Characterization of<i>mWake</i>expression in the murine brain

Bell, Benjamin J.; Wang, Annette A.; Kim, Dong Won; Blackshaw, Seth; Wu, Mark N.

doi:10.1101/2020.05.25.114363

Cited by 3 publications

(2 citation statements)

References 105 publications

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“…Liu et al (9) found that the circadian output molecule wide awake (WAKE) responds to CLK oscillations, reducing excitability in wake-promoting l-LN V s. This pathway upregulates the GABA A receptor resistant to dieldrin (RDL). They also found that the mammalian homolog of WAKE ( 9), mWAKE, is expressed in both the SCN and dorsal medial hypothalamus (99). These data support the notion that mWAKE is conserved in driving wakefulness by modulating firing rates.…”

Section: The Neural Network That Regulates the Circadian Clocksupporting

confidence: 52%

Light/Clock Influences Membrane Potential Dynamics to Regulate Sleep States

et al. 2021

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The circadian rhythm is a fundamental process that regulates the sleep–wake cycle. This rhythm is regulated by core clock genes that oscillate to create a physiological rhythm of circadian neuronal activity. However, we do not know much about the mechanism by which circadian inputs influence neurons involved in sleep–wake architecture. One possible mechanism involves the photoreceptor cryptochrome (CRY). In Drosophila, CRY is receptive to blue light and resets the circadian rhythm. CRY also influences membrane potential dynamics that regulate neural activity of circadian clock neurons in Drosophila, including the temporal structure in sequences of spikes, by interacting with subunits of the voltage-dependent potassium channel. Moreover, several core clock molecules interact with voltage-dependent/independent channels, channel-binding protein, and subunits of the electrogenic ion pump. These components cooperatively regulate mechanisms that translate circadian photoreception and the timing of clock genes into changes in membrane excitability, such as neural firing activity and polarization sensitivity. In clock neurons expressing CRY, these mechanisms also influence synaptic plasticity. In this review, we propose that membrane potential dynamics created by circadian photoreception and core clock molecules are critical for generating the set point of synaptic plasticity that depend on neural coding. In this way, membrane potential dynamics drive formation of baseline sleep architecture, light-driven arousal, and memory processing. We also discuss the machinery that coordinates membrane excitability in circadian networks found in Drosophila, and we compare this machinery to that found in mammalian systems. Based on this body of work, we propose future studies that can better delineate how neural codes impact molecular/cellular signaling and contribute to sleep, memory processing, and neurological disorders.

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Section: The Neural Network That Regulates the Circadian Clocksupporting

confidence: 52%

Light/Clock Influences Membrane Potential Dynamics to Regulate Sleep States

et al. 2021

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“…Brain cell dissociation was performed as described previously (Bell et al 2021;Kim et al 2021) using one female mouse per genotype. Mouse brain matrix (0.5 mm) was used to slice brains into coronal sections, and relative brain regions (cerebral cortex, striatum, midbrain) were dissected into Hibernate-A media with a 2% B-27 and GlutaMAX supplement (0.5 mM final).…”

Section: Cell Dissociation and Scrna-seqmentioning

confidence: 99%

Ptbp1 deletion does not induce glia-to-neuron conversion in adult mouse retina and brain

Hoang

Kim

Appel

et al. 2021

Preprint

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Somatic reprogramming of glia into neurons is a potentially promising approach for the replacement of neurons lost to injury or neurodegenerative disorders. Knockdown of the polypyrimidine tract-binding protein Ptbp1 has been recently reported to induce efficient conversion of retinal Mϋller glia and brain astrocytes into functional neurons. However, genetic analysis of Ptbp1 function in adult glia has not been conducted. Here, we use a combination of genetic lineage tracing, scRNA-Seq, and electrophysiological analysis to show that specific deletion of Ptbp1 in adult retinal Mϋller glia and brain astrocytes does not lead to any detectable level of glia-to-neuron conversion. Few changes in gene expression are observed in glia following Ptbp1 deletion, and glial identity is maintained. These findings highlight the importance of using genetic manipulation and lineage tracing methods in studying cell type conversion.

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A Clock-Driven Neural Network Critical for Arousal

Bell

Liu

Kim

et al. 2020

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Main The function of WAKE is conserved in mammalsWe previously identified the clock-output molecule WIDE AWAKE (WAKE) from a forward genetic screen in Drosophila 4 . WAKE modulates the activity of arousalpromoting clock neurons at night, in order to promote sleep onset and quality 4,5 . The mammalian proteome contains a single ortholog, mWAKE (also named ANKFN1/Nmf9), with 56% sequence similarity and which is enriched in the core region of the master circadian pacemaker suprachiasmatic nucleus (SCN) 4,6 ( Fig. 1a, Extended Data Fig. 1a). To investigate whether the function of WAKE is conserved in mice, we generated a putative null allele of mWAKE (mWAKE (-) ) by CRISPR/Cas9 insertion of 8 base pairs (containing a stop codon and generating a downstream frameshift) in exon 4, which is predicted to be in all splice isoforms of mWAKE ( Fig. 1b). As expected, mWAKE expression, as assessed by quantitative PCR and in situ hybridization (ISH), was markedly reduced in mWAKE (-/-) mice, likely due to nonsense-mediated decay (Fig. 1c, 1d). Given mWAKE expression in the SCN, we first examined locomotor circadian rhythms and found that mWAKE (-/-) mice exhibit a mild but non-significant decrease in circadian period length (Extended Data Fig. 1b, 1c). These results are similar to findings from fly wake mutants and mice bearing the Nmf9 mutation (a previously identified ENU-generated allele of mWAKE) 4,6 .Because we previously demonstrated that WAKE mediates circadian regulation of sleep timing and quality in fruit flies 4,5 , we next assessed sleep in mWAKE (-/-) mice via electroencephalography (EEG). Under light:dark (L:D) conditions, there was no difference in the amount of wakefulness, non-rapid eye movement (NREM), or REM sleep between mWAKE (-/-) mutants and wild-type (WT) littermate controls (Extended Data Fig. 1d). In constant darkness (D:D), there is a modest main effect of genotype on wakefulness (P<0.05) and NREM sleep (P<0.05), and a mild but significant decrease in REM sleep in mWAKE (-/-) mutants (Fig. 1e). Although the amount of wakefulness did not appreciably differ in mWAKE (-/-) mutants compared to controls, there was a change in the distribution of wakefulness at night; mutants spent more daily time in prolonged wake bouts, and some mutants exhibited dramatically long bouts of wakefulness (Extended Data Fig. 1e, 1f).

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Characterization ofmWakeexpression in the murine brain

Cited by 3 publications

References 105 publications

Light/Clock Influences Membrane Potential Dynamics to Regulate Sleep States

Light/Clock Influences Membrane Potential Dynamics to Regulate Sleep States

Ptbp1 deletion does not induce glia-to-neuron conversion in adult mouse retina and brain

A Clock-Driven Neural Network Critical for Arousal

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