Neuromedin U-deficient rats do not lose body weight or food intake

Studies in genetically modified mice establish that essential roles of endogenous neuromedin U (NMU) are anorexigenic function and metabolic regulation, indicating that NMU is expected to be a potential target for anti-obesity agents. However, in central administration experiments in rats, inconsistent results have been obtained, and the essential role of NMU energy metabolism in rats remain unclear. This study aims to elucidate the role of endogenous NMU in rats. We generated NMU knockout (KO) rats that unexpectedly showed no difference in body weight, adiposity, circulating metabolic markers, body temperature, locomotor activity, and food consumption in both normal and high fat chow feeding. Furthermore, unlike reported in mice, expressions of Nmu and NMU receptor type 2 (Nmur2) mRNA were hardly detectable in the rat hypothalamic nuclei regulating feeding and energy metabolism, including the arcuate nucleus and paraventricular nucleus, while Nmu was expressed in pars tuberalis and Nmur2 was expressed in the ependymal cell layer of the third ventricle. These results indicate that the species-specific expression pattern of Nmu and Nmur2 may allow NMU to have distinct functions across species, and that endogenous NMU does not function as an anorexigenic hormone in rats

Anisotropic nanomaterials: structure, growth, assembly, and functions

Comprehensive knowledge over the shape of nanomaterials is a critical factor in designing devices with desired functions. Due to this reason, systematic efforts have been made to synthesize materials of diverse shape in the nanoscale regime. Anisotropic nanomaterials are a class of materials in which their properties are direction-dependent and more than one structural parameter is needed to describe them. Their unique and fine-tuned physical and chemical properties make them ideal candidates for devising new applications. In addition, the assembly of ordered one-dimensional (1D), two-dimensional (2D), and three-dimensional (3D) arrays of anisotropic nanoparticles brings novel properties into the resulting system, which would be entirely different from the properties of individual nanoparticles. This review presents an overview of current research in the area of anisotropic nanomaterials in general and noble metal nanoparticles in particular. We begin with an introduction to the advancements in this area followed by general aspects of the growth of anisotropic nanoparticles. Then we describe several important synthetic protocols for making anisotropic nanomaterials, followed by a summary of their assemblies, and conclude with major applications

A Genome-Wide Association Study of Diabetic Kidney Disease in Subjects With Type 2 Diabetes

dentification of sequence variants robustly associated with predisposition to diabetic kidney disease (DKD) has the potential to provide insights into the pathophysiological mechanisms responsible. We conducted a genome-wide association study (GWAS) of DKD in type 2 diabetes (T2D) using eight complementary dichotomous and quantitative DKD phenotypes: the principal dichotomous analysis involved 5,717 T2D subjects, 3,345 with DKD. Promising association signals were evaluated in up to 26,827 subjects with T2D (12,710 with DKD). A combined T1D+T2D GWAS was performed using complementary data available for subjects with T1D, which, with replication samples, involved up to 40,340 subjects with diabetes (18,582 with DKD). Analysis of specific DKD phenotypes identified a novel signal near GABRR1 (rs9942471, P = 4.5 x 10(-8)) associated with microalbuminuria in European T2D case subjects. However, no replication of this signal was observed in Asian subjects with T2D or in the equivalent T1D analysis. There was only limited support, in this substantially enlarged analysis, for association at previously reported DKD signals, except for those at UMOD and PRKAG2, both associated with estimated glomerular filtration rate. We conclude that, despite challenges in addressing phenotypic heterogeneity, access to increased sample sizes will continue to provide more robust inference regarding risk variant discovery for DKD.Peer reviewe

Negative regulation of neuromedin U mRNA expression in the rat pars tuberalis by melatonin.

The pars tuberalis (PT) is part of the anterior pituitary gland surrounding the median eminence as a thin cell layer. The characteristics of PT differ from those of the pars distalis (PD), such as cell composition and gene expression, suggesting that the PT has a unique physiological function compared to the PD. Because the PT highly expresses melatonin receptor type 1, it is considered a mediator of seasonal and/or circadian signals of melatonin. Expression of neuromedin U (NMU) that is known to regulate energy balance has been previously reported in the rat PT; however, the regulatory mechanism of NMU mRNA expression and secretion in the PT are still obscure. In this study, we examined both the diurnal change of NMU mRNA expression in the rat PT and the effects of melatonin on NMU in vivo. In situ hybridization and quantitative PCR analysis of laser microdissected PT samples revealed that NMU mRNA expression in the PT has diurnal variation that is high during the light phase and low during the dark phase. Furthermore, melatonin administration significantly suppressed NMU mRNA expression in the PT in vivo. On the other hand, 48 h fasting did not have an effect on PT-NMU mRNA expression, and the diurnal change of NMU mRNA expression was maintained. We also found the highest expression of neuromedin U receptor type 2 (NMUR2) mRNA in the third ventricle ependymal cell layer, followed by the arcuate nucleus and the spinal cord. These results suggest that NMU mRNA expression in the PT is downregulated by melatonin during the dark phase and shows diurnal change. Considering that NMU mRNA in the PT showed the highest expression level in the brain, PT-NMU may act on NMUR2 in the brain, especially in the third ventricle ependymal cell layer, with a circadian rhythm

In vivo fluorescence correlation spectroscopy analyses of FMBP-1, a silkworm transcription factor

