Neuron specific quantitation of Gαolf expression and signaling in murine brain tissue.

The heterotrimeric G-protein α subunit, Gαolf, acts to transduce extracellular signals through G-protein coupled receptors (GPCRs) and stimulates adenylyl cyclase mediated production of the second messenger cyclic adenosine monophosphate. Numerous mutations in the GNAL gene, which encodes Gαolf, have been identified as causative for an adult-onset dystonia. These mutations disrupt GPCR signaling cascades in in vitro assays through several mechanisms, and this disrupted signaling is hypothesized to lead to dystonic motor symptoms in patients. However, the cells and circuits that mutations in GNAL corrupt are not well understood. Published patterns of Gαolf expression outside the context of the striatum are sparse, conflicting, often lack cell type specificity, and may be confounded by expression of the close GNAL homolog of GNAS. Here, we use RNAScope in-situ hybridization to quantitatively characterize Gnal mRNA expression in brain tissue from wildtype C57BL/6J adult mice. We observed widespread expression of Gnal puncta throughout the brain, suggesting Gαolf is expressed in more brain structures and neuron types than previously accounted for. We quantify transcripts at a single cell level, and use neuron type specific markers to further classify and understand patterns of GNAL expression. Our data suggests that brain regions classically associated with motor control, initiation, and regulation show the highest expression of GNAL, with Purkinje Cells of the cerebellum showing the highest expression of any neuron type examined. Subsequent conditional Gnal knockout in Purkinje cells led to markedly decreased intracellular cAMP levels and downstream cAMP-dependent enzyme activation. Our work provides a detailed characterization of Gnal expression throughout the brain and the biochemical consequences of loss of Gαolf signaling in vivo in neurons that highly express Gnal.

G α olf Regulates Biochemical Signaling in Neurons Associated with Movement Control and Initiation.

The heterotrimeric G-protein α subunit, Gα olf , acts to transduce extracellular signals through G-protein coupled receptors (GPCRs) and stimulates adenylyl cyclase mediated production of the second messenger cyclic adenosine monophosphate. Numerous mutations in the GNAL gene, which encodes Gα olf , have been identified as causative for an adult-onset dystonia. These mutations disrupt GPCR signaling cascades in in vitro assays through several mechanisms, and this disrupted signaling is hypothesized to lead to dystonic motor symptoms in patients. However, the cells and circuits that mutations in GNAL corrupt are not well understood. Published patterns of Gα olf expression outside the context of the striatum are sparse, conflicting, often lack cell type specificity, and may be confounded by expression of the close GNAL homolog of GNAS . Here, we use RNAScope in-situ hybridization to quantitatively characterize Gnal mRNA expression in brain tissue from wildtype C57BL/6J adult mice. We observed widespread expression of Gnal puncta throughout the brain, suggesting Gα olf is expressed in more brain structures and neuron types than previously accounted for. We quantify transcripts at a single cell level, and use neuron type specific markers to further classify and understand patterns of GNAL expression. Our data suggests that brain regions classically associated with motor control, initiation, and regulation show the highest expression of GNAL , with Purkinje Cells of the cerebellum showing the highest expression of any neuron type examined. Subsequent conditional Gnal knockout in Purkinje cells led to markedly decreased intracellular cAMP levels and downstream cAMP-dependent enzyme activation. Our work provides a detailed characterization of Gnal expression throughout the brain and the biochemical consequences of loss of Gα olf signaling in vivo in neurons that highly express Gnal .

American Association for Anatomy recommendations for the management of legacy anatomical collections.

