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Article

Article
保住, 功
Published: 新潟医学会雑誌 = 新潟医学会雑誌.  101  pp.433-442,  1987-07.  新潟医学会
Full Text Link: http://hdl.handle.net/10191/36532
Abstract: Fabry's disease is an X-linked disorder of glycosphingolipid catabolism. It is uncommon for female heterozygotes to show major clinical manifestations seen in hemizygotes. We have experienced a heterozygotic patient with cardiomyopathy and severe pain in the extremities. To elucidate the difference in clinical expressions between the symptomatic heterozygote and hemizygotes, we examined the spinal and sympathetic ganglia, heart, liver and kidney histochemically and biochemically and observed the accumulation of glycolipids in these organs of the heterozygote, which was similar to the hemizygote. Quantative analyses revealed marked accumulation of ceramide trihexoside (CTH) in the heart of the heterozygote, which was in contrast to predominant accumulation of CTH and ceramide dihexoside (CDH) in the kidney of the hemizygote. Digalactosyl ceramide was shown to be predominantly accumulated in CDH fraction obtained from sympathetic ganglia of the heterozygote. The α-galactosidase activity of the heterozygote was decreased to 16-40% of normal levels in various organs, especially in the heart. Two other heterozygotic patients were identified by low α-galactosidase activity and the cardiac involvement was demonstrated, although they did not have clinically evident cardiomyopathy. The difference in phenotypic expressions between sex can not be explained and further genetic study is necessary to explain it. Read more
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Article

Article
Takano, Hiroko ; Niimura, Sueo
Published: 新潟大学農学部研究報告 = 新潟大学農学部研究報告.  61  pp.27-33,  2008-09.  新潟大学農学部
Full Text Link: http://hdl.handle.net/10191/13064
Abstract: We previously reported in porcine oocytes that the activities of some hydroxysteroid dehydrogenases (HSDs) and the size of lipid droplets in the cytoplasm decreased as nuclear maturation progressed, and that the progression of nuclear maturation, the decrease of HSD activities and the reduction in the size of lipid droplets did not occur in those treated with olomoucine. From these results, we suggested that the changes in the steroid metabolism and the size of lipid droplets in the cytoplasm were closely associated with nuclear maturation. However, whether olomoucine directly acts on the cytoplasm to inhibit such changes could not be determined. In the present investigation, the activities of some HSDs and Sudanophilic lipid droplets were histochemically demonstrated in porcine oocytes in which the nuclear maturation was suppressed and the high cAMP level in the cytoplasm was maintained by the treatment of IBMX or dbcAMP, in order to clarify the relationship between nuclear maturation and the metabolism of steroids and the number of lipid droplets in the cytoplasm. Of the oocytes cultured with IBMX or dbcAMP for 22 hrs, 97 and 100% were in the germinal vesicle (GV) stage, respectively. The percentages of oocytes in the GV stage were significantly higher in both the treated oocytes than in control oocytes. The rates of the treated oocytes showing the activities of Δ^<5>-3β-HSD (using pregnenolone and 17α-hydroxypregnenolone as the substrates), 17β-HSD (estradiol-17β), 20α-HSD (20α-hydroxyprogesterone) and 20β-HSD (17β-hydroxyprogesterone) did not differ from those of control oocytes. Also, there were no differences in the number of lipid droplets of different sizes between IBMX- or dbcAMP-treated oocytes and control oocytes. From these findings, it was suggested that the changes in the steroid metabolism and the size of lipid droplets in the cytoplasm with oocyte maturation depend on the cAMP level in their cytoplasm rather than the progression of nuclear maturation.<br />我々は、ブタ卵母細胞において、核の成熟に伴って細胞質ではいくつかのステロイドの代謝低下と脂質小滴の小型化が起こることを以前報告した。また、核の成熟をオロモウシンで処置して阻止した卵母細胞では、ステロイド代謝の低下および脂質小滴の小型化はみられなかったことから、細胞質でのこれらの変化は核の成熟と密接に関係していることを推察して報告した。しかし、オロモウシンがステロイド代謝の低下および脂質小滴の小型化に直接関係しているのか否かは明らかにできなかった。そこで、オロモウシンとは異なった機構、すなわち細胞質のcAMPレベルを高く維持することによって核の成熟を抑制する作用のあるIBMXとdbcAMPを用いて、ブタの卵母細胞を処置し、ステロイド代謝と脂質小滴の数を組織化学的に観察した。IBMXあるいはdbcAMPで22時間処置した卵母細胞において、核は97および100%で卵核胞期にあり、これらの処置により成熟分裂の再開が抑制されていることが確認された。一方、IBMX あるいはdbcAMPで22時間処置した卵母細胞において、Δ^<5>-3β-HSD活性(基質としてpregnenoloneと17α-hydroxypregnenolone を使用)、17β-HSD 活性(estradiol-17β)、20α-HSD 活性(20α-hydroxyprogesterone)および20β-HSD活性(17β-hydroxyprogesterone)を有するものの割合、ならびに各種大きさの脂質小滴の数は、どちらも対照の卵母細胞におけるそれらと相違なかった。以上の結果から、細胞質におけるステロイド代謝の低下および脂質小滴の小型化は、核の成熟の進行よりも、細胞質のcAMP の量に依存して起こることが推察された。 Read more
3.

