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L M Houdebine

Publications and source records attributed to L M Houdebine.

At least 127 records · Page 7Linked to original sources
PubMed1982

[Role of membrane colchicine binding proteins in the transmission of prolactin message to casein genes in the rabbit mammary gland].

Previous work demonstrated that tubulin binding drugs specifically inhibit the capacity of prolactin to initiate casein and DNA synthesis in the mammary cell. It was concluded that microtubules or other tubulin containing cellular structures were involved in the transmission of the prolactin message to genes. In the present work, it is shown that griseofulvin, an antimitotic drug which alters microtubule structure and function, does not prevent prolactin actions. Autoradiographic studies showed that [3H]colchicine binds preferentially to plasma and Golgi membranes in the mammary cell. Short term cultures of mammary explants with [3H]colchicine demonstrated that the labelled drug binds to membranous cellular structures which were isolated from explants at the end of the culture. Fractions containing plasma and Golgi membranes contained the highest amount of radioactivity. Solubilisation of the membranes by Triton X-100 dissociated the [3H]colchicine from the prolactin receptors as judged by a chromatography of the soluble fraction on a Sepharose 6 B column. On the column, the labelled colchicine remains associated with a molecular entity which may be free tubulin. In all cases, the binding of [3H]colchicine was greatly attenuated by an excess of unlabelled colchicine but was only slightly affected by the competition with lumicolchicine. These results suggest that mammary membranes contain tubulin and that binding of drugs to this molecule inhibits the generation of the prolactin second messengers eliciting the hormonal actions in the mammary cell. This also suggests that microtubules are probably not involved in the mechanism of prolactin action.

Animals↗
PubMed1982

Effects of insulin, cortisol and prolactin on lipid, protein and casein syntheses in goat mammary tissue in organ culture.

The actions of insulin, prolactin and cortisol on protein, lipid and casein syntheses were examined in goat mammary explants in culture. The synthesis of the three products was stimulated by insulin and prolactin associated or not with cortisol. Casein synthesis was stimulated only when prolactin was present in the culture medium. Prolactin alone was able to significantly support the induction of casein synthesis. After preculture in the presence of insulin alone, the tissue lost part of its capacity to respond to the prolactin stimulus but, when precultured in the presence of insulin + cortisol, it retained its total sensitivity. These data suggest that insulin, prolactin and cortisol are involved in the maintenance of goat mammary tissue in culture but that prolactin essentially stimulates milk synthesis.

Adipose Tissue↗
PubMed1982

Correlation between prolactin-receptor interaction, down-regulation of receptors, and stimulation of casein and deoxyribonucleic acid biosynthesis in rabbit mammary gland explants.

Increasing concentrations of PRL were added to culture media of rabbit mammary gland explants. After 24 h of culture, free and total PRL receptors, casein synthesis, casein messenger (m) RNA concentrations and DNA synthesis were estimated. In the PRL concentration range of 0-100 ng/ml, receptors were progressively down-regulated by the hormone, but no occupied desaturable receptors could be detected. At concentrations between 1,000 and 20,000 ng/ml, the PRL receptors were increasingly occupied without being quantitatively down-regulated. There exists a reciprocal correlation between the rate of casein synthesis, casein mRNA concentration, DNA synthesis and the down-regulation of the receptors as a function of PRL concentration. The maximal responses were reached around a PRL concentration of 100 ng/ml. Interestingly, with a large excess of PRL a desensitization process was observed, the responses to the hormone being attenuated. This desensitization was more pronounced for DNA synthesis than for casein synthesis or for casein mRNA accumulation. The data suggest that down-regulation of PRL receptors occurs even at physiological concentrations of PRL. These observations are compatible with the hypothesis that each receptor can generate a limited number of intracellular relays eliciting hormonal action before being irreversibly inactivated and degraded. In addition, high concentrations of PRL induce a refractory state in the mammary gland that is possibly related to a low level of peripheral PRL receptors.

Animals↗
PubMed1982

Hormonal control of casein synthesis in mammary explants from pregnant goats.

The effects of insulin, cortisol, prolactin, 3,3',5-triiodo-L-thyronine (L-T3) and progesterone on the synthesis of total protein and casein in mammary explants from pregnant goats were studied. In the absence of hormones and in the presence of insulin plus cortisol the rate of incorporation of 14C-leucine into proteins that were precipitated with the anti-casein antibody decreased during culture. The addition of prolactin to hormonal combination of insulin and cortisol caused large stimulation of rates of casein synthesis. Maximum incorporation of leucine was attained between 3 and 5 days of culture in the presence of 0.5 microgram ml-1 of prolactin. Prolactin stimulated-casein and total protein synthesis were not consistently affected by the addition of L-T3 or progesterone. The inhibition of DNA synthesis by hydroxyurea or cytosine-arabinofuranoside had no effect on casein synthesis in mammary explants from pregnant goats.

