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bioRxiv2022

Generating an in vitro gut model with physiologically relevant biophysical mucus properties

IntroductionGastrointestinal (GI) in vitro models have received lasting attention as an effective tool to model drug and nutrient absorption, study GI diseases, and design new drug delivery vehicles. A complete model of the GI epithelium should at a minimum include the two key functional components of the GI tract: mucus and the underlying epithelium. Mucus plays a key role in protecting and lubricating the GI tract, poses a barrier to orally administered therapies and pathogens, and serves as the microenvironment for the GI microbiome. These functions are reliant on the biophysical material properties of the mucus produced, including viscosity and pore size. MethodsIn this study, we generated in vitro models containing Caco-2 enterocyte-like cells and HT29-MTX goblet-like cells and determined the effects of coculture and mucus layer on epithelial permeability and biophysical properties of mucus using multiple particle tracking (MPT). ResultsWe found that mucus height increased as the amount of HT29-MTX goblet-like cells increased. Additionally, we found that increasing the amount of HT29-MTX goblet-like cells within culture corresponded to an increase in mucus pore size and mucus microviscosity, measured using MPT. When compared to ex vivo mucus samples from mice and pigs, we found that a 90:10 ratio of Caco-2:HT29-MTX coculture displayed similar mucus pore size to porcine jejunum and that the mucus produced from 90:10 and 80:20 ratios of cells shared mechanical properties to porcine jejunum and ileum mucus. ConclusionsGI coculture models are valuable tools in simulating the mucus barrier and can be utilized for a variety of applications including the study of GI diseases, food absorption, or therapeutic development. BiographyDr. Maisel joined the University of Maryland in January 2019 having done interdisciplinary training in nanotechnology, mucosal immunology, lymphatic immunology, and immunoengineering. She completed her PhD in Biomedical Engineering at the Johns Hopkins University in 2014 after which she was an NIH postdoctoral fellow at the University of Chicago in Molecular Engineering and Immunology. The Maisel Lab uses in vitro modeling, nanotechnology, and immunoengineering approaches to study and develop treatments for diseases at mucosal surfaces. They are interested in designing nanoparticles to take advantage of and study the interface between biological barriers, particularly the lymphatics, interstitial tissue, and mucosal surfaces, and nanoparticles. Dr. Maisel has won a number of awards, including NSF GRFP and NIH F32 fellowships as a trainee, the American Lung Association Dalsemer Award, LAM Foundation Career Development Award, an NSF CAREER Award, and an NIH NIGMS Maximizing Investigator Research Award. Her work has led to numerous high-impact publications, particularly in the field of drug delivery and mucosal immunoengineering, and several patents.

bioengineering↗

bioRxiv2022

The effects of brief heat during early booting on reproductive, developmental and physiological performance in common wheat (Triticum aestivum L.)

Rising temperatures due to climate change threaten agricultural crop productivity. As a cool-season crop wheat is heat sensitive, but often exposed to high temperatures during cultivation. In the current study, a bread wheat panel of spring wheat genotypes, including putatively heat-tolerant Australian and CIMMYT genotypes, was exposed to a 5-day mild (34{degrees}C/28{degrees}C, day/night) or extreme (37{degrees}C/27{degrees}C) heat stress during the sensitive pollen developmental stage. Worsening effects on anther morphology were observed as heat stress increased from mild to extreme. Even under mild heat a significant decrease in pollen viability and grain number per spike from primary spike was observed compared with the control (21{degrees}C/15{degrees}C), with Sunstar and two CIMMYT breeding lines performing well. A heat-specific positive correlation between the two traits indicates the important role of pollen fertility for grain setting. Interestingly, both mild and extreme heat induced development of new tillers after the heat stress, providing an alternative sink for accumulated photosynthates and significantly contributing to the final yield. Measurements of flag leaf maximum potential quantum efficiency of Photosystem II (Fv/Fm) showed an initial inhibition after the heat treatment, followed by a full recovery within a few days. Despite this, model fitting using chlorophyll SPAD measurements showed an earlier onset or faster senescence rate under heat stress. The data presented here provide interesting entry points for further research into pollen fertility, tillering dynamics and leaf senescence under heat. The identified tolerant wheat genotypes can be used to dissect the underlying mechanisms and breed climate-resilient wheat.