Collections of human remains in scientific and private institutions have a long tradition, though throughout history there has often been variable regard for the respect and dignity that these tissues demand. Recent public scandals around the use of human remains, coupled with an increasing community awareness around accountability in such instances, forces scholars to confront the ethical and moral concerns associated with these collections. This includes specific focus on the acquisition, storage, use, and disposition of these remains, which were often collected with no consent and with little knowledge, or concern, about the individual or their respective culture and practices surrounding death and postmortem treatment. As a response, the American Association for Anatomy convened a Legacy Anatomical Collections Task Force to consider these issues and to develop recommendations to assist those working with these tissues in education, research, and museum collections. This has culminated with the development of Recommendations for the Management of Legacy Anatomical Collections. The recommendations provide both an ethical foundation and practical considerations for the use, storage, and disposition of legacy collections of human tissues, and deliver guidance for establishing appropriate management and oversight, investigating provenance, and engaging with communities of care. While these Recommendations are considered a living document which will change over time as ethical principles concerning human tissue evolve, they currently represent 'best practice' recommendations that can guide researchers, teachers, and museum associates as they consider the future of legacy anatomical collections in their care.

Enhancer-promoter interactions are reconfigured through the formation of long-range multiway hubs as mouse ES cells exit pluripotency.

Enhancers bind transcription factors, chromatin regulators, and non-coding transcripts to modulate the expression of target genes. Here, we report 3D genome structures of single mouse ES cells as they are induced to exit pluripotency and transition through a formative stage prior to undergoing neuroectodermal differentiation. We find that there is a remarkable reorganization of 3D genome structure where inter-chromosomal intermingling increases dramatically in the formative state. This intermingling is associated with the formation of a large number of multiway hubs that bring together enhancers and promoters with similar chromatin states from typically 5-8 distant chromosomal sites that are often separated by many Mb from each other. In the formative state, genes important for pluripotency exit establish contacts with emerging enhancers within these multiway hubs, suggesting that the structural changes we have observed may play an important role in modulating transcription and establishing new cell identities.

Increased Wellbeing following Engagement in a Group Nature-Based Programme: The Green Gym Programme Delivered by the Conservation Volunteers.

The wellbeing benefits of engaging in a nature-based programme, delivered by the Voluntary, Community and Social Enterprise sector, were examined in this study. Prior to attending The Conservation Volunteers' Green Gym™, attendees (n = 892) completed demographics, health characteristics and the Warwick Edinburgh Mental Wellbeing Short-Form Scale. Attendees (n = 253, 28.4%) provided a measure on average 4.5 months later. There were significant increases in wellbeing after engaging in Green Gym, with the greatest increases in those who had the lowest starting levels of wellbeing. Wellbeing increases were sustained on average 8.5 months and 13 months later in those providing a follow up measure (n = 92, n = 40, respectively). Attendees who continued to engage in Green Gym but not provide follow up data (n = 318, 35.7%) tended to be more deprived, female and self-report a health condition. Attendees who did not continue to engage in Green Gym (n = 321, 36.0%) tended to be less deprived and younger. These findings provide evidence of the wellbeing benefits of community nature-based activities and social ('green') prescribing initiatives and indicate that Green Gym targets some groups most in need.

Single-cell transcriptional analysis reveals ILC-like cells in zebrafish.

Innate lymphoid cells (ILCs) are important mediators of the immune response and homeostasis in barrier tissues of mammals. However, the existence and function of ILCs in other vertebrates are poorly understood. Here, we use single-cell RNA sequencing to generate a comprehensive atlas of zebrafish lymphocytes during tissue homeostasis and after immune challenge. We profiled 14,080 individual cells from the gut of wild-type zebrafish, as well as of rag1-deficient zebrafish that lack T and B cells, and discovered populations of ILC-like cells. We uncovered a rorc-positive subset of ILCs that could express cytokines associated with type 1, 2, and 3 responses upon immune challenge. Specifically, these ILC-like cells expressed il22 and tnfa after exposure to inactivated bacteria or il13 after exposure to helminth extract. Cytokine-producing ILC-like cells express a specific repertoire of novel immune-type receptors, likely involved in recognition of environmental cues. We identified additional novel markers of zebrafish ILCs and generated a cloud repository for their in-depth exploration.