Evaluation of β‐carotene assimilation in leopard geckos (Eublepharis macularius)

Although leopard geckos (Eublepharis macularius) are commonly kept under human care, their vitamin requirements are largely unknown. Many invertebrate preys display a low vitamin A concentration; thus, gut‐loading insects with vitamin A or carotenoids is a common practice. The objective of this prospective experimental study was to investigate whether dietary supplementation with β‐carotene, including prey gut‐loading, leads to sufficient vitamin A hepatic storage and prevents epithelial squamous metaplasia development in leopard geckos. Ten clinically healthy female leopard geckos were randomly divided in two groups with various supplementations: a group receiving vitamin A supplementation and a group receiving β‐carotene. Insects were gut‐loaded continuously with a supplement containing vitamin A or β‐carotene, depending on the group. Oral supplementation with cod liver oil or carrot juice was administered weekly to each lizard from “vitamin A group” and “carotenoid group” respectively. After 10 weeks of supplementation, surgical hepatic biopsies were obtained in three geckos of each group while the two remaining geckos were euthanized to undergo complete necropsy. Hepatic vitamin A concentration was determined for each lizard (n = 10) by ultra‐performance liquid chromatography. Histopathology revealed hepatocellular vacuolization and vitellogenic follicles in five females. Epithelial squamous metaplasia was not observed in any of the geckos. Hepatic vitamin A concentration was significantly higher in the carotenoid‐supplemented group than in the vitamin A‐supplemented group (p = 0.03). Our results suggest that in leopard geckos, dietary supplementation with β‐carotene allows sufficient vitamin A hepatic storage.     

Differential gene expression of Eph‐ephrin A1 and LEPR‐LEP with high or low number of embryos in pigs during implantation

The objective of this study was to ascertain whether mRNA and protein expressions of implantation‐related genes (erythropoietin‐producing hepatocellular receptor–ligand A1, Eph‐ephrin A1 and leptin receptor–leptin, LEPR‐LEP) differed between pigs with high and low number of embryos, and whether these differences in gene expression might affect embryo implantation. Experimental pig groups (n = 24) for high and low number of embryos were prepared by altering the number of eggs ovulated in pre‐pubertal gilts treated with 1.5 × (High) or 1.0 × (Low) PG600 ([400 IU PMSG + 200 IU hCG]/dose, AKZO‐NOBEL). Gilts expressing oestrus were artificially inseminated twice and maintained in breeding and gestation until the reproductive tract was collected on day 22 of pregnancy. At slaughter, the reproductive tracts from each pregnant gilt from each treatment were immediately processed to collect samples for RNA and protein analysis. Within each gilt, three conceptus points were sampled, one from each horn and then a random conceptus within the tract. At each conceptus point, endometrial attachment site, chorion–allantois and embryo were collected and immediately frozen in liquid nitrogen. Number of corpus luteum (CL) (35.4 vs. 12.6) and total embryo number (18.8 vs. 10.2) were greater in the high‐embryo compared to the low‐embryo group, respectively (p < .05). Real‐time qPCR results showed that Eph‐ephrin A1 mRNA expression was less in the high‐embryo (p < .05) compared to the low‐embryo group. In addition, Western blotting analysis indicated that Eph‐ephrin A1 and LEP protein expression at endometrial attachment site in high‐embryo was less (p < .05) compared to low‐embryo group. It was also noted that mRNA expression of Eph‐ephrin A1 and LEPR‐LEP was greater in pregnant than non‐pregnant gilts (p < .05). Moreover, mRNA expression of Eph‐ephrin A1 (p < .05) and LEPR‐LEP was greatest at endometrial attachment site among all three tissues. There was a positive correlation between expressions of Eph‐ephrin A1, LEPR‐LEP and embryo length with the correlation coefficient 0.31–0.59. For Eph‐ephrin A1, the highest correlation coefficient appeared between Eph A1 expression and normal embryo number, between ephrin A1 expression and embryo length. For LEPR‐LEP, the highest correlation coefficient appeared between LEPR‐LEP expression and ovary weight (0.79 for both, p < .05), followed by embryo length and weight. The results of this study suggest that low expression of Eph‐ephrin A1 and LEPR‐LEP is somehow related to increased embryo number during implantation and that endometrial attachment site might be the main target tissue of these gene products. Yet, the increased expression of Eph‐ephrin A1 and LEPR‐LEP appeared associated with increased embryo growth (length and weight) and ovary weight, Eph‐ephrin A1 and LEPR‐LEP might play roles in the regulation of embryo implantation in pigs.     

