30 research outputs found
FrancisCrickInstitute/Pachnis-lab: January_2022_Nature_Methods
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Molecular profiling of enteric nervous system cell lineages
Yuuki Obata, Álvaro Castaño, Todd Fallesen, Ana Carina Bon-Frauches, Stefan Boeing, Almaz Huseynova, Sarah McCallum, Tiffany Heanue and Vassilis Pachnis
and:
Obata et al. Neuronal programming by microbiota regulates intestinal physiology.
Nature 578, 284–289 (2020). DOI: https://doi.org/10.1038/s41586-020-1975-
An optogenetic method for interrogating YAP1 and TAZ nuclear-cytoplasmic shuttling
No full textThe shuttling of transcription factors and transcriptional regulators into and out of the nucleus is central to the regulation of many biological processes. Here we describe a new method for studying the rates of nuclear entry and exit of transcriptional regulators. A photo-responsive LOV (light–oxygen–voltage) domain from Avena sativa is used to sequester fluorescently labelled transcriptional regulators YAP1 and TAZ (also known as WWTR1) on the surface of mitochondria and to reversibly release them upon blue light illumination. After dissociation, fluorescent signals from the mitochondria, cytoplasm and nucleus are extracted by a bespoke app and used to generate rates of nuclear entry and exit. Using this method, we demonstrate that phosphorylation of YAP1 on canonical sites enhances its rate of nuclear export. Moreover, we provide evidence that, despite high intercellular variability, YAP1 import and export rates correlate within the same cell. By simultaneously releasing YAP1 and TAZ from sequestration, we show that their rates of entry and exit are correlated. Furthermore, combining the optogenetic release of YAP1 with lattice light-sheet microscopy reveals high heterogeneity of YAP1 dynamics within different cytoplasmic regions, demonstrating the utility and versatility of our tool to study protein dynamics
Downward spiral: The impact of out-of-home placement on paternal welfare dependency
No full textAbstractIn this article, we test how out-of-home placement affects men's labor market attachment, and in so doing we provide a novel parallel to existing research on how fatherhood affects men, which focuses almost exclusively on a child's arrival. Using population panel data from Denmark that include all first time fathers whose children were placed in out-of-home care from 1995 to 2005, we find that having a child placed in care is associated with up to a 4 percentage point increase in welfare dependency. Having a child placed in out-of-home care appear to aggravate conditions that likely necessitated the out-of-home placement to begin with, thereby likely necessitating longer duration of out-of-home placements. Thus, out-of-home placements have substantial secondary costs for parents and society
KINESIN-MICROTUBULE INTERACTIONS DURING GLIDING ASSAYS UNDER MAGNETIC FORCE
No full textConventional kinesin is a motor protein capable of converting the chemical energy of ATP into mechanical work. In the cell, this is used to actively transport vesicles through the intracellular matrix. The relationship between the velocity of a single kinesin, as it works against an increasing opposing load, has been well studied. The relationship between the velocity of a cargo being moved by multiple kinesin motors against an opposing load has not been established. A major difficulty in determining the force-velocity relationship for multiple motors is determining the number of motors that are moving a cargo against an opposing load
Decline in Rate of Divorce and Separation Filings in Denmark in 2020 Compared with Previous Years
No full textThe radical changes to everyday life brought on by the coronavirus disease 2019 (COVID-19) pandemic and the closure of nonfamily social spheres in particular may have impacted marriage dynamics. The author provides evidence on the monthly rates of initiation of divorce and separation filings in Denmark for the period from 2016 to 2020 to examine how filing behavior changed during 2020 compared with the four previous years. Because filing precedes divorce, rates reflect more precisely the temporal dynamic of divorce initiation. Rates of initiation of divorce filings declined in 2020 to the lowest level across the period from 2016 to 2020. On average, monthly rates in 2020 were 7 percent lower than 2019 rates and 20 percent lower than 2016 rates. There is little indication that the COVID-19 pandemic had an immediate influence on divorce dynamics, although the filing rate was more depressed during lockdown periods.</p
Neuronal programming by microbiota regulates intestinal physiology
No full textNeural control of the function of visceral organs is essential for homeostasis and health. Intestinal peristalsis is critical for digestive physiology and host defence, and is often dysregulated in gastrointestinal disorders1. Luminal factors, such as diet and microbiota, regulate neurogenic programs of gut motility2-5, but the underlying molecular mechanisms remain unclear. Here we show that the transcription factor aryl hydrocarbon receptor (AHR) functions as a biosensor in intestinal neural circuits, linking their functional output to the microbial environment of the gut lumen. Using nuclear RNA sequencing of mouse enteric neurons that represent distinct intestinal segments and microbiota states, we demonstrate that the intrinsic neural networks of the colon exhibit unique transcriptional profiles that are controlled by the combined effects of host genetic programs and microbial colonization. Microbiota-induced expression of AHR in neurons of the distal gastrointestinal tract enables these neurons to respond to the luminal environment and to induce expression of neuron-specific effector mechanisms. Neuron-specific deletion of Ahr, or constitutive overexpression of its negative feedback regulator CYP1A1, results in reduced peristaltic activity of the colon, similar to that observed in microbiota-depleted mice. Finally, expression of Ahr in the enteric neurons of mice treated with antibiotics partially restores intestinal motility. Together, our experiments identify AHR signalling in enteric neurons as a regulatory node that integrates the luminal environment with the physiological output of intestinal neural circuits to maintain gut homeostasis and health.sponsorship: We thank the Crick Science Technology Platforms, the University of Bern FACSLab and the Bern Clean Mouse Facility for expert support; R. Lasrado and S.-H. Chng for assistance with tissue dissection; M. Shapiro for bioinformatic input; C. Schiering for useful advice; all members of the Pachnis laboratory for insightful comments on the manuscript and discussions; and M. D'Amato for insightful comments on the manuscript. Y.O. was supported by an EMBO long-term fellowship (ALTF 1214-2015), travel grants from Boehringer Ingelheim Fonds and the Society for Mucosal Immunology (SMI); he is currently supported by an HFSP postdoctoral fellowship (LT000176/2016). This work was supported by the Medical Research Council (MRC) and The Francis Crick Institute (which receives funding from the MRC, Cancer Research UK and the Wellcome Trust). V.P. was also funded by BBSRC (BB/L022974) and the Wellcome Trust (212300/Z/18/Z). (EMBO long-term fellowship|ALTF 1214-2015, Boehringer Ingelheim Fonds, Society for Mucosal Immunology (SMI), HFSP postdoctoral fellowship|LT000176/2016, Medical Research Council (MRC), Francis Crick Institute (MRC), Francis Crick Institute (Cancer Research UK), Francis Crick Institute (Wellcome Trust), BBSRC|BB/L022974, Wellcome Trust|212300/Z/18/Z, Wellcome Trust|212300/Z/18/Z, BBSRC|BB/L022974/1)status: Publishe
“Popoffs” Under a Transverse Force Reveal the Number and Location of Active Kinesin Motors During Motility Assays
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