposit of silver or of osmium on the cytoplasmic inclusions mentioned in the first paragraph, but especially on the basiphil strands, which have a particular affinity for silver. At the base of the axon there are non-basiphil threads, which are also blackened by the Golgi methods.


1960 ◽  
Vol s3-101 (53) ◽  
pp. 69-74
Author(s):  
S. K. MALHOTRA

The purpose of this investigation was to repeat as exactly as possible the original work on the ‘apparatus’ of Golgi in the Purkinje cells of the cerebellum of owls, and to re-investigate these cells by modern methods. The tawny owl, Strix aluco, was used instead of the closely-related ‘Strix flammea’ of Golgi. Golgi's ‘osmio-bichromique’technique of 1898 for silver impregnation was used successfully. A reticulum corresponding to the basiphil Netz of Nissl can be seen in the living neurones by interference microscopy. The classical Golgi apparatus of the perikaryon is a deposit of silver or of osmium on this reticulum. The reticulum extends into the axons in the form of thin non-basiphil filaments, which are also blackened by Golgi methods. These findings are in conformity with recent studies of the neurones of other vertebrates.


1961 ◽  
Vol 153 (952) ◽  
pp. 357-366 ◽  

A new method of using the interference microscope is described which enables a continuous record to be obtained of changes in dry mass and concentration during the growth of a single tissue cell. The method has been applied to the analysis of changes of dry mass in the chromosomes, spindle and cell-plate area during mitosis in endosperm. The results of these observations have been compared with earlier studies of birefringence during mitosis. It is shown that birefringence develops under conditions of progressively increasing concentration of macro-molecular material.


1966 ◽  
Vol 25 ◽  
pp. 323-325 ◽  
Author(s):  
B. Garfinkel

The paper extends the known solution of the Main Problem to include the effects of the higher spherical harmonics of the geopotential. The von Zeipel method is used to calculate the secular variations of orderJmand the long-periodic variations of ordersJm/J2andnJm,λ/ω. HereJmandJm,λare the coefficients of the zonal and the tesseral harmonics respectively, withJm,0=Jm, andωis the angular velocity of the Earth's rotation. With the aid of the theory of spherical harmonics the results are expressed in a most compact form.


Author(s):  
J. Quatacker ◽  
W. De Potter

Mucopolysaccharides have been demonstrated biochemically in catecholamine-containing subcellular particles in different rat, cat and ox tissues. As catecholamine-containing granules seem to arise from the Golgi apparatus and some also from the axoplasmic reticulum we examined wether carbohydrate macromolecules could be detected in the small and large dense core vesicles and in structures related to them. To this purpose superior cervical ganglia and irises from rabbit and cat and coeliac ganglia and their axons from dog were subjected to the chromaffin reaction to show the distribution of catecholamine-containing granules. Some material was also embedded in glycolmethacrylate (GMA) and stained with phosphotungstic acid (PTA) at low pH for the detection of carbohydrate macromolecules.The chromaffin reaction in the perikarya reveals mainly large dense core vesicles, but in the axon hillock, the axons and the terminals, the small dense core vesicles are more prominent. In the axons the small granules are sometimes seen inside a reticular network (fig. 1).


Author(s):  
John P. Robinson ◽  
J. David Puett

Much work has been reported on the chemical, physical and morphological properties of urinary Tamm-Horsfall glycoprotein (THG). Although it was once reported that cystic fibrotic (CF) individuals had a defective THG, more recent data indicate that THG and CF-THG are similar if not identical.No studies on the conformational aspects have been reported on this glycoprotein using circular dichroism (CD). We examined the secondary structure of THG and derivatives under various conditions and have correlated these results with quaternary structure using electron microscopy.THG was prepared from normal adult males and CF-THG from a 16-year old CF female by the method of Tamm and Horsfall. CF female by the method of Tamm and Horsfall.


Author(s):  
S.R. Allegra

The respective roles of the ribo somes, endoplasmic reticulum, Golgi apparatus and perhaps nucleus in the synthesis and maturation of melanosomes is still the subject of some controversy. While the early melanosomes (premelanosomes) have been frequently demonstrated to originate as Golgi vesicles, it is undeniable that these structures can be formed in cells in which Golgi system is not found. This report was prompted by the findings in an essentially amelanotic human cellular blue nevus (melanocytoma) of two distinct lines of melanocytes one of which was devoid of any trace of Golgi apparatus while the other had normal complement of this organelle.


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