2000 Fiscal Year Annual Research Report

老化に関る遺伝子・klothoの分子機能の研究

Research Project

Project/Area Number	10308032
Research Institution	KYOTO UNIVERSITY
Principal Investigator	鍋島陽一京都大学, 医学研究科, 教授 (60108024)
Co-Investigator(Kenkyū-buntansha)	吉田松生京都大学, 医学研究科, 助手 (60294138) 藤森俊彦京都大学, 医学研究科, 助手 (80301274) 星野幹雄京都大学, 医学研究科, 助手 (70301273)
Keywords	個体老化 / Klotho遺伝子 / β-glucosidase / ホメオスタシス / 挿入突然変異
Research Abstract	Klothoマウスと原因遺伝子:klothoの解析を行い、以下の結果をえた。 (1)Klotho蛋白には立体構造、結合糖鎖、翻訳後プロセッシングの違いによる4つのタイプがあり、それらはER,Golgiに存在する細胞内タイプ、細胞膜タイプ、細胞外ドメインの中央でプロセスされる分泌型short Form(mKL1よりなる)、膜貫通ドメインでプロセスされる全長に近い分泌型Long Form(mKL1,mKL2を含む)からなる。これらは結合糖鎖や立体構造が異なっており、それぞれを優先的に認識するモノクローナル抗体を作成した。(2)分泌型を高感度に認識する抗体を用いて、Klotho蛋白が血清中に存在することを証明した。(3)Klotho蛋白は活性中心の構造に変異があるとはいえ、構造的にはβ-Glucosidaseに相同性があることから、酵素活性の有無を追求してきたが、糖鎖切断活性を強く支持する結果が得られた。Klotho蛋白は少なくもターゲット分子の糖鎖を修飾対象、あるいは認識対象の一部としていると推定される。(4)Klotho遺伝子座にlacZ遺伝子をノックインしたマウスの解析により、Klotho遺伝子は腎尿細管、脳の脈絡膜、副甲状腺ホルモンを産生する副甲状腺の主細胞で強く発現することを見いだした。(5)Klothoを発現する尿細管細胞ではカルシウムホメオスタシスに関連する重要な遺伝子がそろって発現しており、更に、Klotho変異マウスではカルシウム制御の鍵となる遺伝子の発現変異が観察された。これらの結果はKlotho発現細胞、及び、Klotho蛋白のカルシウムホメオスタシス制御における重要性を強く示唆している。(6)蛋白の全体構造、β-Glucosidase活性中心の構造比較から、klotho遺伝子のホモログ、β-klotho遺伝子を同定した。β-klotho遺伝子は膵臓、肝臓、白色、褐色脂肪細胞で比較的高い発現が確認されている。また、ノックアウトマウスの作成、モノクローナル抗体を作成した。

Research Products

(16 results)

All Other

All Publications (16 results)

