Npas4 is activated by melatonin, and drives the clock gene Cry1 in the ovine pars tuberalis

Seasonal mammalsintegrate changes in the duration of nocturnal melatonin secretion to drive annual physiologic cycles. Melatonin receptors within the proximal pituitary region, the pars tuberalis (PT), are essential in regulating seasonal neuroendocrine responses. In the ovine PT, melatonin is known to influence acute changes in transcriptional dynamics coupled to the onset (dusk) and offset (dawn) of melatonin secretion, leading to a potential interval-timing mechanism capable of decoding changes in day length (photoperiod). Melatonin offset at dawn is linked to cAMP accumulation, which directly induces transcription of the clock gene Per1. The rise of melatonin at dusk induces a separate and distinct cohort, including the clock-regulated genes Cry1 and Nampt, but little is known of the upstream mechanisms involved. Here, we used next-generation sequencing of the ovine PT transcriptome at melatonin onset and identified Npas4 as a rapidly induced basic helix-loop-helix Per-Arnt-Sim domain transcription factor. In vivo we show nuclear localization of NPAS4 protein in presumptive melatonin target cells of the PT (α-glycoprotein hormone-expressing cells), whereas in situ hybridization studies identified acute and transient expression in the PT of Npas4 in response to melatonin. In vitro, NPAS4 forms functional dimers with basic helix loop helix-PAS domain cofactors aryl hydrocarbon receptor nuclear translocator (ARNT), ARNT2, and ARNTL, transactivating both Cry1 and Nampt ovine promoter reporters. Using a combination of 5'-deletions and site-directed mutagenesis, we show NPAS4-ARNT transactivation to be codependent upon two conserved central midline elements within the Cry1 promoter. Our data thus reveal NPAS4 as a candidate immediate early-response gene in the ovine PT, driving molecular responses to melatonin

Circadian clock mechanism driving mammalian photoperiodism

From Springer Nature via Jisc Publications RouterHistory: received 2020-01-16, accepted 2020-07-27, registration 2020-08-04, pub-electronic 2020-08-27, online 2020-08-27, collection 2020-12Publication status: PublishedAbstract: The annual photoperiod cycle provides the critical environmental cue synchronizing rhythms of life in seasonal habitats. In 1936, Bünning proposed a circadian-based coincidence timer for photoperiodic synchronization in plants. Formal studies support the universality of this so-called coincidence timer, but we lack understanding of the mechanisms involved. Here we show in mammals that long photoperiods induce the circadian transcription factor BMAL2, in the pars tuberalis of the pituitary, and triggers summer biology through the eyes absent/thyrotrophin (EYA3/TSH) pathway. Conversely, long-duration melatonin signals on short photoperiods induce circadian repressors including DEC1, suppressing BMAL2 and the EYA3/TSH pathway, triggering winter biology. These actions are associated with progressive genome-wide changes in chromatin state, elaborating the effect of the circadian coincidence timer. Hence, circadian clock-pituitary epigenetic pathway interactions form the basis of the mammalian coincidence timer mechanism. Our results constitute a blueprint for circadian-based seasonal timekeeping in vertebrates

Extensive functional genomics information from early developmental time points for pig and chicken

The global Functional Annotation of Farm Animal Genomes initiative (FAANG) aims to improve animal breeding by improved genomic prediction via integration of functional genomics information. The GENESWitCH project has produced extensive functional genomics information for a variety of important tissues at early embryonic timepoints for both chickens and pigs. These datasets will be integrated to produce both tissue and time-point specific transcript, gene, and regulatory annotation for both species. In this paper, we describe the aims of the project, and the initial release of both raw and processed data

XAS and GIXRD Study of Co Sites in CoAl2O4 Layers Grown by MOCVD

The chemical environment of Co sites in CoAl2O4 layers grown by metal-organic chemical vapor deposition has been investigated by X-ray absorption spectroscopy (XAS) and X-ray diffraction (XRD). It is shown that the air- or [O2 + H2O]-annealing at 500 °C of the layers deposited at low temperature induce a partial crystallization with the formation of (Co1-2ηAl2)(Co2Al2η)(Co 2ηAl2(1-η)))O4 spinel (η = 0.2-0.27). Nevertheless, slightly more than half of Co remains in an amorphous phase, and XAS data are consistent with the formation of a medium-range ordered Co3O4 phase, especially upon air-annealing. Layers grown at higher temperatures (600-650 °C) exhibit a similar but more complex structure, since the presence of an additional medium-range ordered phase (likely, CoAl2O4) is also revealed. The air-annealing at high temperature (800 °C) generates blue, almost completely crystalline, CoAl2O4 layers. Optical properties of deposited layers are discussed by referring to the outcomes of structural results. In particular, the optical absorption spectrum results negligibly affected by the presence of the amorphous phase, while absorptions present in the 300-500 nm range, responsible for the green layer color and evident in samples annealed in an oxidizing atmosphere or grown at high temperature, are likely caused by the octahedrally coordinated Co ions of the partially inverted spinel (Co;- 2ηAl2η)(Co2(1-η)Al 2(1-η))O4 -))O4 phase. Despite the XRD analysis that ultimately demonstrates the presence of octahedrally coordinated Co ions, whose oxidation state in the spinel phase is in majority Co(II), the occurrence of Co(III) species with an octahedral environment cannot be ruled out