The effect of ethanol upon early development in mice and rats. XIV. The effect of acute intoxication with beer and wine upon preimplantation development in rats

The preimplantation effect of acute intoxication during the preimplantation period of pregnancy (i.p. or i.v. injection on day 4) with beer and wine has been investigated on female albino rats (Wistar strain). The following criteria were applied for checking preimplantation development: mean number of embryos/animal; topographical distribution of the embryos; developmental stage attained; occurrence of pathological embryonic forms. The control on day 5 of pregnancy revealed no significant effect upon the developmental criteria used. The data obtained are compared with our own previous results.     

The effect of ethanol upon early development in mice and rats. VIII. The effect of chronic consumption of some beverages upon preimplantation development in rats

The effects of chronic consumption of some beverages (plum-brandy 24% and cognac 20%) upon preimplantation development in rats were studied. The control of possible effects was performed on day 5 by usual flushing, examination and photographying of oviductal and uterine embryos. In order to evaluate the effect of the beverage applied, the following criteria were used: mean litter size, migration of the embryos from the oviduct to the uterus, the developmental stage attained by the pre-implantation embryos and the appearance of pathological embryos. The main results were the following: both beverages applied influenced the preimplantation development; with respect to the developmental rate and to the induction of pathological changes, the effect of both beverages was similar (retardation and an increased, number of pathological morulae and blastocysts); a different action could be detected as to the mean litter size and to the migration of preimplantation embryos: plum-brandy reduced more substantially the mean litter size, whereas cognac had a more marked retarding effect upon the migration of embryos from the oviduct to the uterus: all the changes detected show a more or less marked "litter-effect". The present data were compared with the corresponding effects of chronic ethanol administration observed previously in our laboratory. No obvious potentiating effect of beverage congeners could be established. The findings are discussed in connection with other experimental models of alcohol embryo and fetopathy.     

The effect of ethanol upon early development in mice and rats. I. In vivo effect upon preimplantation and early postimplantation stages

The preimplantation and early postimplantation effect of chronic alcohol consumption (at least a month before mating and during pregnancy until killing) and of acute ethanol intoxication during the preimplantation period (i.v. infection of ethanol) was studied on albino rats (Wistar) and albino mice (RAP). The main results were as follows: Chronic alcoholization. Rats: significant retardation of preimplantation development and in early postimplantation stages; a tendency of lowering of the mean litter size. Mice: significant increase of the number of preimplantation pathological forms; a tendency of lowering of the mean litter size. Pathological changes show, both in rats and mice, an obvious "litter effect". Acute ethanol intoxication. Rats: significant retardation in some litters, normal or even advanced development in others. This effect differs from the previously reported effect of acute ethanol intoxication during early postimplantation stages. The results obtained attest the prenatal noxious effect of chronic ethanol consumption in both species used and of acute ethanol intoxication during preimplantation development upon early postimplantation development in rats. Within the limits of extrapolation possibilities, they represent a risk signal for other species (including human).     

The effect of ethanol upon early development in mice and rats. VI. In vivo effect of acetaldehyde upon preimplantation stages in rats

In completion of the previously outlined "experimental alcohol blastopathy", the role of acetaldehyde in the induction of preimplantation pathological changes in rat embryos has been controlled. Two experimental models were used: the direct administration of acetaldehyde by gavage and the blockage of acetaldehyde metabolization by ANTALCOL (an aldehyde-dehydrogenase blocking compound). The main results were as follows: The exogenous acetaldehyde in the blood of pregnant animals has an obvious effect upon the developmental rate during the late preimplantation period (retarding segmentation, blastulation), and in one of the experimental models upon the oviductal-uterine migration rate. The increase of the blood acetaldehyde level by blockage of its further metabolization has a more marked effect as compared with the direct intravenous administration of the substance. According to our previous observations the intravenous application of ethanol on the same day (day 4) has no such effect. The direct noxious influence upon the developing preimplantation embryos (fragmentation) of the increased level of acetaldehyde obtained by ANTALCOL treatment is similar but more marked than this effect obtained previously by ethanol administration. The same effect observed after the direct administration of the substance is less marked than the effect of ANTALCOL treatment but more marked than the effect of intravenous ethanol administration. These results attest that acetaldehyde may contribute (alone or together with the effect of ethanol) to the induction of "experimental alcohol blastopathy". The less marked action of the substance proper introduced into the blood stream may be due--in our opinion--to its possible alteration during the period between distillation and application.     

