Niemann-Pick type C2 (NPC2) is a type of small soluble protein involved in lipid metabolism and triglyceride accumulation in vertebrates and arthropods. Recent stud- ies have determined that NPC2 also participates i...Niemann-Pick type C2 (NPC2) is a type of small soluble protein involved in lipid metabolism and triglyceride accumulation in vertebrates and arthropods. Recent stud- ies have determined that NPC2 also participates in chemical communication of arthropods. In this work, two novel NPC2 proteins (MmedNPC2a and MmedNPC2b) in Microplitis mediator were identified. Real-time quantitative PCR (qPCR) analysis revealed that Mmed- NPC2a was expressed higher in the antennae than in other tissues of adult wasps compared with MmedNPC2b. Subsequent immunolocalization results demonstrated that NPC2a was located in the lymph cavities of sensilla placodea. To further explore the binding charac- terization of recombinant MmedNPC2a to 54 candidate odor molecules, a fluorescence binding assay was performed. It was found MmedNPC2a could not bind with selected fatty acids, such as linoleic acid, palmitic acid, stearic acid and octadecenoic acid. How- ever, seven cotton volatiles, 4-ethylbenzaldehyde, 3,4-dimethylbenzaldehyde, fl-ionone, linalool, m-xylene, benzaldehyde and trans-2-hexen-l-al showed certain binding abilities with MmedNPC2a. Moreover, the predicted 3D model of MmedNPC2a was composed of seven r-sheets and three pairs of disulfide bridges. In this model, the key binding residues for oleic acid in CjapNPC2 of Camponotus japonicus, Lue68, Lys69, Lys70, Phe97, Thr103 and Phe127, are replaced with Phe85, Ser86, His87, Leu113, Tyr119 and Ile143 in MmedNPC2a, respectively. We proposed that MmedNPC2a in M. mediator may play roles in perception of plant volatiles.展开更多
AMP-activated kinase (AMPK), a protein that regulates energy balance and metabolism, has recently been identified in boar spermatozoa where regulates key functional sperm processes essential for fertilization. This ...AMP-activated kinase (AMPK), a protein that regulates energy balance and metabolism, has recently been identified in boar spermatozoa where regulates key functional sperm processes essential for fertilization. This work's aims are AMPK identification, intracellular localization, and their role in human spermatozoa function. Semen was obtained from healthy human donors. Sperm AMPK and phospho-Thr172-AMPK were analyzed by Western blotting and indirect immunofluorescence. High- and low-quality sperm populations were separated by a 40%-80% density gradient. Human spermatozoa motility was evaluated by an Integrated Semen Analysis System (ISAS) in the presence or absence of the AMPK inhibitor compound C (CC). AMPK is localized along the human spermatozoa, at the entire acrosome, midpiece and tail with variable intensity, whereas its active form, phospho-Thr172-AMPK, shows a prominent staining at the acrosome and sperm tail with a weaker staining in the midpiece and the postacrosomal region. Interestingly, spermatozoa bearing an excess residual cytoplasm show strong AMPK staining in this subcellular compartment. Both AMPK and phospho-Thr172-AMPK human spermatozoa contents exhibit important individual variations. Moreover, active AMPK is predominant in the high motility sperm population, where shows a stronger intensity compared with the low motility sperm population. Inhibition of AMPK activity in human spermatozoa by CC treatment leads to a significant reduction in any sperm motility parameter analyzed: percent of motile sperm, sperm velocities, progressivity, and other motility coefficients. This work identifies and points out AMPK as a new molecular mechanism involved in human spermatozoa motility. Further AMPK implications in the clinical efficiency of assisted reproduction and in other reproductive areas need to be studied.展开更多
β-连环蛋白(CTNNB1,β-catenin)是一种存在于细胞膜、细胞质和细胞核的多种功能性胞内蛋白,在动物胚胎发育和器官发生中起着重要作用。为了研究β-catenin在淇河鲫(Carassius auratus in Qihe river)体内各组织中的表达模式,本实验用...β-连环蛋白(CTNNB1,β-catenin)是一种存在于细胞膜、细胞质和细胞核的多种功能性胞内蛋白,在动物胚胎发育和器官发生中起着重要作用。为了研究β-catenin在淇河鲫(Carassius auratus in Qihe river)体内各组织中的表达模式,本实验用免疫组化法检测β-catenin在孵化后10天淇河鲫的眼、脑、鳃、软骨、肠、肝、脾、肾和孵化后120天的卵巢组织的表达情况。结果显示,β-catenin在肠的杯状细胞,肾小管上皮细胞,鳃小片的泌氯细胞,肝组织的肝细胞,脾脏的巨噬细胞,脑组织的中脑和间脑的支持细胞,卵巢组织的Ⅰ期和Ⅱ期卵母细胞,软骨的软骨基质和软骨膜以及眼组织的全视网膜、色素层和晶状体中表达。以上结果暗示,淇河鲫β-catenin可能参与了器官的发育过程。这些数据将为了解β-catenin调控硬骨鱼器官发生的分子机制提供参考。展开更多
