scholarly journals CHANGES IN NUCLEAR HISTONES DURING FERTILIZATION, AND EARLY EMBRYONIC DEVELOPMENT IN THE PULMONATE SNAIL, Helix aspersa

1960 ◽  
Vol 8 (1) ◽  
pp. 69-81 ◽  
Author(s):  
David P. Bloch ◽  
Howard Y. C. Hew
Keyword(s):  
Developmental Stages ◽  
Picric Acid ◽  
Polar Body ◽  
Helix Aspersa ◽  
Bromphenol Blue ◽  
Fast Green ◽  
Specific Staining ◽  
Trichloracetic Acid ◽  
Basic Proteins ◽  
Snail Helix Aspersa

Calf thymus histories comprising two fractions, one rich in lysine, the other having roughly equal amounts of lysine and arginine, Loligo testes histones rich in arginine, and salmine, are compared with respect to their amino acid compositions, and their staining properties when the proteins are fixed on filter paper. The three types of basic proteins; somatic, arginine-rich spermatid histones, and protamine can be distinguished on the following basis. Somatic and testicular histones stain with fast green or bromphenol blue under the same conditions used for specific staining of histones in tissue preparations. The former histones lose most or all of their stainability after deamination or acetylation. Staining of the arginine-rich testicular histones remains relatively unaffected by this treatment. Protamines do not stain with fast green after treatment with hot trichloracetic acid, but are stained by bromphenol blue or eosin after treatment with picric acid. These methods provide a means for the characterization of nuclear basic proteins in situ. Their application to the early developmental stages of Helix aspersa show the following: After fertilization the protamine of the sperm is lost, and is replaced by faintly basic histones which differ from adult histones in their inability to bind fast green, and from protamines, by both their inability to bind eosin, and their weakly positive reaction with bromphenol blue. These "cleavage" histones are found in the male and female pronuclei, the early polar body chromosomes, and the nuclei of the cleaving egg and morula stages. During gastrulation, the histone complement reverts to a type as yet indistinguishable from that of adult somatic cells.

scholarly journals CYTOCHEMISTRY OF CYTOPLASMIC AND INTRANUCLEAR INCLUSIONS INDUCED BY BOVINE PARAINFLUENZA 3 VIRUS (SF-4) IN HUMAN CELL CULTURES

1966 ◽  
Vol 14 (2) ◽  
pp. 187-195 ◽  
Author(s):  
FREDERICK H. KASTEN ◽  
A. E. CHURCHILL
Keyword(s):  
Picric Acid ◽  
Methyl Green ◽  
Bromphenol Blue ◽  
Intranuclear Inclusions ◽  
Fast Green ◽  
Metaphosphoric Acid ◽  
Basic Proteins ◽  
Azure B ◽  
Human Cell Cultures ◽  
Schiff Reaction

Infection of human IIEp-2 cells with myxovirus bovine parainfluenza 3 (SF-4) induces the formation of cytoplasmic and intranuclear inclusions. Both types of inclusions were shown to contain some RNA in late stages (gallocyanin chromalum, azure B bromide, pyronin, RNase), abundant protein at all stages (acidic fast green, mercuric bromphenol blue) primarily with lysine-rich residues (naphthol yellow S, lysine blockage) and lesser amounts of arginine (Sakaguchi) and tyrosine (diazotization coupling). Inclusions failed to stain for histones (alkaline fast green, ammoniacal silver), basic proteins (alkaline Biebrich scarlet, picric acid-eosin). DNA (Feulgen, fluorescent-Feulgen, methyl green, gallocyanin chromalum, azure B bromide) and polysaccharides (fluorescent-PAS, Hale). There was a slight staining for histones with metaphosphoric acid-gallocyanin and basic proteins with picric acid-bromphenol blue or -eosin and a negative reaction for proteins with the fluorescent modification of the N-Bromosuccinimide-Schiff reaction. Acridine orange fluorochroming revealed a false positive reaction for DNA (green fluorescence) which was due to protein.

scholarly journals Cell Shrinkage caused by Fixatives and Paraffin-wax Embedding in Ordinary Cytological Preparations

1953 ◽  
Vol s3-94 (26) ◽  
pp. 125-139
Author(s):  
K.F. A. ROSS
Keyword(s):  
Cell Size ◽  
Picric Acid ◽  
Linear Shrinkage ◽  
Helix Aspersa ◽  
Volume Shrinkage ◽  
Paraffin Wax ◽  
Size Distributions ◽  
Cell Shrinkage ◽  
Saline Solutions ◽  
Snail Helix Aspersa