plant biology↗

bioRxiv2022

In vivo physiology of foveal retinal ganglion cells in Macaca fascicularis

The primate fovea is specialized for high acuity chromatic vision, with the highest density of cone photoreceptors and a disproportionately large representation in visual cortex. The unique visual properties conferred by the fovea are conveyed to the brain by retinal ganglion cells, the somas of which lie at the margin of the foveal pit. Microelectrode recordings of these centermost retinal ganglion cells have been challenging due to the fragility of the fovea in the excised retina. Here we overcome this challenge by combining high resolution fluorescence adaptive optics ophthalmoscopy with calcium imaging to optically record functional responses of foveal retinal ganglion cells in the living eye. We use this approach to study the chromatic responses and spatial transfer functions of retinal ganglion cells using spatially uniform fields modulated in different directions in color space and monochromatic drifting gratings. We recorded from over 350 cells across three Macaca fascicularis primates over a time period of weeks to months. We find that the majority of the L vs. M cone opponent cells serving the most central foveolar cones have spatial transfer functions that peak at high spatial frequencies (20-40 c/deg), reflecting strong surround inhibition that sacrifices sensitivity at low spatial frequencies but preserves the transmission of fine detail in the retinal image. In addition, we fit to the drifting grating data a detailed model of how ganglion cell responses draw on the cone mosaic to derive receptive field properties of L vs. M cone opponent cells at the very center of the foveola. The fits are consistent with the hypothesis that foveal midget ganglion cells are specialized to preserve information at the resolution of the cone mosaic. By characterizing the functional properties of retinal ganglion cells in vivo through adaptive optics, we characterize the response characteristics of these cells in situ.

neuroscience↗

bioRxiv2022

Cross-generational heritability analysis of physiological traits in Porites astreoides across an inshore-offshore gradient in the Lower Florida Keys

Estimating the heritable genetic variation in fitness-related traits is key to projecting the adaptive evolution of organisms in response to a changing environment. While heritability studies on reef-building corals to date support adaptive capacity, little is known about the dynamics of trait heritability across life stages in which distinct selective pressures can have long-lasting effects both within and across generations. In this study, we obtained heritability estimates for energetic and thermal stress response traits in larval, recruit, and adult Porites astreoides from two populations in the Lower Florida Keys. To induce bleaching phenotypes among individual families, larvae were exposed to a 4-day thermal stress at 32 {degrees}C, whereas adults and recruits received the same treatment for 22 days. Origin-dependent tolerance was observed in two life stages where offshore recruits lost more symbiont cells under heat than inshore recruits compared to their respective controls and heat-treated offshore adults suffered a greater loss in total protein content. Surprisingly, larvae appeared to be largely insensitive to heat regardless of origin. Broad sense heritability (H2) estimates varied greatly among traits and life stages, which may reflect changes in the relative importance of genetic and environmental variation throughout development. Notably, more than 80% of the variation in larval chlorophyll a concentration was attributed to genetic factors. The overall moderate to high H2 estimates measured here suggest these corals have considerable potential to adapt to environmental change.

ecology↗

bioRxiv2022

Mechanistic Neural Mass Modeling: from physiology to pathophysiological brain activity

