Royan, M;Siddique, K;Nourizadeh-lillabadi, R;Weltzien, F;Henkel, C;FONTAINE, R;
| DOI: 10.2139/ssrn.4142092
In fish, prolactin-producing cells (lactotropes) are located in the anterior part of the pituitary and play an essential role in osmoregulation. However, small satellite lactotrope populations have been described in other parts of the pituitary in several species. The functional and developmental backgrounds of these extra populations are not known. We recently described two distinct prolactin-expressing cell types in Japanese medaka, a salinity tolerant fish, using single cell transcriptomics. In this study, we thus characterize the two transcriptomically distinct lactotrope cell types and explore the hypothesis that they represent the spatially distinct cell populations found in other species. Single cell RNA sequencing shows that one of the two lactotrope cell types exhibits an expression profile similar to that of stem cell populations. Using in situ hybridization, we show that the medaka pituitary often develops additional small satellite lactotrope cell groups, like in other teleost species. These satellite clusters arise early during development and grow in cell number throughout life regardless of the animal’s sex. Surprisingly, there seems to be no correspondence between the stem cell-like lactotropes and these newly emerging lactotrope populations. Instead, our data support a scenario in which the stem cell-like lactotropes are an intrinsic stage in the development of every spatially distinct lactotrope cluster. In addition, lactotrope activity in the medaka pituitary decreases when environmental salinity increases in the two spatially distinct lactotrope clusters, supporting their role in osmoregulation. However, this decrease appears weaker in the satellite lactotrope cell groups, suggesting that these lactotropes are differentially regulated.
The Journal of comparative neurology
Takanami, K;Oti, T;Kobayashi, Y;Hasegawa, K;Ito, T;Tsutsui, N;Ueda, Y;Carstens, E;Sakamoto, T;Sakamoto, H;
PMID: 35686563 | DOI: 10.1002/cne.25376
Gastrin-releasing peptide (GRP) and its receptor (GRPR) have been identified as itch mediators in the spinal and trigeminal somatosensory systems in rodents. In primates, there are few reports of GRP/GRPR expression or function in the spinal sensory system and virtually nothing is known in the trigeminal system. The aim of the present study was to characterize GRP and GRPR in the trigeminal and spinal somatosensory system of Japanese macaque monkeys (Macaca fuscata). cDNA encoding GRP was isolated from the macaque dorsal root ganglion (DRG) and exhibited an amino acid sequence that was highly conserved among mammals and especially in primates. Immunohistochemical analysis demonstrated that GRP was expressed mainly in the small-sized trigeminal ganglion and DRG in adult macaque monkeys. Densely stained GRP-immunoreactive (ir) fibers were observed in superficial layers of the spinal trigeminal nucleus caudalis (Sp5C) and the spinal cord. In contrast, GRP-ir fibers were rarely observed in the principal sensory trigeminal nucleus and oral and interpolar divisions of the spinal trigeminal nucleus. cDNA cloning, in situ hybridization, and Western blot revealed substantial expression of GRPR mRNA and GRPR protein in the macaque spinal dorsal horn and Sp5C. Our Western ligand blot and ligand derivative stain for GRPR revealed that GRP directly bound in the macaque Sp5C and spinal dorsal horn as reported in rodents. Finally, GRP-ir fibers were also detected in the human spinal dorsal horn. The spinal and trigeminal itch neural circuits labeled with GRP and GRPR appear to function also in primates.
