颈动脉注射腺苷对去缓冲神经大鼠延髓腹外侧头端区神经元放电的影响

在28只切断双侧缓冲神经的Sprague－Dawley大鼠,应用细胞外记录方法,观察了72个自发放电单位中颈动脉注射腺苷对延髓腹外侧头端（RVLM）区神经元自发放电活动的影响。所得结果如下：（1）颈动脉注射腺苷（25μg/kg）,31个单位的放电频率由23．5±3．0下降至（16．5±2．6）spikes／s（P＜0．001）,血压和心率无明显变化（P＞0．05）;（2）在24个单位中,应用非选择性腺苷受体拮抗剂8-苯茶碱（8-phenyltheophylline,15μg/kg）和选择性腺苷A1受体持抗剂8-环戊-1,3-二丙基黄嘌呤（8－cyclopentyl-1,3－dipropylxanthine,50μg/kg）均可完全阻断腺苷的抑制效应;（3）在应用ATP敏感性钾通道阻断剂格列苯脲（500μg/kg）的12个单位中,腺苷的上述效应亦被消除。以上结果提示,腺苷对RVLM区神经元自发放电有抑制作用,而此作用与A1受体介导的ATP敏感性钾通道开放有关。     

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在45只切断双侧缓冲神经的Sprague-Dawley 大鼠,应用细胞外记录方法, 观察了颈动脉内注射腺苷对76 个最后区 (AP) 神经元自发放电活动的影响。所得结果如下(1) 在记录到的42个自发放电单位中, 颈动脉内注射腺苷(25 μg/kg) 引起其中29个单位的放电频率由6.26±0.75 下降至4.74±0.76 spikes/s (P<0.01), 6 个单位放电频率由4.13±0.77增加至4.72±0.83 spikes/s (P<0.05),另外7个单位放电频率无明显变化, 而血压和心率在实验中无变化; (2)在应用非选择性腺苷受体拮抗剂8-苯茶碱(8-phenyltheophylline, 15 μg/kg) 的10个单位, 腺苷对放电的抑制效应可被完全阻断; (3) 应用选择性腺苷A     

颈动脉内注射腺苷对去缓冲神经大鼠最后区神经元放电的影响

在45只切断双侧缓冲神经的SpragueDawley大鼠,应用细胞外记录方法,观察了颈动脉内注射腺苷对76个最后区(AP)神经元自发放电活动的影响。所得结果如下:(1)在记录到的42个自发放电单位中,颈动脉内注射腺苷(25μg/kg)引起其中29个单位的放电频率由626±075下降至474±076spikes/s(P<001),6个单位放电频率由413±077增加至472±083spikes/s(P<005),另外7个单位放电频率无明显变化,而血压和心率在实验中无变化;(2)在应用非选择性腺苷受体拮抗剂8苯茶碱(8phenyltheophylline,15μg/kg)的10个单位,腺苷对放电的抑制效应可被完全阻断;(3)应用选择性腺苷A1受体拮抗剂8环戊1,3二丙基黄嘌呤(8cyclopentyl1,3dipropylxanthine,50μg/kg)亦可有效地阻断腺苷对12个单位的抑制效应;(4)应用ATP敏感性钾通道阻断剂格列苯脲(500μg/kg)的12个单位,腺苷的上述效应也被消除。以上结果提示,腺苷对AP区神经元自发放电有抑制作用,而此作用与A1受体介导的ATP敏感性钾通道开放有关。     