The effect of ethanol upon early development in mice and rats. V. In vivo effect of acute preimplantation intoxication with or without previous chronic alcoholization

Albino rats (Wistar) and albino mice (RAP) were either injected intravenously with ethanol during the preimplantation period (day 4 and 3, respectively) or injected in the same way after a previous chronic alcoholization (peroral consumption of 20% ethanol for 50-60 and 32-35 days, respectively before mating, adding the days until killing). The control of possible effects was performed on day 5 (rats) and 4 (mice) by usual flushing, examination and photographing of oviductal and uterine embryos. A group of albino rats, with chronic alcoholization, was controlled for late, fetal effects (resorption rate, skeletal control, possible ocular anomalies). The main results obtained were as follows: Acute ethanol intoxication. Rats: significant increase of pathological, fragmented preimplantation embryos with a marked litter effect. Mice: no deleterious effect upon preimplantation development. Chronic alcoholization + acute ethanol intoxication. Rats: significant retardation of the preimplantation development rate and a significant increase of the number of pathological, fragmented embryos with a marked litter effect. Mice: demonstrable advance of preimplantation development and migration rate. Chronic alcoholization--late fetal control in rats: the increase of resorption rate; the more frequent absence of sacral vertebrae; very rare rib anomalies and the absence of ocular malformations.     

The effect of ethanol upon early development in mice and rats. II. In vitro effect upon early postimplantation rat embryos

The direct effect of ethanol upon in vitro cultured 9.5 day rat embryos was investigated (2, 4, 8 and 10% ethanol added to the culture medium). The main effects recorded were as follows: 1. Significant increase of the number of "dying" embryos (beating heart without yolk sac circulation); 2. No significant increase of mortality; 3. Significant increase of the number of living embryos with deficient blood circulation; 4. Significant retardation of coiling in living embryos with a significant dose-effect relation, when the effects of 20/00 and 80/00 ethanol were compared; 5. Lowering of the mean somite number in living embryos; 6. Various macro- and macroscopical pathological changes (mainly necrotic areas in the central nervous system).     

The effect of ethanol upon early development in mice and rats. III. In vivo effect of acute ethanol intoxication upon implantation and early postimplantation stages in mice

Female albino mice (RAP strain) were injected intravenously with absolute alcohol diluted aa by a.d., on days 2, 3 and 4 of pregnancy respectively. Macroscopically pregnant and empty uteri were controlled by microscopic examination. The developmental rate of postimplantation embryos was controlled by a previously reported biometrical method. The acute ethanol intoxication did not influence the postimplantation developmental rate (except for the appearance and development of the allantoic bud). In some of the uteri of the treated females (mainly of those treated on day 4), free blastocysts were found in the uterine lumen on day 9 of pregnancy, supposedly due to the deleterious effect of ethanol intoxication on central and local factors of implantation. Regressive changes of the decidua and consecutive embryonic death found in two litters may be caused by the same deleterious effect or (and) by inflammatory changes present in the endometrium of some control and treated females. The acute ethanol intoxication lowered the mean litter size in all the experimental groups.     

The effect of ethanol upon early development in mice and rats. IV. The effect of acute ethanol intoxication of day 4 of pregnancy upon implantation and early postimplantation development in mice

Acute ethanol intoxication in albino mice (RAP) induced by intravenous administration of ethanol on day 4 of pregnancy delayed or inhibited implantation in about 25 per cent of the cases. The noxious action upon the implantation process showed a clear-cut "litter effect" and the mean litter was not affected by the experimental intervention. In very early postimplantation stage (day 6 of pregnancy) a statistically significant advance of some main morphogenetic indices was detected in treated specimens. As a possible explanation of this finding, a "selection" of more resistant and viable embryos by the acute ethanol intoxication is presumed. The data discussed in the present paper, together with authors' previous findings suggest a possible noxious action of acute ethanol intoxication during preimplantation stages upon implantation.     

The effect of chronic oral alcoholization upon late fetal development in mice

The late fetal effect of chronic alcoholization in two strains of mice (RAP and RAP female x CBAT6 male) was controlled. Unilateral ocular anomalies (retinal folding, various degrees of ocular disorganization) were detected in 16% of the alcoholized RAP female x CBAT6 male group. The findings are discussed in connection with other experimental models of alcohol embryo and fetopathy.     

The influence of chronic ethanol consumption on the distribution of thiopental in rats.

Ethanol was administered chronically for 14 days to male Sprague-Dawley rats. Day 15 was ethanol-free. On day 16 the rats received 25 mg of thiopental per kilogram (intravenously). One minute after the injection, the ethanol-treated rats showed lower blood levels of thiopental and higher liver levels of the drug than control rats given sucrose in place of ethanol. Samples of blood drawn 5 and 10 min after injection showed no significant difference in thiopental levels between the ethanol and control groups. The ethanol-treated group slept for a significantly shorter period of time. It is concluded that chronic ethanol consumption for 14 days decreases the pharmacological effects of thiopental and alters its initial distribution in the body.     