Connexin mRNA distribution in adult mouse kidneys
Pflugers Archiv : European journal of physiology
Geis, L;Boudriot, FF;Wagner, C;
PMID: 34365513 | DOI: 10.1007/s00424-021-02608-0
Kidneys are thought to express eight different connexin isoforms (i.e., Cx 26, 30, 32, 37, 40, 43, 45, and 46), which form either hemichannels or gap junctions serving to intercellular communication and functional synchronization. Proper function of connexins has already been shown to be crucial for regulation of renal hemodynamics and renin secretion, and there is also growing evidence for connexins to play a role in pathologic conditions such as renal fibrosis or diabetic nephropathy. Therefore, exact intrarenal localization of the different connexin isoforms gains particular interest. Until now intrarenal expression of connexins has mainly been examined by immunohistochemistry, which in part generated conflicting results depending on antibodies and fixation protocols used. In this work, we used fluorescent RNAscope as an alternative technical approach to localize renal connexin mRNAs in healthy mouse kidneys. Addition of RNAscope probes for cell type specific mRNAs was used to assign connexin mRNA signals to specific cell types. We hereby found Cx26 mRNA strongly expressed in proximal tubules, Cx30 mRNA was selectively detected in the urothelium, and Cx32 mRNA was found in proximal tubules and to a lesser extent also in collecting ducts. Cx37 mRNA was mainly associated with vascular endothelium, Cx40 mRNA was largely found in glomerular mesangial and less in vascular endothelial cells, Cx43 mRNA was sparsely expressed by interstitial cells of all kidney zones, and Cx45 mRNA was predominantly found in smooth muscle cell layers of both blood vessels and ureter as well as in mesangial and interstitial (fibroblastic) cells. Cx46 mRNA could not be detected. In summary our results essentially confirm previous data on connexin expression in the renal vasculature and in glomeruli. In addition, they demonstrate strong connexin gene expression in proximal tubules, and they suggest significant connexin expression in resident tubulointerstitial cells.
Emerging infectious diseases
Temeeyasen, G;Sharafeldin, TA;Lin, CM;Hause, BM;
PMID: 35076011 | DOI: 10.3201/eid2802.211681
In 1978, canine parvovirus type 2 originated from spillover of a feline panleukopenia-like virus, causing a worldwide pandemic of enteritis and myocarditis among canids. In 2020, the virus was identified in pigs in South Dakota, USA, by PCR, sequencing, in situ hybridization, and serology. Genetic analysis suggests spillover from wildlife.
Progneaux, A;Evrard, C;De Glas, V;Fontaine, A;Dotreppe, C;De Vuyst, E;Nikkels, AF;García-González, V;Dumoutier, L;Lambert de Rouvroit, C;Poumay, Y;
PMID: 36645024 | DOI: 10.1111/exd.14749
Atopic dermatitis (AD) is a Th2-type inflammatory disease characterized by an alteration of epidermal barrier following the release of IL-4 and IL-13. These cytokines activate type II IL-4Rα/IL-13Rα1 receptors in the keratinocyte. Whilst IL-2Rγ, that forms type I receptor for IL-4, is only expressed in haematopoietic cells, recent studies suggest its induction in keratinocytes, which questions about its role. We studied expression of IL-2Rγ in keratinocytes and its role in alteration of keratinocyte function and epidermal barrier. IL-2Rγ expression in keratinocytes was studied using both reconstructed human epidermis (RHE) exposed to IL-4/IL-13 and AD skin. IL-2Rγ induction by type II receptor has been analyzed using JAK inhibitors and RHE knockout (KO) for IL13RA1. IL-2Rγ function was investigated in RHE KO for IL2RG. In RHE, IL-4/IL-13 induce expression of IL-2Rγ at the mRNA and protein levels. Its mRNA expression is also visualized in keratinocytes of lesional AD skin. IL-2Rγ expression is low in RHE treated with JAK inhibitors and absent in RHE KO for IL13RA1. Exposure to IL-4/IL-13 alters epidermal barrier, but this alteration is absent in RHE KO for IL2RG. A more important induction of IL-13Rα2 is reported in RHE KO for IL2RG than in not edited RHE. These results demonstrate IL-2Rγ induction in keratinocytes through activation of type II receptor. IL-2Rγ is involved in the alteration of the epidermal barrier and in the regulation of IL-13Rα2 expression. Observation of IL-2Rγ expression by keratinocytes inside AD lesional skin suggests a role for this receptor subunit in the disease.
