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Biomedical subjects

L M Houdebine

Publications and source records attributed to L M Houdebine.

At least 145 records · Page 8Linked to original sources
PubMed1980

Effects of lysomotropic agents, and of microfilament- and microtubule-disrupting drugs on the activation of casein-gene expression by prolactin in the mammary gland.

The organ-culture technique was used to investigate the effects of lysomotropic agents (NH4Cl and chloroquine) and of modifiers of microfilaments (cytochalasin B) and microtubules (colchicine) on the induction of casein synthesis and the accumulation of casein mRNA by prolactin in the rabbit mammary gland. Neither chloroquine nor NH4Cl altered the lactogenic action of prolactin. Cytochalasin B attenuated the response to prolactin in terms of casein synthesis. However, this drug did not hamper the accumulation of casein mRNA. Colchicine exhibited a marked specific inhibitory effect on the induction of casein synthesis. It also prevented the accumulation of casein mRNA. These results suggest that a putative degradation of the internalized prolactin--receptor complex by lysosomes is not strictly involved in prolactin action. In addition, the integrity of the microfilaments seems unnecessary in the process of casein-gene activation by prolactin. By contrast, the integrity of the microtubule network seems absolutely necessary to ensure the transmission of prolactin information to the nucleus.

Ammonium Chloride↗
PubMed1980

[Effect of ouabain on the lactogenic action of prolactin and on the level of mammary prolactin receptors].

Ouabain added to the culture medium of rabbit mammary gland inhibits prolactin action on the initiation of lactose and casein synthesis. The degree of inhibition is a function of the ouabain concentration in the medium. Likewise, ouabain blocks the accumulation of casein mRNA supported by prolactin. In addition, ouabain provokes a rapid disappearance of prolactin receptors. Conversely prolactin keeps its capacity to enhance the concentration of casein mRNA and the parallel casein synthesis when K+ ions are totally absent from the culture medium. These results suggest that although prolactin induces a modification of the K+/Na+ ratio in the mammary cell and ouabain prevents this effect of prolactin, the inhibitory action of ouabain on lactogenesis can be explained essentially by its effect on the hormone receptors.

Animals↗
PubMed1980

[Effect of indomethacin on the lactogenic action of prolactin].

The action of indomethacin on the lactogenic activity of prolactin has been evaluated usind the technique of rabbit mammary gland organ culture. Indomethacin is totally unable to inhibit prolactin action as estimated by lactose synthetase activity and casein synthesis. These data suggest, as opposed to previous works, that prostaglandins are not involved in the mechanism of prolactin action on lactogenesis.

Animals↗
PubMed1980

Role of prolactin in the transcription of beta-casein and 28-S ribosomal genes in the rabbit mammary gland.

Isolated mammary nuclei were incubated in the presence of HgCTP and the neosynthesized RNA was isolated with a SH-Sepharose column. The concentration of beta-casein mRNA and 28-S ribosomal RNA in the neosynthesized RNA fractions was evaluated using [3H]DNA probes complementary to beta-casein mRNA and 28-S rRNA respectively. In the unstimulated pseudopregnant rabbit, the transcription of both genes was easily detectable. Injections of prolactin progressively enhanced the transcription rate of both genes and preferentially the beta-casein gene. A comparison between the transcription rates and the accumulation of the corresponding gene products in the cell revealed that there is a good correlation between these two parameters for the 28-S rRNA gene. By contrast, the acceleration of beta-casein gene transcription by prolactin is unable to account for the simultaneous accumulation of beta-casein mRNA, indicating that prolactin is a potent stabilizer of casein mRNA. Injections of CB154 into lactating rabbits (a drug which suppresses the secretion of prolactin by hypophysis), induced a rapid drop of beta-casein mRNA concentration and a slow decline of beta-casein gene transcription. Simultaneously the drug was responsible for a marked and rapid decrease of 28-S rRNA gene transcription, while the concentration of the rRNA remained elevated. During weaning the transcription of the beta-casein gene and, to a lower degree, the transcripton of the 28-S rRNA gene proceeded more slowly and this phenomenon was accompanied by a progressive decline of the RNA concentrations.

Animals↗
PubMed1980

Effect of various lysosomotropic agents and microtubule disrupting drugs on the lactogenic and the mammogenic action of prolactin.