Article

Article
新村, 末雄 ; 大樌, 香苗
Published: 新潟大学農学部研究報告 = 新潟大学農学部研究報告.  58  pp.39-43,  2005-08.  新潟大学農学部
Full Text Link: http://hdl.handle.net/10191/657
Abstract: 体外で成熟過程にあるブタの卵母細胞、1細胞期から拡張胚盤胞期までの体外受精に由来するブタの卵子と初期胚について、グルコース-6-リン酸脱水素酵素(G-6-PDH)の活性を組織化学的に検出した。G-6-PDH活性は、胞状卵胞から採取直後の卵母 細胞では強く、この強い活性は培養後44時間の卵母細胞まで維持された。また、この酵素の活性は、媒精後の受精卵子でも強かったが、2細胞期から16細胞期の胚ではやや弱まり、弱度ないし強度となった。桑実胚期以降、酵素活性はさらに弱まり、活性を示さない胚も出現するとともに、拡張胚盤胞では活性はまったくみられなかった。以上の結果と従来のステロイド代謝能の結果とを考え合わせると、成熟過程にあるブタの卵母細胞と発生過程にあるブタの初期胚は、G-6-PDHの作用によって産生したNADPHをステロイドの生合成のために利用していることが推察された。The activity of gucose-6-phosphate dehydrogenase (G-6-PDH) in porcine oocytes and embryos was histochemically examined by the Rudolph and Klein method. Strong activity of G-6-PDH was observed in oocytes cultured for maturation and in fertilized oocytes, and blue diformazan granules produced by the enzyme reaction were spread evenly throughout the cytoplasm. In embryos, the activity somewhat decreased at the 2-cell stage, and the such activity was maintained up to the 16-cell stage. The activity weakened in embryos at the morula and early blastocyst stages, and some of them showed no enzyme activity. The enzyme activity completely disappeared from expanded blastocysts. In cleaved embryos, diformazan granules were distributed throughout the cytoplasm of blastomeres, while the amount of the granules differed among blastomeres. In early blastocysts, the granules were distributed in the cytoplasm of inner-cell-mass cells, but not in the cytoplasm of trophoblasts. The results obtained from this investigation and former studies concerning steroid metabolism seem to suggest that porcine oocytes and preimplantation embryos utilize NADPH produced by G-6-PDH for biosynthesis of steroids. Read more
4.