[Publications] Yamashita T.,Yoshitake H.,Tsuji K.,Kawaguchi N.,Nabeshima Y.,Noda M.: "Retardation in bone resorption after bone marrow ablation in Klotho mutant mice."Endocrinology. 141. 438-445 (2000)
[Publications] Kato Y.,Arakawa E.,Kinoshita S.,Shirai A.,Furuya A.,Yamano K.,Nabeshima Y. et al: "Establishment of the anti-Klotho monoclonal antibodies and detection of Klotho protein in kidneys."Biochem.Biophys.Res.Comm.. 267. 597-602 (2000)
[Publications] Suga T.,Kurabayashi M.,Sando Y.,Ohyama Y.,Maeno T.,Maeno Y.,Aizawa H.,Matsumura Y.,Kuwaki T.,Kuro-o M.,Nabeshima Y.,Nagai R.: "Disruption of the klotho gene causes pulmonary emphysema in mice defect in maintenance of pulmonary integrity during postnatal life."Am.J.Respir.,Cell Mol,Biol.. 22. 26-33 (2000)
[Publications] Yamashita T.,Nabeshima Y.,Noda M.: "High-resolution μCT (micro-computed tomography) analyses of the abnormal trabecular structures in Klotho gene mutant mice."J.Endocrinology. 164(2):. 239-245 (2000)
[Publications] Seiji Okada,Toru Yoshida,Zhang Hong,Genichiro Ishii,Masahiko Hatano,Makoto Kuro-o,Yoko Nabeshima,Yoichi Nabeshima,and Takeshi Tokuhisa: "Impairment of B lymphopoiesis in precocious aging (klotho) mice."Int.Immunol.. 12. 861-971 (2000)
[Publications] Shiraki-iida T.,Iida A.,Nabeshima Y.,Anazawa H.,Nishikawa S.,Noda M.,Kuro-o M.,Nabeshima Y.: "Improvement of multiple pathophysiological phenotypes of klotho (kl/kl) mice by adenovirus-mediated expression of the klotho gene. J.Gene Medicine"J.Gene Medicine. 2. 233-242 (2000)
[Publications] Utsugi T,Ohno T,Ohyama Y,Uchiyama T,Saito Y,Matsumura Y,Aizawa H,Itoh H,Kurabayashi M,Kawazu S,Tomono S,Oka Y,Suga T,Kuro-o M,Nabeshima Y,Nagai R.: "Decreased insulin production and increased insulin sensitivity in the klotho mutant mouse, a novel animal model for human ageing."Metabolism. 49. 1118-1123 (2000)
[Publications] Sone M.Suzuki E.,Hoshino M.,Hou D.,Kuromi H.,Fukata M.,Kuroda S.,Kaibuchi K.,Nabeshima Y.,Hama C.: "Synaptic development is controlled in the peri-active zones of Drosophila synapses."Development. 127. 4157-4168 (2000)
[Publications] Nakimoto K.,Kuro-o M.,Yanaka N.,Omori K.,Komuro I.,Imai Y.,Nabeshima Y.: "Isolation and characterization of Na+/Ca++ exchanger gene and splicing isoforms in mice"DNA Sequence. 11. 75-81 (2000)
[Publications] Yoshida N.,Yoshida S.,Araie M.,Hannda H.,Nabeshima Y: "Ets family transcription factors ESE-1 is expressed in corneal epithelial cells and is involved in their differentiation."Mechan.Dev.. 97. 27-34 (2000)
[Publications] Ito S.,Kinoshita S.,Shiraishi N.,Nakagawa S.,Sekine S.,Fujimori T.,Nabeshima Y.: "Molecular cloning and expression anlysis of a novel gene βklotho, which encodes a novel Klotho family protein."Mechan.Dev.. 98. 115-119 (2000)
[Publications] Wakimoto K.,Kobayashi K.,Kuro-o M.,Yanaka N.,Yao A.,Kita S.,Iwamoto T.,Azuma S.,Toyota H.,Omori K.,Shigekawa M.,Imahie H.,Imai Y.,Nabeshima Y.,Komurol.: "Targeted disruptioin of the mice Na+/Ca2+ exchanger gene leads to the defects in the heart beats and blood circulation in embryo."J.Biol.Chem. 275. 36991-36998 (2000)
[Publications] Takebayashi H.,Yoshida S.,Kominami R.,Nakafuku M.,Nabeshima Y.: "Olig family of basic helix-loop-helix factors includes olig 1 and Olig 2 implicated in oligodendrogenesis and the third paralogous gene, Olog3."Mechan.Dev.. 99. 143-148 (2000)
[Publications] Koh N.,Fujimori T.,Tamori A.,Nishiguchi S.,Shiomi S.,Nakatani T.,Sugimura K.,Kishimoto T.,Kuroki T.,Nabeshima Y.: "Severely reduced expression of Klotho gene in human chronic renal failure kidney."Biochem.Biophys.Res.Comm.. 280. 1015-1020 (2001)
[Publications] Mori K.,Yahata K.,Mukoyama M.,Suganami T.,Makino H.,Nagae T.,Masuzaki H.,Ogawa Y.,Sugawara A.,Nabeshima Y.,Nakao K.: "Disruption of Klotho gene causes an abnormal energy homeostasis in mice."Biochem.Biophys.Res.Comm. (in press). (2001)
[Publications] Yamashita T.,Sekiya I.,Kawaguchi N.,Nabeshima Y.,Noda M.: "klotho-deficient mice are resistant to bone loss induced by unloading due to sciatic neurectomy."J.Endcrinology. (in press). (2001)

2000 Fiscal Year Annual Research Report

老化に関る遺伝子・klothoの分子機能の研究

Principal Investigator

鍋島 陽一 京都大学, 医学研究科, 教授 (60108024)