Brain rhythms emerge from the mean-field activity of networks of neurons. There have been many efforts to build mathematical and computational embodiments in the form of discrete cell-group activities - termed neural masses - to understand particular the origins of evoked potentials, intrinsic patterns of activities such as theta, regulation of sleep, Parkinsons disease related dynamics, and instabilities such as seizure dynamics. As originally utilized, standard neural masses convert input through a sigmoidal function to a firing rate, and firing rate through a synaptic alpha function to other masses. Here we define a process to build almost from first principles (i.e. Hodgkin Huxley type models) mean field models of different neuron types that duplicate the stability, firing rate, and associated bifurcations as function of relevant slow variables - such as extracellular potassium - and synaptic current; and whose output is both firing rate and impact on the slow variables - such as transmembrane potassium flux. Small networks composed of just excitatory and inhibitory masses demonstrate expected dynamical states including stable firing, runaway excitation and depolarization block, and these transitions change in biologically observed ways with changes in extracellular potassium and excitatory-inhibitory balance.

neuroscience↗

bioRxiv2022

Vertical gradients in physiological function and resource allocation of white spruce diverge at the northern- and southern-most range extremes

Light availability drives vertical canopy gradients in photosynthetic functioning and carbon (C) balance, yet patterns of variability in these gradients remain unclear. We measured light availability, photosynthetic CO2 and light response curves, foliar C, nitrogen (N) and pigment concentrations, and the photochemical reflectance index (PRI) on upper and lower canopy needles of white spruce trees (Picea glauca) at the species northern and southern range extremes. We combined our photosynthetic data with previously published respiratory data to compare and contrast canopy C balance between latitudinal extremes. We found steep canopy gradients in irradiance, photosynthesis, and leaf traits at the southern range limit, but a lack of variation across canopy positions at the northern range limit. Thus, unlike many tree species from tropical to mid-latitude forests, high latitude trees may not require vertical gradients of metabolic activity to optimize photosynthetic C gain. Consequently, accounting for self-shading is less critical for predicting gross primary productivity at northern relative to southern latitudes. Northern trees also had a significantly smaller net positive leaf C balance than southern trees suggesting that, regardless of canopy position, low photosynthetic rates coupled with high respiratory costs may ultimately constrain the northern range limit of this widely distributed boreal species. SUMMARY STATEMENTCanopy gradients in photosynthetic capacity of white spruce diminish at high compared to low latitudes. Low carbon balance in high latitude trees may determine the extent of northern treeline.

ecology↗

bioRxiv2022

In vitro generation of human embryonic stem cell-derived heart organoids possessing physiological ion currents

Recently, methods for in vitro organogenesis have been broadly developed due to their strong potential for human applications in medicine. In the present study, we optimized the culturing method of human heart organoids (hHOs) from embryonic stem cells (ESCs) in the presence of the highly concentrated laminin-entactin and fibroblast growth factor 4. The resulting hHOs showed distinctive cardiac morphology with atrium- and ventricle-like chambers composed of cardiac cells as well as expressed the integral proteins of gap junctions and ion channels. In fact, isolated cardiomyocytes from these hHOs exhibited Na and Ca currents by patch clamp analysis. These results indicated that the present method will provide a powerful tool for cardiac safety assessment of newly developed drugs as an in vitro human ESC-derived test system. One-Sentence SummaryFGF4 and ECM contribute to the generation of human HOs with heart compartments and electrophysiological properties.

bioengineering↗

bioRxiv2022

Serine Racemase is a Cysteine Racemase and Physiologic Down Regulator of Insulin Promoter Methylation

