Advanced analysis of a cryptochrome mutation's effects on the robustness and phase of molecular cycles in isolated peripheral tissues of Drosophila

Table 2 Effects of the cry^b mutation on BG-luc and tim-luc reporter activity in antennal specimens under light-dark cycles (LD 12:12), constant darkness (DD and constant temperature (HH)^1,2

Genotype³	N_a⁴	Number⁵ Rhythmic (statistical significance)	Period (Mean ± SEM)	Rhythmicity⁶ Index (Mean ± SEM)	Rhythmicity Statistic (Mean ± SEM)	Rhythm Amplitude (Mean ± SEM)	Phase	Activity Counts (MeanfSEM)
LD
BG-luc; cry ⁺	36	33 (23)	24.2 ± 0.3	0.49 ± 0.03	3.0 ± 0.2	0.17 ± 0.01	2.8, 0.9	335 ± 33
BG-luc; cry ^b	41	16 (3)	25.8 ± 1.0	0.12 ± 0.06	0.8 ± 0.4	0.09 ± 0.02	8.2, 0.9	299 ± 65
tim-luc; cry ⁺	20	16 (9)	25.0 ± 0.3	0.36 ± 0.05	2.3 ± 0.3	0.15 ± 0.02	2.1,0.9	309 ± 30
tim-luc; cry ^b	36	11 (1)	26.5 ± 1.0	0.23 ± 0.05	1.4 ± 0.3	0.09 ± 0.01	9.2, 1.0	477 ± 105
DD
BG-luc; cry ⁺	38	33(4)	25.7 ± 0.4	0.20 ± 0.03	1.2 ± 0.2	0.12 ± 0.01	5.5, 1.0	404 ± 27
BG-luc; cry ^b	31	14(1)	26.8 ± 1.3	0.03 ± 0.04	0.2 ± 0.2	0.02 ± 0.00	10.5,0.3	350 ± 75
tim-luc; cry ⁺	23	22(0)	26.4 ± 0.4	0.12 ± 0.03	0.7 ± 0.2	0.08 ± 0.00	5.4, 0.8	269 ± 40
tim-luc; cry ^b	37	8(0)	24.3 ± 2.2	-0.1 ± 0.09	-0.6 ± 0.5	0.04 ± 0.00	7.9, 0.5	625 ± 109
HH
BG-luc; cry ⁺	112	94(5)	25.4 ± 0.3	0.12 ± 0.01	0.7 ± 0.1	0.08 ± 0.00	-0.1,0.6	487 ± 29
BG-luc; cry ^b	107	28(2)	25.4 ± 0.5	0.14 ± 0.03	0.9 ± 0.2	0.06 ± 0.00	0.9, 0.6	624 ± 98
tim-luc; cry ⁺	112	74(0)	25.3 ± 0.3	0.14 ± 0.01	0.8 ± 0.1	0.09 ± 0.00	0.2, 0.7	902 ± 73
tim-luc; cry ^b	80	37(4)	25.8 ± 0.2	0.11 ± 0.02	0.7 ± 0.1	0.08 ± 0.00	-1.4,0.5	674 ± 90

1) The experimental conditions are described in Krishnan et al [23] and in Materials and Methods. 2) Ambient conditions are light-dark cycles with 12 hours of light and 12 hours of darkness (LD 12:12) or constant darkness at 25°C or constant temperature in which the specimens were first synchronized to temperature cycles in constant darkness consisting of 12 hours of 18°C followed by 12 hours of 27°C before they were evaluated under a constant high temperature of 27°C (HH). See Materials and Methods for further details. Also see Krishnan et al. [23]3) All headings in Table 2 are defined as in Table 1. 4) The analysis was applied to samples previously described in Krishnan et al [23]. The number of specimens analyzed (N_a) in LD and DD is unchanged, while the number of specimens in HH has increased by 123 additional cry⁺ antennal pairs and 62 additional cry^b specimens (see Materials and Methods for more details) 5) We asked whether cry^b affects rhythmicity in antennal pairs for each ambient condition. Chi-squared tests showed significant effects of the mutation on rhythmicity with either BG-luc or tim-luc in all cases (p < .001). 6) The cry^b mutation significantly reduced the RI value for each reporter in LD and DD (t-test, p < .001) but not HH (p > .05).

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