Comparative studies of the structure and regeneration of tissues in different vertebrates make it possible to get ideas from the evolution and regeneration potential of tissues. In this study are considered the reactive changes in the liver of larvae of the amphibian Rana temporaria to CC14 of different concentrations. Tadpoles of different sizes and different growth were placed once each day every day in a vessel with a determined solution of CC14 in water. The liver of tadpoles has a distinctive structure. It is distinguished by structure not only from the liver of higher vertebrates, but also from the liver of the adult frogs. The liver of the latter has an insignificant amount of fat and a whole series of other characters not typical of the liver of the tadpole. Placing the tadpoles in a solution of CCl4 with a concentration of 0.1% did not produce in the liver noticeable morphological changes. The author concludes that the absence of degenerative changes in the liver tadpoles, in spite of a high percentage of death of the experimental animals, tells of the well-known resistance of their liver to the influence of CC14

Grenberg, T.F.

English

Aquatic Commons

Freshwater Biological Association 
FBA Translation (New Series) No. 144 
Title: Infleunce of carbon tetrachloride on the liver of tadpoles of 
brown frog (Rana temporaria). 
Author(s) J.F. Grenberg 
Reference: Trudy leningr.sanit.-gig.med. Inst. 82, 32-36 
Original language: Russian 
Date of publication of original: 1967 
Translator: J.E.M. Horne 
Date of publication of translation: 1981 
Number of pages of translation: 6 
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FBA TRANSLATION (NEW SERIES) No.144 
Influence of carbon tetrachloride on the liver of tadpoles of 
brown frog [Rana temporaria]. T.F.Grenberg (1967) 
Trudy leningr. sanit.- gig, med. Inst. 82, 32-36. ( 1967) 
[translated by J.E.M. Horne] 
Comparative studies of the structure and regeneration of tissues 
in different vertebrates make it possible to get ideas from the 
evolution and regeneration potential of tissues. The majority of 
works carried out on these lines, are devoted to the study of 
organs and tissues of adult animals. (N.I. Grigor'ev 1949, 1951; 
L.D. Liozner & V.F. Sidorova, 1950; R.P.. Zhenevskaya, 1954; L.D. 
Liozner 1961, and others). Study of the regeneration potential 
in ontogenesis in different representatives of the vertebrates 
undoubtedly augments our idea on the rules of development of 
tissues and organs. 
In our work are considered the reactive changes in the 
liver of larvae of the amphibian Rana temporaria, of embryos 
of white rats and newborn rats after exposure to CC14 of 
different concentrations. 
Tadpoles of different sizes and different growth were 
placed once each day every day in a vessel with a determined 
solution of CC14 in water. In the solutions were used the 
following doses of the toxin: 0.02, 0.05, 0.08, 0.1, 0.5 and 1.0g 
per 100 cm3 of water. The time the tadpoles stayed in the 
solution ranged from 5 to 15 rnin. depending on the concentration 
of the solution. 
Entire larvae or prepared liver were fixed according to 
Karnua and in 10% neutral formalin. One part of the material was 
poured into paraffin-celloid and the other was used for 
obtaining sections on a freezing microtome. The prepared 
sections were stained with iron hematoxylin, according to 
Mallory, and gallocyanin. Polysaccharides were determined with 
the aid of SHIK reaction. The fat was stained with sudan III 
with the coloured hematoxylin of Bemer. 
In view of the fact that up to the moment of fixation 
of the tadpoles the. overwhelming majority of them attained a 
length of 25 - 30 mm, it is necessary to adduce the description 
of the normal structure of the liver of tadpoles of this length. 
The liver constitutes secretory tubules which branch 
and anastomose among themselves in different directions. On a 
transverse section the secretory tubules consist of 3 or 4 
hepatic cells that correspond with the number of cells comprising 
. the secretory tubules of adult representatives of tailed and 
tailess amphibians (N.I. Grigor'ev, 1951). 
2 
Since the hepatic cells in most cases are very 
large, the nucleus does not always fall on the section and gives 
rise to the impression that some secretory tubules comprise cells 
without nuclei (especially in longitudinal section). Hepatic 
cells show polar differentiation, therefore in the secretory 
tubule is produced a peculiar 'radial' differentiation. In the 
