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1.
《中国畜牧兽医》2020,(8)
为了研究4F2hc在奶牛乳腺中的表达模式及调控方式,进一步明确氨基酸在奶牛乳腺上皮细胞中的跨膜转运过程,本研究采用Western blotting和实时荧光定量PCR技术检测了4F2hc在泌乳期和干奶期奶牛乳腺组织中的表达变化;在体外培养的泌乳期奶牛乳腺上皮细胞中添加亮氨酸,采用Western blotting和实时荧光定量PCR技术检测其对奶牛乳腺上皮细胞中4F2hc表达的影响;采用雷帕霉素抑制剂抑制mTOR信号通路,使用Western blotting方法检测mTOR信号抑制后奶牛乳腺上皮细胞中4F2hc表达以及乳蛋白合成的变化。结果显示,在泌乳期的奶牛乳腺组织中4F2hc的mRNA和蛋白表达水平均显著或极显著高于干奶期(P0.05,P0.01);在体外培养的奶牛乳腺上皮细胞中添加亮氨酸可以极显著提高乳腺上皮细胞中4F2hc的mRNA和蛋白质表达水平(P0.01);亮氨酸刺激可以激活细胞内的mTOR信号通路(P0.05),而雷帕霉素处理则可以显著抑制mTOR信号分子的磷酸化并极显著抑制亮氨酸诱导的4F2hc的表达(P0.05,P0.01),进而极显著抑制β-Casein的合成(P0.01)。以上研究结果表明,4F2hc基因的表达与奶牛乳腺的泌乳活性之间呈正相关,亮氨酸可以通过激活mTOR信号通路来调节4F2hc基因的表达,进而影响乳蛋白的合成。 相似文献
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为研究L型氨基酸转运载体1（L type amino acid transporter 1,LAT1）对奶牛乳腺中β-酪蛋白表达的作用,本试验采用PCR技术体外扩增奶牛LAT1基因并构建LAT1真核表达载体,采用脂质体转染技术将LAT1真核表达载体转染奶牛乳腺上皮细胞,并于转染48 h后采用Western blotting技术检测LAT1过表达后乳腺上皮细胞中LAT1、4F2hc和β-酪蛋白的表达变化。荧光显微镜检测结果显示,LAT1真核表达载体成功转染奶牛乳腺上皮细胞;Western blotting检测结果显示,LAT1过表达的奶牛乳腺上皮细胞中LAT1极显著增加（P<0.01）,β-酪蛋白的表达显著升高（P<0.05）,4F2hc表达变化不显著（P>0.05）。这些结果提示LAT1对奶牛乳腺上皮细胞乳蛋白合成具有促进作用。 相似文献
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《中国畜牧兽医》2016,(10)
为研究L型氨基酸转运载体1(L type amino acid transporter 1,LAT1)对奶牛乳腺中β-酪蛋白表达的作用,本试验采用PCR技术体外扩增奶牛LAT1基因并构建LAT1真核表达载体,采用脂质体转染技术将LAT1真核表达载体转染奶牛乳腺上皮细胞,并于转染48h后采用Western blotting技术检测LAT1过表达后乳腺上皮细胞中LAT1、4F2hc和β-酪蛋白的表达变化。荧光显微镜检测结果显示,LAT1真核表达载体成功转染奶牛乳腺上皮细胞;Western blotting检测结果显示,LAT1过表达的奶牛乳腺上皮细胞中LAT1极显著增加(P0.01),β-酪蛋白的表达显著升高(P0.05),4F2hc表达变化不显著(P0.05)。这些结果提示LAT1对奶牛乳腺上皮细胞乳蛋白合成具有促进作用。 相似文献