Article

Article
新村, 末雄 ; 斎藤, 千智
Published: 新潟大学農学部研究報告 = 新潟大学農学部研究報告.  56  pp.17-24,  2003-08.  新潟大学農学部
Full Text Link: http://hdl.handle.net/10191/25126
Abstract: cAMPの分解酵素であるホスホジエステラーゼの活性を阻害する作用を有する3-イソブチル-1-メチルキサンチン(IBMX)で処置したマウス卵母細胞について, 核の成熟状態を観察するとともに, 20α-ヒドロキシステロイド脱水素酵素(20α-H SD)の活性を調べ, 卵母細胞の核の成熟と細胞質での20α-ヒドロキシプロゲステロン代謝との関係の有無を検討した。成熟分裂を再開した卵母細胞の割合は, 50μMの濃度のIBMXで処置したものでは3.0%であり, IBMX処置していない対照の卵母細胞の100%(こ比べ, 有意に低かった。一方, 20α-HSD活性は, IBMX処置した卵母細胞と対照の無処置卵母細胞のすべてに認められ, 両者の間で活性に相違はみられなかった。以上の結果から, マウス卵母細胞の核の成熟と細胞質での20α-ヒドロキシプロゲステロン代謝との間には関係のないことが確認された。<br />The state of nuclear maturation and the activity of 20α-hydroxysteroid dehydrogenase (20α-HSD) were observed in mouse oocytes treated with 3-isobutyl-1-methylxanthine (IBMX), an inhibitor of 3', 5'-cyclic nucleotide phosphodiesterase, and the relationship between nuclear maturation and 20α-hydroxyprogesterone metabolism in the cytoplasm was examined. Nuclei of the oocytes cultured with 50μM IBMX for 8 hrs were almost in the germinal vesicle stage (97%), while those of control oocytes cultured without IBMX were in the metaphase I to telophase I stages, mostly metaphase I stage (68%). The percentage of oocytes in the germinal vesicle stage was significantly higher in the IBMX-treated group than in control group. On the other hand, the activity of 20α-HSDwas demonstrated in all oocytes of both IBMX-treated and non-treated groups. From the present findings, it was confirmed in mouse oocytes that the 20α-hydroxyprogesterone metabolism in the cytoplasm is not related to nuclear maturation. Read more
5.

Article

Article
Niimura, Sueo ; Kawakami, Shin-ya
Published: 新潟大学農学部研究報告 = 新潟大学農学部研究報告.  55  pp.91-97,  2003-03.  新潟大学農学部
Full Text Link: http://hdl.handle.net/10191/25112
Abstract: The activities of △^5-3β-hydroxysteroid dehydrogenase (△^5-3β-HSD) (pregnenolone and 17α-hydroxypregnenolone as the subs trates), 20α-HSD (20α-hydroxyprogesterone as the substrate) and 20β-HSD (17α-hydroxyprogesterone and 20β-hydroxyprogesterone as the substrates) were histochemically demonstrated in mouse eggs and embryos, and the progestagen production in those during the early development was examined. In unfertilized mouse eggs, the activities of such HSDs were observed in 88 to 98% of those. The percentages of embryos showing the activities of such HSDs did not change during the early development. It was also histochemically confirmed that the activity of △^5-3β HSD with DHA as the substrate, which is involved in steroid biosynthesis, was observed in 86 to 100% of the eggs and embryos from the un fertilized to blastocyst stages. From the present findings, it was suggested that the progestagens such as progesterone, 17α-hydroxyprogesterone, 17α,20β-dihydroxyprogesterone, 20α-hydroxyprogesterone and 20β-hydroxyprogesterone are constantly synthesized in mouse eggs and embryos during the early development.<br />マウスの未受精卵子および前核期から胚盤抱期までの初期胚について、△^5-3β-HSD(基質としてpregnenoloneおよび17α-hydroxypregnenoloneを使用)、 20α-HSD (20α-hydroxyprogesterone)および20β-HSD (17α-hydroxyprogesterone、 20β-hydroxyprogesterone)の活性を組織化学的に検出し、プロジェスタジェン合成能を検肘した。未受精期の卵子において、これらHSDの活性は88ないし98%のものに認められた。また、前核期から胚盤抱期までの初期胚においても、 83%以上でこれらのHSD活性が検出された。一方、ステロイド生合成の指標となる△^5-3β-HSD (DHA)の活性は、 86%以上の卵子と初期胚に常に観察された。これらのことから、発生初期のマウスの卵子と胚は、常にprogesterone、 17α-hydroxyprogesterone. 17α,20β-hydroxyprogesterone、 20α-hydroxyprogesteroneおよび20β-hydroxyprogesteroneの合成を行っており、これらプロジェスタジェンの合成能は、胚の初期発生に伴って変化しないことが考えられた。 Read more
6.