Research Products

[Publications] Yamashita T.,Yoshitake H.,Tsuji K.,Kawaguchi N.,Nabeshima Y.,Noda M.: "Retardation in bone resorption after bone marrow ablation in Klotho mutant mice."Endocrinology. 141. 438-445 (2000)

[Publications] Kato Y.,Arakawa E.,Kinoshita S.,Shirai A.,Furuya A.,Yamano K.,Nabeshima Y. et al: "Establishment of the anti-Klotho monoclonal antibodies and detection of Klotho protein in kidneys."Biochem.Biophys.Res.Comm.. 267. 597-602 (2000)

[Publications] Yamashita T.,Nabeshima Y.,Noda M.: "High-resolution μCT (micro-computed tomography) analyses of the abnormal trabecular structures in Klotho gene mutant mice."J.Endocrinology. 164(2):. 239-245 (2000)

[Publications] Seiji Okada,Toru Yoshida,Zhang Hong,Genichiro Ishii,Masahiko Hatano,Makoto Kuro-o,Yoko Nabeshima,Yoichi Nabeshima,and Takeshi Tokuhisa: "Impairment of B lymphopoiesis in precocious aging (klotho) mice."Int.Immunol.. 12. 861-971 (2000)

[Publications] Shiraki-iida T.,Iida A.,Nabeshima Y.,Anazawa H.,Nishikawa S.,Noda M.,Kuro-o M.,Nabeshima Y.: "Improvement of multiple pathophysiological phenotypes of klotho (kl/kl) mice by adenovirus-mediated expression of the klotho gene. J.Gene Medicine"J.Gene Medicine. 2. 233-242 (2000)

[Publications] Sone M.Suzuki E.,Hoshino M.,Hou D.,Kuromi H.,Fukata M.,Kuroda S.,Kaibuchi K.,Nabeshima Y.,Hama C.: "Synaptic development is controlled in the peri-active zones of Drosophila synapses."Development. 127. 4157-4168 (2000)

[Publications] Nakimoto K.,Kuro-o M.,Yanaka N.,Omori K.,Komuro I.,Imai Y.,Nabeshima Y.: "Isolation and characterization of Na+/Ca++ exchanger gene and splicing isoforms in mice"DNA Sequence. 11. 75-81 (2000)

[Publications] Yoshida N.,Yoshida S.,Araie M.,Hannda H.,Nabeshima Y: "Ets family transcription factors ESE-1 is expressed in corneal epithelial cells and is involved in their differentiation."Mechan.Dev.. 97. 27-34 (2000)

[Publications] Ito S.,Kinoshita S.,Shiraishi N.,Nakagawa S.,Sekine S.,Fujimori T.,Nabeshima Y.: "Molecular cloning and expression anlysis of a novel gene βklotho, which encodes a novel Klotho family protein."Mechan.Dev.. 98. 115-119 (2000)

[Publications] Takebayashi H.,Yoshida S.,Kominami R.,Nakafuku M.,Nabeshima Y.: "Olig family of basic helix-loop-helix factors includes olig 1 and Olig 2 implicated in oligodendrogenesis and the third paralogous gene, Olog3."Mechan.Dev.. 99. 143-148 (2000)

[Publications] Koh N.,Fujimori T.,Tamori A.,Nishiguchi S.,Shiomi S.,Nakatani T.,Sugimura K.,Kishimoto T.,Kuroki T.,Nabeshima Y.: "Severely reduced expression of Klotho gene in human chronic renal failure kidney."Biochem.Biophys.Res.Comm.. 280. 1015-1020 (2001)

[Publications] Mori K.,Yahata K.,Mukoyama M.,Suganami T.,Makino H.,Nagae T.,Masuzaki H.,Ogawa Y.,Sugawara A.,Nabeshima Y.,Nakao K.: "Disruption of Klotho gene causes an abnormal energy homeostasis in mice."Biochem.Biophys.Res.Comm. (in press). (2001)

[Publications] Yamashita T.,Sekiya I.,Kawaguchi N.,Nabeshima Y.,Noda M.: "klotho-deficient mice are resistant to bone loss induced by unloading due to sciatic neurectomy."J.Endcrinology. (in press). (2001)

鍋島陽一京都大学, 医学研究科, 教授 (60108024)