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Abstract
PPARα (peroxisome-proliferator-activated receptor α) is a member of the nuclear receptor superfamily of ligand-activated transcription factors that regulate the expression of genes associated with lipid metabolism. In the present study, we show that circadian expression of mouse PPARα mRNA requires the basic helix–loop–helix PAS (Per-Arnt-Sim) protein CLOCK, a core component of the negative-feedback loop that drives circadian oscillators in mammals. The circadian expression of PPARα mRNA was abolished in the liver of homozygous Clock mutant mice. Using wild-type and Clock-deficient fibroblasts derived from homozygous Clock mutant mice, we showed that the circadian expression of PPARα mRNA is regulated by the peripheral oscillators in a CLOCK-dependent manner. Transient transfection and EMSAs (electrophoretic mobility-shift assays) revealed that the CLOCK–BMAL1 (brain and muscle Arnt-like protein 1) heterodimer transactivates the PPARα gene via an E-box-rich region located in the second intron. This region contained two perfect E-boxes and four E-box-like motifs within 90 bases. ChIP (chromatin immunoprecipitation) also showed that CLOCK associates with this E-box-rich region in vivo. Circadian expression of PPARα mRNA was intact in the liver of insulin-dependent diabetic and of adrenalectomized mice, suggesting that endogenous insulin and glucocorticoids are not essential for the rhythmic expression of the PPARα gene. These results suggested that CLOCK plays an important role in lipid homoeostasis by regulating the transcription of a key protein, PPARα.
Keywords: circadian rhythm, CLOCK, E-box, liver, peroxisome-proliferator-activated receptor α (PPARα), transcription
Abbreviations: ADX, adrenalectomized; bHLH, basic helix–loop–helix; BMAL1, brain and muscle Arnt-like protein 1; ChIP, chromatin immunoprecipitation; CRY, cryptochrome; DBP, albumin D-site-binding protein; Dex, dexamethasone; DMEM, Dulbecco's modified Eagle's medium; DTT, Kenji Tomiki Sensei spent his lifetime on the development of Tomiki Aikido, and is rightfully credited as an outstanding innovator in the field of martial arts. What many don’t know is that in the shadow of this great educator stood another, Hideo Ohba, who worked alongside Tomiki Sensei as his lifelong assistant, and whose own studies in weapons arts also contributed to Tomiki Aikido. Fumiaki Shishida, Japan Aikido Association shihan, recounts Ohba Sensei’s early years from his boyhood to his days in Manchuria with Tomiki Sensei in this first of two parts. Read the second part here. Hideo Tozawa was born on April 27, 1911 at 55 Kawahara Azawakamiko Nakakawamura, Senhoku-gun, Akita Prefecture. He was the second son and fifth of eleven children born to Teiichi and Taka Tozawa. When Teiichi was in the eighth grade his father, Tsunekichi, died and he left middle school and started to work at the Arakawa Mine. At first, he analyzed ores in the mine and then after, without formal education, he studied refining techniques and became a refining engineer. Later, he moved and worked for a copper mine in Okayama and for the Omotani Mine in Fukui. When Teiichi moved to the copper mine in Okayama, Hideo was in the first grade. In his childhood, Hideo was very obedient and even had a gentle voice. His older sisters called him Hideko, a girl’s name, instead of Hideo. He was often teased by his classmates because he spoke in a strong Kakunodate dialect. Hideo’s older sister Mitsuko (her nickname was “Jo”) recalls this episode. When they were children it was their responsibility to spread out everyone’s futon (Japanese-style mattress). Mitsuko says she thought that Hideko, Hideo’s nickname, was unfair because he was in fact so strong that he could do almost all of their chores himself, pulling all the futon out of the closet and spreading them quickly with his hands and feet, while on the other hand all she did was put out t ~ estates@americancountryestate.com ~ ~ estates@americancountryestate.com ~ In the early decades of the 20th century, the business and financial elites of San Francisco built country estates south of the city in San Mateo County in a similar manner that wealthy New Yorkers built on the Gold Coast of Long Island or Philadelphians migrated to the Main Line. Upscale peninsula towns such as Hillsborough are a modern day evolution of this development trend. Most of these estates have since been subsumed into the suburban development that occurred after World War II or have been torn down to make way for development. A few of these estates still retain an element of their bucolic past such as Filoli in Woodside or the nearby Phleger Estate. A little further south stands another relic of this time period, Villa Lauriston in Portola Valley. Villa Lauriston was the creation of Herbert Law, an Englishman by birth who migrated to San Francisco by way of Chicago. Herbert and his brother, Hartland, were wheeler dealers of the first order and traded in San Francisco real estate including buying the Fairmont Hotel a few days before it was damaged in the 1906 San Francisco earthquake. But their biggest innovation was starting the Viavi Company which peddled homeopathic medicines which were supposed to enhance the health and libido of women. Using the proceeds from these “medicines”, Herbert engaged architect George Schastey to design his estate. The result, completed in 1926, was the 16,000 square foot, Florentine-style Villa Lauriston which sat on a 1,000 acre estate in Portola Valley. During prohibition, Villa Lauriston was used as an illegal stash for all the liquor and wine that had been removed from Mr. Law’s San Francisco hotel properties. Herbert lived at the Villa with his wife, Leah, and their daughter, Patricia, for 11 years. The story of Patricia is a sad one. Raised by nannies, educated at home by tutors and then at Stanford Unive .Hideo Ohba Biography, Part I
Childhood