TIM PER
TIM e PER
2 6 1 0 14 18 22 2 6 1 0 14 18 22
Zeitgeber Time (hr)
TIM
FIG.
7.
Nuclear localization ofPER
is impaired in rit. Horizontal section of fly headsstained with X-gal
(A-D
andE).
The spatial pattern of the per gene expression was monitored using the per-lacZ fusion gene. Horizontal head sections in wild-type and rit flies kept at 24OC (A
andB)
or at
30°C (C
andD)
were stained for 2 hrs at37°C.
The per expression were in retina (ret), optic lobes (lamina; la, medulla; me, and lobula (lo) and central brain (br) in wild-type(A
andC).
Nucleiin photoreceptor cells are clearly stained (black arrow heads) in wild-type but not in rit both at 24 and
30°C.
The stainings of lateral neurons (LNs) are indicated by black arrow. The expression levelin PER-BGAL
fusion protein in rit is lower than wild-type. Such a weakPER
localization in nucleiwas rescued by the excess of per+ gene dosage in +/w+Y; rit even at
30°C (E).
Scale bar represent80
j.lm.- p.
65-24°C 30°C
""""
ret
wild-
, � ,type
Ia brme lo
A c
rit
B D
+fw+y; rit
E
FIG. 8 PER localization in lateral neurons at 30°C. Horizontal sections of fly heads obtained at ZT 19, 21 and 23.5 were stained with anti-PER antibody coupled to FIT C (green color). Counter staining of nuclei was done with propidium iodide (red color) after RNase treatment When PER staining overlaps with nuclear staining, yellow color is observed. For symbols in this figure, see the legend ofFig.7. Scale bars in A and
D
represent 80 and 10 Jlm, respectively. In wild-type (A, Band C), signals of PER are observed comparable to the X-gal staining in Fig. 7C. The 8 X magnification of lateral neurons (LNs) were represented(D,
E, F). At ZT 19, PER signal is observed incytoplasm, while both at ZT 21 and 23.5 PER is in nuclei. In
rit (G,
H and I), the similar staining pattern to that in wild-type can be observed, although the strength of PER signal is weaker especially in the LNs(J,
K and L ). Thus, images when PER signals were amplified are illustrated(M,
N and 0). PER is in cytoplasm at ZT19(J
andM).
At ZT21, there are three patterns of PERstaining surrounding a nucleus (the center and right upper comer in N), overlapping in a nucleus of the center of a broad PER staining area (middle left inN) and just overlapping in nuclei (the inset in N). AT ZT23.5, PER enters in nuclei (0) else stays in cytoplasm (the inset in 0).
- p.
67-wild
type
rit
ZT19 ZT21 ZT23.5
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