Various drugs added to the culture medium of rabbit mammary gland were assayed for their capacity to affect the lactogenic and the mammogenic activities of prolactin. Three lysosomotropic agents NH4Cl, chloroquine and methylamine which were previously demonstrated to inhibit the degradation of the hormone-receptor complex after its internalization (down-regulation) did not prevent the initiation of casein synthesis, of lactose synthetase activity and of DNA synthesis. Five microtubule disrupting drugs, colchicine, colcemid, vinblastin, podophyllotoxin and nocodazole inhibited the induction of casein and DNA synthesis by prolactin whereas two inactive analogues, trimethylcolchicinic acid and lumicolchicine had no effect. None of these drugs exhibited any general cytotoxic effect as judged by the capacity of the tissue to incorporate 14C aminoacids into total proteins and 3H-uridine into total RNA. The microtubule disrupting drugs did not greatly reduce the rate of casein synthesis in the cultured mammary tissue explanted from lactating rabbits. The data suggest that the down-regulation of prolactin receptor is not strictly required for the two considered prolactin activities. By contrast, the integrity of microtubules, or at least of structures in which tubulin is involved, is necessary to ensure a normal transmission of the prolactin information responsible for the initiation of milk and DNA synthesis. In addition, the fact that the lactogenic and the mammogenic activities of prolactin are affected by the same drugs suggests that these two properties of the hormone are mediated by cellular mechanisms which have at least one common step.

Ammonium Chloride↗
PubMed1980

Lactogenesis induced by prostaglandin F2 alpha in pregnant rabbits.

PGF2 alpha may have a primarily luteolytic action and hence diminish Pg secretion. This triggers the release of PRL and other hormones of the lactogenic complex with the consequent production of milk substances in the mammary gland. Later, when the uterine tissue is depleted of Pg, pregnancy is terminated.

Animals↗
PubMed1979

[Role of lysosomes, microtubules and microfilaments in the mechanism of the lactogenic action of prolactin in the rabbit mammary gland].

The lysosomotropic agents, ammonium chloride and chloroquine, added to the culture medium of pseudopregnant Rabbit mammary gland, did not inhibit the initiation of casein synthesis by prolactin. By contrast, they considerably reduced the down-regulation of prolacting receptors. Converse-y, colchicine totally blocked the lactogenic action of prolactin without altering the down-regulation of the receptor. Cytochalasin B inhibited only partly the lactogenic action of prolactin while it has no effect on the down-regulation of the receptor. These data suggest that the degradation of the prolactin-receptor complex in lysosome is not a compulsory step in the mechanism of prolactin action. The integrity of microtubules but not of microfilaments is required for prolactin to initiate casein synthesis. These elements of the cytoskeleton are not strictly involved in the down-regulation of the receptor.

Ammonium Chloride↗
PubMed1979

Role of prolactin the the expression of casein genes in the virgin rabbit.

Prolactin was injected into mature virgin rabbits. Casein mRNA, already present prior to the treatment was enhanced by prolactin. This accumulation of casein mRNA was not accompanied by a parallel significant synthesis of casein. Th effect of prolactin took place without a previous cellular multiplication. The mammary cell is therefore sensitive to prolactin before the first pregnancy but prolactin then displays only an abortive effect on the expression of casein genes.

Animals↗
PubMed1979

Role of glucocorticoids and progesterone in the development of rough endoplasmic reticulum involved in casein biosynthesis.

Hydrocortisone acetate injected into pseudopregnant rabbits induced casein synthesis and a parallel accumulation of casein mRNA. These effects were not accompanied by any enrichment of total RNA in the mammary cell. Hydrocortisone acetate did not favour the attachment of polysomes to endoplasmic reticulum. Casein mRNA concentration was enhanced in free and membrane-bound polysomes. After long treatments, the concentration of casein mRNA reached a plateau in membrane bound polysomes whereas it continued to be accumulated in free polysomes, suggesting that a substantial part of casein synthesis is then carried out by free polysomes. Progesterone injected with high doses of prolactin was unable to prevent the stimulatory action of prolactin on the synthesis of casein, the accumulation of casein mRNA and mammary gland growth, as judged by DNA content. By contrast, the increase in the total RNA content of mammary gland was still significantly reduced by progesterone. In addition, progesterone inhibited almost completely the formation of membrane-bound polysomes and the anchorage of casein mRNA to endoplasmic reticulum. From these data, it was concluded that the formation of the endoplasmic reticulum is not a prerequisite for the initiation of casein synthesis. Glucocorticoids do not play a major role in the formation of the endoplasmic reticulum and the Golai apparatus and in the binding of casein synthesizing polysomes to membranes. Progesteronne is capable of inhibiting preferentially and gradually the stimulation of cellular functions requiring the most potent prolactin stimulation.