内毒素对大鼠小肠黏膜组织的影响及多价阳离子A的保护效应

旨在揭示大肠埃希菌内毒素(ET)对大鼠小肠黏膜的结构、绒毛长度、上皮内淋巴细胞(IEL)的数量和分布的影响,并探讨多价阳离子A(CA)对上述指标的保护效应。选用72只140g~150g SPF级SD大鼠随机分为3组,即对照组、ET组和CA保护组,经相应处理后分别在3、4、8、12h采集十二指肠、空肠组织作为检测样本,制备病理组织切片,HE染色并利用图像分析系统进行分析。ET组十二指肠和空肠绒毛长度在3、4、8、12h均显著低于对照组和CA保护组(P0.01),ET组十二指肠、空肠IEL数量在3、4、8、12h均显著低于对照组(P0.01);CA保护组十二指肠和空肠IEL数量在4、8、12h均显著高于ET组(P0.01)。结果显示,ET在不同程度上能够破坏小肠黏膜的正常组织结构,降低小肠绒毛长度,减少IEL的数量,从而影响小肠正常的吸收和免疫功能,而CA则能明显降低ET所导致的毒性作用,发挥其保护效应。     

干热河谷印楝树干液流密度动态变化特征研究

采用TDP技术对金沙江干热河谷元谋段主要造林树种印楝的树干液流密度动态变化进行了观测,得到3点主要结论:①无论干热季节或湿润季节,印楝树干液流密度昼夜变化的规律性较强,呈明显的单峰曲线;②由干热季节转向湿润季节时,印楝树干液流密度呈下降趋势,5年生印楝在5月、7月及10月的典型晴天里的树干液流密度日变化平均值分别为12.11、7.48及6.90cm3·cm-2·h-1;③在干热季节里印楝蒸腾耗水具有相当的被动性,而在湿润季节时其蒸腾耗水则表现出主动性和平氨性,反映了印楝在不同季节里的耗水机制差异.     

Effect of Psammaplin A on in vitro Development of Bovine Aging Oocytes

To investigate the effect of histone deacetylation inhibitor Psammaplin A (PsA) on the development of bovine aging oocytes in vitro,oocytes were randomly divided into control group,aging group and 50 mmol/L PsA treated aging group (PsA group).Immunofluorescence staining and JC-1 were used to detect the blastocyst rate of bovine oocytes after parthenogenetic activation,the number of cells in blastocysts,apoptosis,reactive oxygen species (ROS),glutathione (GSH) and mitochondrial membrane potential intensity of embryos.The results showed that the blastocyst rate of the aging group was significantly lower than that of PsA and control groups (P<0.05).The blastocyst rate of PsA group was not significantly different from that of control group (P>0.05).The number of cells in the blastocysts of control group and PsA group were significantly higher than that of aging group (P<0.05).The number of cells in the blastocysts of PsA group was not significantly different from that of control group (P>0.05).The apoptosis rate in aging group was significantly higher than that of control and PsA groups (P<0.05),the apoptosis rate of PsA group was significantly higher than that of control group (P<0.05).The GSH level of MⅡ oocytes in aging group was significantly lower than that of control and PsA groups (P<0.05).There was no significant difference in GSH level between control and PsA groups (P>0.05).The ROS level of the embryos in aging group was significantly higher than that of control and PsA groups (P<0.05).The ROS level in PsA group was significantly higher than that of control group (P<0.05).The mitochondrial membrane potential of early embryos of aging group 4-8 cells was significantly lower than that of control and PsA groups (P<0.05).The mitochondrial membrane potential intensity of control group was significantly higher than that of PsA group (P<0.05).In summary,PsA could effectively delay the aging of bovine oocytes and improve the quality of oocytes.     

质核互作型不育系阜CMS5A大豆的选育及应用

阐述了质核互作型不育系阜CMS5A大豆的选育经过,总结了其特征特性,并分析了其应用成效,以供参考。     

日粮中添加β-胡萝卜素对牦牛皮下脂肪颜色及组织中维生素A含量的影响

试验旨在探究日粮中添加不同水平β-胡萝卜素（β-C）对牦牛皮下脂肪颜色及组织维生素A含量的影响。试验选取12头2~3岁牦牛为研究对象，随机分为4个组，每组3个重复。各处理组日粮分别在基础日粮中添加0（对照）、720（低剂量）、1 440（中剂量）、1 620 mg/d（高剂量）β-C，饲养90 d后，分别测定牦牛脂肪、肌肉色度、背膘厚度及不同组织中β-C和代谢产物维生素A的含量，结果表明：日粮中添加不同水平β-C，牦牛背部、胸部脂肪及半腱肌的黄度值（b^*）显著高于对照组（P<0.05），亮度值（L^*）和红度值（a^*）没有显著变化（P>0.05）；牦牛背膘厚度在中剂量组达到最大值；对照组牦牛血清中β-C及维生素A含量显著低于各试验组（P<0.05），且维生素A含量随β-C添加量的上升呈上升趋势（P<0.05）；牦牛皮下脂肪、腹腔脂肪、心脏、肝脏、脾脏、肺脏、肾脏及空肠等组织器官中β-C含量在中剂量组均达到最大值，除肝脏中维生素A含量较高外，其他组织中维生素A含量在各处理组间差异不显著（P<0.05）。综上所述，日粮添加β-C影响牦牛皮下脂肪颜色，且在中剂量组牦牛皮下脂肪、心脏、肝脏、脾脏及空肠等组织器官中β-C含量达到最大值，故推荐其最佳摄入量为1 440 mg/d；在日粮中添加β-C影响牦牛肝脏和血清中维生素A含量，对其余组织器官中维生素A含量没有影响。     