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为了研究昆明小鼠乳腺发育4个时期即青春期、妊娠期、泌乳期和退化期的同源异型盒基因(Hex)蛋白表达规律及组织定位,试验采用免疫组织化学法进行检测。结果表明:昆明小鼠乳腺组织结构呈周期性变化,青春期至妊娠期Hex蛋白逐渐增加,泌乳期达到高峰,退化期逐渐下降至类似青春期水平;Hex蛋白主要定位于细胞质中,泌乳期在细胞核中也有少部分出现;昆明小鼠乳腺发育的4个阶段均有Hex蛋白表达,且与乳腺周期性的变化一致。说明Hex蛋白参与乳腺的发育和泌乳调控过程。 相似文献
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为了研究4F2hc在奶牛乳腺中的表达模式及调控方式，进一步明确氨基酸在奶牛乳腺上皮细胞中的跨膜转运过程，本研究采用Western blotting和实时荧光定量PCR技术检测了4F2hc在泌乳期和干奶期奶牛乳腺组织中的表达变化；在体外培养的泌乳期奶牛乳腺上皮细胞中添加亮氨酸，采用Western blotting和实时荧光定量PCR技术检测其对奶牛乳腺上皮细胞中4F2hc表达的影响；采用雷帕霉素抑制剂抑制mTOR信号通路，使用Western blotting方法检测mTOR信号抑制后奶牛乳腺上皮细胞中4F2hc表达以及乳蛋白合成的变化。结果显示，在泌乳期的奶牛乳腺组织中4F2hc的mRNA和蛋白表达水平均显著或极显著高于干奶期（P<0.05，P<0.01）；在体外培养的奶牛乳腺上皮细胞中添加亮氨酸可以极显著提高乳腺上皮细胞中4F2hc的mRNA和蛋白质表达水平（P<0.01）；亮氨酸刺激可以激活细胞内的mTOR信号通路（P<0.05），而雷帕霉素处理则可以显著抑制mTOR信号分子的磷酸化并极显著抑制亮氨酸诱导的4F2hc的表达（P<0.05，P<0.01），进而极显著抑制β-Casein的合成（P<0.01）。以上研究结果表明，4F2hc基因的表达与奶牛乳腺的泌乳活性之间呈正相关，亮氨酸可以通过激活mTOR信号通路来调节4F2hc基因的表达，进而影响乳蛋白的合成。 相似文献
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《畜牧兽医学报》2015,(8)
旨在研究山羊y+L碱性氨基酸转运系统中4F2hc、y+LAT1和y+LAT2mRNA表达的组织特异性及其在不同年龄山羊小肠中的发育性表达规律。试验选取健康、体重相近的1日龄、6月龄、8月龄、10月龄、12月龄陕北白绒山羊各3只,共15只,屠宰后采集心、肝、肾、大脑、背最长肌、体侧皮肤、瘤胃、十二指肠、空肠、回肠和结肠组织,利用实时荧光定量PCR技术检测4F2hc、y+LAT1和y+LAT2mRNA在1日龄山羊上述组织和全部山羊十二指肠、空肠和回肠的表达情况。结果表明,(1)4F2hc、y+LAT1和y+LAT2mRNA在1日龄山羊多种组织中均有表达,且有一定的组织特异性。其中,4F2hc mRNA在皮肤表达量最高,y+LAT1 mRNA在回肠表达量最高,y+LAT2mRNA在大脑和回肠表达量最高。(2)4F2hc mRNA在山羊十二指肠、空肠和回肠表达量均以1日龄山羊为最高,其他月龄山羊相同肠段之间表达量无显著差异(P0.05);y+LAT1mRNA在十二指肠、空肠和回肠表达量随山羊年龄的增加均呈现逐渐降低的趋势;y+LAT2mRNA在十二指肠、空肠和回肠表达量随山羊年龄的增加呈现先升高后降低的趋势,其在10月龄山羊表达量最高;y+LAT2mRNA在回肠表达量以1日龄山羊为最低,其他月龄山羊之间差异不显著(P0.05)。结果说明,山羊y+L碱性氨基酸转运系统的主要转运部位为小肠;该系统中轻链y+LAT1和重链4F2hc构成的异二聚体转运蛋白可能发挥更为主要的转运作用;4F2hc和y+LAT2基因可能分别在山羊皮肤和大脑中发挥重要的生理功能;4F2hc、y+LAT1和y+LAT2基因在山羊小肠中受肠段和发育阶段的影响,具有不同的发育性表达规律。 相似文献
