Disaccharide combinations and the expression of enolase3 and plasma membrane Ca2+ ATPase isoform in sturgeon sperm cryopreservation          下载免费PDF全文

P Li  H Du  XM Qiao  ZG Liu  QW Wei 《Reproduction in domestic animals》2018,53(2):472-483

Acipenser sinensis and Acipenser dabryanus are critically endangered species, so germplasm conservation via cryopreservation of sperm is necessary. Disaccharides can act as membrane‐impermeable cryoprotectants, and enolase3 (ENO3) and plasma membrane Ca²⁺ ATPase isoform (PMCA2) are proteins associated with sperm quality. We considered seven characteristics of sperm quality in cultured brood stock from A. sinensis and A. dabryanus. We tested use of sucrose or trehalose alone and in combination at different concentrations for cryopreservation of A. dabryanus sperm. A low concentration of sucrose plus trehalose (S₁₅T₁₅) was optimal. Mixing of the extender with sucrose, lactose, or trehalose alone or with pairwise mixtures revealed that a mixture of lactose and trehalose (L₁₅T₁₅) gave the best results for both A. sinensis and A. dabryanus. Enolase3 and PMCA2 expression levels were measured in cryopreserved A. sinensis sperm via Western blotting. Relative ENO3 and PMCA2 expression levels were examined, and the relationship between disaccharide composition, sperm quality and protein expression was explored in A. sinensis. The results showed that relative ENO3 and PMCA2 expression levels were the highest at L₁₅T₁₅ in cryopreserved A. sinensis sperm. There were significant positive correlations between ENO3 expression and percentage membrane integrity, and between PMCA2 expression and sperm motility parameters (percentage of motile sperm, curvilinear velocity, straight‐line velocity and average path velocity; p < .05) in cryopreserved A. sinensis sperm. Our results indicate the optimal disaccharide combination and concentrations for cryopreservation of A. sinensis and A. dabryanus sperm and suggest that ENO3 and PMCA2 expression levels could serve as a valuable indicator of sperm quality in A. sinensis.  相似文献   

高效液相色谱-蒸发光散射检测法测定茶叶中单糖和双糖   总被引：1，自引：0，他引：1  

袁勇  黄建安  李银花  陈金华  袁玲  刘仲华 《茶叶科学》2010,30(6):435-439

建立了高效液相色谱-蒸发光散射检测器（HPLC-ELSD）测定茶叶中单糖和双糖的检测方法。采用Shodex NH₂P-50-4E（4.6 mm×250 mm,5 μm）色谱柱,以水和乙腈为流动相进行梯度洗脱,梯度洗脱程序为：0~14 min,85%乙腈;14~16 min,85%~70%乙腈;16~28 min,70%乙腈;28~32 min,70%~85%乙腈,流速：1.0 mL/min,柱温：20℃,雾化管温度为30℃,漂移管温度为70℃,氮气压力为310.275 Pa。鼠李糖、阿拉伯糖、木糖、果糖、甘露糖、葡萄糖、蔗糖、麦芽糖在3.202~184.828 μg范围内程良好的线性关系,8种糖的加标回收率在90.97%~105.07%之间,最低检测限（S/N=3）范围在6.976~1376.297 ng。该方法具有操作简便,分离效果好等特点。  相似文献   

Inheritance of the concentration of water-soluble carbohydrates and its relationship with the concentrations of fibre and crude protein in herbage of cocksfoot (Dactylis glomerata L.)     

Y. Sanada  T. Takai  T. Yamada 《Grass and Forage Science》2007,62(3):322-331

The concentration of water‐soluble carbohydrates (WSC) of cocksfoot is lower than that of other temperate grasses. Increasing the WSC concentration in cocksfoot is important in increasing its digestibility and preference by ruminants. The genetic variation in mono‐ and disaccharides, fructans and total WSC concentrations and their inter‐relationships with crude protein (CP) and fibre concentrations of cocksfoot (Dactylis glomerata L.) at the vegetative growth stage in half‐sib cross populations were assessed in two experiments conducted under spaced planting and sward conditions. There was a wide range in the means for concentrations of fructan, WSC and neutral‐detergent fibre (NDF) in parents and progeny whereas there was a narrow range of the means for concentrations of mono‐ and disaccharides, CP and acid‐detergent fibre (ADF). Mean concentrations of mono‐ and disaccharides showed the smallest range. Mean squares of entries in parents and progenies were significant for mono‐ and disaccharides, fructan and total WSC concentrations in all populations. The entry × year interactions for fructan and total WSC concentrations were significant in the parents and the half‐sib progeny. However, the entry × year interactions for mono‐ and disaccharide concentrations in progeny were not significant. Concentration of WSC was under genetic control as mean squares of the concentration values were significant and variance components for all traits were significantly larger than zero. There were negative genetic correlations between WSC and ADF and NDF concentrations. Narrow‐sense heritabilities (h_n) estimated from variance components of progeny for total WSC concentrations were 0·59 and 0·53 in sward and spaced planting conditions respectively. The h_n of fructan and WSC concentrations were similar in both sward and spaced planting conditions, whereas that of mono‐ and disaccharide concentrations varied from 0·20 to 0·69. It was found that the genetic variation of the total WSC concentration in cocksfoot depended mainly on genetic variation in fructan concentration. These results suggested that the forage quality of cocksfoot at the vegetative growth stage was influenced by an additive gene effect and could be improved genetically by recurrent selection.  相似文献   

青钱柳叶提取物对Caco-2细胞二糖酶活性的抑制作用     

王丽玲  柏明娥  刘本同  王衍彬  徐高福 《浙江林业科技》2019,(1):28-34

采用Caco-2细胞模型,分析了青钱柳Cyclocarya paliurus叶水提物、醇提后的水提物、95%醇提物、乙酸乙酯萃取物和正丁醇萃取物5种不同提取物对Caco-2细胞活力的影响及对Caco-2细胞上麦芽糖酶、蔗糖酶、乳糖酶3种二糖酶活性的抑制作用。结果表明,5种不同提取物中除95%醇提物对Caco-2细胞活力产生一定影响外,其余4种提取物对Caco-2细胞基本无毒性,在200 mg·L^-1浓度提取物范围内培养24～72 h后细胞仍能保持较高活力;对3种二糖酶活性抑制作用效果最好的为醇提后的水提物,尤其是对蔗糖酶和乳糖酶的抑制作用更为明显,最高的抑制率分别达到76.35%和60.13%,其次为乙酸乙酯萃取物,对乳糖酶的抑制率最高达54.41%。从青钱柳叶不同方法提取物的制备流程说明多糖类或中等极性的黄酮甙类有可能是青钱柳叶中抑制二糖酶活性的主要组成成分。  相似文献