Article

Article
Niimura, Sueo ; Kawakami, Shin-ya
Published: 新潟大学農学部研究報告 = 新潟大学農学部研究報告.  55  pp.27-34,  2002-08.  新潟大学農学部
Full Text Link: http://hdl.handle.net/10191/25111
Abstract: The activities of hydroxysteroid dehydrogenases (HSDs) were histochemically demonstrated in mouse oocytes in the process of maturation in vivo and in vitro, and the changes in steroid metabolism during meiotic maturation were examined. In mouse oocytes soon after collection, the activities of △5-3β- HSD, 17β-HSD and 20β-HSD were observed in 87 to 97% of those, while the activity of 20α-HSD was not. The percentages of oocytes showing the activities of △5-3β-HSD (using DHA, pregnenolone and 17α- hydroxypregnenolone as the substrates), 17β -HSD (estradiol-17β and testosterone) and 20β -HSD (17α- hydroxyprogesterone and 20β-hydroxyprogesterone) did not change during maturation in vivo and in vitro. The oocytes with the activity of 20α-HSD (20α hydroxyprogesterone) appeared 4 hrs after the hCG injection or after culturing for 4 hrs and the rates of those reached 92 and l00%, respectively, 14 hrs after the hCG injection or after cultunng for 14 hrs. In the oocytes cultured for 8 hrs with olomoucine, an inhibitor of cychn-dependent kmase, nuclei were almost in the germinal vesicle stage, and the activity of 20α-HSD (20α hydroxyprogesterone) was observed in 84% of the treated oocytes. On the other hand, 81% of control oocytes also showed such the HSD activity, showing no difference from the rate of the treated oocytes. From the present findings, it was suggested that the metabolic abilities of progesterone, Viahydroxyprogesterone, 17α,20β dihydroxyprogesterone, 20β -hydroxyprogesterone, estradiol-17/7 and androgen are constantly present in mouse oocytes in the process of maturation in vivo and in vitlo. And it was also suggested that the metabolic ability of 20α-hydroxyprogesterone in mouse oocytes varies with maturation, while the change in the metabolic ability of such steroid is not related to nuclear maturation.<br />体内および体外で成熟過掛こあるマウス卵母細胞について、各種ヒドロキシステロイド脱水索酵素(HSD)の活性を組織化学的に検出し、卵母細胞の成熟に伴うステロイド代謝能の変化を検肘した。採取直後の卵母細胞において、 87ないし97%で△^5-3β-HSD、 17β-HSDおよび20β-HSDの活性が検出されたが、 20α-HSDの活性は検出されなかった。一方、 △^5-3β-HSD(基質としてDHA、 pregnenoloneおよび17α hydroxypregnenoloneを使用)、 17β-HSD (estradiol-17βとtestosterone)および20β-HSD O7α hydroxyprogesteroneと20β-hydroxyprogesterone)の活性を示す卵母細胞の割合は、体内および体外での成熟の経過に伴う変化を示さなかったが、成熟に伴って、20α-HSD (20α-hydroxyprogesterone)の活性を示す卵母細胞は出現し、その割合は高くなった。また、 8時間オロモウシン処置した卵母細胞において、核はほとんどで卵核胞期にあるとともに、 20α-HSD (20α-hydroxyprogesterone)の活性を示すものの割合は84%であり、オロモウシン処置していない対照の卵母細胞の81%と相違なかった。以上の結果から、マウス卵母細胞のprogesterone、 17α-hydroxyprogesterone、 17α,20β-dihydroxyprogesterone、 20β-hydroxyprogesterone、 estradiol-17βおよびandrogenの代謝能は、体内および体外での成熟に関わらず変化しないことが示唆された。また、マウス卵母細胞では、成熟に伴って20α- hydroxyprogesteroneの代謝能は出果して高まるが、この代謝能の変化は、核の成熟とは関係ないことが推察された。 Read more
7.