pH降低和尼古丁对家兔颈动脉体化学感受器活动的影响

在29只家兔的颈动脉体-窦神经标本,记录了55个窦神经中化学感受性传入单位的电活动。结果如下：（1）灌流液pH降低时,所有单位的放电均增加,频率由0．86±0．21增至1．75±040imp／s（P＜0.001）。（2）在灌洗液中加入尼古丁（1,3,6,10和15μg/ml）后,10个单位的自发放电频率,由0．60±0.21分别增加到0．96±0．21,1．19±0.30,1．24±0．29,1．48±0．41和1．82±0．39imp／s,呈明显的剂量依赖性肝（r=0．94,P＜O．001）。简箭毒可阻断尼古丁的作用,但对自发放电无显著影响。（3）当以酸性Locke氏溶液灌流标本时,19个单位的放电频率由0.95±0．34增至1.84±0．55imp／s;在酸性灌流液中加入尼古丁后,放电频率由0.96±0．25增至1．53±0．24imp／s;与单纯酸刺激时相比,放电频率增加幅度减小（P＜0．05）。（4）17个自发放电频率为1.08±0．33imp／s的单位,换以酸性Locke氏液灌流后,放电频率增加到1．64±0．43imp／s;加筒箭毒后,由对照的1.01±0.34增至1．70±0．43imp／s;灌流液pH降低时筒箭毒对化学感受性单位放电频率的变化无显著影响。上述结果提示：（1）ACh在颈动脉体化学感受性功能方面可能仅起调质的作用。（2）pH降低时颈动脉体化学感受性功能活动增强与N-型胆碱能受体     

腺苷易化大鼠颈动脉窦压力感受器的活动

在36只麻醉大鼠,以隔离灌流颈动脉窦区记录窦神经传入放电的方法观察了腺苷（adenosine,Ado）对颈动脉窦压力感受器传入放电的影响。所得结果如下：（1）以75μmol／LAdo隔离灌流大鼠左侧颈动脉窦区时,窦内压-窦神经传入放电积分（ISP－ISNA）关系曲线向左上方移位,曲线最大斜率（PS）由（18．75±0．12）％／kPa增至（22．21±0．11）％／kPa（P＜0．001）,最大积分值（PIV）由（209．83±2．57）％增至（239．17±1．75）％;阈压（TP）和饱和压（SP）则分别从（8．57±0．24）和（22．99±0．34）下降至（7．15±0．23）kPa（P＜0．001）和（21．21±0,43）kPa（P＜0．01）。再分别以125和175μmol／LAdo灌流,机能曲线进一步向左上方移位,PS、TP和SP的变化均呈明显的剂量依赖性。（2）用腺苷选择性A1受体拮抗剂8－cyclopentyl－1,3－dipropylxanthine（0．134mmol／L）预处理后,Ado的上述效应即被阻断。（3）先给予KATP通道阻断剂格列苯脲（10μmol／L）亦可取消腔苷对窦神经传入放电的影响。结果表明,在体隔离灌流大鼠颈动脉窦区条件下,Ado对颈动脉窦压力感受器活动有易化作用,此作用似与腺苷A1受体介导的KATP通道开放有关。     

腺苷抑制海马神经元自发放电和谷氨酸所致癫痫样放电

应用细胞外记录单位放电技术,在大鼠海马脑片上观察腺苷（ａｄｃｎｏｓｉｎｅ,Ａｄｏ）对ＣＡ１区神经元自发和谷氨酸所致癫痫样放电的影响。实验结果如下：⑴２０个海刀ＣＡ１神经元在给予Ａｄｏ（０．０１－０．１μｍｏｌ／Ｌ）时自发放电频率降低,且呈明显的剂量依赖性;⑵在２２个ＣＡ１单位,应用腺苷受体非选择性拮抗剂８－苯茶碱（８－ｐｈｅｎｙｌ－ｔｈｅｏｐｈｙｌｌｉｎｅ,８－ＰＴ,０．５ｍｍｏｌ／Ｌ）和腺苷Ａ１     