PPF1 is a vegetative growth related gene that encodes a putative membrane protein having high homology with Arabidopsis chloroplast thylakoid protein ALB3. Immunoelectron microscopic assay showed that PPF1 was mainly ...PPF1 is a vegetative growth related gene that encodes a putative membrane protein having high homology with Arabidopsis chloroplast thylakoid protein ALB3. Immunoelectron microscopic assay showed that PPF1 was mainly localized in the thylakold membrane and was highly expressed in well-developed chloroplasts of short day (SD) grown G2 pea while having a very low abundance in chloroplasts of long day (LD) grown plants two weeks after flowering. Comparison of the leaf senescence processes in transgenic Arabidopsis and wild type plants revealed that overexpression of PPF1 delayed leaf senescence, while the depression of its Arabidopsts homologue (ALB3) with PPF1 antisense mRNA accelerated leaf senescence obviously. Ultrastructural analysis of transgenic Arabidopsis plants showed that when PPF1 was overexpressed in Arabidopsis, the chloroplasts were bigger and had much more grana and stroma thylakoid membranes than those of wild type plants. On the contrary, when PPF1 was expressed in antisense orientation to reduce the level of PPF1 homologue in Arabidopsis, the transgenic plants had smaller chloroplasts With less grana. and poorly developed thylakoid membrane systems. These results suggested that the developmental status of chloroplasts was positively correlated with the level of PPF1 or its Arabidopsts homologue, ALB3. Our results suggested that PPF1 gene might regulate plant development by controlling chloroplast development.展开更多
Intercellular connections and communications play very important roles during organisms growth and development. Although plasmodesmata and gap junction as intercellular channels are different in anatomy and structure,...Intercellular connections and communications play very important roles during organisms growth and development. Although plasmodesmata and gap junction as intercellular channels are different in anatomy and structure, they exhibit striking similarities in their functions. It is suggested that they have some similar biochemical regulation mechanism. Connexins are the transfer of cytoplasmic material by conformational changes.展开更多
Abstract: Arabinogalactan proteins (AGPs) and pectins were detected in the floral buds of cucumber (Cucumis sativus L.) during its sex determination using the following monoclonal antibodies: MAC 207 (recognizes AGP e...Abstract: Arabinogalactan proteins (AGPs) and pectins were detected in the floral buds of cucumber (Cucumis sativus L.) during its sex determination using the following monoclonal antibodies: MAC 207 (recognizes AGP epitopes); JIM 8 (recognizes a subset of AGP epitopes); and JIM 5 and JIM 7 (epitopes of pectins esterified to various degrees). In the stem apex meristem (SAM) of the cucumber, epitopes of MAC 207, JIM 7, and JIM 5 were localized in the cells from second to third peripheral layers when the sex organ primodium began to differentiate; epitopes of MAC 207 and JIM 5 were also detected in the ragged edge cells. A very dense labeling signal with MAC 207 was observed in the carpel and pistil primodium. The AGP epitopes recognized by JIM 8 were localized in the anther of the male flower and the anther-like portion of the stagnant stamen of the female flower. This suggests that the AGPs and pectins in the SAM of the cucumber are closely associated with the differentiation of the SAM, from meristematic cells to floral primodium. The subset of AGPs recognized by JIM 8 may play an important role in stamen formation.展开更多
[Objective] To observe the distribution of hepatitis E virus (HEV) in livestock in Hailongjiang Province. [Method] The serum samples collected from two regions of Heilongjiang Province included 719 samples from pigs...[Objective] To observe the distribution of hepatitis E virus (HEV) in livestock in Hailongjiang Province. [Method] The serum samples collected from two regions of Heilongjiang Province included 719 samples from pigs older than 3 months old, 840 samples from pigs younger than 3 months old, 505 bovine serum samples and 515 ovine serum samples. The samples were detected by enzyme-linked immunosorbent assay ( ELISA). [ Result] The pigs had the highest level of antibodies against HEV, and the positive rate of HEV was 89.71%, indicating pigs were more likely to be infected by HEV than other livestock populations. HEV was detected in 59 porcine serum samples, three bovine serum samples and one ovine serum sample collected from Heilongjiang Province. The positive rate of HEV in pigs was higher in Heilongjiang Province than in Jinlin Province and Liaoning Province. [ Conclusion] HEV infection was wide in pigs in Heilongjiang Province, and the infection rate was higher in the old pigs than in the young pigs.展开更多
基金This work was supported by the China National ‘973' Basic Research Program (2012CBl14104) and the Na- tional Natural Science Foundation of China (31272048, 31471778, and 31672038).
