本研究在应用免疫组织化学技术观察了享廷顿蛋白相关蛋白1(huntingtin associated protein 1，HAP1)在大鼠脑内分布特征的基础上，比较了下丘脑HAP1表达的年龄变化，观察了下丘脑视上核、室旁核HAP1与加压素(vasopressin，VP)间的关系，...本研究在应用免疫组织化学技术观察了享廷顿蛋白相关蛋白1(huntingtin associated protein 1，HAP1)在大鼠脑内分布特征的基础上，比较了下丘脑HAP1表达的年龄变化，观察了下丘脑视上核、室旁核HAP1与加压素(vasopressin，VP)间的关系，HAP1在神经垂体中定位，并观察了禁水对下丘脑和神经垂体中HAP1表达的影响。展开更多
目的探讨慢性复合应激大鼠肾上腺髓质细胞亨廷顿蛋白相关蛋白1(Huntingtin-associated protein 1,HAP-1)表达的变化及其意义。方法36只大鼠随机分为两组:慢性复合应激组和对照组。应激组动物进行6周的垂直旋转、睡眠剥夺、捆绑(6h/d)和...目的探讨慢性复合应激大鼠肾上腺髓质细胞亨廷顿蛋白相关蛋白1(Huntingtin-associated protein 1,HAP-1)表达的变化及其意义。方法36只大鼠随机分为两组:慢性复合应激组和对照组。应激组动物进行6周的垂直旋转、睡眠剥夺、捆绑(6h/d)和夜间光照等慢性复合性应激试验;实验结束后,所有动物采用免疫组织化学、Western-blot以及RT-PCR等方法检测肾上腺髓质细胞内HAP-1蛋白和mRNA水平的变化。结果与对照组相比,慢性复合应激组的大鼠肾上腺髓质中HAP-1的表达明显增强(P<0.05),HAP-1 mRNA水平明显升高(P<0.05)。结论6周慢性复合性应激大鼠HAP-1在肾上腺髓质区的表达加强,mRNA水平提高。提示HAP-1在慢性复合应激促进肾上腺功能中可能发挥一定作用。展开更多
目的观察亨廷顿蛋白相关蛋白1(huntingtin-associated protein 1,HAP1)在正常大鼠脊髓发育过程中的表达变化。方法采用免疫组织化学ABC法。结果免疫组织化学结果显示:在胚胎期HAP1阳性细胞主要分布在脊髓腹侧,背侧数量较少;随着胚龄的增...目的观察亨廷顿蛋白相关蛋白1(huntingtin-associated protein 1,HAP1)在正常大鼠脊髓发育过程中的表达变化。方法采用免疫组织化学ABC法。结果免疫组织化学结果显示:在胚胎期HAP1阳性细胞主要分布在脊髓腹侧,背侧数量较少;随着胚龄的增加,HAP1阳性细胞在脊髓灰质中分布广泛,腹侧阳性细胞体积较小,分布较密集,背侧阳性细胞体积较大,分布较稀疏。在生后早期HAP1阳性细胞呈蝶形广泛分布在脊髓灰质内,阳性细胞体积较小,分布比胚胎期稀疏;在成年期HAP1阳性细胞主要分布在脊髓背角灰质内,背角浅层的HAP1阳性细胞体积较小,数量较少,细胞为圆形,越靠近深层阳性细胞数量越少,HAP1阳性细胞有明显的胞体和突起,胞体为圆形或椭圆形,突起细长。结论 HAP1的表达贯穿脊髓发育过程中,HAP1的表达随着胚龄的增加而发生规律性变化;生后大鼠脊髓内的HAP1阳性细胞分布与脊髓灰质的分化一致。展开更多
As a critical organelle in Toxoplasma gondii,an important zoonotic pathogen,the apicoplast plays a vital role in survival by hosting essential metabolic pathways.Its unique four-membrane structure provides both stabil...As a critical organelle in Toxoplasma gondii,an important zoonotic pathogen,the apicoplast plays a vital role in survival by hosting essential metabolic pathways.Its unique four-membrane structure provides both stability and a relatively independent metabolic compartment.To identify previously uncharacterized apicoplast membrane proteins,we undertook a proteomics analysis of the apicoplast using a TurboID proximity biotinylation approach with our previously discovered apicoplast membrane proteins—AMT1,AMT2,HAP1,and HAP9.The identified apicoplast membrane proteins in this study,along with 10 previously discovered apicoplast membrane proteins,were subjected to phenotypic and functional analyses.This demonstrated that the major facilitator superfamily transport HAP1 and the ATP-binding cassette transporter HAP9 are both apicoplast proteins essential for in vitro parasite growth in host cell monolayers and in vivo mouse infection.Depletion of the transporters caused similar defects in parasite replication,endogenous biotinylation of ACC1 and stability of the apicoplast.However,transmission electron microscopy showed that depletion of HAP1 and HAP9 caused different structural defects in the apicoplast:a lucent intra-membrane space in the HAP1-depleted apicoplast,and a lucent matrix and loosened membranes in the HAP9 depleted apicoplast.This study thus provides critical insights into the physiological and structural roles of these newly identified transporters in the apicoplast of T.gondii and,potentially,in closely related protists containing the apicoplast.展开更多