Pratta, M;Burke, L;DiPersio, J;Maziarz, R;Feldman, P;Brodeur, T;Timmers, C;Ivanova, O;Barbour, A;Morariu-Zamfir, R;Frigault, M;
| DOI: 10.1182/blood-2022-169382
PRATTA:_Incyte Corporation:_ Current Employment, Current equity holder in private company, Current holder of _stock options_ in a privately-held company. BURKE:_Incyte Corporation:_ Current Employment, Current equity holder in private company, Current holder of _stock options_ in a privately-held company. DIPERSIO:_BioLineRx, Ltd.:_ Research Funding; _Macrogenics:_ Research Funding; _NeoImmune Tech:_ Research Funding; _Amphivena Therapeutics:_ Research Funding; _hC Bioscience, Inc.:_ Membership on an entity's Board of Directors or advisory committees; _RiverVest Venture Partners:_ Consultancy, Membership on an entity's Board of Directors or advisory committees; _Incyte:_ Consultancy, Research Funding; _WUGEN:_ Current equity holder in private company, Research Funding; _CAR-T cell Product with Washington University and WUGEN:_ Patents & Royalties; _VLA-4 Inhibitor with Washington University and Magenta Therapeutics:_ Patents & Royalties; _Magenta Therapeutics:_ Current equity holder in private company, Membership on an entity's Board of Directors or advisory committees. MAZIARZ:_ASTCT:_ Membership on an entity's Board of Directors or advisory committees; _CRISPR Therapeutics:_ Consultancy, Honoraria; _Novartis:_ Other: Support for research on CART; _Allovir:_ Other: Support for research on Allo HCT costs of care of infectious related complications; _Orca Bio:_ Other: Support for research analysis and for medical writing. FELDMAN:_Incyte Corporation:_ Current Employment, Current equity holder in private company, Current holder of _stock options_ in a privately-held company. BRODEUR:_Incyte Corporation:_ Current Employment, Current equity holder in private company, Current holder of _stock options_ in a privately-held company. TIMMERS:_Incyte Corporation:_ Current Employment, Current equity holder in private company, Current holder of _stock options_ in a privately-held company. IVANOVA:_Incyte Corporation:_ Current Employment, Current equity holder in private company, Current holder of _stock options_ in a privately-held company. BARBOUR:_Incyte Corporation:_ Ended employment in the past 24 months; _Karyopharm:_ Current Employment, Current equity holder in publicly-traded company. MORARIU-ZAMFIR:_Incyte Corporation:_ Current Employment, Current equity holder in private company, Current holder of _stock options_ in a privately-held company. FRIGAULT:_Cytoagents:_ Consultancy; _Iovance:_ Consultancy; _Novartis:_ Consultancy, Research Funding; _Kite/Gilead:_ Consultancy, Research Funding; _Arcellx:_ Research Funding; _JnJ/Legend:_ Consultancy; _BMS:_ Consultancy.
Intercellular Arc Signaling Regulates Vasodilation
The Journal of neuroscience : the official journal of the Society for Neuroscience
de la Peña, JB;Barragan-Iglesias, P;Lou, TF;Kunder, N;Loerch, S;Shukla, T;Basavarajappa, L;Song, J;James, DN;Megat, S;Moy, JK;Wanghzou, A;Ray, PR;Hoyt, K;Steward, O;Price, TJ;Shepherd, J;Campbell, ZT;
PMID: 34326146 | DOI: 10.1523/JNEUROSCI.0440-21.2021
Injury responses require communication between different cell types in the skin. Sensory neurons contribute to inflammation and can secrete signaling molecules that affect non-neuronal cells. Despite the pervasive role of translational regulation in nociception, the contribution of activity-dependent protein synthesis to inflammation is not well understood. To address this problem, we examined the landscape of nascent translation in murine dorsal root ganglion (DRG) neurons treated with inflammatory mediators using ribosome profiling. We identified the activity-dependent gene, Arc, as a target of translation in vitro and in vivo Inflammatory cues promote local translation of Arc in the skin. Arc-deficient male mice display exaggerated paw temperatures and vasodilation in response to an inflammatory challenge. Since Arc has recently been shown to be released from neurons in extracellular vesicles (EVs), we hypothesized that intercellular Arc signaling regulates the inflammatory response in skin. We found that the excessive thermal responses and vasodilation observed in Arc defective mice are rescued by injection of Arc-containing EVs into the skin. Our findings suggest that activity-dependent production of Arc in afferent fibers regulates neurogenic inflammation potentially through intercellular signaling.SIGNIFICANCE STATEMENTNociceptors play prominent roles in pain and inflammation. We examined rapid changes in the landscape of nascent translation in cultured dorsal root ganglia (DRGs) treated with a combination of inflammatory mediators using ribosome profiling. We identified several hundred transcripts subject to rapid preferential translation. Among them is the immediate early gene (IEG) Arc. We provide evidence that Arc is translated in afferent fibers in the skin. Arc-deficient mice display several signs of exaggerated inflammation which is normalized on injection of Arc containing extracellular vesicles (EVs). Our work suggests that noxious cues can trigger Arc production by nociceptors which in turn constrains neurogenic inflammation in the skin.
