Amh regulate female folliculogenesis and fertility in a dose-dependent manner through Amhr2 in Nile tilapia Publication date: 1 January 2020 Source: Molecular and Cellular Endocrinology, Volume 499 Author(s): Xingyong Liu, Hesheng Xiao, Mimi Jie, Shengfei Dai, Xin Wu, Minghui Li, Deshou Wang Abstract In the present study, Amh was found to be abundantly expressed in the granulosa cells of the primary growth follicles, and Amhr2 in the granulosa cells, oogonia and phase I oocytes in tilapia by immunohistochemistry. In addition, Amh and Amhr2 were also found to be expressed in the brain and pituitary. Heterozygous mutation of either amh or amhr2 resulted in increased primary growth follicles and decreased fertility, and homozygous mutation resulted in hypertrophic ovaries with significantly increased primary follicles and failed transition from primary to vitellogenic follicles. Expression of gnrh3 in the brain, fsh and lh in the pituitary and serum E2 concentration were significantly decreased in both mutants. Significantly increased apoptosis of follicle cells was observed in both mutants. However, administration of E2 failed to rescue the folliculogenesis defects of the mutants. Our results suggested that Amh acts in a dose-dependent manner by binding Amhr2 in tilapia.

Lessons from animal models of endocrine disorders caused by defects of protein folding in the secretory pathway Publication date: 1 January 2020 Source: Molecular and Cellular Endocrinology, Volume 499 Author(s): Yoshiaki Morishita, Peter Arvan Abstract Most peptide hormones originate from secretory protein precursors synthesized within the endoplasmic reticulum (ER). In this specialized organelle, the newly-made prohormones must fold to their native state. Completion of prohormone folding usually occurs prior to migration through the secretory pathway, as unfolded/misfolded prohormones are retained by mechanisms collectively known as ER quality control. Not only do most monomeric prohormones need to fold properly, but many also dimerize or oligomerize within the ER. If oligomerization occurs before completion of monomer folding then when a poorly folded peptide prohormone is retained by quality control mechanisms, it may confer ER retention upon its oligomerization partners. Conversely, oligomerization between well-folded and improperly folded partners might help to override ER quality control, resulting in rescue of misfolded forms. Both scenarios appear to be possible in different animal models of endocrine disorders caused by genetic defects of protein folding in the secretory pathway. In this paper, we briefly review three such conditions, including familial neurohypophyseal diabetes insipidus, insulin-deficient diabetes mellitus, and hypothyroidism with defective thyroglobulin.

Prenatally androgenized female rats develop uterine hyperplasia when adult Publication date: 1 January 2020 Source: Molecular and Cellular Endocrinology, Volume 499 Author(s): Silvana Rocío Ferreira, Alicia Alejandra Goyeneche, María Florencia Heber, Giselle Adriana Abruzzese, Carlos Marcelo Telleria, Alicia Beatriz Motta Abstract Prenatal hyperandrogenization (PH) is hypothesized as one of the main factors contributing to the development of polycystic ovary syndrome (PCOS). In this study, we aimed to investigate the impact of prenatal exposure to androgen excess on the uterus when animals reach their adulthood. We found that PH altered the morphology of the uteri that show a hyperplastic morphology with increased total uterine thickness as well as luminal epithelium thickness, with both enhanced and altered distribution of glands as compared with controls. Morphological alterations were associated with an unbalanced homeostasis as assessed by the expression of regulators of cell cycle progression and cell death dynamics. PH also causes disturbances in the cell cycle of the uterine tissue and dysregulates cell death and survival pathways leading to the development of uterine hyperplasia. These findings suggest that PH may have a deleterious effect on the uterus.

Generation of a lentiviral vector system to efficiently express bioactive recombinant human prolactin hormones Publication date: 1 January 2020 Source: Molecular and Cellular Endocrinology, Volume 499 Author(s): Hernán Alarcón, Elena Bonzon-Kulichenko, Rocío Peinado, Filip Lim, Jesús Vázquez, Antonio Rodríguez Abstract The contribution of the pleiotropic hormone Prolactin (PRL) to several physiological and pathological processes is still unknown. To clarify the role of PRL in these processes during the last decade, different human PRL antagonists have been produced to either partially or fully block the wild type hormone activity. In this work, we have cloned these wild type and antagonist sequences in lentivectors (LV) to express them as recombinant self-processing polypeptides by employing a P2A sequence (hPRL-P2A-GFP). We show that these LVs can efficiently transduce and express the hPRL proteins in different cell types and that the P2A sequence does not affect their activities. Additionally, we have tested their activities in paracrine and autocrine cell culture experiments. Our results demonstrate that these recombinant hPRL-P2A proteins are bioactive in both paracrine and autocrine modes, highlighting the potential usefulness of these hPRL-containing LVs for determining the contribution of hPRL to different biological processes. Graphical abstract