Fibroin modulator-binding protein 1 (FMBP-1) is a silkworm transcription factor that has a unique DNA-binding domain called the one score and three amino acid peptide repeat (STPR). Here we used fluorescence correlation spectroscopy (FCS) to analyze the diffusion properties of an enhanced green fluorescent protein-tagged FMBP-1 protein (EGFP-FMBP-1) expressed in posterior silk gland (PSG) cells of Bombyx mori at the same developmental stage as natural FMBP-1 expression. EGFP-FMBP-1 clearly localized to cell nuclei. From the FCS analyses, we identified an immobile DNA-bound component and three discernible diffusion components. We also used FCS to observe the movements of wild-type and mutant EGFP-FMBP-1 proteins in HeLa cells, a simpler experimental system. Based on previous in vitro observation, we also introduced a single amino acid substitution in order to suppress stable FMBP-1-DNA binding; specifically, we replaced the ninth Arg in the third repeat within the STPR domain with Ala. This mutation completely disrupted the slowest diffusion component as well as the immobile component. The diffusion properties of other FMBP-1 mutants (e.g. mutants with N-terminal or C-terminal truncations) were also analyzed. Based on our observations, we suggest that the four identifiable movements might correspond to four distinct FMBP-1 states: (a) diffusion of free protein, (b) and (c) two types of transient interactions between FMBP-1 and chromosomal DNA, and (d) stable binding of FMBP-1 to chromosomal DNA

NMU receptor type 2 mRNA expression in the rat brain.

<p>(A) Microphotograph of NMU mRNA expression detected by ISH in the fixed frozen sections of the rat brain. Staining signal was detected in the EC of the 3V (arrows). Upper part of the EC of 3V did not show NMUR2 expression (arrowheads). (B) Enlarged view of A. The staining signal was found in the EC of the 3V (arrows), but not in the bottom of the 3V, adjacent to the median eminence (arrowheads). (C) Sense RNA probes to NMUR2 generated no specific signal. Scale bar: 200 µm. PT, pars tuberalis; ARC, arcuate nucleus; 3V, third ventricle; EC, ependymal cell layer of the third cerebroventricle. (D) qPCR analysis of NMUR2 in the PT, PD, SCN, ARC, EC, and SC at ZT6 and ZT18. NMUR2 expression was detected in the EC at the highest level, followed by the ARC and SC, and was detected in the SCN at a considerably lower level. NMUR2 expression was not detected in the PT and PD. NMUR2 expression did not change at either time point in the SCN, ARC, EC, and SC.</p

NMU mRNA-expressing cell types in the PT.

<p>Microphotograph of NMU mRNA expression detected by double staining by HNPP fluorescent ISH for NMU mRNA and by fluorescent IHC for TSH in fixed, frozen 5-µm sections. (A) Microphotograph of NMU mRNA expression detected by HNPP fluorescent ISH (red). NMU mRNA-expressing cells were widely distributed in the PT. (B) Microphotograph of TSH detected by fluorescent IHC staining. Many TSH-immunostained cells were distributed throughout the PT (green). (C) Microphotograph of merged pictures of double staining for NMU mRNA expression, which was detected by HNPP fluorescent ISH and TSH detected by fluorescent IHC. Yellow color represents the merged signal for NMU mRNA and TSH production. Although almost all NMU-expressing cells showed TSH immunoreactivity (arrows, yellow), cells exhibiting only TSH immunostaining were also found (arrowheads, green). (D) Magnified image of the boxed area in C. NMU mRNA-expressing and TSH producing cells (arrows, yellow) and cells producing only TSH (arrowheads, green) exist. Scale bars: 50 µm. PT, pars tuberalis; ME, median eminence.</p

Effects of fasting on NMU mRNA expression in PT.

<p>qPCR analysis of NMU in the PT performed for the sham and fasting groups at ZT6 and ZT18. The NMU mRNA expression level was not changed when the sham group was compared with the fasting group at both sampling times, ZT6 (fasting from ZT6 to ZT6 for 48 h) and ZT18 (fasting from ZT18 to ZT18 for 48 h), and the diurnal change was maintained even under fasting conditions (high at ZT6, low at ZT18, <i>P</i><0.01; ZT6 control vs. ZT18 control, ZT6 fasting vs. ZT18 fasting).</p

Effects of melatonin on NMU mRNA expression in the PT, PD, and SCN.

<p>(A–D) Microphotograph of NMU mRNA expression detected by ISH in the PT of control and melatonin-treated groups at ZT6 and ZT18. (A, B) Staining signal in the PT was decreased in the melatonin-treated group compared to the sham group at ZT6. (C, D) In the ZT18 samples, a similar staining intensity was observed in the sham and melatonin-treated groups. Scale bar: 200 µm. PT, pars tuberalis; ARC, arcuate nucleus; 3V, third ventricle. (E) qPCR analysis of NMU in the PT was performed for the sham and melatonin groups at ZT6 and ZT18 using LMD samples. Two-way ANOVA revealed a significant interaction between melatonin treatment and time point (<i>P</i><0.05) and that the NMU mRNA expression level of the ZT6 melatonin group was significantly higher than that of the ZT6 melatonin, ZT18 control, and ZT18 melatonin groups (one way ANOVA, <i>P</i><0.01). (F, G) qPCR analysis for NMU in the SCN and PD performed for the sham and melatonin-treated rats at ZT6 and ZT18. NMU mRNA expression in the SCN and PD were considerably lower and not influenced by time or melatonin treatment.</p