secretory tubules of tadpoles is contained a very large amount of 
fat. The fatty inclusions in the shape of large and small drops 
fill almost all of the cell, leaving free only a small part of 
. the cytoplasm. The latter has a kind of fine veins and strands 
connecting with the periphery of the cell and in its apical 
part. In the apical part of the cell the cytoplasm is free from 
fat and here forms a fine fringe next to the secretory fringe. 
The nuclei are situated as a rule in the basal part of the cell. 
They contain more or less significant lumps of chromatin and a 
few very large nucleoli. (see fig., a). 
Processing of the preparations with gallocyanin, 
providing a specific colouring for nucleic acid, showed that 
cytoplasmic RNA has a fine-grained character and also is 
diffusely distributed throughout all the cytoplasm. Nuclear RNA 
is found in the nucleoles. DNA surrounds the nucleus as a ring 
and isolated within it in a kind of clump of different sizes. 
(see fig., b.) 
The SHIk-reaction showed that glycogen in the cells of 
the liver is disposed unevenly. In some cells the clump is 
bigger, in others smaller. Glycogen has a granular character. 
The largest granules are concentrated chiefly in the basal part 
of the cells and consequently on the periphery of the secretory 
tubules. The intervals between the secretory tubules constitute 
the blood capillaries, in which are found greater or lesser 
quantities of the regular elements of the blood. Among them 
occur a very large quantity of mitotic dividing erythrocytes and 
leukocytes. Cambial elements of blood-formation are arranged 
chiefly on the periphery of the liver between the fine 
connective-tissue capsule and the secretory tubules in the form 
of isolated islets. 
Thus, the liver of tadpoles has a distinctive 
structure. It is distinguished by structure not only from the 
liver of higher vertebrates, but also from the liver of the adult 
frogs. The liver of the latter has an insignificant amount of 
fat and a whole series of other characters not typical of the 
liver of the tadpole. 
Placing the tadpoles in a solution of CCl4 with a 
concentration of 0.1% did not produce in the liver noticeable 
morphological changes, although it should be recorded that for 
many tadpoles these doses appeared lethal. Placing the tadpoles 
in 0.5% and 1.0% solutions of' CC14 produced certain changes in 
the secretory tubules and the hepatic cells. 
In 0.5% solution of CCI4 were placed tadpoles during 
the course of two weeks for 5 minutes every day. We studied 
3 . 
preparations of the liver of the tadpoles under different periods 
of experiment. 
After 3-fold action of the toxin in the secretory 
tubules of the liver no apparent aberration from normal could be 
recorded. Not until after 14-fold action of 0.5% solution of 
CC14 was it possible to see certain changes in the cytochemical 
structure of the secretory tubules. 
As before, in the cells occur a great quantity of fatty 
inclusions. If the diameter of the secretory tubules is not 
changed, then the diameter of their lumen sharply varies. 
Tubules are found with a diameter 3 or 4 times reduced in 
comparison with the normal. Along with these are tubules with 
the usual normal diameter of lumen. Insignificantly reduced is 
the quantity of glycogen. Large granules are found rarely. In 
the character of its arrangement RNA shows some changes. Fibres 
of ergastoplasm become more thick and RNA is converted from fine-
grain into an aggregate of rods and "wormlets" (see fig., b). 
Similar changes in RNA were found by G.I. Roskin (1951) in cells 
of the pancreas with the action of pilocarpine on them. N.I. 
Grigor'ev reckons such transformation of RNA as a sign of the 
regenerative ability of cells. In each case, evidently, such 
disposition of RNA involves somehow energetic accumulation, 
indicative of active resistance of cells to harmful influences. 
The blood vessels in the liver after the 14-fold' action 
of CC14 are very expanded, and the secretory tubules have the 
character of broadly-looped nets. 
Placing the tadpoles in a 1% solution of CC14 
produced more noticeable changes in the liver, although as before 
degeneration of the hepatic cells was not evident. In such a 
solution the tadpoles failed to survive more than 5 minutes. 
This dose seemed lethal for the majority of the tadpoles, 
therefore from very great numbers of tadpoles at the start of the 
experiment, at the end of it remained alive an insignificant 
number. Such massive death of experimental animals, naturally, 