文摘本研究在应用免疫组织化学技术观察了享廷顿蛋白相关蛋白1(huntingtin associated protein 1，HAP1)在大鼠脑内分布特征的基础上，比较了下丘脑HAP1表达的年龄变化，观察了下丘脑视上核、室旁核HAP1与加压素(vasopressin，VP)间的关系，HAP1在神经垂体中定位，并观察了禁水对下丘脑和神经垂体中HAP1表达的影响。
文摘目的探讨慢性复合应激大鼠肾上腺髓质细胞亨廷顿蛋白相关蛋白1(Huntingtin-associated protein 1,HAP-1)表达的变化及其意义。方法36只大鼠随机分为两组:慢性复合应激组和对照组。应激组动物进行6周的垂直旋转、睡眠剥夺、捆绑(6h/d)和夜间光照等慢性复合性应激试验;实验结束后,所有动物采用免疫组织化学、Western-blot以及RT-PCR等方法检测肾上腺髓质细胞内HAP-1蛋白和mRNA水平的变化。结果与对照组相比,慢性复合应激组的大鼠肾上腺髓质中HAP-1的表达明显增强(P<0.05),HAP-1 mRNA水平明显升高(P<0.05)。结论6周慢性复合性应激大鼠HAP-1在肾上腺髓质区的表达加强,mRNA水平提高。提示HAP-1在慢性复合应激促进肾上腺功能中可能发挥一定作用。
文摘目的观察亨廷顿蛋白相关蛋白1(huntingtin-associated protein 1,HAP1)在正常大鼠脊髓发育过程中的表达变化。方法采用免疫组织化学ABC法。结果免疫组织化学结果显示:在胚胎期HAP1阳性细胞主要分布在脊髓腹侧,背侧数量较少;随着胚龄的增加,HAP1阳性细胞在脊髓灰质中分布广泛,腹侧阳性细胞体积较小,分布较密集,背侧阳性细胞体积较大,分布较稀疏。在生后早期HAP1阳性细胞呈蝶形广泛分布在脊髓灰质内,阳性细胞体积较小,分布比胚胎期稀疏;在成年期HAP1阳性细胞主要分布在脊髓背角灰质内,背角浅层的HAP1阳性细胞体积较小,数量较少,细胞为圆形,越靠近深层阳性细胞数量越少,HAP1阳性细胞有明显的胞体和突起,胞体为圆形或椭圆形,突起细长。结论 HAP1的表达贯穿脊髓发育过程中,HAP1的表达随着胚龄的增加而发生规律性变化;生后大鼠脊髓内的HAP1阳性细胞分布与脊髓灰质的分化一致。
基金funded by the National Key Research and Development Project of China(Grant No.2022YFD1800202).
文摘As a critical organelle in Toxoplasma gondii,an important zoonotic pathogen,the apicoplast plays a vital role in survival by hosting essential metabolic pathways.Its unique four-membrane structure provides both stability and a relatively independent metabolic compartment.To identify previously uncharacterized apicoplast membrane proteins,we undertook a proteomics analysis of the apicoplast using a TurboID proximity biotinylation approach with our previously discovered apicoplast membrane proteins—AMT1,AMT2,HAP1,and HAP9.The identified apicoplast membrane proteins in this study,along with 10 previously discovered apicoplast membrane proteins,were subjected to phenotypic and functional analyses.This demonstrated that the major facilitator superfamily transport HAP1 and the ATP-binding cassette transporter HAP9 are both apicoplast proteins essential for in vitro parasite growth in host cell monolayers and in vivo mouse infection.Depletion of the transporters caused similar defects in parasite replication,endogenous biotinylation of ACC1 and stability of the apicoplast.However,transmission electron microscopy showed that depletion of HAP1 and HAP9 caused different structural defects in the apicoplast:a lucent intra-membrane space in the HAP1-depleted apicoplast,and a lucent matrix and loosened membranes in the HAP9 depleted apicoplast.This study thus provides critical insights into the physiological and structural roles of these newly identified transporters in the apicoplast of T.gondii and,potentially,in closely related protists containing the apicoplast.
