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. 2017 Jun 15;6:e22861. doi: 10.7554/eLife.22861

Figure 2. Developmental ablation of ipRGCs results in a lengthened circadian period.

(A) Representative actograms of wild-type, Opn4LacZ/LacZ, Opn4DTA/+, Opn4DTA/LacZ, Opn4DTA/DTA and Opn4aDTA/aDTA mice under 12:12-LD cycle and constant darkness (DD). White background indicates light and grey background indicates darkness. (B) The circadian period length of all tested genotypes (wild-type: n = 18, Opn4LacZ/LacZ: n = 17, Opn4DTA/+: n = 8, Opn4DTA/LacZ: n = 7, Opn4DTA/DTA: n = 7, and Opn4aDTA/aDTA: n = 18). (C) For phase-shifting experiments, a subset of wild-type, Opn4LacZ/LacZ, Opn4aDTA/aDTA were assessed, and the total mice analyzed was as follows: wild-type (n = 7), Opn4LacZ/LacZ (n = 8), Opn4DTA/+ (n = 8), Opn4DTA/LacZ (n = 7), Opn4DTA/DTA (n = 7), and Opn4aDTA/aDTA (n = 8). As expected from substantial ipRGC ablation, phase shifting was significantly reduced in Opn4DTA/LacZ, Opn4DTA/DTA and Opn4aDTA/aDTA mice. (D) Representative actograms of wild-type mice enucleated at either P0 (n = 8) or P60 (n = 7), under 12:12-LD and DD. (E) The circadian period length for both enucleation groups. Only mice enucleated at P0 exhibited a lengthened circadian period (B and C) One-way ANOVA with Bonferroni's multiple comparisons test and adjusted p values. (E) Two-way ANOVA, Bonferroni's multiple comparisons test and adjusted p values. Error bars represent s.e.m. for all graphs. See also Figure 2—figure supplements 1–2.

DOI: http://dx.doi.org/10.7554/eLife.22861.005

Figure 2.

Figure 2—figure supplement 1. Additional actograms for Opn4DTA/LacZ and Opn4DTA/DTA mice.

Figure 2—figure supplement 1.

(A) Actograms of all tested Opn4DTA/LacZ. (B) Actograms of all tested Opn4DTA/DTA mice.
DOI: http://dx.doi.org/10.7554/eLife.22861.006
Figure 2—figure supplement 2. Expression pattern of transcription factors critical for SCN development.

Figure 2—figure supplement 2.

In situ hybridization for several developmentally important transcription factors, Rora, Six3, and Lhx1 as well as the functionally important neuropeptide vasopressin (AVP) in control (Opn4DTA/+) and Opn4DTA/DTA mice to test whether the lengthened circadian period exhibited by Opn4DTA/DTA (but not Opn4DTA/+) mice could be attributed to gross changes in the SCN. Each row represents one animal. The expression patterns of these genes were similar between these two groups of mice.
DOI: http://dx.doi.org/10.7554/eLife.22861.007