👤 Damasia Becu-Villalobos

Dairy heifers with gastrointestinal nematodes have reduced growth rates, and delayed age at puberty and milk production onset related to late mammary gland development. IGF1 and Notch signaling systems are important in this process, and an altered profile of serum IGF1 has been associated with the detrimental effect of the nematodes on parenchymal development. In this context, we aimed to study the molecular mechanisms involved in bovine mammary gland development around pre and postpuberty, focusing on proliferative and angiogenic processes that involve the Notch and IGF1 pathways. We used mammary tissue samples from pre and pubertal heifers, treated or untreated with anthelmintics, and MAC-T bovine mammary epithelial cells in vitro. Anthelminthic treatment effectively lowered EPG in feces. Mammary glands from treated heifers had increased proliferation rate (measured by PCNA) and angiogenic marker expression (VEGF and CD34), as well as increased αSMA area compared to age-matched control parasitized heifers. These changes were preceded by increased expression of Notch targets at 20 wk of age (HES1, HEY2, and HEY1), indicating a possible interaction. Similarly, IGF1R expression was increased at 30 weeks of age. To study the crosstalk between systems, bovine MAC-T cells were treated with DAPT (50 μM) to inhibit Notch signaling. DAPT decreased the proliferation of cells as evidenced by a decrease in PCNA, pERK, CYCYLIN D1; and the wound healing capacity of HMEC cells was impaired in the presence of the supernatants of DAPT-treated cells. Furthermore, DAPT decreased IGF1 and increased IGF1R mRNA levels in MAC-T cells. On the other hand, cells treated with 10 ng/mL IGF1 Increased their proliferation (MTS assay), and induced a strong tendency to increase Notch target genes (HEY1, and HES1). Furthermore, IGF1 treatment tampered the decrease in the proliferation rate induced by DAPT. Finally, a positive correlation between the IGF1R and Notch target genes (HEY1, and HES1) further suggested a relation between these two signaling systems in the bovine mammary gland. In conclusion, pubertal delay related to parasitosis is counteracted by anthelminthic treatments, which increase serum IGF1, mammary cell proliferation, and angiogenesis. We postulate the Notch pathway, mainly through the HEY1 target gene, which is modulated by the IGF1 system, may regulate both proliferative and angiogenic processes favoring normal development of the bovine mammary gland during puberty. In addition, we demonstrate that the interaction between the Notch and the IGF1 pathways may affect cell proliferation. Show less

The development of the mammary gland of cows during pre-weaning and puberty will condition its future productive capacity and warrants special study. In this respect, Notch signaling regulates tissue development and fate by modifying cell proliferation and differentiation and has been involved in stem cell maintenance, but has not been extensively studied in the developing mammary glands in cows. We therefore investigated Notch receptor expression and localization, as well as the expression of Notch ligands and target genes in the mammary gland of Holstein heifers in pre- and post-pubertal stages. Notch receptors 1 to 4 were detected by immunohistochemistry in the parenchyma and stroma of the developing gland. The subcellular localization of the four receptors was predominantly cytoplasmic except for NOTCH4, which was mostly nuclear. The membrane and the active intracellular domains of NOTCH paralogues were identified by western blot. NOTCH1 and NOTCH2 active domains increased during pubertal stages while NOTCH3 and NOTCH4 active domains decreased, suggesting strikingly different involvement of NOTCH paralogues in bovine mammary gland development and differentiation. The mRNA expression levels of the target genes HEY1 and HEY2 increased during peri-puberty whereas no variation of HES1 mRNA levels was observed. The mRNA levels of the Notch ligands JAGGED1 and DELTA1 also increased gradually during development. In conclusion, Notch signaling system dynamically varies throughout the development of the mammary gland during puberty pointing to specific time involvement of each component. Show less

Pituitary adenomas are among the most frequent intracranial neoplasms and treatment depends on tumor subtype and clinical features. Unfortunately, non responder cases occur, then new molecular targets are needed. Notch system component expression and activation data are scarce in pituitary tumorigenesis, we therefore aimed to characterize Notch system in pituitary tumors of different histotype. In human pituitary adenomas we showed Show less

We studied the impact of high prolactin titers on liver and adipocyte gene expression related to glucose and insulin homeostasis in correlation with obesity onset. To that end we used mutant female mice that selectively lack dopamine type 2 receptors (D2Rs) from pituitary lactotropes (lacDrd2KO), which have chronic high prolactin levels associated with increased body weight, marked increments in fat depots, adipocyte size, and serum lipids, and a metabolic phenotype that intensifies with age. LacDrd2KO mice of two developmental ages, 5 and 10 mo, were used. In the first time point, obesity and increased body weight are marginal, although mice are hyperprolactinemic, whereas at 10 mo there is marked adiposity with a 136% increase in gonadal fat and a 36% increase in liver weight due to lipid accumulation. LacDrd2KO mice had glucose intolerance, hyperinsulinemia, and impaired insulin response to glucose already in the early stages of obesity, but changes in liver and adipose tissue transcription factors were time and tissue dependent. In chronic hyperprolactinemic mice liver Prlr were upregulated, there was liver steatosis, altered expression of the lipogenic transcription factor Chrebp, and blunted response of Srebp-1c to refeeding at 5 mo of age, whereas no effect was observed in the glycogenesis pathway. On the other hand, in adipose tissue a marked decrease in lipogenic transcription factor expression was observed when morbid obesity was already settled. These adaptive changes underscore the role of prolactin signaling in different tissues to promote energy storage. Show less