The effect of chronic ethanol consumption on temperature-dependent physical properties of liver mitochondrial membranes

We have reported that the monovalent ionophore monensin causes undersulfated chondroitin sulfate biosynthesis in cultured chondrocytes. In order to clarify the mechanism of this diminished sulfation, we have measured the rate of incorporation of sulfate into chondrocytes and assayed the cellular ATP levels. We have also measured sulfatase activity, the incorporation of ³⁵SO₄ into 3′-phosphoadenosine 5′-phospho[³⁵S]sulfate and endogenous sulfotransferase activity in the cell-free extracts. We find that: (1) The incorporation of ³⁵SO₄ into the free sulfate pool in chondrocytes was not inhibited by monensin. (2) The ATP levels of monensin-treated chondrocytes were the same as control cells. (3) There was no sulfatase activity in both control and monensin-treated chondrocytes. (4) Enzymatic analyses revealed that ³⁵SO₄ incorporation into 3′-phosphoadenosine 5′-phospho[³⁵S]sulfate and subsequent sulfotransferase activity were not inhibited in the presence of monensin. At present the most tenable hypothesis to account for monensin causing undersulfated chondroitin sulfate synthesis is that the ionophore impairs the access of proteoglycans to the sulfotransferases in the luminal walls of the Golgi structures.     

The effect of acute and chronic ethanol administration on serum corticosterone concentration in rats

The effect of intraperitoneally (i.p.) and intragastrically (i.g.) administered ethanol solution, and the influence of voluntary ethanol uptake (20% v/v) on adrenocortical activity of adult male rats was studied. Both i.p. and i.g. ethanol administration resulted in a significant activation of adrenocortical mechanisms, while voluntary ethanol uptake failed to induce elevation of serum corticosterone concentration. No difference was found in blood ethanol concentration among these groups. The responsiveness of adrenocortical mechanisms was also tested in rats which were given the free choice between ethanol solution (5% v/v) and tap-water for three weeks. Unavoidable electric foot-shocks, as stressor, resulted in an elevation of serum corticosterone concentration in control animals, but this response was found to be significantly reduced in chronically ethanol drinking rats.     

Chronic ethanol consumption depresses hypothalamic-pituitary-adrenal function in aged rats.

In separate experiments, nine (n = 20) and fifteen (n = 12) month old rats were treated with either 6% ethanol or 12% sucrose (to balance caloric intake) in the drinking water to examine the effect of chronic ethanol consumption on the hypothalamic-pituitary-adrenal axis of aged rats. Rats were maintained on these treatment regimens for thirty days and were killed by decapitation. Blood was collected and plasma concentrations of adrenocorticotropin (ACTH) and corticosterone were determined by radioimmunoassay. Adrenal glands were cleaned, quartered and used to test in vitro responsiveness to ACTH. Anterior pituitary glands from all 15 month old rats and one half of the nine month old rats were collected, frozen and extracted for measurement of tissue ACTH concentration. The remaining anterior pituitary glands from the nine month old rats were challenged with corticotropin releasing hormone (CRH) to test in vitro responsiveness. In nine month old rats, chronic ethanol consumption decreased plasma ACTH and corticosterone (P less than 0.05). Pituitary ACTH concentrations were unchanged in treated nine month old rats, but the amount of pituitary ACTH released in response to CRH was decreased (P less than 0.05) in rats consuming ethanol. In vitro responsiveness of the adrenal gland to ACTH in nine month old rats consuming ethanol was unchanged (P greater than 0.05). Plasma ACTH and corticosterone concentrations were also decreased in 15 month old rats chronically consuming ethanol (P less than 0.05). No differences were noted in responsiveness of the adrenal gland or in the amount of pituitary ACTH due to ethanol consumption in 15 month old rats (P greater than 0.05). The results of these experiments indicate that chronic ethanol consumption decreases hypothalamic-pituitary-adrenal function in aged rats.     

Cytogenetic analysis of oocytes and early preimplantation embryos from XO mice.

The cytoplasm of early sea urchin embryos contains nonribosomal, high molecular weight RNA both associated with ribosomes in polysomes and free of ribosomes in particles termed free RNP. In a 1-hr labeling period, 50% of the newly synthesized RNA enters the pool of ribosome-free RNP particles during the cleavage stages, and this percentage decreases until less than 20% of the new RNA in the mesenchyme blastula stage is found in the free RNP. mRNA from both polysomes and free RNP contain poly(A)(+) and poly(A)(?) species. During the cleavage stages only 8–10% of the RNA from each fraction is polyadenylated; however, in the blastula, 40–50% of the nonhistone polysomal RNA is polyadenylated while only 22–30% of the free RNP RNA is polyadenylated. At any developmental stage, the poly(A)(+)RNA from the free RNA and polysomes have identical sedimentation profiles; this is also the case for the poly(A)(?)RNA except for the absence of the 9 S histone mRNA from the free RNP. Changes in poly(A)(+)RNA content and sedimentation profiles during development occur simultaneously in the free RNP and the polysomes. Kinetic studies of these two RNP populations as well as nuclear RNP show that the bulk of the free RNP are not unusually stable cytoplasmic components. The free RNP decay with a half-life of about 40 min while nuclear RNA and polysomal RNA display half-lives of about 12 and 65 min, respectively. Further, the rate of synthesis of the free RNP is not consistent with their being the only precursors for polysomes. Our estimates of the rates of synthesis for nuclear RNA, polysomes, and free RNP are, respectively, 1.1 × 10^?15, 2.2 × 10^?16, and 5.0 × 15^?17 g/min/nucleus. The data on free RNP is discussed in terms of translational regulation of protein synthesis in the developing sea urchin.