McMeekin LJ, Li Y, Fox SN, Rowe GC, Crossman DK, Day JJ, Li Y, Detloff PJ, Cowell RM.
PMID: 29491012 | DOI: 10.1523/JNEUROSCI.0848-17.2018
Multiple lines of evidence indicate that a reduction in the expression and function of the transcriptional coactivator peroxisome proliferator activated receptor gamma coactivator-1α (PGC-1α) is associated with neurodegeneration in diseases such as Huntington Disease (HD). Polymorphisms in the PGC-1α gene modify HD progression, and PGC-1α expression is reduced in striatal medium spiny neurons (MSNs) of HD patients and mouse models. However, neither the MSN-specific function of PGC-1α nor the contribution of PGC-1α deficiency to motor dysfunction is known. We identified novel PGC-1α-dependent transcripts involved in RNA processing, signal transduction and neuronalmorphology and confirmed reductions in these transcripts in male and female mice lacking PGC-1α specifically in MSNs, indicating a cell-autonomous effect in this population. MSN-specific PGC-1α deletion caused reductions in previously identified neuronal and metabolic PGC-1α-dependent genes, without causing striatal vacuolizations. Interestingly, these mice exhibited a hypoactivity with age, similar to several HD animal models. However, these newly identified PGC-1α-dependent genes were upregulated with disease severity and age in knockin HD mouse models independent of changes in PGC-1α transcript, contrary to what would be predicted from a loss-of-function etiological mechanism. These data indicate that PGC-1α is necessary for MSN transcriptional homeostasis and function with age and that, while PGC-1α loss in MSNs does not replicate an HD-like phenocopy, its downstream genes are altered in a repeat-length and age-dependent fashion. Understanding the additive effects of PGC-1α gene functional variation and mutant huntingtin on transcription in this cell type may provide insight into the selective vulnerability of MSNs in HD.SIGNIFICANCE STATEMENTReductions in PGC-1α-mediated transcription have been implicated in the pathogenesis of Huntington Disease (HD). We show that while PGC-1α-dependent transcription is necessary to maintain MSN function with age, its loss is insufficient to cause striatal atrophy in mice. We also highlight a set of genes that can serve as proxies for PGC-1α functional activity in the striatum for target engagement studies. Furthermore, we demonstrate that PGC-1α-dependent genes are upregulated in a dose and age-dependent fashion in HD mouse models, contrary to what would be predicted from a loss-of-function etiological mechanism. However, given this role for PGC-1α in MSN transcriptional homeostasis, it is important to consider how genetic variation in PGC-1α could contribute to mutant-huntingtin-induced cell death and disease progression.
Jaligama S, Patel VS, Wang P, Sallam A, Harding J, Kelley M, Mancuso SR, Dugas TR, Cormier SA.
PMID: 29724254 | DOI: 10.1186/s12989-018-0255-3
Abstract
BACKGROUND:
Pollutant particles containing environmentally persistent free radicals (EPFRs) are formed during many combustion processes (e.g. thermal remediation of hazardous wastes, diesel/gasoline combustion, wood smoke, cigarette smoke, etc.). Our previous studies demonstrated that acute exposure to EPFRs results in dendritic cell maturation and Th17-biased pulmonary immune responses. Further, in a mouse model of asthma, these responses were enhanced suggesting exposure to EPFRs as a risk factor for the development and/or exacerbation of asthma. The aryl hydrocarbon receptor (AHR) has been shown to play a role in the differentiation of Th17 cells. In the current study, we determined whether exposure to EPFRs results in Th17 polarization in an AHR dependent manner.
