Intersectional Targeting Reveals Heterogeneous Nature of Glp1r- and Lepr expressing POMC Neurons

The anorexigenic pro-opiomelanocortin (Pomc)-expressing neurons of the arcuate nucleus of the hypothalamus (ARC) are constituents of the melanocortin system, and their function is pivotal in control of systemic energy homeostasis. Adjacency to the median eminence and a fenestrated blood-brain barrier exposes the neurons of the melanocortin system to hormones and stimuli from the periphery. These signals are integrated and serve as a basis for the exertion of optimal responses in the regulation of homeostatic processes. Defined by the expression of POMC, it has been assumed that these neurons are a uniform group; however, cumulative electrophysiological and single-cell sequencing evidence hint at both functional and molecular heterogeneity of POMC neurons. Deciphering this heterogeneity is crucial in understanding how a wide range of stimuli are integrated and unraveling the intricacies in metabolic control mediated by different POMC subpopulations. In the present study, we utilize intersectional Cre/Dre-dependent transgenic mouse models to successfully target distinct leptin receptor (Lepr) and glucagon-like peptide receptor (Glp1r) expressing POMC neurons. This system enabled a detailed description of anatomical distribution, translational profiling, and functional characterization of these two POMC subpopulations. Our experiments reveal that POMCLepr+ and POMCGlp1r+ neurons exhibit distinct anatomical distribution patterns within the ARC by using tissue clearing and three-dimensional image assessment. In addition, electrophysiological recordings from these neurons illustrated variations in their key intrinsic properties. Molecular profiling revealed a differential expression in receptors for energy state communicating hormones and neurotransmitters. Finally, employing the Cre/Dre-dependent activatory chemogenetic system for both neuronal groups demonstrated differential regulation of the two subpopulations with regards to food intake. Collectively, this work reveals heterogeneity of critical metabolism-regulatory POMC neurons

Neurocalcin Delta Suppression Protects against Spinal Muscular Atrophy in Humans and across Species by Restoring Impaired Endocytosis

This document is the Accepted Manuscript version of the following article: Riessland et al., 'Neurocalcin Delta Suppression Protects against Spinal Muscular Atrophy in Humans and across Species by Restoring Impaired Endocytosis', The American Journal of Human Genetics, Vol. 100 (2): 297-315, first published online 26 January 2017. The final, published version is available online at doi: http://dx.doi.org/10.1016/j.ajhg.2017.01.005 © 2017 American Society of Human Genetics.Homozygous SMN1 loss causes spinal muscular atrophy (SMA), the most common lethal genetic childhood motor neuron disease. SMN1 encodes SMN, a ubiquitous housekeeping protein, which makes the primarily motor neuron-specific phenotype rather unexpected. SMA-affected individuals harbor low SMN expression from one to six SMN2 copies, which is insufficient to functionally compensate for SMN1 loss. However, rarely individuals with homozygous absence of SMN1 and only three to four SMN2 copies are fully asymptomatic, suggesting protection through genetic modifier(s). Previously, we identified plastin 3 (PLS3) overexpression as an SMA protective modifier in humans and showed that SMN deficit impairs endocytosis, which is rescued by elevated PLS3 levels. Here, we identify reduction of the neuronal calcium sensor Neurocalcin delta (NCALD) as a protective SMA modifier in five asymptomatic SMN1-deleted individuals carrying only four SMN2 copies. We demonstrate that NCALD is a Ca(2+)-dependent negative regulator of endocytosis, as NCALD knockdown improves endocytosis in SMA models and ameliorates pharmacologically induced endocytosis defects in zebrafish. Importantly, NCALD knockdown effectively ameliorates SMA-associated pathological defects across species, including worm, zebrafish, and mouse. In conclusion, our study identifies a previously unknown protective SMA modifier in humans, demonstrates modifier impact in three different SMA animal models, and suggests a potential combinatorial therapeutic strategy to efficiently treat SMA. Since both protective modifiers restore endocytosis, our results confirm that endocytosis is a major cellular mechanism perturbed in SMA and emphasize the power of protective modifiers for understanding disease mechanism and developing therapies.Peer reviewedFinal Accepted Versio