维生素A调节动物一氧化氮的生成及其相关机制

动物机体由于旺盛的新陈代谢或在受到外界有害因素刺激的情况下会产生大量一氧化氮(NO),进而损伤细胞中的蛋白质、类脂膜和DNA,引发细胞炎症并阻断细胞信号通路,使机体产生氧化应激。维生素A可以有效地调控NO的生成,提高机体抗氧化水平并清除自由基,预防细胞炎症及氧化应激的发生。本文主要综述了维生素A对动物NO生成的调节作用及其相关机理的研究进展,对今后深入研究其调节机理、科学补充维生素A及提高机体的抗氧化功能具有一定参考价值。     

高粱光敏色素A基因的克隆、蛋白结构与长短日照诱导表达模式

光敏色素是一类红光/远红光受体,对植物光形态建成、外部形态和生理功能等方面起着重要的调节作用。为明晰光敏色素A在调控高粱光形态建成发育过程中的作用,本研究比较分析光温敏感程度不同的高粱品种Btx623、Rio、Tx430和LH645的PhyA基因序列和PHYA蛋白的保守结构域,利用qRT-PCR明确长短日照条件处理下PhyA基因的表达模式。结果表明,Btx623、Rio和Tx430之间PhyA基因序列存在非同义突变,LH645在PhyA基因的第7个外显子上插入了5 bp碱基序列;高粱PhyA蛋白中含有1个PAS-2结构域、1个GAF结构域、1个Phytochrome结构域、2个PAS结构域、1个组氨酸激酶A结构域和1个类似组氨酸激酶的ATP激酶结构;遗传进化树分析表明,单子叶和双子叶植物PhyA蛋白明显聚为2大类,高粱PhyA、谷子PhyA和玉米PhyA1、PhyA2聚为一个亚类,相互间亲缘关系较近,与水稻PhyA同源性相对较远;在短日照(SD)条件下各时期PhyA基因转录水平变化显著,Rio PhyA基因转录水平较Btx623和Tx430变化明显,LH645 PhyA基因转录变化不显著,在长日照(LD)条件下Rio PhyA基因转录水平低于Btx623和Tx430,LH645 PhyA基因转录水平变化不显著。以上结果表明,PhyA受光周期诱导,推测其在高粱光形态建成中起重要调控作用。本研究为进一步解析PhyA基因功能及其在调控高粱光形态建成发育过程中的作用提供理论基础。     

m6A甲基化在鸡肌肉生长发育中的表达研究

试验旨在研究RNA m6A修饰相关基因去甲基化酶Alk B同源蛋白5（Alk B homologue 5，ALKBH5）、去甲基化酶肥胖相关蛋白（fat mass and obesity-associated protein，FTO）、甲基转移酶样蛋白3（methyltransferase like 3，METTL3）、甲基转移酶样蛋白14（methyltransferase like 14，METTL14）和成肾细胞瘤1-结合蛋白（Wilms’tumor 1-associating protein，WTAP）在鸡骨骼肌发育过程中的表达，分析其与骨骼肌m6A甲基化水平的相关性。首先，利用实时荧光定量PCR技术检测m6A甲基化相关基因在金茅花鸡12（E12）、14（E14）、16（E16）、18（E18）胚龄和1日龄腿肌和胸肌组织中mRNA表达水平，以及其在鸡成肌细胞50%、100%增殖期和1、2、3、4、5 d分化期的mRNA表达水平；随后，利用m6A甲基化试剂盒检测金茅花鸡E12和1日龄腿肌和胸肌组织中m6A甲基化修饰水平，与m6A甲基化相关基因表达水平进行相关性分析。结果显示，m6A去甲基化基因ALKBH5和FTO mRNA表达水平在骨骼肌发育过程中显著上调（P<0.05），即在E12、E14低表达，E16、E18逐渐上调，1日龄达到最高。m6A甲基化写入基因METTL14、METTL3和WTAP mRNA表达水平在E12、E14、E16逐渐上升，E18下降，随后至1日龄表达量回升。在细胞增殖过程中，ALKBH5、FTO、METTL14、METTL3和WTAP基因表达均上调；在细胞分化过程中ALKBH5和FTO基因表达水平显著上调（P<0.05），在分化第5天达到最高。METTL14、METTL3和WTAP基因mRNA表达水平在细胞诱导分化的1、2、3、4 d表达量呈下降趋势，而在诱导分化的第5天有所回升。甲基化水平检测结果显示，腿肌和胸肌m6A甲基化水平变化趋势一致，均在胚胎发育过程中显著下降（P<0.05），至1日龄达到最低。相关性分析结果显示，鸡骨骼肌RNA m6A甲基化水平与m6A去甲基化修饰基因ALKBH5、FTO mRNA表达水平呈显著负相关（P<0.05）。综合以上试验结果，推测m6A甲基化修饰与鸡骨骼肌发育相关，而去甲基化基因ALKBH5、FTO可能通过调控RNA m6A甲基化水平，影响鸡骨骼肌发育。本研究结果为进一步研究m6A甲基化修饰调控鸡骨骼肌生长发育的功能和分子机制提供理论依据。