Upregulation of adamts9 by gonadotropin in preovulatory follicles of zebrafish Publication date: 1 January 2020 Source: Molecular and Cellular Endocrinology, Volume 499 Author(s): Dong Teng Liu, Wan Shu Hong, Shi Xi Chen, Yong Zhu Abstract Previously we had identified adamts9 as a downstream target of Pgr, which is essential for ovulation in zebrafish. The primary goal of this study is to determine whether human chorionic gonadotropin (hCG, LH analog) also regulate adamts9 expression prior to ovulation. The expression of adamts9 was induced by hCG in a dose and time dependent manner in zebrafish preovulatory follicles in vitro. Interestingly, the stimulatory effect of hCG on adamts9 expression was not blocked in pgr−/− follicles but blocked in lhcgr−/−. This effect of hCG was via Lhcgr and its associated cAMP and PKC signaling pathways. Reduced fecundity and reduced expression of adamts9 were also found in lhcgr−/− females in vivo. Therefore, we have provided the first evidence of gonadotropin (hCG) regulated adamts9 in zebrafish, which could be important for ovulation.

The challenging use and interpretation of blood biomarkers of exposure related to lipophilic endocrine disrupting chemicals in environmental health studies Publication date: 1 January 2020 Source: Molecular and Cellular Endocrinology, Volume 499 Author(s): German Cano-Sancho, Philippe Marchand, Bruno Le Bizec, Jean-Philippe Antignac Abstract The use of exposure biomarkers has been growing during the last decades, being considered the ‘gold-standard’ approach for individual exposure assessment to environmental chemicals. However, lipophilic endocrine disrupting chemicals (LEDC) have specific physicochemical and biological properties implying particular analytical challenges and interpretative caveats. The epidemiological literature is therefore afflicted by methodological inconsistencies and results divergences, in part due to recognised sources of exposure measurement error and misinterpretation of results. The aim of the present review is to identify external and endogenous sources of variability and uncertainty associated with the LEDC blood biomarkers in epidemiological studies. The dynamic nature of blood and an overview of the known mechanisms of transport, storage and partition of LEDCs in the organism are first described. The external sources of variability and uncertainty introduced at pre-analytical and analytical level are subsequently presented. Subsequently, we present some specific cases where the dynamics of lipids and LEDCs may be substantially modified and thus, the interpretation of biomarkers can be particularly challenging. The environmental obesogens as source of biomarkers variability is also discussed in the light of the most recent findings. Finally, different modelling approaches (statistical and pharmacokinetic models) proposed to improve the use and interpretation of biomarkers are appraised.

Diiodothyronines regulate metabolic homeostasis in primary human hepatocytes by modulating mTORC1 and mTORC2 activity Publication date: 1 January 2020 Source: Molecular and Cellular Endocrinology, Volume 499 Author(s): Davide Gnocchi, Ewa C.S. Ellis, Helene Johansson, Mats Eriksson, Giovannella Bruscalupi, Knut R. Steffensen, Paolo Parini Abstract Until three decades, ago 3,5-diiodothyronine (3,5-T2) and 3,3′-diiodothyronine (3,3′-T2) were considered products of thyroid hormone catabolism without biological activity. Some metabolic effects have been described in rodents, but the physiological relevance in humans and the mechanisms of action are unknown. Aim of this work was to investigate the role and the mechanisms of action of 3,5-T2 and 3,3′-T2 in the regulation of metabolic homeostasis in human liver. We used primary human hepatocytes freshly isolated from donors and grown on Matrigel as the golden standard in vitro model to study human hepatic metabolism. Results show that diiodothyronines in the range of plasma physiological concentrations reduced hepatic lipid accumulation, by modulating the activity of the mTORC1/Raptor complex through an AMPK-mediated mechanism, and stimulated the mTORC2/Rictor complex-activated pathway, leading to the down regulation of the expression of key gluconeogenic genes. Hence, we propose that diiodothyronines act as key regulators of hepatic metabolic homeostasis in humans. Graphical abstract

