Can ID Repetitive Elements Serve as Cis-acting Dendritic Targeting Elements? An In Vivo Study

Dendritic localization of mRNA/RNA involves interaction of cis-elements and trans-factors. Small, non-protein coding dendritic BC1 RNA is thought to regulate translation in dendritic microdomains. Following microinjections into cultured cells, BC1 RNA fused to larger mRNAs appeared to impart transport competence to these chimeras, and its 5′ ID region was proposed as the cis-acting dendritic targeting element. As these ID elements move around rodent genomes and, if transcribed, form a long RNA stem-loop, they might, thereby, lead to new localizations for targeted gene products. To test their targeting ability in vivo we created transgenic mice expressing various ID elements fused to the 3′ UTR of reporter mRNA for Enhanced Green Fluorescent Protein. In vivo, neither ID elements nor the BC1 RNA coding region were capable of transporting EGFP RNA to dendrites, although the 3′ UTR of α-CaMKII mRNA, an established cis-acting element did produce positive results. Other mRNAs containing naturally inserted ID elements are also not found in neuronal dendrites. We conclude that the 5′ ID domain from BC1 RNA is not a sufficient dendritic targeting element for mRNAs in vivo

Two primate-specific small non-protein-coding RNAs in transgenic mice: neuronal expression, subcellular localization and binding partners

In a rare occasion a single chromosomal locus was targeted twice by independent Alu-related retroposon insertions, and in both cases supported neuronal expression of the respective inserted genes encoding small non-protein coding RNAs (npcRNAs): BC200 RNA in anthropoid primates and G22 RNA in the Lorisoidea branch of prosimians. To avoid primate experimentation, we generated transgenic mice to study neuronal expression and protein binding partners for BC200 and G22 npcRNAs. The BC200 gene, with sufficient upstream flanking sequences, is expressed in transgenic mouse brain areas comparable to those in human brain, and G22 gene, with upstream flanks, has a similar expression pattern. However, when all upstream regions of the G22 gene were removed, expression was completely abolished, despite the presence of intact internal RNA polymerase III promoter elements. Transgenic BC200 RNA is transported into neuronal dendrites as it is in human brain. G22 RNA, almost twice as large as BC200 RNA, has a similar subcellular localization. Both transgenically expressed npcRNAs formed RNP complexes with poly(A) binding protein and the heterodimer SRP9/14, as does BC200 RNA in human. These observations strongly support the possibility that the independently exapted npcRNAs have similar functions, perhaps in translational regulation of dendritic protein biosynthesis in neurons of the respective primates

Evaluation of the role of perceived quality and satisfaction of beneficiaries about the health care services and benefits of community clinics in Bangladesh

Introduction Bangladesh provides free healthcare to its citizens through 10,723 community clinics which provide one-stop healthcare services that is vital in ensuring primary healthcare. Measuring beneficiary' contentment is crucial to improve the quality of care and the perceived service quality have a significant impact on how often people use those health services. This study focuses on perceived quality and satisfaction of beneficiaries about the primary health care services and benefits of community clinics in Bangladesh. Methods A quantitative descriptive survey was conducted from March 2019 to April 2019 in the catchment area of 80 community clinics located at 16 Upazila under eight districts of eight divisions in Bangladesh. The survey compiled local data on client’s experience on health care service of community clinics. Besides, Sixteen Upazilas from eight districts were randomly selected for conducting interviews. Results Data was provided by a total of 760 female participants, among them majority (41%) belonged to the age group of 18-24 years old, and this very group showed more satisfaction than others (Odds Ratio 1.44). Besides, childless married women were also found to be content with the community clinic services compared to the remaining types of clients (Odds Ratio 1.64). However, gender, education, and economic perspective were positive aspects of getting service from community clinics. Conclusions Although there is a challenge balancing psychosocial and medical care, promoting client-oriented care with a focus on the cultural factors of the area is vital. This can be done through community-focused training together with explaining written prescriptions to the beneficiary, including the signs, symptoms, treatment, and referral points. The study findings will enable responsible authority to improve quality of primary health care services, realizing beneficiary’ ideas of community clinic service quality