心室内注射腺苷对肾交感神经传出活动的影响

在切断两侧缓冲神经和迷走神经的麻醉大鼠,观察心室内注射腺苷及其同系物对肾交感神经传出活动的影响。心室内冲击注射腺昔（0．5μmol／kg,0．1ml）时,肾交感神经传出放电（RSNA）增加41．9±6．08％（P＜0．001）,并引起平均动脉血压（MAP）先短暂升高1．39±0．19kPS（P＜0．001）和随后下降3．74±0．64kPa（P＜0．001）,以及心率（HR）减慢95±14bpm（P＜0．001）。为了确定何种受体亚型介导腺苷所致RSNA增加,又分别应用了选择性腺昔A1受体激动剂[（-）-N6－（2－Phenylisopropyl）adenosine,R－PIA]和A2受体激动剂[5＇－（N－ethylcarboxamido）adenosine,NECA]。无论R－PIA（0．05μmol／kg,0．1ml）或NECA（0．05μmol／kg,0．1ml）均使MAP下降和HR减慢,且作用持续时间较腺苷的明显延长;R－PIA使RSNA增加31．6±5．21％（P＜0．001）,但NECA对RSNA无明显影响。选择性腺苷A1受体拮抗剂（8－cyclopentyl－1,3－dipropylxanthine,DPCPX）可完全抑制腺苷对RSNA的兴奋效应。切除双侧星状神经节后,腺苷兴奋RSNA的效应消失。以上结果提示,腺苷可通过A1受体激活心交感神经传入纤维,反射地增强肾交感神经的传出活动。     

辣椒素对大鼠延髓腹外侧头端区神经元电活动的影响

在35只切断两侧缓冲神经的麻醉大鼠,应用细胞外记录的电生理学方法,观察颈总动脉注射辣椒素(capsaicin)对延髓腹外侧头端区(RVLM)巨细胞旁外侧核(PGL)自发电活动的影响。所得结果如下:(1)颈动脉注射辣椒素(10μmol,01ml),MAP由1074±013升至1256±021kPa(P<0001);HR由374±4增至395±5bpm(P<0001);30个PGL神经元自发放电单位的放电频率由126±07增至209±11spikes/s(P<0001)。(2)在10个放电单位,应用辣椒素受体阻断剂钌红(rutheniumred;200mmol,01ml)后,明显抑制辣椒素的上述效应。以上结果提示,辣椒素可能通过激活RVLM神经元上的辣椒素受体,进而兴奋PGL神经元     

帕金森病大鼠中缝背核5-羟色胺能神经元电活动的变化

本实验采用玻璃微电极细胞外记录法,观察了帕金森病(Parkinson’s disease,PD)大鼠中缝背核(dorsal raphe nucleus, DRN)5-羟色胺(5-hydroxytryptamine,5-HT)能神经元电活动的变化。在大鼠右侧中脑黑质致密部内微量注射6-羟多巴胺(6- hydroxydopamine,6-OHDA)制作PD模型。结果显示,对照组和PD组大鼠DRN中5-HT能神经元的放电频率分别是(1．76±0．11)spikes／s(n=24)和(2．43±0．17)spikes／(n=21),PD组大鼠的放电频率显著高于对照组(P<0．001)。在对照组大鼠,92％(22／24)的神经元呈规则放电,8％(2／24)为爆发式放电;在PD组大鼠,具有规则、不规则和爆发式放电的神经元比例分别为9％(2／21)、43％(9／21)和48％(10／21),爆发式放电的5-HT能神经元比例明显高于对照组(P<0．001)。在对照组大鼠,DRN内局部注射5-HT1A拮抗剂WAY-100635(3μg/200nL)显著增加5-HT能神经元的放电频率而不影响其放电形式(n=19,P<0．002);而WAY-100635不改变PD组大鼠5-HT能神经元的放电频率和放电形式(n=17,P>0．05)。结果提示,用6-OHDA损毁黑质致密部造成的PD模型大鼠中神经元5-HT1A受体功能失调,并且DRN参与PD的病理生理学机制。     

肾动脉内注射腺苷兴奋肾神经传入纤维的自发活动

应用记录肾神经传入纤维多单位和单位放电的方法,观察肾动脉内注射腺苷对麻醉家兔肾神经传入纤维自发放电活动的影响。结果表明：(1)肾动脉内注射50,100和200nmol／kg腺苷可呈剂量依赖性地兴奋肾神经传入纤维的活动,而动脉血压不变。(2)肾动脉内预先应用选择性腺苷A1受体阻断剂DPCPX(160nmol/kg),可部分阻断腺苷对肾神经传入纤维的兴奋作用。(3)静脉应用一氧化氮合酶抑制剂L-NAME(0．1mmol／kg)预处理,延长并增强了肾神经传入纤维对腺苷的反应。以上结果提示,肾动脉内应用腺苷可兴奋肾传入纤维的自发放电活动,一氧化氮作为抑制性因素参与腺苷诱导的肾神经传入纤维兴奋。     