made difficult substitute experiments for a long time. 
/ / 
The twofold action of the solution did not induce any 
kind of change in the structure of the tadpole liver. 
The threefold action of 1.0% CC14 led to the 
formation of different structure of the cells in the secretory 
tubules. Together with the secretory tubules, the cells of which 
are rich in fatty inclusions, in the liver are a great quantity 
of secretory tubules, the cells of which contain significantly 
less fat, and some cells with almost none. On the other hand, in 
the cytoplasm such cells contain large quantities of RNA. Rough 
fibres of ergastoplasm, in the form of rods are distributed in the 
cytoplasm of the hepatic cells. In the secretory tubules, thanks 
to the radial arrangement of these rods, is produced a general 
"radial" striation. Glycogen is disposed in the cells diffusely, 
irregularly by clumps throughout all the cytoplasm. 
4 
Fourfold and sixfold action of 1.0% solution of CC14 
induces more noticeable changes. The diameter of the secretory 
tubules is lessened, the fatty inclusions and various' vacuoles 
disappear. The cells are filled with basophile cytoplasm. The 
same cells become smaller, with the result that in the secretory 
tubules can be seen the dense distribution of the nuclei. The 
vascular system is expanded. The fine-grained glycogen is 
diffusely arranged in the cells. The RNA has as before a kind of 
rough rods. 
The common form of secretory tubules is changed more 
strongly. Increased are the diameter of the tubules and the 
diameter of their lumens. In the cells appear small vacuoles of 
fat. The cells are rich in cytoplasm. The quantity of glycogen 
in the cytoplasm of the cells is significantly reduced in 
comparison with the normal, and it is disposed diffusely in the 
form of minute granules. 
Protein synthesis is intensified and the quantity of 
RNA is increased. The blood capillaries are widened, in them are 
observed multiplying the regular elements of the blood, indeed, 
not so intensively as in the normal. 
All recorded changes of the liver, originating in the 
result of the effect of CC14, do not bear a degenerative 
character. Here are observed oscillations in fat, polysaccharide 
and protein metabolism. These reactive changes tell of the 
stability of the liver epithelium of tadpoles in relation to the 
toxin at the concentrations in question. 
From what has been stated it is possible to draw some 
preliminary conclusions. 
1. Cells of the liver of tadpoles of 25-30 mm length have polar 
differentiation which appears in the form of a cytoplasmic fringe 
in the apical part, in the disposition of clumps of glycogen and 
organelles. 
2. Reactive changes of the cells of the liver tadpoles with the 
effect of 0.5 - 1.0% solutions of CC14 manifest themselves in 
the dissipation of cell fat and polysaccharides and the 
accumulation of RNA. 
3. The absence of degenerative changes in the liver tadpoles, in 
spite of a high percentage of death of the experimental animals, 
tells of the well-known resistance of their liver to the 
influence of CC14. 
References 
Grigor'ev N.I. Trudy gistdl.Konf. (Leningrad, 5-9 June 1947). 
1949, 80-82. 
Grigor'ev N.I. Structure of the liver of different vertebrates 
and its changes in conditions of reparative 
regeneration. Trudy 5 Vses. s"ezda anat.,gist.i. 
embr., Leningrad, 1951. 
Zhenevskaya, R.P. Regenerating processes in the liver of verte-
brate animals. Trudy Instituta morphol. zhivotn. im. 
A.N. Severtsova, 2, 1964. . 
Liozner, L.D. Byull.eksp.biol.i med. 1961, 52, No.8, 98-101 
Liozner, L.D. & Sidorova V.F. Dokl.AN SSSR, 72, No.2, 1950, 
425-27 
Roskin G. I. Trudy 5 Vses. s"ezda anat., gist. i embr. , 1951, 
424-27 
5 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
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Please note that these translations were produced to assist the scientific staff of the 
FBA (Freshwater Biological Association) in their research. These translations were 
done by scientific staff with relevant language skills and not by professional 
translators. 
 
 
 


Influence of carbon tetrachloride on the liver of tadpoles of brown frog (Rana temporaria) [Translation from: Trudy leningt.sanit.-gig.med.Inst. 82, 32-36, 1967]

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Influence of carbon tetrachloride on the liver of tadpoles of brown frog (Rana temporaria) [Translation from: Trudy leningt.sanit.-gig.med.Inst. 82, 32-36, 1967]

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