Defining G protein-coupled receptor peptide ligand expressomes and signalomes in human and mouse islets

Introduction

Islets synthesise and secrete numerous peptides, some of which are known to be important regulators of islet function and glucose homeostasis. In this study, we quantified mRNAs encoding all peptide ligands of islet G protein-coupled receptors (GPCRs) in isolated human and mouse islets and carried out in vitro islet hormone secretion studies to provide functional confirmation for the species-specific role of peptide YY (PYY) in mouse islets.

Materials and methods

GPCR peptide ligand mRNAs in human and mouse islets were quantified by quantitative real-time PCR relative to the reference genes ACTB, GAPDH, PPIA, TBP and TFRC. The pathways connecting GPCR peptide ligands with their receptors were identified by manual searches in the PubMed, IUPHAR and Ingenuity databases. Distribution of PYY protein in mouse and human islets was determined by immunohistochemistry. Insulin, glucagon and somatostatin secretion from islets was measured by radioimmunoassay.

Results

We have quantified GPCR peptide ligand mRNA expression in human and mouse islets and created specific signalomes mapping the pathways by which islet peptide ligands regulate human and mouse GPCR signalling. We also identified species-specific islet expression of several GPCR ligands. In particular, PYY mRNA levels were ~ 40,000-fold higher in mouse than human islets, suggesting a more important role of locally secreted Pyy in mouse islets. This was confirmed by IHC and functional experiments measuring insulin, glucagon and somatostatin secretion.

Discussion

The detailed human and mouse islet GPCR peptide ligand atlases will allow accurate translation of mouse islet functional studies for the identification of GPCR/peptide signalling pathways relevant for human physiology, which may lead to novel treatment modalities of diabetes and metabolic disease.

     

Prion protein cleavage fragments regulate adult neural stem cell quiescence through redox modulation of mitochondrial fission and SOD2 expression

Neurogenesis continues in the post-developmental brain throughout life. The ability to stimulate the production of new neurones requires both quiescent and actively proliferating pools of neural stem cells (NSCs). Actively proliferating NSCs ensure that neurogenic demand can be met, whilst the quiescent pool makes certain NSC reserves do not become depleted. The processes preserving the NSC quiescent pool are only just beginning to be defined. Herein, we identify a switch between NSC proliferation and quiescence through changing intracellular redox signalling. We show that N-terminal post-translational cleavage products of the prion protein (PrP) induce a quiescent state, halting NSC cellular growth, migration, and neurite outgrowth. Quiescence is initiated by the PrP cleavage products through reducing intracellular levels of reactive oxygen species. First, inhibition of redox signalling results in increased mitochondrial fission, which rapidly signals quiescence. Thereafter, quiescence is maintained through downstream increases in the expression and activity of superoxide dismutase-2 that reduces mitochondrial superoxide. We further observe that PrP is predominantly cleaved in quiescent NSCs indicating a homeostatic role for this cascade. Our findings provide new insight into the regulation of NSC quiescence, which potentially could influence brain health throughout adult life.     

Structural,compositional and mineralogical characterization of carbonatitic copper sulfide: Run of mine,concentrate and tailings

The aim of this study was to determine the structural, compositional, and mineralogical composition of carbonatitic copper sulfide concentrator plant streams. Three samples, each from a different stream(run of mine(ROM), concentrate, and tailings) of a copper concentrator were characterized using various techniques, including stereomicroscopy, X-ray fluorescence, X-ray diffraction, Fourier transform infrared(FTIR) spectroscopy, scanning electron microscopy(SEM) in conjunction with energy-dispersive X-ray spectroscopy(EDS), and optical microscopy. The results reveal that each stream possesses its own unique compositional features. Carbonate minerals associated with calcite and dolomite, followed by quartz, remain the major minerals in both the ROM and tails streams. In the ROM stream, chalcopyrite appears to occur as veins within the carbonatite-hosting ore body. Mineral phase mutation was discovered in the tails stream because magnetite formerly identified in the ROM as the primary iron oxide had evolved into hematite. This metamorphosis was likely promoted by the concentration process. The concentration process was effective, upgrading the chalcopyrite content from 2 wt% in the ROM stream to 58 wt% in the concentrate stream; it was accompanied by bornite(4 wt%), anilite(3 wt%), and digenite(2.5 wt%). In addition, the concentrate stream exhibited properties distinctive from those of the other streams. The FTIR analysis showed the existence of a sulfide group related to the chalcopyrite mineral. Free chalcopyrite grains were observed in the concentrate by SEM analysis, and their mineral presence was supported by the EDS analysis results. All characterization techniques corresponded well with each other regarding the structure, chemistry, and composition of the samples.     