文摘Niemann-Pick type C2 (NPC2) is a type of small soluble protein involved in lipid metabolism and triglyceride accumulation in vertebrates and arthropods. Recent stud- ies have determined that NPC2 also participates in chemical communication of arthropods. In this work, two novel NPC2 proteins (MmedNPC2a and MmedNPC2b) in Microplitis mediator were identified. Real-time quantitative PCR (qPCR) analysis revealed that Mmed- NPC2a was expressed higher in the antennae than in other tissues of adult wasps compared with MmedNPC2b. Subsequent immunolocalization results demonstrated that NPC2a was located in the lymph cavities of sensilla placodea. To further explore the binding charac- terization of recombinant MmedNPC2a to 54 candidate odor molecules, a fluorescence binding assay was performed. It was found MmedNPC2a could not bind with selected fatty acids, such as linoleic acid, palmitic acid, stearic acid and octadecenoic acid. How- ever, seven cotton volatiles, 4-ethylbenzaldehyde, 3,4-dimethylbenzaldehyde, fl-ionone, linalool, m-xylene, benzaldehyde and trans-2-hexen-l-al showed certain binding abilities with MmedNPC2a. Moreover, the predicted 3D model of MmedNPC2a was composed of seven r-sheets and three pairs of disulfide bridges. In this model, the key binding residues for oleic acid in CjapNPC2 of Camponotus japonicus, Lue68, Lys69, Lys70, Phe97, Thr103 and Phe127, are replaced with Phe85, Ser86, His87, Leu113, Tyr119 and Ile143 in MmedNPC2a, respectively. We proposed that MmedNPC2a in M. mediator may play roles in perception of plant volatiles.
文摘AMP-activated kinase (AMPK), a protein that regulates energy balance and metabolism, has recently been identified in boar spermatozoa where regulates key functional sperm processes essential for fertilization. This work's aims are AMPK identification, intracellular localization, and their role in human spermatozoa function. Semen was obtained from healthy human donors. Sperm AMPK and phospho-Thr172-AMPK were analyzed by Western blotting and indirect immunofluorescence. High- and low-quality sperm populations were separated by a 40%-80% density gradient. Human spermatozoa motility was evaluated by an Integrated Semen Analysis System (ISAS) in the presence or absence of the AMPK inhibitor compound C (CC). AMPK is localized along the human spermatozoa, at the entire acrosome, midpiece and tail with variable intensity, whereas its active form, phospho-Thr172-AMPK, shows a prominent staining at the acrosome and sperm tail with a weaker staining in the midpiece and the postacrosomal region. Interestingly, spermatozoa bearing an excess residual cytoplasm show strong AMPK staining in this subcellular compartment. Both AMPK and phospho-Thr172-AMPK human spermatozoa contents exhibit important individual variations. Moreover, active AMPK is predominant in the high motility sperm population, where shows a stronger intensity compared with the low motility sperm population. Inhibition of AMPK activity in human spermatozoa by CC treatment leads to a significant reduction in any sperm motility parameter analyzed: percent of motile sperm, sperm velocities, progressivity, and other motility coefficients. This work identifies and points out AMPK as a new molecular mechanism involved in human spermatozoa motility. Further AMPK implications in the clinical efficiency of assisted reproduction and in other reproductive areas need to be studied.