Animals↗
PubMed1979

Stimulation of milk synthesis in the rabbit by fish pituitary extract.

The lactogenic properties of extracts of the pituitary glands of salmon and trout were evaluated by using the organ culture technique with rabbit mammary explants. Crude extracts and fractions obtained after chromatography on Ultrogel and selected for their capacity to compete with ovine prolactin in a rabbit mammary gland radioreceptor assay were added to the culture medium. The criteria of lactogenesis were lactose synthetase activity, casein synthesis, measurements of the concentration of casein messenger RNA and the histology of mammary glands. All these tests led to the conclusion that salmon and trout pituitary glands contain a prolactin-like principle capable of initiating milk synthesis in the rabbit mammary cell.

Animals↗
PubMed1978

Role of prolactin and glucocorticoids in the expression of casein genes in rabbit mammary gland organ culture. Quantification of casein mRNA.

Milk synthesis is initiated solely by prolactin in the pseudopregnant rabbit and glucocorticoids potentiate this action of prolactin. In organ culture, prolactin, in the presence or in the absence of insulin, enhances casein synthesis and cortisol (inactive alone) amplifies this action. Measurements of casein mRNA concentration in total cellular RNA, by hybridization with DNA complementary to casein mRNA, revealed that the stimulation of casein synthesis by the glucocorticoid is accompanied by an increase in the amount of casein mRNA. A systematic comparison of variations of these two parameters indicated that the major effect of glucocorticoids on lactogenesis in the rabbit at this stage of mammary gland development is mediated through an increase in the quantity of casein mRNA available for translation. No simultaneous control of casein mRNA translation by cortisol was observed.

Animals↗
PubMed1978

Role of spermidine in casein gene expression in the rabbit.

Spermidine concentration in rabbit mammary gland was estimated during pregnancy, lactation and after the induction of milk synthesis by prolactin and glucocorticoids in vivo and in vitro. It was observed that mammogenesis and lactogenesis during preganancy and the initiation of milk secretion at parturition are accompanied by an enhancement of spermidine concentration in the mammary gland. By contrast, the initiation of these phenomena by hormone injections does not require such variations of spermidine concentration. In organ culture, a slight increase in spermidine concentration was obtained under the influence of an hormonal combination including insulin, prolactin and cortisol. Spermidine added to the culture medium was unable to mimic cortisol action. An amplification of casein synthesis and a parallel increase of casein mRNA concentration was provoked by cortisol even when spermidine synthesis was blocked. Thus, one of the major actions of glucocorticoids during lactogenesis in the rabbit is not mediated through an increase in spermidine concentration in the mammary gland.

Animals↗
PubMed1978

Stabilization of casein mRNA by prolactin and glucocorticoids.

Prolactin injected into pseudopregnant rabbits led to a parallel enhancement of casein synthesis and casein mRNA concentration. When this stimulation was followed by a withdrawal of prolactin obtained by injections of bromocriptine, the rate of casein synthesis progressively diminished. In the presence of endogenous prolactin after the initial stimulation, the decline of casein synthesis was delayed. Hydrocortisone acetate injected with bromocriptine after the initial stimulation by prolactin was able to maintain a high rate of casein synthesis. Measurements of casein mRNA concentration by hybridization with casein cDNA indicated that in all cases the amount of casein mRNA was correlated with the magnitude of casein synthesis. This suggests that the lactogenic hormones, prolactin and glucocorticoids, which were previously demonstrated to be responsible for the enhancement of casein mRNA concentration are involved in their stabilization.

Animals↗
PubMed1977

Effects of glucocorticoids on casein gene expression in the rabbit.

Milk protein synthesis is initiated by prolactin and a glucocorticoid. In the rabbit, prolactin alone is sufficient. However, glucocorticoids potentiate the action of prolactin. The stimulatory effect of glucocorticoids was evaluated after injections of hydrocortisone acetate alone or associated with prolactin by measurements of (a) the total RNA and DNA content of mammary glands, (b) the lactose synthetase activity, (c) casein synthesis, and (d) the concentration of casein mRNA in total cellular RNA and in polysomal RNA by hybridization with its cDNA. The glucocorticoid, totally inactive alone, proved to have a stimulatory effect proportional to the dose injected when prolactin was present. This effect was more evident with low doses of prolactin. Glucocorticoids proceeded by amplifying the capacity of prolactin to enhance the concentration of casein mRNA available for translation. A parallel effect of glucocorticoids on translation of casein mRNA was suspected. Glucocorticoids injected with low doses of prolactin were unable to mimic all the effects of high doses of prolactin alone.

Animals↗