A series of measurements was made of the diameters of the cells and nuclei of the primary spermatocytes of the snail Helix aspersa in preparations made by embedding in paraffin wax after fixation in a number of different cytological fixatives. These were compared with similar measurements made on the living spermatocytes mounted in suitable saline solutions, and the shrinkage that the cells underwent as a result of fixation and embedding in each case was calculated from the medians of the size-distributions recorded. The results showed that the amount of shrinkage varies to some extent according to the fixative employed--picric acid gave the greatest linear cell-shrinkage (about 40 per cent.) and Sanfelice's fluid by far the least (about 13 per cent.); but with this one exception the differences in final cell-size were not very great, and represent a linear shrinkage to about 67 per cent, and a volume shrinkage to about 30 per cent, of the original dimensions of both cell and nucleus.

scholarly journals CHANGING NUCLEAR HISTONE PATTERNS DURING DEVELOPMENT I. FERTILIZATION AND EARLY CLEAVAGE IN THE CRAB, EMERITA ANALOGA

1968 ◽  
Vol 16 (7) ◽  
pp. 473-479 ◽  
Author(s):  
JACK C. VAUGHN
Keyword(s):  
Developmental Stages ◽  
Sperm Nucleus ◽  
Bromphenol Blue ◽  
Early Cleavage ◽  
Fast Green ◽  
Adult Type ◽  
Emerita Analoga ◽  
Cleavage Stages ◽  
Nuclear Histone ◽  
Early Developmental

Application of cytochemical techniques to the early developmental stages of the decapod crab, Emerita analoga, shows the following. After fertilization, the sperm nucleus, which apparently contains no basic proteins prior to this stage, becomes associated with a class of weakly basic histones, which differ from adult type histones in their apparent inability to bind alkaline fast green and from protamines by their inability to bind bromphenol blue following acetylation. This class of histones only persists until the late pronucleus stage, by which time the chromosomes contain a class of histones that are indistinguishable from adult histones by qualitative cytochemical techniques. No further changes in the nuclear histones are detected in the zygote or early cleavage stages. These changing histone patterns during early crab development are discussed with reference to other similar studies in other organisms.

The Evaporation of Water from Helix Aspersa

1964 ◽  
Vol 41 (4) ◽  
pp. 783-792
Author(s):  
JOHN MACHIN
Keyword(s):  
Air Flow ◽  
Complex Form ◽  
Ambient Air ◽  
Helix Aspersa ◽  
Flat Surfaces ◽  
Wind Speeds ◽  
Low Wind Speeds ◽  
Rate Of Evaporation ◽  
Snail Helix Aspersa

1. The construction and use of a wind-tunnel apparatus is described in which measurements of evaporation under controlled conditions of temperature, humidity and air flow can be made. 2. Two mathematical formulae, applicable to evaporation in relatively low wind speeds, are described. It is suggested that a promising approach to evaporation from moist-skinned animals is provided by the application of Leighly's formula: E = K(p0-pd)c(v/x)n, where the rate of evaporation (E) is expressed in terms of the vapour pressure at the evaporating surface (p0) and in the ambient air (pd), the wind speed (v) and the length of the evaporating surface parallel to the wind (x). The constant, K, is calculated independently and the terms n and c are left for empirical determination. 3. Values of n and c for different types of evaporating surface are given together with the method used in their calculation. Those relating to flat evaporators and to the snail, Helix aspersa, are shown to differ significantly. 4. In general n increases and c decreases as the amount of air disturbance caused by the snail increases. 5. The fact that n for flat surfaces is in good agreement with previously established theory is taken as evidence that Leighly's formula may be validly applied. 6. The combined determination of n and c is introduced as a convenient assessment of a complex form in terms of air flow and evaporation.

The use of methyl green as a histochemical reagent

1965 ◽  
Vol s3-106 (73) ◽  
pp. 3-13
Author(s):  
JOHN R. BAKER ◽  
ELIZABETH G. M. WILLIAMS
Keyword(s):  
Connective Tissue ◽  
Malachite Green ◽  
Helix Aspersa ◽  
Common Snail ◽  
Methyl Green ◽  
Dilute Solution ◽  
Dilute Solutions ◽  
The Common ◽  
Snail Helix Aspersa

The cation of methyl green carriea two poaitive charges, that of malachite green only one; but the two dyes behave towards tissue-constituents in almost exactly the same way. These dyes are not specific for chromatin. They colour certain objects that are devoid of DNA, even when they are used in very dilute solution. The granules of cells called Körnchenzellen in the connective tissue of the common snail, Helix aspersa, are strongly coloured by both dyes from very dilute solutions, and thus provide a striking instance of the unspecificity of these dyes. Malachite green, which is stable and free from contamination by metachromstic impurities, can advantageously replace the methyl green commonly used in mixtures with pyronine. It is suggested that pyronine may have a greater capacity for penetrating into close-textured objects, such ss nucleoli and ribosomes, than methyl and malachite greens.

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