文摘β-连环蛋白(CTNNB1,β-catenin)是一种存在于细胞膜、细胞质和细胞核的多种功能性胞内蛋白,在动物胚胎发育和器官发生中起着重要作用。为了研究β-catenin在淇河鲫(Carassius auratus in Qihe river)体内各组织中的表达模式,本实验用免疫组化法检测β-catenin在孵化后10天淇河鲫的眼、脑、鳃、软骨、肠、肝、脾、肾和孵化后120天的卵巢组织的表达情况。结果显示,β-catenin在肠的杯状细胞,肾小管上皮细胞,鳃小片的泌氯细胞,肝组织的肝细胞,脾脏的巨噬细胞,脑组织的中脑和间脑的支持细胞,卵巢组织的Ⅰ期和Ⅱ期卵母细胞,软骨的软骨基质和软骨膜以及眼组织的全视网膜、色素层和晶状体中表达。以上结果暗示,淇河鲫β-catenin可能参与了器官的发育过程。这些数据将为了解β-catenin调控硬骨鱼器官发生的分子机制提供参考。
文摘PPF1 is a vegetative growth related gene that encodes a putative membrane protein having high homology with Arabidopsis chloroplast thylakoid protein ALB3. Immunoelectron microscopic assay showed that PPF1 was mainly localized in the thylakold membrane and was highly expressed in well-developed chloroplasts of short day (SD) grown G2 pea while having a very low abundance in chloroplasts of long day (LD) grown plants two weeks after flowering. Comparison of the leaf senescence processes in transgenic Arabidopsis and wild type plants revealed that overexpression of PPF1 delayed leaf senescence, while the depression of its Arabidopsts homologue (ALB3) with PPF1 antisense mRNA accelerated leaf senescence obviously. Ultrastructural analysis of transgenic Arabidopsis plants showed that when PPF1 was overexpressed in Arabidopsis, the chloroplasts were bigger and had much more grana and stroma thylakoid membranes than those of wild type plants. On the contrary, when PPF1 was expressed in antisense orientation to reduce the level of PPF1 homologue in Arabidopsis, the transgenic plants had smaller chloroplasts With less grana. and poorly developed thylakoid membrane systems. These results suggested that the developmental status of chloroplasts was positively correlated with the level of PPF1 or its Arabidopsts homologue, ALB3. Our results suggested that PPF1 gene might regulate plant development by controlling chloroplast development.
基金Project supported in part by the China National Natural Science Foundation for Young Scientists.
文摘Intercellular connections and communications play very important roles during organisms growth and development. Although plasmodesmata and gap junction as intercellular channels are different in anatomy and structure, they exhibit striking similarities in their functions. It is suggested that they have some similar biochemical regulation mechanism. Connexins are the transfer of cytoplasmic material by conformational changes.
文摘Abstract: Arabinogalactan proteins (AGPs) and pectins were detected in the floral buds of cucumber (Cucumis sativus L.) during its sex determination using the following monoclonal antibodies: MAC 207 (recognizes AGP epitopes); JIM 8 (recognizes a subset of AGP epitopes); and JIM 5 and JIM 7 (epitopes of pectins esterified to various degrees). In the stem apex meristem (SAM) of the cucumber, epitopes of MAC 207, JIM 7, and JIM 5 were localized in the cells from second to third peripheral layers when the sex organ primodium began to differentiate; epitopes of MAC 207 and JIM 5 were also detected in the ragged edge cells. A very dense labeling signal with MAC 207 was observed in the carpel and pistil primodium. The AGP epitopes recognized by JIM 8 were localized in the anther of the male flower and the anther-like portion of the stagnant stamen of the female flower. This suggests that the AGPs and pectins in the SAM of the cucumber are closely associated with the differentiation of the SAM, from meristematic cells to floral primodium. The subset of AGPs recognized by JIM 8 may play an important role in stamen formation.
基金supported by the Foundation of Jilin Provincial Science and Technology Department (20100950)
文摘[Objective] To observe the distribution of hepatitis E virus (HEV) in livestock in Hailongjiang Province. [Method] The serum samples collected from two regions of Heilongjiang Province included 719 samples from pigs older than 3 months old, 840 samples from pigs younger than 3 months old, 505 bovine serum samples and 515 ovine serum samples. The samples were detected by enzyme-linked immunosorbent assay ( ELISA). [ Result] The pigs had the highest level of antibodies against HEV, and the positive rate of HEV was 89.71%, indicating pigs were more likely to be infected by HEV than other livestock populations. HEV was detected in 59 porcine serum samples, three bovine serum samples and one ovine serum sample collected from Heilongjiang Province. The positive rate of HEV in pigs was higher in Heilongjiang Province than in Jinlin Province and Liaoning Province. [ Conclusion] HEV infection was wide in pigs in Heilongjiang Province, and the infection rate was higher in the old pigs than in the young pigs.