一氧化氮抑制血管紧张素Ⅱ和内皮素-1诱导的心肌细胞原癌基因c-fos表达

在原代培养的新生大鼠心肌细胞上, 探讨一氧化氮 (NO)对血管紧张素Ⅱ (AⅡ)和内皮素-1 (ET-1)诱导的心肌细胞肥大和原癌基因c-fos表达的影响.用Bradford 法测定心肌细胞总蛋白含量 (作为心肌细胞肥大的指标); 用基因特异性引物和 SuperScript一步法进行逆转录聚合酶链式反应 (RT-PCR), 检测大鼠心肌细胞原癌基因c-fos的表达 (以GAPDH为内标).结果显示, AⅡ和ET-1分别作用5 d和3 d后, 心肌细胞总蛋白含量显著增加; 硝普钠 (NO供体)可抑制AⅡ或ET-1诱导的心肌细胞总蛋白增加.AⅡ,ET-1和PMA (蛋白激酶C激动剂)均可诱导心肌细胞原癌基因c-fos的表达; L-精氨酸可抑制AⅡ,ET-1和PMA诱导心肌细胞原癌基因c-fos的表达, L-NAME (NOS抑制剂)可抑制L-精氨酸的这一作用; 硝普钠对可抑制AⅡ,ET-1和PMA诱导心肌细胞原癌基因c-fos的表达.结果表明, NO可抑制AⅡ或ET-1诱导的心肌细胞肥大和原癌基因c-fos表达, 其作用机制可能与蛋白激酶C这一环节有关.     

Structural Analysis of Peptides of PSⅡ Light-harvesting Complexes in Siphonous Green Algae,Codium fragile

Peptide composition and arrangement of 4 major light harvesting complexes LHCP1-3 and LHCP3′isolated from siphonous green algae （Codium fragile (Sur.) Hariot.） were investigated. LHCP1 showed five main peptides, 34.4, 31.5, 29.5, 28.2 and 26.5 kD in SDS PAGE, the 34.4 and 31.5 kD peptides were never found in higher plants. LHCP3 contained the other four kinds of LHCP1 peptides except 34.4 kD, while LHCP3′consisted of only 28.2 and 26.5 kD peptides. We found that 34.4, 28.2 and 26.5 kD peptides were easy to decompose from LHCP 1 when subjected to SDS PAGE without pretreatment. They might be located at the exterior of LHCP1, while the 31.5 and 29.5 kD peptides were at the central part. The 28.2 and 26.5 kD peptides often occurred in CPa, the center complex of PSⅡ. They are possibly the LHCⅡ peptides tightly associated with CCⅡ. According to the results described above, a peptide map of LHCP1 was sketched.     

Can intracellular signalling pathways predict developmental abnormalities?

Abstract

Sensitivity of the adenylyl cyclase/c-fos protooncogene cascade to β-adrenergic agonists and glucocorticoids in foetal rat

We examined whether measurements of adenylyl cyclase and its control of c-fos protooncogene mRNA expression in mid-gestation foetal rat could be used to detect developmental effects of apparently unrelated compounds: terbutaline, a β-adrenergic receptor stimulant, and dexamethasone, a glucocorticoid hormone. On gestational day 14, acute administration of terbutaline to pregnant rats resulted in sixfold induction of c-fos mRNA within 1 h; the same increase was obtained when a membrane-permeable analogue of cAMP was given. Treatment with dexamethasone on gestational days 11,12 and 13 produced the same increase in c-fos mRNA on gestational day 14 as had been seen with acute terbutaline or CAMP; no further increase could be obtained with acute CAMP treatment in the dexamethasone-pretreated animals. Adenylyl cyclase activity was evaluated on gestational day 14. Animals treated with dexamethasone showed a 50% enhancement of basal enzyme activity that reflected a parallel increase in total catalytic activity as measured with forskolin-Mn²⁺. Dexamethasone also increased adenylyl cyclase activity in the presence of a β-agonist but to a lesser extent than the increase in total catalytic activity. These results indicate that cell signalling pathways mediating the expression of the genes that control cell differentiation are a likely target for structurally and mechanistically unrelated drugs and chemicals and may therefore be useful as early biomarkers of abnormal development.     