D-amino acids are being recognized in mammals as important molecules with function. This is a first identification of endogenous D-cysteine in mammalian pancreas. D-cysteine is synthesized by serine racemase (SR) and SR-/- mice produce 6-10 fold higher levels of insulin in the pancreas and plasma including higher glycogen and ketone bodies in the liver. The excess insulin is stored as amyloid in secretory vesicles and exosomes. In glucose stimulated insulin secretion studies in mouse and human islets, equimolar amount of D-cysteine showed higher inhibition of insulin secretion compared to D-serine, another closely related stereoisomer synthesized by SR. In mouse models of diabetes (STZ and NOD) and human pancreas, the diabetic state showed increased expression of D-cysteine compared to D-serine followed by increased expression of SR. SR-/- mice show decreased cAMP in the pancreas followed by reduced phosphorylation of CREB (S133), lower DNA methyltransferase enzymatic and promoter activities resulting in decreased methylation of the Ins1 promoter. D-cysteine is efficiently metabolized by D-amino acid oxidase and transported by ASCT2 and Asc1. Dietary supplementation with methyl donors restored the high insulin levels and low DNMT enzymatic activity in SR-/- mice. Our data show that endogenous D-cysteine in the mammalian pancreas is a regulator of insulin secretion. HighlightsO_LISerine Racemase also functions as a cysteine racemase. C_LIO_LILack of Serine Racemase results in significantly high levels of insulin in the pancreas, plasma and larger islets. C_LIO_LID-cysteine shows greater inhibition of insulin secretion compared to D-serine. C_LIO_LIEndogenous D-cysteine signals via cyclic AMP that mediates downstream CREB-DNMT1 interaction. C_LIO_LICREB-DNMT1 interaction results in hypomethylation of Ins1 promoter that can be rescued by high methyl donor dietary supplementation rescuing high insulin levels. C_LI

biochemistry↗

bioRxiv2022

A general framework for the combined morphometric, transcriptomic, and physiological analysis of cells using metric geometry

High-resolution imaging-based single-cell profiling has transformed the study of cells in their spatial context. However, the lack of quantitative methods that can summarize the great diversity of complex cell shapes found in tissues and infer associations with other single-cell data modalities limits current analyses. Here, we report a general computational framework for the multi-modal analysis and integration of single-cell morphological data. We build upon metric geometry to construct cell morphology latent spaces, where distances in these spaces indicate the amount of physical deformation needed to change the morphology of one cell into that of another. Using these spaces, we integrate morphological data across technologies and leverage associated single-cell RNA-seq data to infer relations between morphological and transcriptomic cellular processes. We apply this framework to imaging and multi-modal data of neurons and glia to uncover genes related to neuronal plasticity. Our approach represents a strategy for incorporating cell morphological data into single-cell omics analyses.

bioinformatics↗

bioRxiv2022

Lasting consequences on physiology and social behavior following cesarean delivery in prairie voles

Cesarean delivery is associated with diminished plasma levels of several birth-signaling hormones, such as oxytocin and vasopressin. These same hormones have been previously shown to exert organizational effects when acting in early life. For example, our previous work found a broadly gregarious phenotype in prairie voles exposed to oxytocin at birth. Meanwhile, cesarean delivery has been previously associated with changes in social behavior and metabolic processes related to oxytocin and vasopressin. In the present study, we investigated the long-term neurodevelopmental consequences of cesarean delivery in prairie voles. After cross-fostering, vole pups delivered either via cesarean or vaginal delivery were studied throughout development. Cesarean-delivered pups responded to isolation differently in terms of their vocalizations (albeit in opposite directions in the two experiments), huddled in less cohesive groups under warmed conditions, and shed less heat. As young adults, we observed no differences in anxiety-like or alloparental behavior. However, in adulthood, cesarean-delivered voles of both sexes failed to form partner preferences with opposite sex conspecifics. In a follow-up study, we replicated this deficit in partner-preference formation among cesarean-delivered voles and were able to normalize pair-bonding behavior by treating cesarean-delivered vole pups with oxytocin (0.25 mg/kg) at delivery. Finally, we detected minor differences in regional oxytocin receptor expression within the brains of cesarean-delivered voles, as well as microbial composition of the gut. Gene expression changes in the gut epithelium indicated that cesarean-delivered male voles have altered gut development. These results speak to the possibility of unintended developmental consequences of cesarean delivery, which currently accounts for 32.9% of deliveries in the U.S. and suggest that further research should be directed at whether hormone replacement at delivery influences behavioral outcomes in later life.

neuroscience↗

bioRxiv2022

Linking physiology to ecosystem function: how vulnerable are different functional groups to climate change?