Elucidating the diel and seasonal calling behaviour of Elachistocleis matogrosso (Anura: Microhylidae)

ABSTRACT

Acoustic monitoring provides the opportunity to study ecological processes that are difficult to assess with traditional surveys. Elachistocleis matogrosso is an anuran species, described in 2010, for which limited biological information is available. This study investigated the calling activity of the species in the north-eastern portion of the Pantanal, Brazil, a wetland area with marked seasonality between the dry and wet seasons. The calling activity of E. matogrosso was monitored using automated digital recorders in combination with automated signal recognition software over two different annual cycles. The species was vocally active only during the wet season (October – April), with a peak in November-December during the 2013–2014 annual cycle and in February-March during the 2015–2016 annual cycle. The peak calling activity occurred at dusk. This species has nocturnal habits and an explosive breeding activity. The detection of the species was intermittent, which suggests that environmental predictors or site-specific conditions might play an important role in species detection. Moreover, this intermittent occupancy indicated that surveys that employ traditional field techniques would likely fail to detect this species. We describe an effective protocol for detecting E. matogrosso with acoustic monitoring, which requires recording during 20 days in February from 17:01 to 05:00. Our procedure would be easy to adapt to other anuran species, and it could be used for investigating new localities and assessing population changes over time.     

Quantum reality: A pragmaticized neo-Kantian approach

Despite remarkable efforts, it remains notoriously difficult to equip quantum theory with a coherent ontology. Hence, Healey (2017, 12) has recently suggested that “quantum theory has no physical ontology and states no facts about physical objects or events”, and Fuchs et al. (2014, 752) similarly hold that “quantum mechanics itself does not deal directly with the objective world”. While intriguing, these positions either raise the question of how talk of ‘physical reality’ can even remain meaningful, or they must ultimately embrace a hidden variables-view, in tension with their original project. I here offer a neo-Kantian alternative. In particular, I will show how constitutive elements in the sense of Reichenbach (1920) and Friedman (1999, 2001) can be identified within quantum theory, through considerations of symmetries that allow the constitution of a ‘quantum reality’, without invoking any notion of a radically mind-independent reality. The resulting conception will inherit elements from pragmatist and ‘QBist’ approaches, but also differ from them in crucial respects. Furthermore, going beyond the Friedmanian program, I will show how non-fundamental and approximate symmetries can be relevant for identifying constitutive principles.     

The role of heme oxygenase-1 in hematopoietic system and its microenvironment

Cellular and Molecular Life Sciences - Hematopoietic system transports all necessary nutrients to the whole organism and provides the immunological protection. Blood cells have high turnover,...     

Evidence of G-protein-coupled receptor and substrate transporter heteromerization at a single molecule level

G-protein-coupled receptors (GPCRs) can constitute complexes with non-GPCR integral membrane proteins, while such interaction has not been demonstrated at a single molecule level so far. We here investigated the potential interaction between the thyrotropin receptor (TSHR) and the monocarboxylate transporter 8 (MCT8), a member of the major facilitator superfamily (MFS), using fluorescence cross-correlation spectroscopy (FCCS). Both the proteins are expressed endogenously on the basolateral plasma membrane of the thyrocytes and are involved in stimulation of thyroid hormone production and release. Indeed, we demonstrate strong interaction between both the proteins which causes a suppressed activation of G_q/11 by TSH-stimulated TSHR. Thus, we provide not only evidence for a novel interaction between the TSHR and MCT8, but could also prove this interaction on a single molecule level. Moreover, this interaction forces biased signaling at the TSHR. These results are of general interest for both the GPCR and the MFS research fields.