Functional analysis suggested that variation, in circadian genes including BMAL1
, CRY2, CSNK1E, NPAS2, PER3, REV1, and downstream transcription factors KLF10 and SENP3 through disruption of hormonal pathways or changes in light/dark schedules, is associated with ovarian cancer.
The passage 5 BMSCs were seeded in 6-well plastic plates at an initial density of 5 x [10.sup.5] cells per well in basic media, and the Bmal1
shRNA lentivirus (Arntl shRNA, synthesized by Hanheng bio, Shanghai, China) (group 2, Bmal1
repression group, Lenti-Bmal1 shRNA) or the GPF-expressing lentivirus (pHBLV-U6-ZsGreen-Puro) (group 3, Lenti-GFP-Puro) was added per well at a multiplicity of infection (MOI) of 20.
In contrast with the complex phenotypic effects of the germline deletion of Bmal1
, the phenotypic effects resulting from the genetic disruption of other core clock genes are less severe.
The researchers, who worked for more than three years on this series of studies, did not see any negative consequences of increasing Bmal1
in the muscle.
Furthermore, eliminating BMAL1
from their muscles caused a severe disruption in normal sleep patterns, leading to a heightened need for sleep, deeper sleep and a diminished ability to recover.
The activation of melanopsin receptors by the light induces proteins like CLOCK, BMAL1
, REV-ERB and PERIOD1.
Real-time PCR was conducted on the 7500 real-time PCR system (Applied Biosystems, Foster City, CA, USA) with the following primers: 18S (forward: 5'-TCAACACGGGAAACCTCAC-3', reverse: 5'-CGCTCCACCAACTAAGAAC-3'), Bmal1
(forward: 5'-CTTGTCTGTAAAACTTGCCTGTGAC-3', reverse: 5'-GTAGATCAGAGGGCGACGGCTA-3'), Clock (forward: 5'-GTAGGTTTCCAGTCCTGTCG-3', reverse: 5'-TGGGGTCTATGCTTCCTGGT-3'); Per2 , (forward: 5'-AGGATCCAAGAACGGCACAG-3', reverse: 5'-CGGACCTGGCTTCAGTTCAT-3'); Rora (forward: 5'-CCAAACTTGACAGCATCTCGA-3', reverse: 5'-GAAGGCTGCAAGGGCTTTTTCAGGA-3').
Fang et al., "Bmal1
and fi-cell clock are required for adaptation to circadian disruption, and their loss of function leads to oxidative stress-induced fi-cell failure in mice," Molecular and Cellular Biology, vol.
All PER and BMAL1
genes were also affected by glucocorticoids in human peripheral blood mononuclear cells .
When the researchers repeated the experiment in mice lacking Bmal1
, they found high levels of virus replication regardless of the time of infection.
A capacidade reprodutiva de camundongos, cuja expressao da proteina BMAL1
e ausente em todos os tecidos, e que sao conhecidos como camundongos BMAL1
knockouts (KO), foi analisada e ambos os sexos demonstraram serem estereis.