RESULTS:
Exposure to EPFRs resulted in Th17 and IL17A dependent pulmonary immune responses including airway neutrophilia. EPFR exposure caused a significant increase in pulmonary Th17 cytokines such as IL6, IL17A, IL22, IL1β, KC, MCP-1, IL31 and IL33. To understand the role of AHR activation in EPFR-induced Th17 inflammation, A549 epithelial cells and mouse bone marrow-derived dendritic cells (BMDCs) were exposed to EPFRs and expression of Cyp1a1 and Cyp1b1, markers for AHR activation, was measured. A significant increase in Cyp1a1 and Cyp1b1 gene expression was observed in pulmonary epithelial cells and BMDCs in an oxidative stress and AHR dependent manner. Further, in vivo exposure of mice to EPFRs resulted in oxidative stress and increased Cyp1a1 and Cyp1b1 pulmonary gene expression. To further confirm the role of AHR activation in pulmonary Th17 immune responses, mice were exposed to EPFRs in the presence or absence of AHR antagonist. EPFR exposure resulted in a significant increase in pulmonary Th17 cells and neutrophilic inflammation, whereas a significant decrease in the percentage of Th17 cells and neutrophilic inflammation was observed in mice treated with AHR antagonist.
CONCLUSION:
Exposure to EPFRs results in AHR activation and induction of Cyp1a1 and in vitro this is dependent on oxidative stress. Further, our in vivo studies demonstrated a role for AHR in EPFR-induced pulmonary Th17 responses including neutrophilic inflammation.
Wang, W;Xu, M;Yue, J;Zhang, Q;Nie, X;Jin, Y;Zhang, Z;
PMID: 35894166 | DOI: 10.1210/endocr/bqac115
Increasing incidence of metabolic disturbances has become a severe public healthcare problem. Ion channels and receptors in the paraventricular nucleus (PVN) of the hypothalamus serve vital roles in modulating neuronal activities and endocrine functions, which are linked to the regulation of energy balance and glucose metabolism. In this study, we found that acid-sensing ion channel 1a (ASIC1a), a Ca2+-permeable cationic ion channel was localized in the PVN. Knockdown of ASIC1a in this region led to significant body weight gain, glucose intolerance, and insulin resistance. Pharmacological inhibition of ASIC1a resulted in an increase in food intake and a decrease in energy expenditure. Our findings suggest ASIC1a in the PVN as a potential new target for the therapeutic intervention of metabolic disorders.
Fienko, S;Landles, C;Sathasivam, K;Gomez-Paredes, C;McAteer, S;Milton, R;Osborne, G;Jones, S;Phillips, J;Kordasiewicz, H;Bates, G;
| DOI: 10.1136/jnnp-2022-ehdn.3
RESULTS Microscopic analysis revealed that the full-length _HTT_ mRNA (_FL-HTT_) was retained in RNA nuclear clusters together with the incompletely spliced _HTT1a_ transcript. These clusters were not observed in zQ175 HD mouse model where, instead, _FL-Htt_ and _Htt1a_ mRNAs were detected as mostly cytoplasmic molecules. Immunohistochemistry showed a progressive appearance of aggregated HTT in nuclei in the cortex, striatum, hippocampus and cerebellum. HTRF indicated that the level of exon 1 HTT was highest in the cerebellum. Soluble mutant exon 1 HTT decreased with age, with concomitant increase in aggregated HTT. In YAC128 MEFs, _HTT1a_ was detected and ASOs targeting _HTT_ were efficient in lowering _HTT_ levels in this model system.
Investigative Ophthalmology & Visual Science
Yang, H;Yuan, M;Gaurang, P;Sun, A;
RESULTS : In rodent eye (both rat and mouse), CFH mRNA is strongly expressed in the retinal pigment epithelium with some expression also found in inner nuclear (INL) and retinal ganglion cell (RGC) layers of the retina. C3 mRNA is expressed mainly in RGC, INL of retina, ciliary body, corneal epithelium with some expression is also found in rodent retinal pigment epithelium layer. However, in human eye, CFH and C3 mRNA are strongly expressed in the choroid. Some expression is also found in RGC, INL layer of retina, ONH, sclera, cornea endothelial and stroma; and ciliary body. There is no C3 or CFH signal detected in RPE cells.