人膀胱癌中某些癌基因的研究

本实验应用Northern和斑点印渍杂交技术探测了人膀胱癌细胞株中c-myc、c-fos、erbB等癌基因的表达,以及TPA对这些癌基因表达的调控,发现BIU-87细胞有这些癌基因的表达,并能被TPA所增强,同时也发现人膀胱癌组织有c-myc、c-fos、erbB、N-ras基因的高表达。提示蛋白激酶C的激活可以诱导某些癌基因的表达。多种癌基因的表达异常可能在膀胱癌中起重要作用。     

Characterization of the Responses of Purkinje Cells to Neurotrophin Treatment

Abstract: The ability of the neurotrophins nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and neurotrophin-4/5 (NT-4/5) to promote neuronal survival and phenotypic differentiation was examined in dissociated cultures from embryonic day 16 rat cerebellum. BDNF treatment increased the survival of neuron-specific enolase-immunopositive cells by 250 and 400% after 8 and 10 days in culture, respectively. A subpopulation of these neurons, the Purkinje cells, identified by calbindin staining, was increased to an equivalent extent, ∼200%, following BDNF, NT-4/5, or NT-3 treatment. The number of GABAergic neurons, identified by GABA immunoreactivity, was greatly increased by treatment with BDNF (470%) and moderately by NT-4/5 (46%), whereas NT-3 was without effect. NGF failed to increase the number of either Purkinje cells or GABAergic neurons. Addition of BDNF within 48 h of cell plating was required to obtain a maximal increase in Purkinje cell number after 8 days. In contrast, the NT-3 responses were nearly equivalent even if treatment was delayed for 96 h after plating. BDNF, NT-4/5, and NT-3, but not NGF, induced the rapid expression of the immediate early gene c- fos . Immunocytochemical double-labeling with antibodies to c-fos and calbindin was used to identify Purkinje cells that responded to neurotrophin treatment by induction of c-fos. After 4 days in vitro, both BDNF and NT-3 induced the formation of c-fos protein in calbindin-immunopositive neurons, whereas NT-4/5 did not. The latter results suggest that although BDNF and NT-4/5 have been shown to act through a common receptor, TrkB, it appears that the effects of BDNF and NT-4/5 are not identical.     

反义c－fos和c－jun抑制IL－1诱导阿片肽分泌

白细胞介素-1(IL-1)及阿片肽作为神经调质参与了神经细胞兴奋性毒性作用.以大鼠大脑皮层神经细胞为研究对象,探讨了IL-1、阿片肽和c-fos、c-jun表达产物之间的关系.结果表明,IL-1β能诱导大脑皮层神经细胞c-fos、c-jun mRNA瞬时短暂表达,15min增高,30min达高峰,c-fos mKNA 2h回至基线水平,c-jun mRNA 8h回至基线水平;联合应用c-fos、c-jun反义寡核苷酸能部分抑制IL-1诱导的大脑皮层神经细胞脑啡肽及β-内啡肽分泌增加,呈一定量效关系,相应意义寡核苷酸无抑制作用.提示IL-1促进大脑皮层神经细胞脑啡肽及β-内啡肽分泌作用部分受Fos和Jun蛋白调控.     

电针刺激下即刻早期基因和前脑啡肽基因在中枢神经系统中的表达及定位研究

采用免疫组织化学、Ｎｏｒｔｈｅｒｎ杂交与原位杂交等方法,观察到２／１５Ｈｚ电针刺激可诱导大鼠中枢神经系统内即刻早期基因ｃ－ｆｏｓ、ｃ－ｊｕｎ的ｍＲＮＡ与免疫阳性物质以及前脑啡肽ｍＲＮＡ的生成。Ｎｏｒｔｈｅｒｎ杂交显示,电针在１－２ｈ内引起即刻早期基因ｃ－ｆｏｓｍＲＮＡ的表达,继之以ＰＥＮＫ ｍＲＮＡ的表达,在４８ｈ内方达顶点;形态学结果表明,电针诱导ｃ－ｆｏｓ、ｃ－ｊｕｎ与ＰＥＮＫ ｍＲＮＡ及其蛋