Animals↗
PubMed1982

Effect of bovine growth hormone on development of goat mammary tissue in organ culture.

The effect of bovine growth hormone (bGH) on DNA, protein and casein synthesis in goat mammary explants was studied. Growth hormone was unable to stimulate DNA synthesis or potentiate insulin-stimulated DNA synthesis either in high or low oxygen concentrations. In the presence of insulin and cortisol bGH had no effect on the synthesis of explant cytosol proteins. Two preparations of bGH were tested for their effect on casein synthesis. The preparation NIH-GH-B17 at concentration 50 micrograms ml-1 increased casein synthesis similarly as about 0.5 micrograms ml-1 of prolactin. Our preparation, prepared by the method of Prusík and Braun [1967], at concentration 50 micrograms ml-1 had effect lower than 0.05 micrograms ml-1 of prolactin. The lactogenic activity of NIH-GH-B17 was decreased by treatment with specific antisera to bovine prolactin. When submaximum concentration of prolactin (0.05 microgram ml-1) was added, bGH at concentration as low as 0.5 microgram ml-1 had synergistic effect on prolactin stimulated casein synthesis.

Animals↗
PubMed1981

Prolactin induces release of a factor from membranes capable of stimulating beta-casein gene transcription in isolated mammary cell nuclei.

Crude microsomes from lactating rabbit mammary gland were incubated with prolactin. The incubation mixture was centrifuged and the supernatant was incubated with isolated mammary cell nuclei from lactating rabbits treated for 4 days by bromocryptin to antagonize prolactin and to deinduce casein gene transcription. Nuclei were incubated with HgCTP, and the newly synthesized mercurated RNA was isolated on SH-Sepharose columns. The content of beta-casein mRNA sequences in the fraction eluted with 2-mercaptoethanol was estimated with a [(3)H]cDNA probe obtained from partially purified beta-casein mRNA. The supernatant markedly stimulated beta-casein gene transcription but not 28S rRNA transcription. The same effect was obtained with other lactogenic hormones such as human growth hormone and ovine placental lactogen but was not observed with bovine growth hormone, insulin, parathyroid hormone, luteotropic hormone, or epidermal growth factor. Prolactin and human growth hormone were totally inactive when added directly to nuclei. The factor stimulating beta-casein gene transcription was also generated by membranes containing prolactin receptors such as those from liver, ovary, adrenals, and brain but not by membranes from heart, lung, and muscle, which do not bind prolactin. The factor stimulated beta-casein transcription when added to mammary nuclei from pseudopregnant or bromocryptin-treated lactating rabbits, in which the transcription rate is submaximal, but was ineffective on mammary nuclei prepared from untreated fully lactating rabbits. The factor was unable to induce beta-casein gene transcription in nuclei isolated from rabbit liver and reticulocytes. The factor did not stimulate the transcription of globin genes in nuclei isolated from reticulocytes or the transcription of mammary "housekeeping" genes evaluated by a cDNA probe prepared from total mRNA isolated from an unstimulated mammary gland. The transcription of beta-casein genes was abolished by adding alpha-amanitin to the medium in the presence or in the absence of the factor, indicating that the generation of mercurated beta-casein mRNA sequences depended upon the transcriptional activity of RNA polymerase II. The addition of the factor to the incubation mixture did not enhance total and alpha-amanitin-sensitive RNA synthesis. These data suggest that the binding of prolactin to its receptor in vitro induces the formation of a second messager, which specifically stimulates the transcription of prolactin-sensitive genes.

Animals↗
PubMed1981

Prolactin-like activity of anti-prolactin receptor antibodies on casein and DNA synthesis in the mammary gland.

