mCry1 and Bmal1 mRNA, and mCRY1 protein expression
profiles of wild-type (solid line), young mPer1-/- mCry2-/- (dotted line), and old
mPer1-/- mCry2-/- (dashed line) mice.
(A) Diurnal expression of mCry1 in the SCN in LD. The black
and white bars on the X-axis indicate dark and light phases,
respectively. (Right panels) Representative micrographs of SCN probed
with the mCry1 antisense probe at time points of minimal(ZT0) and
maximal(ZT12) expression. Bar, 200 μm. (B) Circadian expression of
mCry1 in the SCN on the fourth day in DD. The gray and black bars on
the X-axis indicate subjective day and night, respectively.
(C) Diurnal variation of mCRY1 immunoreactivity in the SCN in LD.
Quantification was performed by counting immunoreactive nuclei in the area of
the SCN. In old double mutants, oscillation of mCRY1 immunoreactivity is
significantly dampened (p < 0.05), with constantly high numbers of
immunoreactive nuclei throughout the LD cycle. (Right panels)
Representative micrographs of immunostained SCN at time points of minimal(ZT0)
and maximal(ZT12) immunoreactivity. Bar, 100 μm. (D) Diurnal
expression of mCRY1 protein in the SCN of wild-type (solid line),
mPer1-/- (dotted line), and mCry2-/- (hatched line) mice. (E) Diurnal variation of Bmal1 mRNA
expression in the SCN in LD. In old double mutants, Bmal1 cycling is
significantly dampened (p < 0.05). (F) Circadian
expression of Bmal1 mRNA in the SCN on the fourth day in DD.
(G) Diurnal time course of Avp mRNA expression in the SCN in
LD. (H) Diurnal time course of Dbp mRNA expression in the
SCN in LD. Tissue was visualized by Hoechst dye nuclear staining (blue);
silver grains are artificially colored (red) for clarification. All data
presented are mean ± S.D. for three different experiments.