O_LIThe resilience of ecosystem function under global climate change is governed by individual species vulnerabilities and the functional groups they contribute to (e.g. decomposition, primary production, pollination, primary, secondary and tertiary consumption). Yet it remains unclear whether species that contribute to different functional groups, which underpin ecosystem function, differ in their vulnerability to climate change. C_LIO_LIIt is important to examine if functional group vulnerability differs across space (e.g. tropical vs temperate latitudes) to determine if some regions will be more vulnerable to loss of ecosystem function than others, and to examine whether localized effects of particular community compositions override global patterns of functional group vulnerability. C_LIO_LIWe used existing upper thermal limit data across a range of terrestrial species (N = 1,743) to calculate species warming margins (degrees distance between a species upper thermal limit and the maximum environmental temperature they inhabit), as a metric of climate change vulnerability, to determine whether species that comprise different functional groups exhibit differential vulnerability to climate change, and if vulnerability trends change across geographic space. C_LIO_LIWe found that primary producers had the broadest warming margins across the globe ( = 21.85 {degrees}C) and that tertiary consumers had the narrowest warming margins ( = 4.37 {degrees}C), where vulnerability tended to increase with trophic level. C_LIO_LISpecies that contribute towards primary production were more vulnerable in low-latitude than mid-latitude regions, but warming margins across all other functional groups did not differ across regions when evolutionary history was considered. However, when evolutionary history was excluded from the analyses (as closely related species often play similar functional roles within ecosystems demonstrating true variation in functional group warming margins) we found that pollinators are more vulnerable in mid-latitude regions and that primary producers are more vulnerable in low-latitude environments. C_LIO_LIThis study provides a critical first step in linking individual species vulnerabilities with whole ecosystem responses to climate change. C_LI

ecology↗

bioRxiv2022

Tailoring and optimizing fatty acid production by oleaginous yeasts through the systematic exploration of their physiological fitness

BackgroundThe use of palm oil for our current needs is unsustainable. Replacing palm oil with oils produced by microbes through the conversion of sustainable feedstocks is a promising alternative. However, there are major technical challenges that must be overcome to enable this transition. Foremost among these challenges is the stark increase in lipid accumulation and production of higher content of specific fatty acids. Therefore, there is a need for more in-depth knowledge and systematic exploration of the oil productivity of the oleaginous yeasts. In this study, we cultivated Cutaneotrichosporon oleaginosus and Yarrowia lipolytica at various C/N ratios and temperatures in a defined medium with glycerol as carbon source and urea as nitrogen source. We ascertained the synergistic effect between various C/N ratios of a defined medium at different temperatures with Response Surface Methodology (RSM) and explored the variation in fatty acid composition through Principal Component Analysis. ResultsBy applying RSM, we determined a temperature of 30 {degrees}C and a C/N ratio of 175 g/g to enable maximal oil production by C. oleaginosus and a temperature of 21 {degrees}C and a C/N ratio of 140 g/g for Y. lipolytica. We increased production by 71 % and 66 % respectively for each yeast compared to the average lipid accumulation in all tested conditions. Modulating temperature enabled us to steer the fatty acid compositions. Accordingly, switching from higher temperature to lower cultivation temperature shifted the production of oils from more saturated to unsaturated by 14 % in C. oleaginosus and 31 % in Y. lipolytica. Higher cultivation temperatures resulted in production of even longer saturated fatty acids, 3 % in C. oleaginosus and 1.5 % in Y. lipolytica. ConclusionsIn this study, we provided the optimum C/N ratio and temperature for C. oleaginosus and Y. lipolytica by RSM. Additionally, we demonstrated that lipid accumulation of both oleaginous yeasts was significantly affected by the C/N ratio and temperature. Furthermore, we systematically analyzed the variation in fatty acids composition and proved that changing the C/N ratio and temperature steer the composition. We have further established these oleaginous yeasts as platforms for production of tailored fatty acids.

systems biology↗

bioRxiv2022

Surface productivity gradients govern changes in abundance and physiological status of deep ocean prokaryotes across the tropical and subtropical Atlantic