تأثیر شستشوي پوسته تخم مرغ با آب سرد بر کیفیت فیزیکو شیمیایی آن طی مدت نگهداري در یخچال

زمینه: شستشوي پوسته تخممرغ روشی ساده و کم هزینه بوده و توسط بسیاري از افراد جهت تمیز کردن پوسته تخم مرغ مورد استفاده قرار میگیرد. هدف: تأثیر شستشوي پوسته تخم مرغ بر کیفیت فیزیکو شیمیایی آن بررسی شد. سفیده، شاخصزرده، درصد کاهش وزن و ارتفاع آلبومین طی مدت زمان نگه داري pH مواد و روشها: تعداد 45 نمونه تخم مرغ از لحاظ میزان در یخچال (روزهاي 15 ،10 ،5 ،1 و 20 ) بررسی شدند. تخم مرغها به سه گروه ؛ شاهد شامل تخم مرغ هاي شسته نشده و گر وه آزمایش تخم مرغهاي شستشو شده با آب سرد و تخم مرغهاي شستشو شده با اسکاچ همراه با یک قطره مایع ظرفشویی تقسیم شدند. هر یک از آزمایشها سفیده، شاخصزرده، درصد کاهش وزن و ارتفاع آلبومین) در سه تکرار انجام و میانگین آنها به روش تحلیل واریانس یک pH (شامل اندازهگیري انجام شد. P<0/ طرفه تجزیه و تحلیل شد. مقایسه میانگینها با آزمون چنددامنهاي دانکن در سطح احتمال 05 افزایش و وزن، ارتفاع آلبومین و شاخصزرده کاهش یافتند. در خصوصتغییر وزن، بین روش شستشو با آب pH ، یافتهها: در همه تخم مرغها در تمام (Haugh Unit) HU ارتفاع آلبومین، شاخصزرده و شاخص .(P<0/ سرد و شستشو با اسکاچ اختلاف آماري معناداري وجود داشت ( 05 تیمارها طی مدت نگهداري روند نزولی داشتند اما اختلاف معنیداري مشاهده نشد. نتیجهگیري: در تمامی تخم مرغهاي مورد بررسی کلیه عوامل فیزیکو شیمیایی تغییر کردند اما تغییرات فیزیکو شیمیایی محسوسی در خواص فیزیکو شیمیایی تخم مرغهاي شستشو شده نسبت به تخم مرغهاي شستشو نشده مشاهده نشد

Effects of cold-water egg shell washing on Salmonella contamination in the shell and its contents

Background: Egg shell washing is one of the simple and inexpensive methods using by many people to clean egg shells. Objective: In this study, the effects of egg shell washing (with 5° C water) on salmonella contamination of shell and its contents was assessment during refrigerated storage for 20 d. Methods: The 60 eggs samples chosen from Qazvin markets. Egg samples were divided into control (without washing) and cold-washed group with cold water. Salmonella contamination in egg samples was studied before and after washing during cold storage (1, 5, 10, 15, 20 d). Findings: Salmonella was not isolated in any of the treatments after washing with cold water during refrigerated storage.Washing with cold water followed by storage in the refrigerator led to destroying of Salmonella contamination in eggs. Also, no contamination was found in any of egg contents. Conclusion: Cold water washing, as a simple and low-cost method at the community level can be used to improve the health of eggs and shells. Keywords: Eggs, Health, Salmonella, Washing, Shelf lif

Effects of cold-water egg shell washing on Salmonella contamination in the shell and its contents

