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The Regulatory Factor ZFHX3 Modifies Circadian Function in SCN via an AT Motif-Driven Axis.

Parsons MJ, Brancaccio M, Sethi S, Maywood ES, Satija R, Edwards JK, Jagannath A, Couch Y, Finelli MJ, Smyllie NJ, Esapa C, Butler R, Barnard AR, Chesham JE, Saito S, Joynson G, Wells S, Foster RG, Oliver PL, Simon MM, Mallon AM, Hastings MH, Nolan PM - Cell (2015)

Bottom Line: Using RNA sequencing, we found minimal effects on core clock genes in Zfhx3(Sci/+) SCN, whereas the expression of neuropeptides critical for SCN intercellular signaling was significantly disturbed.Lentiviral transduction of SCN slices showed that the ZFHX3-mediated activation of AT motifs is circadian, with decreased amplitude and robustness of these oscillations in Zfhx3(Sci/+) SCN slices.In conclusion, by cloning Zfhx3(Sci), we have uncovered a circadian transcriptional axis that determines the period and robustness of behavioral and SCN molecular rhythms.

View Article: PubMed Central - PubMed

Affiliation: MRC Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, UK.

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Circadian Gene Expression in Zfhx3Sci/+ and Zfhx3+/+ SCN across the Light:Dark Cycle, Related to Figure 1(A–F) mRNA expression for (A) Per1, (B) Per2, (C) Cry1, (D) Cry2, (E) Bmal1, and (F) Dbp1 showed no significant differences in the SCN of Zfhx3Sci/+ (gray lines) compared to Zfhx3+/+ (black lines) at multiple time points throughout the day (n = 4, p > 0.2, ANOVA).
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figs2: Circadian Gene Expression in Zfhx3Sci/+ and Zfhx3+/+ SCN across the Light:Dark Cycle, Related to Figure 1(A–F) mRNA expression for (A) Per1, (B) Per2, (C) Cry1, (D) Cry2, (E) Bmal1, and (F) Dbp1 showed no significant differences in the SCN of Zfhx3Sci/+ (gray lines) compared to Zfhx3+/+ (black lines) at multiple time points throughout the day (n = 4, p > 0.2, ANOVA).

Mentions: Using ex vivo organotypic slices from Zfhx3Sci/+ or Zfhx3+/+ animals on a PER2::LUC background, we found that Zfhx3Sci/+ SCN had a shorter circadian period and a decreased amplitude of fusion protein expression relative to wild-type (p < 0.05, t test) (Figures 1F–1H). Similar decreases were found in individual neurons from Zfhx3Sci/+ SCN slices compared to wild-type neurons (Figures S1A and S1B). Furthermore, the period distribution was broader, and RAE increased in individual Zfhx3Sci/+ neurons imaged across the SCN circuit (Figures S1C and S1D) (p < 0.05, t test). There were no period differences in organotypic lung slices (Figure S1E), suggesting a central oscillator specific effect. These ex vivo SCN findings mirror the differences seen in locomotor behavior and suggest that photic inputs are not necessary for the expression of the short-period phenotype. Conversely, there were no significant differences in mRNA expression patterns of core circadian genes in the SCN of Zfhx3Sci/+ and Zfhx3+/+ animals sampled at six time points across the light:dark cycle (Figure S2), suggesting that any differences in clock gene expression are masked by the light:dark cycle. Collectively, these data predict that the mutation disrupts a Zfhx3-dependent and canonical TTFL-independent effect on SCN circadian period.


The Regulatory Factor ZFHX3 Modifies Circadian Function in SCN via an AT Motif-Driven Axis.

Parsons MJ, Brancaccio M, Sethi S, Maywood ES, Satija R, Edwards JK, Jagannath A, Couch Y, Finelli MJ, Smyllie NJ, Esapa C, Butler R, Barnard AR, Chesham JE, Saito S, Joynson G, Wells S, Foster RG, Oliver PL, Simon MM, Mallon AM, Hastings MH, Nolan PM - Cell (2015)

Circadian Gene Expression in Zfhx3Sci/+ and Zfhx3+/+ SCN across the Light:Dark Cycle, Related to Figure 1(A–F) mRNA expression for (A) Per1, (B) Per2, (C) Cry1, (D) Cry2, (E) Bmal1, and (F) Dbp1 showed no significant differences in the SCN of Zfhx3Sci/+ (gray lines) compared to Zfhx3+/+ (black lines) at multiple time points throughout the day (n = 4, p > 0.2, ANOVA).
© Copyright Policy - CC BY
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4537516&req=5

figs2: Circadian Gene Expression in Zfhx3Sci/+ and Zfhx3+/+ SCN across the Light:Dark Cycle, Related to Figure 1(A–F) mRNA expression for (A) Per1, (B) Per2, (C) Cry1, (D) Cry2, (E) Bmal1, and (F) Dbp1 showed no significant differences in the SCN of Zfhx3Sci/+ (gray lines) compared to Zfhx3+/+ (black lines) at multiple time points throughout the day (n = 4, p > 0.2, ANOVA).
Mentions: Using ex vivo organotypic slices from Zfhx3Sci/+ or Zfhx3+/+ animals on a PER2::LUC background, we found that Zfhx3Sci/+ SCN had a shorter circadian period and a decreased amplitude of fusion protein expression relative to wild-type (p < 0.05, t test) (Figures 1F–1H). Similar decreases were found in individual neurons from Zfhx3Sci/+ SCN slices compared to wild-type neurons (Figures S1A and S1B). Furthermore, the period distribution was broader, and RAE increased in individual Zfhx3Sci/+ neurons imaged across the SCN circuit (Figures S1C and S1D) (p < 0.05, t test). There were no period differences in organotypic lung slices (Figure S1E), suggesting a central oscillator specific effect. These ex vivo SCN findings mirror the differences seen in locomotor behavior and suggest that photic inputs are not necessary for the expression of the short-period phenotype. Conversely, there were no significant differences in mRNA expression patterns of core circadian genes in the SCN of Zfhx3Sci/+ and Zfhx3+/+ animals sampled at six time points across the light:dark cycle (Figure S2), suggesting that any differences in clock gene expression are masked by the light:dark cycle. Collectively, these data predict that the mutation disrupts a Zfhx3-dependent and canonical TTFL-independent effect on SCN circadian period.

Bottom Line: Using RNA sequencing, we found minimal effects on core clock genes in Zfhx3(Sci/+) SCN, whereas the expression of neuropeptides critical for SCN intercellular signaling was significantly disturbed.Lentiviral transduction of SCN slices showed that the ZFHX3-mediated activation of AT motifs is circadian, with decreased amplitude and robustness of these oscillations in Zfhx3(Sci/+) SCN slices.In conclusion, by cloning Zfhx3(Sci), we have uncovered a circadian transcriptional axis that determines the period and robustness of behavioral and SCN molecular rhythms.

View Article: PubMed Central - PubMed

Affiliation: MRC Harwell, Harwell Science and Innovation Campus, Oxfordshire OX11 0RD, UK.

Show MeSH