Transcriptional activation by hepatocyte nuclear factor-4 in a cell-free system derived from rat liver nuclei

Hepatocyte nuclear factor-4 (HNF4) regulates gene expression by binding to direct repeat motifs of the RG(G/T)TCA sequence separated by one nucleotide (DR1). In this study we demonstrate that endogenous HNF4 present in rat liver nuclear extracts, as well as purified recombinant HNF4, activates transcription from naked DNA templates containing multiple copies of the DR1 element linked to the adenovirus major late promoter. Recombinant HNF4 also activates transcription from the rat cellular retinol binding protein II (CRBPII) promoter in vitro. The region between –105 and –63 bp of this promoter is essential for HNF-mediated transactivation. The addition of a peptide containing the LXXLL motif abolished HNF4-mediated transactivation in vitro suggesting that LXXLL-containing protein factor(s) are involved in HNF4-mediated transactivation in rat liver nuclear extracts. This is the first report on transactivation by HNF4 in a cell-free system derived from rat liver nuclei

Serine 27, a Human Retinoid X Receptor \alpha Residue, Phosphorylated by Protein Kinase A Is Essential for CyclicAMP-Mediated Downregulation of RXRαRXR\alpha Function

Retinoid X Receptor \alpha $(RXR \alpha$), a member of the steroid-thyroid hormone receptor super family, is phosphorylated in vitro by protein kinase A (PKA) and this phosphorylation is inhibited in presence of PKA inhibitory peptide. Analysis of various deletion mutants of $RXR \alpha$ indicate that the amino-terminal A/B domain is the target for PKA phosphorylation. An $RXR \alpha$ mutant in which serine residue 27 is mutated to alanine is no longer phosphorylated by PKA. In vivo transfection experiments in COS cells indicate that cyclicAMP represses retinoic acid-mediated transcriptional activation of $RXR\alpha$ and this repression is mediated by serine 27. These results indicate that serine 27 of $RXR\alpha$ is an unique target for phosphorylation by PKA in vitro and it has an important role in the crosstalk between $RXR\alpha$ and cyclicAMP signalling pathways

Serine 27, a human retinoid X receptor α residue, phosphorylated by protein kinase A is essential for cyclicAMP-mediated downregulation of RXRα function

Retinoid X Receptor α (RXRα), a member of the steroid-thyroid hormone receptor super family, is phosphorylated in vitro by protein kinase A (PKA) and this phosphorylation is inhibited in presence of PKA inhibitory peptide. Analysis of various deletion mutants of RXRα indicate that the amino-terminal A/B domain is the target for PKA phosphorylation. An RXRα mutant in which serine residue 27 is mutated to alanine is no longer phosphorylated by PKA. In vivo transfection experiments in COS cells indicate that cyclicAMP represses retinoic acid-mediated transcriptional activation of RXRα and this repression is mediated by serine 27. These results indicate that serine 27 of RXRα is an unique target for phosphorylation by PKA in vitro and it has an important role in the crosstalk between RXRα and cyclicAMP signalling pathways

The histone acetyltransferase MOF activates hypothalamic polysialylation to prevent diet-induced obesity in mice

Overfeeding causes rapid synaptic remodeling in hypothalamus feeding circuits. Polysialylation of cell surface molecules is a key step in this neuronal rewiring and allows normalization of food intake. Here we examined the role of hypothalamic polysialylation in the long-term maintenance of body weight, and deciphered the molecular sequence underlying its nutritional regulation. We found that upon high fat diet (HFD), reduced hypothalamic polysialylation exacerbated the diet-induced obese phenotype in mice. Upon HFD, the histone acetyltransferase MOF was rapidly recruited on the St8sia4 polysialyltransferase-encoding gene. Mof silencing in the mediobasal hypothalamus of adult mice prevented activation of the St8sia4 gene transcription, reduced polysialylation, altered the acute homeostatic feeding response to HFD and increased the body weight gain. These findings indicate that impaired hypothalamic polysialylation contribute to the development of obesity, and establish a role for MOF in the brain control of energy balance