miR-181a promotes porcine granulosa cell apoptosis by targeting TGFBR1 via the activin signaling pathway Publication date: 1 January 2020 Source: Molecular and Cellular Endocrinology, Volume 499 Author(s): Jia-Qing Zhang, Bin-Wen Gao, Hong-Xia Guo, Qiao-Ling Ren, Xian-Wei Wang, Jun-Feng Chen, Jing Wang, Zi-Jing Zhang, Qiang Ma, Bao-Song Xing Abstract Activin/Smad3 signaling plays a pivotal role in follicle development and atresia. However, the precise mechanisms underlying this process are not yet fully understood. Herein, we identified miR-181a as a central component of activin/Smad3-mediated follicle atresia. miR-181a was strikingly upregulated in porcine atretic follicles, which induced the apoptosis of porcine granulosa cells (GCs) in vitro. Furthermore, the transforming growth factor-β type 1 receptor (TGFBR1) was confirmed as a direct target of miR-181a by bioinformatics analysis and luciferase assays. Transfection with an miR-181a agomir repressed the TGFBR1 mRNA and protein levels. In addition, TGFBR1 overexpression repressed GC apoptosis, whereas TGFBR1 inhibition promoted GC apoptosis. miR-181a overexpression downregulated the phosphorylation of Smad3 and blocked the activation of TGF-β signaling. Moreover, activin A downregulated miR-181a expression and upregulated the TGFBR1 and p-Smad3 protein levels. Collectively, these data suggest that miR-181a regulates porcine GC apoptosis by targeting TGFBR1 via the activin signaling pathway.

Targeting the pentose phosphate pathway increases reactive oxygen species and induces apoptosis in thyroid cancer cells Publication date: 1 January 2020 Source: Molecular and Cellular Endocrinology, Volume 499 Author(s): Chien-Liang Liu, Yi-Chiung Hsu, Jie-Jen Lee, Ming-Jen Chen, Chi-Hsin Lin, Shih-Yuan Huang, Shih-Ping Cheng Abstract The pentose phosphate pathway (PPP) plays an important role in the biosynthesis of ribonucleotide precursor and NADPH. Cancer cells frequently increase the flux of glucose into the PPP to support the anabolic demands and regulate oxidative stress. Consistently, metabolomic analyses indicate an upregulation of the PPP in thyroid cancer. In the present study, we found that the combination of glucose-6-phosphate dehydrogenase (G6PD) and transketolase inhibitors (6-aminonicotinamide and oxythiamine) exerted an additive or synergistic effect on cell growth inhibition in thyroid cancer cells. Targeting PPP significantly increased cellular reactive oxygen species (ROS) and induced endoplasmic reticulum (ER) stress and apoptosis. Suppressed cell viability could be partially rescued with treatment with the ROS scavenger or apoptosis inhibitor but not ER-stress inhibitor. Taken together, dual PPP blockade leads to pharmacologic additivity or synergism and causes ROS-mediated apoptosis in thyroid cancer cells.

Postnatal knockout of beta cell insulin receptor impaired insulin secretion in male mice exposed to high-fat diet stress Publication date: 1 January 2020 Source: Molecular and Cellular Endocrinology, Volume 499 Author(s): Amanda Oakie, Liangyi Zhou, Sydney Rivers, Christy Cheung, Jinming Li, Rennian Wang Abstract The presence of insulin receptor (IR) on insulin-secreting beta cells suggests an autocrine regulatory role for insulin in its own signalling. Congenital beta cell-specific IR knockout (βIRKO) mouse studies have demonstrated the development of age-dependent glucose intolerance. We investigated the role of beta cell IR signalling specifically during postnatal life following undisturbed prenatal pancreatic development and maturation. We utilized a tamoxifen-inducible mouse insulin 1 promoter (MIP) driven Cre recombinase IR knockout mouse model (MIP-βIRKO) to achieve partial knockout of IR in islets and determine the functional role of beta cell IR in adult mice fed a control normal diet (ND) or 60% high-fat diet (HFD). At 24 weeks of age, MIP-βIRKO ND mice maintained glucose tolerance, insulin release, and unchanged beta cell mass when compared to control ND mice. In contrast, 24-week-old MIP-βIRKO mice demonstrated significant glucose intolerance and lower insulin release after 18 weeks of HFD feeding. A reduction in beta cell soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) protein expression, phosphorylated AktS473 and P70S6K1T389, and glucose transporter 2 (GLUT2) expression were also identified in MIP-βIRKO HFD islets. Overall, the postnatal knockout of beta cell IR in HFD-fed mice resulted in decreased expression of beta cell glucose-sensing and exocytotic proteins and a reduction in intracellular signalling. These findings highlight that IR expression in the adult islet is required to maintain beta cell function under hyperglycemic stress.