Prokaryotes represent a major fraction of marine biomass and play a key role in the global carbon cycle. We studied the vertical profiles (from surface down to the bathypelagic realm) of abundance, cytometric signatures, and activity of prokaryotic communities along a productivity gradient in the subtropical and tropical Atlantic to assess whether there is a vertical linkage between surface productivity regimes and deep ocean prokaryotic communities. We found that latitudinal changes in the vertical patterns of cytometric variables were coupled with surface productivity: higher prokaryotic abundances and viabilities, and smaller cell sizes were observed below highly productive surface waters, an effect reaching down to the bathypelagic layer. On the contrary, leucine uptake rates in deep waters showed no clear relationship with surface productivity. Changes in resource and energy allocation to growth vs. maintenance in hostile environments, cell-size-dependent metabolic requirements and variability in leucine to carbon conversion may all be part of the array of factors involved in controlling prokaryotic activity patterns that were measured. Our work adds to the recent findings that highlight the importance of vertical connectivity for prokaryotic communities in the dark ocean.

microbiology↗

bioRxiv2022

Robust capability of renal tubule fatty acid uptake from apical and basolateral membranes in physiology and disease

Excess lipid accumulation is associated with obesity-related chronic kidney disease, but the mechanisms of fatty acid (FA) uptake have been poorly understood. To this end, we investigated how FAs are taken up by tubular epithelial cells (TECs) in mice by using in vivo FA tracing and histological methods. Immunohistochemistry showed that CD36, which is a well-known FA transporter, was abundantly expressed on the basolateral side of proximal TECs (PTECs). The uptake of 125I-BMIPP (a radiolabeled FA tracer) was significantly reduced in CD36-knockout kidneys at 1 min after injection. In vivo imaging with multiphoton microscopy revealed that BODIPY-C12 (a fluorescence-labeled FA tracer) accumulated on both the basolateral and apical sides of PTECs. Numerous lipid droplets accumulated in PTECs after accelerated lipolysis. Furthermore, PTEC-specific injury via diphtheria toxin (DT) injection in transgenic mice expressing the DT receptor resulted in a compensatory increase in lipid accumulation in downstream TECs. Importantly, urinary FAs were undetectable, even in mice and humans with remarkable albuminuria. Our data demonstrate that renal TECs take up FAs from blood (CD36-dependent) and primary urine (CD36-independent) and can store excess FAs as neutral lipids. The results further show that renal tubules have hitherto largely unappreciated mechanisms by which the excretion of FAs into the urine is avoided.

biochemistry↗

bioRxiv2022

When mechanical engineering inspired from physiology improves postural-related somatosensory processes

Being the first stimulated by the relative movement of foot skin and the underneath moving support surface, the plantar tactile receptors (i.e., mechanoreceptors) play an important role in the sensorimotor transformation giving rise to a postural reaction. In this light, a biomimetic surface, i.e., complying with the characteristics of the mechanoreceptors and the skin dermatoglyphs (i.e., pattern of the ridges) should facilitate the cortical processes in response to the somatosensory stimulation involved in the balance recovery motor control. Healthy young adults (n = 21) were standing still either on a biomimetic surface or on two control surfaces (i.e., grooved or smooth), when a sudden but low acceleration of the supporting surface along the lateral direction was triggered. A shorter and more robust evoked somatosensory response (i.e., SEP) was observed when participants were standing on the biomimetic surface. As well, a lower oscillatory response in the theta (5-7 Hz) time-frequency domain in the left posterior parietal cortex (PPC) was observed with the biomimetic surface. The greater shear forces induced by the interaction between the feet and the biomimetic surface during the platform motion was likely at the origin of the increased SEP. Besides, the decrease of theta power suggests that the balance task became less challenging. This interpretation was tested in a second experiment by adding a cognitive task, which should be less detrimental for the postural reaction when standing on a biomimetic surface. Consistent with this hypothesis, a more efficient postural reaction (i.e., shorter latency and greater amplitude) was observed when the cognitive task was performed while standing on the biomimetic surface.