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《畜牧兽医科技信息》2015,(8)
试验选取青春期、妊娠期、泌乳期(分产高品质奶与产低品质奶)与退化期等几个时期的奶牛乳腺组织,提取RNA,实时荧光定量PCR检测各组织中I型和II型肽转运蛋白的表达情况。结果表明,I型肽转运蛋白的表达在青春期较低,在妊娠期与泌乳期显著上升,至泌乳期达到顶峰,在退化期下降至青春期水平;II型肽转运蛋白在各个时期表达无明显变化;在泌乳期高品质乳组织中,I型肽转运蛋白的表达显著高于低品质乳组织,且在这两组样品中,I型肽转运蛋白的表达均显著高于II型肽转运蛋白的表达。结果提示,在奶牛泌乳过程中,I型肽转运蛋白与泌乳的调节有显著的正相关性,而II型肽转运蛋白与泌乳的调节无明显关系。 相似文献
8.
《中国畜牧杂志》2020,(4)
本研究旨在探讨仔猪饲喂不同浓度支链氨基酸(BCAA)日粮对空肠、腰肌、咬肌以及腿肌氨基酸转运载体表达的影响。试验选取18头(10.0±0.9)kg杜×长×大三元杂交断奶仔猪,按体重和性别随机分为3个处理组,每个处理6头猪。3个处理组分别饲喂BCAA缺乏日粮(L-BCAA)、BCAA正常日粮(N-BCAA)和BCAA过量日粮(E-BCAA)。试验期24 d,包括3 d适应期和21 d正试期。结果表明:与N-BCAA组相比,仔猪饲喂L-BCAA日粮会显著增加肠道中性氨基酸转运载体(ASCT2、SNAT2、B~0AT和rBAT)、碱性氨基酸转运载体(y+LAT、CAT1和4F2hc)和小肽转运载体(Pept1)的表达,可显著提高腰肌中(PAT1、PAT2、SNAT2、LAT1和CAT1)、咬肌中(4F2hc和EAAT1)氨基酸转运载体的表达,而显著降低咬肌中(ASCT2、PAT1和PAT2)、腿肌中(PAT1、PAT2和4F2hc)氨基酸转运载体的表达;而仔猪饲喂E-BCAA日粮可显著降低腰肌中(ASCT2、SNAT4、y+LAT1、rBAT、EAAC1)、咬肌中(ASCT2、PAT1、PAT2、EAAC1)、腿肌中(PAT1、PAT2、4F2hc、rBAT、EAAT1、EAAC1)氨基酸转运载体的表达。综上所述,仔猪饲喂L-BCAA日粮可增加肠道氨基酸转运载体的表达,以增强对氨基酸的吸收,但过量添加BCAA会抑制肌肉中氨基酸转运载体的表达。 相似文献
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To investigate the effect of essential amino acids on LAT1 expression in lactation mammary gland, the lactation mammary acini were cultured and LAT1 and 4F2hc expression were detected by Western blotting. The results showed that essential amino acids upregulated LAT1 and 4F2hc expression significantly in lactation mammary acini of dairy cow. It revealed that LAT1 was the mainly amino acid transporter in lactation mammary gland. The expression of LAT1 and 4F2hc was induced by essential amino acids. 相似文献
12.
