Sandrine Cassy

Chicken leptin: properties and actions

Chicken leptin cDNA shows a high homology to mammalian homologous, with an expression localized in the liver and adipose tissue. It is noteworthy, that the hepatic expression is most likely associated with the primary role that this organ plays in lipogenic activity in avian species. As in mammals, chicken leptin expression is regulated by hormonal and nutritional status. This regulation is tissue-specific and with a high sensitivity in the liver compared to adipose tissue. The blood leptin levels are regulated by the nutritional state with high levels in the fed state compared to the fasted state. The recombinant chicken leptin markedly inhibits food intake as reported in mammals, suggesting the presence of an hypothalamic leptin receptor. The chicken leptin receptor has been identified and all functional motifs are highly conserved compared to mammalian homologous. Chicken leptin receptor is expressed in the hypothalamus but also in other tissues such as pancreas, where leptin inhibits insulin secretion and thus may have a key role in regulating nutrient utilization in this species.

Broiler breeder paradox: a project report

A first product of this European project was a tool for scoring chick quality. A link was established between chick quality and embryonic physiological parameters such as heat production. Eggs from broiler breeders that give rise to rapidly growing broilers have a different embryonic development that may need modified incubation conditions. Within genotypes, better chick quality induces better liveability and faster growth in broiler production. Extensive data on broiler breeder production were generated. Feed restriction was necessary to maintain welfare and reproduction at acceptable levels in standard broiler breeders. The dw-experimental genotype (E) was more tolerant to ad libitum feeding. If an alternative to feed restriction had to be found, the use of a dw genotype with less severe feed restriction could be adopted. Fibre per se, or partial feed restriction during the growing period, only compensated to a small extent for the negative effects of early fast growth on later reproduction. The results of the project on behaviour did not support welfare concerns on feed restriction. They confirmed the hypothesis that environmental pecking is a displacement activity rather than a sign of frustration. Factors other than central control by pituitary hormones seem to be involved in the modulation of the laying rate. The local (intra-ovarian) role of growth factors such as Insulin-like Growth Factors (IGFs), Bone Morphogenetic Proteins (BMPs) and leptin are known to modulate the effect of gonadotrophins on ovarian function. For both BMPs and IGFs, feed restriction enhanced the interaction between growth factors and gonadotrophins as well as the proliferation of granulosa cells in vitro. Future genetic selection of broiler breeder production might aim at uncoupling the control of growth factors in the ovary from the selection for rapid growth to maintain or increase the growth rate of chicks without further penalising the already poor reproductive performance of broiler breeders. Putative quantitative trait loci for ovulation rate were identified in the project and may eventually facilitate selection by breeding companies for birds that could be fed enough feed to optimise their welfare.

Increase in prolactin receptor (PRL-R) mRNA level in the mammary gland after hormonal induction of lactation in virgin ewes☆

In order to examine the hormonal regulation of the prolactin-receptor (PRL-R) gene expression during mammary gland development, ewes were treated to induce lactation via an estrogen-progesterone-hydrocortisone and ovine growth hormone treatment. In situ hybridization analysis was used and revealed that sex steroids increased PRL-R mRNA levels in the mammary gland. Using RNase protection assay we showed that the estradiol + progesterone treatment increased both the levels of the long and the short forms of PRL-R mRNA. Addition of hydrocortisone increased the level of alphaS1-casein transcripts and the level of the ratio of the long to the short form of the PRL-R mRNA. This ratio can be further enhanced by addition of ovine growth hormone to the latter treatment. This suggests a role of hydrocortisone and ovine growth hormone in the alternative splicing that leads to the preferential expression of the long form of the PRL-R mRNA. In conclusion, the present experiments suggest that estrogen, progesterone and hydrocortisone are the major regulators of the PRL-R gene expression during pregnancy and prepare the mammary gland for its differentiation.

Cellular localization and evolution of prolactin receptor mRNA in ovine endometrium during pregnancy.

In this study, we have investigated the expression of the prolactin receptor gene in ovine endometrium during oestrus cycle and pregnancy. Using reverse transcription‐PCR analysis, we provided evidence that the prolactin receptor gene is specifically transcribed in this tissue. As shown by Northern blot analysis, the level of the prolactin receptor transcripts increased dramatically during late pregnancy. In situ hybridization experiments revealed that prolactin receptor mRNA was specifically expressed in the glandular compartment and confirmed the dramatic increase of its expression that occurs at the end of pregnancy. Taken together, these findings are consistent with a putative role of prolactin and/or related molecules in the regulation of the proliferation of the glandular compartment and/or in the control of the secretory activity of the endometrium.

Structure and expression of goat GLYCAM1 gene: lactogenic-dependent expression in ruminant mammary gland and interspecies conservation of the proximal promoter.

A macroarray approach used to list genes differentially expressed in goat mammary gland (gestation vs. lactation), other than milk protein genes, allowed us to detect the Glycosylation-dependent Cell Adhesion Molecule 1 (GLYCAM1) gene. GLYCAM1, a member of the glycoprotein mucin family, is a component of the milk fat globule membrane (MFGM). Its complete cDNA and gene sequences were determined and it was mapped by fluorescent in situ hybridization (FISH) on goat and cattle chromosome 5 (CHI5q21 and BTA5q21), and on sheep chromosome 3 (OAR3q21). Northern blot analyses confirmed its differential expression during the development and differentiation of the mammary gland of ruminants with a significantly higher mRNA amount during lactation than during pregnancy. An experimental in vivo induction model for lactation, developed by Kann et al., showed that the expression of GLYCAM1 is hormonally regulated in the mammary gland of ewes. Interspecies comparison of the gene promoter revealed the evolutionary conservation of a short proximal nucleotide sequence encompassing several transcription factor binding sites that could mediate the above-mentioned hormonal regulation.