Characterisation of energy metabolism during lactation in cellular and mouse models

产妇代谢性疾病如糖尿病和肥胖与泌乳不足和母乳喂养失败有关。该项目的目标是进行发现科学，以表征参与能量代谢的分子和组织如何促进母乳的最佳供应。该项目的第一部分将研究脂联素受体1 (AdiporR1)，这是一种在妊娠和哺乳期人类乳腺上皮细胞中高度表达的激素受体。AdiporR1检测脂肪细胞衍生激素脂联素的变化，据报道，脂联素可以上调骨骼肌等组织中的线粒体生物发生。AdiporR1在乳腺中的作用尚不确定，我们将使用表达AdiporR1的人乳腺上皮细胞系MCF-10A进行研究。用重组脂联素刺激培养的MCF-10A细胞，分别用线粒体DNA拷贝数和酶活性测定评估线粒体丰度和功能。将对MCF-10A细胞中的AdiporR1进行RNA沉默，以确定这是否会消除脂联素对线粒体的任何观察到的影响。如果AdipoR1显示影响乳腺上皮细胞功能，那么AdipoRon，一种市售的口服活性脂联素受体激动剂，将在怀孕期间和产后早期给予雌性野生型小鼠，以评估其对哺乳的影响，通过断奶前的幼鼠生长来衡量。该项目的第二部分将确定哪些激素参与调节哺乳期间参与能量代谢的组织。年龄匹配的怀孕和哺乳期成年雌性小鼠将通过评估葡萄糖耐量和胰岛素敏感性、血浆脂质和体脂肪组成进行代谢谱分析。将获得血浆用于测量关键的生乳激素和代谢激素。相关组织，如脂肪组织，骨骼肌，胰岛和乳腺将从这些小鼠解剖rna测序。我们将在这些组织中进行激素受体和信号蛋白表达的全球调查，并将其与从妊娠到哺乳期过渡期间发生的激素和代谢变化相关联。因此，这些研究旨在确定母体组织如何适应母乳喂养的代谢需求，并为了解糖尿病和肥胖等疾病如何导致泌乳不足提供基础。

Maternal metabolic diseases such as diabetes and obesity are associated with lactation insufficiency and breastfeeding failure. The goal of this project is to undertake discovery science to characterise how molecules and tissues involved in energy metabolism contribute to the optimal supply of breast milk. The first part of this project will investigate the adiponectin receptor 1 (AdiporR1), a highly expressed hormone receptor in human mammary epithelial cells during pregnancy and lactation. AdiporR1 detects alterations in an adipocyte-derived hormone known as adiponectin, which has been reported to upregulate mitochondrial biogenesis in tissues such as skeletal muscle. The role of AdiporR1 in the mammary gland is uncertain, and this will be investigated using the MCF-10A human mammary epithelial cell line, which expresses AdiporR1. Cultured MCF-10A cells will be stimulated with recombinant adiponectin, and mitochondrial abundance and function assessed using mitochondrial DNA copy number and enzyme activity assays, respectively. RNA silencing of AdiporR1 in MCF-10A cells will be performed to determine whether this abrogates any observed mitochondrial effects of adiponectin. If AdipoR1 is shown to influence mammary epithelial cell function, then AdipoRon, a commercially available orally active adiponectin receptor agonist will be administered to female wild-type mice during pregnancy and the early post-partum period to assess the effect on lactation, as measured by pup growth prior to weaning. The second part of this project will identify which hormones are involved in regulating tissues involved in energy metabolism during lactation. Age-matched pregnant and lactating adult female mice will undergo metabolic profiling by assessment of glucose tolerance and insulin sensitivity, plasma lipids and body fat composition. Plasma will be obtained for measurement of key lactogenic and metabolic hormones. Relevant tissues e.g. adipose tissue, skeletal muscle, pancreatic islets, and mammary glands will be dissected from these mice for RNA-sequencing. A global survey of hormone receptor and signalling protein expression will be undertaken in these tissues, and correlated with hormonal and metabolic changes occurring during the transition from pregnancy to lactation. Thus, these studies aim to determine how maternal tissues adapt to the metabolic demands of breastfeeding, and also to provide a basis for understanding how diseases such as diabetes and obesity lead to lactation insufficiency.