LIN Lin FAN Hui-quan XING Wei-nan YANG Yang HOU Xiao-ming ZHANG Chang-shun LIN Ye 《中国畜牧兽医》2015,42(5):1240-1244
To investigate the relationship between the expression of SYK and dairy cow mammary gland development and lactation, the expression of SYK in lactating dairy cow mammary gland with high or low quality milk and dry period Holstein dairy cow mammary gland was detected by Western blotting and laser confocal microscope.The results showed that SYK expression in dry period mammary gland was significant higher than that in lactating mammary gland (P<0.05).There was no SYK differential expression detected between lactating mammary gland with high quality milk and low quality milk (P>0.05).SYK was mainly located in the cytoplasm of ductal epithelial cells in dry period mammary gland.In lactating mammary gland, SYK was existed in acinar epithelial cells.All these results revealed that SYK was a regulator in mammary epithelial cell proliferation and differentiation.It participated in mammary gland reconstitution in dry period. 相似文献
13.
为探讨脾源性酪氨酸激酶(spleen tyrosine kinase,SYK)的表达与奶牛乳腺发育和泌乳功能之间的关系,试验采用Western blotting和激光共聚焦显微技术对泌乳期高乳品质、低乳品质及干乳期的中国荷斯坦奶牛乳腺组织中SYK的表达含量和表达部位的变化进行研究。结果表明,干乳期奶牛乳腺组织中SYK的表达显著高于泌乳期奶牛乳腺组织(P<0.05),泌乳期高乳品质、低乳品质奶牛乳腺组织中SYK的表达差异不显著(P>0.05);在干乳期SYK主要在乳腺导管上皮细胞的胞质中表达,而在泌乳期SYK在腺泡上皮细胞中表达。结果提示SYK是乳腺上皮细胞增殖与分化的调节因子,主要参与干乳期乳腺组织的重建过程。 相似文献
14.
Park JC Kim YB Yoon JH Kim HJ Kim SM Kanai Y Endou H Kim DK 《Anatomia, histologia, embryologia》2004,33(2):119-124
Certain amino acid transport systems play an important role in supplying organic nutrients to each cell and for cell proliferation during tooth development. However, the mechanisms responsible for such actions are unclear. This study demonstrated for the first time that LAT1 and 4F2hc are expressed during tooth development in prenatal and postnatal rats, and that the transporters show cell-specific expression in ameloblasts, which are the epithelium-derived dental cells. LAT1 and 4F2hc expression was not observed in other dental cells of the developing teeth such as odontoblasts and cementoblasts. Overall, these results suggest that LAT1 and 4F2hc might play an important role in enamel formation. 相似文献
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D. T. Lincoln F. Sinowatz E. El-Hifnawi R. L. Hughes M. Waters 《Anatomia, histologia, embryologia》1995,24(2):107-115
A panel of monoclonal antibodies to the growth hormone (GH) receptor/binding protein was used to demonstrate the existence and detail the expression of GH receptors in ductal and alveolar epithelial cells from rat and rabbit mammary glands by immunohistochemistry. Intense immunoreactivity was present in membrane, cytoplasm and some nuclei of epithelial cells during proliferation and lactation. Receptor expression decreased during weaning and was absent or weak in regressive mammary glands. Immunoreactivity was weak in ductal epithelial cells from virgin adult animals. Pronounced expression of GH receptor/binding protein was observed with two monoclonal antibodies and lesser reactivity was seen with others, paralleling their affinities for the receptor. The cytoplasmic presence of this putatively plasma membrane located GH receptor is accounted for by the existence of a soluble form on the GH receptor, namely the growth hormone binding protein derived from the membrane receptor by cleavage. Primary localization of the receptor in proliferating and lactating epithelial cells suggests that the rat and rabbit mammary gland is a GH target tissue. This finding is in contradiction to both classical GH action and the somatomedin hypothesis and challenges the widely held view that GH has no direct influence on mammary growth and function. 相似文献
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Ichihara N Asari M Kasuya T Susaki E Matsui K Nishita T Amasaki H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(11):1167-1170