neuroscience↗

bioRxiv2022

Absence of atrial smooth muscle in the heart of the loggerhead sea turtle (Caretta caretta): a re-evaluation of its role in diving physiology

Contraction of atrial smooth muscle in the hearts of semi-aquatic emydid turtles regulates ventricular filling, and it has been proposed that it could regulate stroke volume during characteristic rapid transitions in cardiac output associated with diving. For this hypothesis to be supported, atrial smooth muscle should be widely distributed in diving Testudines. To further understand the putative function and evolutionary significance of endocardial smooth muscle in Testudines, we studied the hearts of loggerhead sea turtles, Caretta caretta (n=7), using immunohistochemistry and histology. Surprisingly, we found no evidence of prominent atrial smooth muscle in C. caretta. However, smooth muscle was readily identified in the sinus venosus. Our results suggest atrial smooth muscle does not contribute to the diving capabilities of C. caretta, indicating that the possible roles of smooth muscle in emydid turtle hearts requires a re-evaluation. In sea turtles, the sinus venosus may instead contribute to regulate cardiac filling.

zoology↗

bioRxiv2022

A truncating mutation of Magel2 in the rat modelled for the study of Schaaf-Yang and Prader-Willi syndromes alters select behavioral and physiological outcomes

Truncating mutations of the maternally imprinted, paternally expressed MAGEL2 gene are the predicted genetic cause of several rare neurodevelopmental disorders including Schaaf-Yang (SYS), Chitayat-Hall and Opitz Trigonocephaly C syndromes. MAGEL2 is also deleted or inactivated in Prader-Willi syndrome (PWS). Previous studies in mice have utilized Magel2 gene deletion models to examine the consequences of its absence. In this study, we report the generation, molecular validation, and phenotypic characterization of a novel rat model with a truncating Magel2 mutation generating a mutant peptide sequence more closely modeling variants associated with SYS-causing mutations. Within the hypothalamus, a brain region wherein mouse and human MAGEL2 is paternally-expressed, we demonstrate at the level of transcript and peptide detection that Magel2 in the rat exhibits a paternal, parent-of-origin effect. In the evaluation of behavioral features across several domains, juvenile Magel2 mutant rats display select alterations in anxiety-like behavior and sociability measures. Moreover, the analysis of peripheral organ systems detected alterations in body composition, cardiac structure and function, and breathing irregularities in Magel2 mutant rats. Several of these findings are concordant with reported mouse phenotypes, signifying the conservation of MAGEL2 function across rodent species for specific behavioral outcome measures. We conclude that our comprehensive analysis demonstrating impairments across multiple domains demonstrates the tractability of this model system for the study of truncating MAGEL2 mutations.

neuroscience↗

bioRxiv2022

Physiological and functional heterogeneity in the mouse locus coeruleus

Neurons in the locus coeruleus (LC) have been traditionally viewed as a homogenous population. Recent studies begin to reveal their heterogeneity at multiple levels, ranging from molecular compositions to projection targets. To further uncover variations of neuronal properties in the LC, we took a genetic-based tagging approach to identify these neurons. Our data revealed diverse spike waveforms among neurons in the LC region, including a considerable fraction of narrow-spiking units. While all wide-spiking units possessed the regular waveform polarity (negative-positive deflection), the narrow units can be further divided based on opposing waveform polarities. Under anesthesia, wide units emitted action potential at a higher rate than the narrow units. Under wakefulness, only one subtype of narrow units exhibited fast-spiking phenotype. These neurons also had long latencies to optogenetic stimulation. In-situ hybridization further supported the existence of a small population of putative GABAergic neurons in the LC core. Together, our data reveal characteristic differences among neurons in the LC region, and suggest that a fraction of electrophysiologically-identified narrow-spiking neurons can be fast-spiking interneurons, and their fast-spiking feature is masked by anesthesia.

neuroscience↗