Prolactin receptors were partially purified from rabbit mammary gland membranes by using an affinity chromatography technique. Antibodies against this prolactin receptor preparation were obtained in guinea pig and sheep. Both antisera were able to inhibit the binding of 125I-labeled ovine prolactin to rabbit mammary gland membranes. When added to culture media of rabbit mammary explants, the anti-prolactin receptor antiserum inhibited the capacity of prolactin to initiate casein synthesis and casein mRNA accumulation as a function of the antiserum concentration. However, in the absence of prolactin, both antisera (guinea pig and sheep) at moderate concentrations were capable of mimicking prolactin action on casein gene expression and on DNA synthesis. At higher concentrations, the anti-prolactin receptor antibodies inhibited their own actions. Several characteristics of the prolactin effect were also observed with the anti-prolactin receptor antibody: the stimulatory effect of the antibody was amplified by glucocorticoids; colchicine, which was capable of blocking prolactin action, also prevented the induction by the antibody. Lysosomotropic agents, which do not interfere with prolactin action, did not alter the response observed with the antibody. These results indicate that an anti-prolactin receptor antibody can mimic two major actions of prolactin obtained in mammary explant culture and suggests that the prolactin molecule is not required beyond the initial binding to its receptor.

Animals↗
PubMed1981

Role of progesterone and glucocorticoids in the transcription of the 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 measured using [3H]cDNA probes complementary to beta-casein mRNA and 28-S rRNA respectively. Prolactin injected into pseudopregnant animals accelerates the transcription of both genes and increases the stability of the beta-casein mRNA but not of the 28-S rRNA. Progesterone injected simultaneously with prolactin reduced considerably all these effects of prolactin, with a lower efficiency when the highest doses of prolactin were injected. These observations suggest that progesterone attenuates the transfer of prolactin information related to the lactogenesis into the mammary cell. Glucocorticoids injected with prolactin amplify the prolactin action on the expression of the beta-casein gene but not of the 28-S rRNA genes. In the absence of prolactin (a situation obtained by injecting simultaneously CB 154, a drug which inhibits pituitary prolactin secretion) glucocorticoids exhibit no effect. In the lactating rabbit glucocorticoids do not delay significantly the drop of beta-casein gene transcription rate provoked by weaning or by prolactin withdrawal obtained by injecting CB 154. A comparison of the beta-casein mRNA accumulation and the transcription rate of the beta-casein gene indicates that glucocorticoids act essentially by amplifying the activation of beta-casein transcription supported by prolactin, but not by enhancing the stability of the beta-casein mRNA.

Animals↗
PubMed1981

Induction of casein synthesis by prolactin and inhibition by progesterone in the pseudopregnant rabbit treated by colchicine without any simultaneous variations of casein mRNA concentration.

Prolactin injected into pseudopregnant rabbits induced casein synthesis and it provoked an accumulation of casein mRNA and of ribosomal RNA. Colchicine, which has been shown to block the prolactin signal, totally prevented the accumulation of beta-casein mRNA, when injected with the hormone. However, the drug did not hamper the initiation of casein synthesis and the accumulation of ribosomal RNA. The effect of prolactin injected with colchicine on casein synthesis was totally abrogated by progesterone administered simultaneously and it was essentially unmodified by glucocorticoids. These results suggest that prolactin controls casein gene expression by supporting the accumulation of casein mRNAs and by stimulating the translation of these mRNAs, through independent mechanisms. Progesterone which is known to prevent the accumulation of casein mRNAs is also a potent inhibitor of casein mRNA translation whereas glucocorticoids exert their effect essentially by favouring the accumulation of casein mRNA but not by modifying their translation efficiency.

Animals↗
PubMed1981

[Control of protein synthesis in eukaryote cells].

In the present report, the following aspects of the control of gene expression in eukaryote cells has been reviewed: 1) the mechanism of mRNA formation in relation to recent findings on introns in genes; 2) the correlation between the concentration of mRNAs in cells and the rate of synthesis of the related proteins; 3) the control of specific transcription; 4) the stabilization of specific mRNAs; 5) the involvement of genome rearrangement as a possible mode of controlling gene expression; 6) the mechanism of hormonal control of specific gene expression.

Animals↗
PubMed1980

Effects of lysosomotropic agents, cytochalasin B and colchicine on the "down-regulation" of prolactin receptors in mammary gland explants.

Effects of lysosomotropic agents and drugs which disrupt the cytoskeleton on prolactin receptor levels were studied in organ culture of rabbit mammary glands. As we have shown previously, prolactin is able to induce a down-regulation of its own receptor levels. In the present experiments, lysosomotropic agents, chloroquine, ammonium chloride and methylamine, in the presence of prolactin are capable of almost completely preventing this down-regulation. Interestingly, these lysosomotropic agents alone could increase the level of prolactin receptors, which confirms that the degradation of prolactin receptors occurs in the lysosomes and that this is a rapid process which can proceed in the absence of prolactin. The almost complete lack of effect of microfilament (cytochalasin B) or microtubule (colchicine) disrupting drugs seems to indicate that the cytoskeleton is not involved in the down-regulation process.

Animals↗