The localization of bovine carbonic anhydrase isozyme VI (CA-VI) was examined immunohistochemically in bovine mammary glands during early lactation period (after 2-3 days of postpartum) and dry period (at about 2 months preparturition in adults), and young calves (at 30 and 150 days after birth) using specific CA-VI antiserum. The immunoreaction for anti-CA-VI antiserum was very weak in the mammary glands in young (prepubescent) calves. In dry period, CA-VI was also weakly expressed in secretory epithelial (acinar) and ductal cells. In contrast, the reaction was intense in mammary gland cells in early lactation period. Dot blotting analysis indicated that anti-CA-VI reacted positively to beastings and mature saliva, but weakly or not at all to milk during the dry period or calf saliva, respectively. The intense expression of CA-VI in the mammary glands in early lactation period might compensate for low levels of secretion from functionally and structurally immature salivary glands in young calves. 相似文献
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Binbing Ling 《Research in veterinary science》2010,89(2):200-202
Glucose, fatty acids, and l-carnitine are important substrates that support mammary epithelial cell metabolism, biosynthetic capacity, and milk yield and composition. Our study investigated the effects of LPS-induced inflammation on the expression of several glucose, fatty acid, and l-carnitine transporters in the lactating rat mammary gland at different lactation stages. Day 4, 11, and 18 lactating rats (n = 3/treatment) were administered LPS (1 mg/kg) or saline by intraperitoneal (IP) injection. Fold differences in the mRNA expression of glucose transporters Glut1, Glut8 and Sglt1, fatty acid transporters Fatp1, Fatp4 and Fabp3, and l-carnitine transporters Octn1, Octn2, and Octn3 were determined using the Comparative CT method. The mRNA expression levels of all transporters evaluated, except Fatp4 and Octn2 were markedly higher in mammary gland at lactation day 11 compared to lactation day 4. LPS caused a marked decrease in transporter mRNA expression at each lactation stage except for Octn3 and Fatp1, which were markedly increased with LPS administration at lactation day 4, and Sglt1, which was slightly increased at day 11 of lactation. Our results suggest LPS-induced inflammation generally downregulates glucose, fatty acid, and l-carnitine transporter expression. Whether such changes lead to reductions in transporter substrate availability to the lactating mammary epithelial cell requires investigation since decreases in the availability of these nutrients may significantly impact mammary epithelial function and milk quality and yield. 相似文献
18.
Localization of the sheep FcRn in the mammary gland 总被引：5，自引：0，他引：5
Mayer B Zolnai A Frenyó LV Jancsik V Szentirmay Z Hammarström L Kacskovics I 《Veterinary immunology and immunopathology》2002,87(3-4):327-330
Among the multiple functions, which have been identified for the neonatal Fc receptor (FcRn), we study its role in the IgG transport in the mammary gland during the colostrum formation. For this reason, we have obtained several mammary gland biopsies from a pregnant sheep around parturition. The presence of the FcRn heavy chain mRNA was detected exclusively in the acinar and ductal epithelial cell by in situ hybridization (ISH). We detected strong signal in samples harvested 24 and 10 days prepartum; however, in samples we collected postpartum was barely detectable. Immunohistochemistry confirmed our ISH data. The cytoplasm of the epithelial cells of the acini and ducts in the mammary gland biopsies stained homogeneously before parturition, although a remarkable difference was observed in the pattern after lambing. The signal indicated uneven distribution of the FcRn alpha chain in the epithelial cells 1 and 5 days postpartum, since the apical sides of the epithelial cells were highlighted. The presence of the FcRn in the acinar and ductal epithelial cells and the obvious change of its distribution before and after parturition suggest that FcRn plays an important role in the IgG transport during colostrum formation. FcRn expression was also found in the lamb duodenal crypt epithelial cells, which have been previously demonstrated to secrete IgG1 in newborn ruminants, suggesting secretory role of the FcRn in ruminant epithelial cells. 相似文献