Background: Egg shell washing is one of the simple and inexpensive methods using by many people to clean egg shells. Objective: In this study, the effects of egg shell washing (with 5° C water) on salmonella contamination of shell and its contents was assessment during refrigerated storage for 20 d. Methods: The 60 eggs samples chosen from Qazvin markets. Egg samples were divided into control (without washing) and cold-washed group with cold water. Salmonella contamination in egg samples was studied before and after washing during cold storage (1, 5, 10, 15, 20 d). Findings: Salmonella was not isolated in any of the treatments after washing with cold water during refrigerated storage.Washing with cold water followed by storage in the refrigerator led to destroying of Salmonella contamination in eggs. Also, no contamination was found in any of egg contents. Conclusion: Cold water washing, as a simple and low-cost method at the community level can be used to improve the health of eggs and shells

Gut-brain communication by distinct sensory neurons differently controls feeding and glucose metabolism

Sensory neurons relay gut-derived signals to the brain, yet the molecular and functional organization of distinct populations remains unclear. Here, we employed intersectional genetic manipulations to probe the feeding and glucoregulatory function of distinct sensory neurons. We reconstruct the gut innervation patterns of numerous molecularly defined vagal and spinal afferents and identify their downstream brain targets. Bidirectional chemogenetic manipulations, coupled with behavioral and circuit mapping analysis, demonstrated that gut-innervating, glucagon-like peptide 1 receptor (GLP1R)-expressing vagal afferents relay anorexigenic signals to parabrachial nucleus neurons that control meal termination. Moreover, GLP1R vagal afferent activation improves glucose tolerance, and their inhibition elevates blood glucose levels independent of food intake. In contrast, gut-innervating, GPR65-expressing vagal afferent stimulation increases hepatic glucose production and activates parabrachial neurons that control normoglycemia, but they are dispensable for feeding regulation. Thus, distinct gut-innervating sensory neurons differentially control feeding and glucoregulatory neurocircuits and may provide specific targets for metabolic control

Dopamine-inhibited POMCDrd2+neurons in the ARC acutely regulate feeding and body temperature

Dopamine acts on neurons in the arcuate nucleus (ARC) of the hypothalamus, which controls homeostatic feeding responses. Here we demonstrate a differential enrichment of dopamine receptor 1 (Drd1) expression in food intake-promoting agouti related peptide (AgRP)/neuropeptide Y (NPY) neurons and a large proportion of Drd2-expressing anorexigenic proopiomelanocortin (POMC) neurons. Owing to the nature of these receptors, this translates into a predominant activation of AgRP/NPY neurons upon dopamine stimulation and a larger proportion of dopamine -inhibited POMC neurons. Employing intersectional targeting of Drd2-expressing POMC neurons, we reveal that dopamine-mediated POMC neuron inhibition is Drd2 dependent and that POMCDrd2+ neurons exhibit differential expression of neuropeptide signaling mediators compared with the global POMC neuron population, which manifests in enhanced somatostatin responsiveness of POMCDrd2+ neurons. Selective chemogenetic activation of POMCDrd2+ neurons uncovered their ability to acutely suppress feeding and to preserve body temperature in fasted mice. Collectively, the present study provides the molecular and functional characterization of POMCDrd2+ neurons and aids our understanding of dopamine-dependent control of homeostatic energy-regulatory neurocircuits

The Fat Mass and Obesity-Associated Protein (FTO) Regulates Locomotor Responses to Novelty via D2R Medium Spiny Neurons

Variations in the human FTO gene have been linked to obesity and altered connectivity of the dopaminergic neurocircuitry. Here, we report that fat mass and obesity-associated protein (FTO) in dopamine D2 receptor-expressing medium spiny neurons (D2 MSNs) of mice regulate the excitability of these cells and control their striatopallidal globus pallidus external (GPe) projections. Lack of FTO in D2 MSNs translates into increased locomotor activity to novelty, associated with altered timing behavior, without impairing the ability to control actions or affecting reward-driven and conditioned behavior. Pharmacological manipulations of dopamine D1 receptor (D1R)- or D2R-dependent pathways in these animals reveal altered responses to D1- and D2-MSN-mediated control of motor output. These findings reveal a critical role for FTO to control D2 MSN excitability, their projections to the GPe, and behavioral responses to novelty