Article

Article
新村, 末雄
Published: 新潟大学農学部研究報告 = 新潟大学農学部研究報告.  48  pp.37-42,  1996-03.  新潟大学農学部
Full Text Link: http://hdl.handle.net/10191/17324
Abstract: 卵核胞期のウシの卵胞卵子,卵胞卵子を体外で成熟させた未受精卵子とこれを体外で受精させたのち培養して発生させた精子侵入期の卵子から孵化胚盤胞期までの胚について,グリコゲソ含量とα-Glycerophosphate dehydrogenase( α-GDH)活性を組織化学的に調べた。グリコゲン穎粒は,1細胞期の卵子から拡張胚盤胞期までの胚では細胞質に常に多量含まれていたが,孵化胚盤胞では減少した。またα-GDH活性は,1細胞期の卵子から拡張胚盤胞期までの胚では中等度であったが,孵化胚盤胞では弱かった。<br />The amount of glycogen granules and the activity of α-glycerophosphate dehydrogenase (α-GDH) were histochemically examined in bovine eggs matured in vitro, and in embryos from the 2-cell to hatched blastocyst stages developed in vitro. For the demonstration of glycogen, eggs and embryos were treated with the periodic acid-Schiff with or without a preceding salivary test. The activity of α-GDH was demonstrated by the method of BARKA and ANDERSON. The amount of glycogen granules was abundant in 1-cell eggs at the germinal vesicle, unfertilized and sperm-penetrated stages, and in embryos from the 2-cell to expanded blastocyst stages, but less in hatched blastocysts. The activity ofα-GDH was moderate in eggs and embryos at the stages from 1-cell to expanded blastocyst, but weak in hatched blastocysts. Read more
8.

Article

Article
Ishihara, Akihiko ; Itoh, Kazuo ; Itoh, Minoru ; Hirofuji, Chiyoko ; Hayashi, Hitomi
Published: Acta medica et biologica = Acta medica et biologica.  46  pp.1-6,  1998-03.  Niigata University School of Medicine
Full Text Link: http://hdl.handle.net/10191/33071
Abstract: Fiber type distributions of the fast-twitch extensor digitorum longus muscle in rats were examined after exposure to hypobaric-hypoxia for 5 weeks from the postnatal age of 5 weeks or for 10 weeks from the postnatal age of 10 weeks. Muscle fibers were classified into fast-twitch oxidative glycolytic (FOG), fast-twitch glycolytic (FG), or slow-twitch oxidative (SO). The percentage of FOG fibers was increased and that of FG fibers was decreased in the hypoxic rats at 10 weeks of age, while there was no difference in the percentage of SO fibers between the normoxic and hypoxic rats. There was no difference in the fiber type distribution between the normoxic and hypoxic rats at 20 weeks of age. These results indicate an age-specific response of hypobarichypoxia on the fiber types of the rat extensor digitorum longus muscle. Read more
9.

Article

Article
Ishihara, Akihiko ; Itoh, Kazuo ; Itoh, Minoru ; Hirofuji, Chiyoko ; Hayashi, Hitomi
Published: Acta medica et biologica = Acta medica et biologica.  44  pp.221-225,  1996-12.  Niigata University School of Medicine
Full Text Link: http://hdl.handle.net/10191/33185
Abstract: The histochemical type shift of fibers from fast-twitch oxidative glycolytic (FOG) to slow-twitch oxidative (SO) in the rat soleus muscle occurs during postnatal development. Our recent experiments demonstrated that the type shift of fibers from FOG to SO is inhibited by hypobaric-hypoxia. Furthermore, the present study has shown that hypobaric-hypoxia inhibits the growth-related type shift of fibers from FOG to SO in the rat soleus muscle, irrespective of the duration of exposure to hypobaric-hypoxia. Read more