腺苷对缺氧时海马神经元内游离Ca^2＋的影响

实验用Ｆｌｕｏ－３负载细胞,在激光扫描共聚焦显微镜下直接监测缺氧后分散培养的大鼠海马ＣＡ１区神经元内游离Ｃａ＾２＋浓度（［Ｃａ＾２＋］ｉ）的变化,观察腺苷对这种变化的影响并初步探讨其作用机制。结果发现,急性缺氧使海马神经元［Ｃａ＾２＋］ｉ显著升高;腺苷（１００μｍｏｌ／Ｌ）明显抑制缺氧引起的［Ｃａ＾２＋］ｉ增高,腺苷Ａ１受体拮抗剂ＣＰＴ以及Ｋ＾＋通道阻断剂４－ＡＰ和ＡＴＰ敏感性Ｋ＾＋通道阻断剂ｇｌ     

F1F0-ATPase from Escherichia coli with mutant F0 subunits. Partial purification and immunoprecipitation of F1F0 complexes

Previously identified mutations in subunits a and b of the F0 sector of the F1F0-ATPase from Escherichia coli are further characterized by isolating detergent-solubilized, partially purified F1F0 complexes from cells bearing these mutations. The composition of the various F1F0 complexes was judged by quantitating the amount of each subunit present in the detergent-solubilized preparations. The composition of the F0 sectors containing altered polypeptides was determined by quantitating the F0 subunits that were immunoprecipitated by antibodies directed against the F1 portion. In this way, the relative amounts of F0 subunits (a, b, c) which survived the isolation procedure bound to F1 were determined for each mutation. This analysis indicates that both missense mutations in subunit a (aser206----leu and ahis245----tyr) resulted in the isolation of F1F0 complexes with normal subunit composition. The nonsense mutation in subunit a (atyr235----end) resulted in isolation of a complex containing the b and c subunits. The bgly131----asp mutation in the b subunit results in an F0 complex which does not assemble or survive the isolation. The isolated F1F0 complex containing the mutation bgly9----asp in the b subunit was defective in two regards: first, a reduction in F1 content relative to F0 and second, the absence of the a subunit. Immunoprecipitations of this preparation demonstrated that F1 interacts with both c and mutant b subunits. A strain carrying the mutation, bgly9----asp, and the compensating suppressor mutation apro240----leu (previously shown to be partially unc+) yielded an F1F0 ++ complex that remained partially defective in F1 binding to F0 but normal in the subunit composition of the F0 sector. The assembly, structure, and function of the F1F0-ATPase is discussed.     

The Structural and Functional Connection between the Catalytic and Proton Translocating Sectors of the Mitochondrial F 1 F 0 -ATP Synthase

The structural and functional connection between the peripheral catalytic F₁ sector and theproton-translocating membrane sector F₀ of the mitochondrial ATP synthase is reviewed. Theobservations examined show that the N-terminus of subunit , the carboxy-terminal and centralregion of F₀I-PVP(b), OSCP, and part of subunit d constitute a continuous structure, the lateralstalk, which connects the peripheries of F₁ to F₀ and surrounds the central element of thestalk, constituted by subunits and . The ATPase inhibitor protein (IF₁) binds at one sideof the F₁F₀ connection. The carboxy-terminal segment of IF₁ apparently binds to OSCP. The42L-58K segment of IF₁, which is per se the most active domain of the protein, binds at thesurface of one of the three / pairs of F₁, thus preventing the cyclic interconversion of thecatalytic sites required for ATP hydrolysis.     

北美CHIHUAHUAN荒漠啮齿动物群落动态III．变量的频次分布和预测

本文用х²方和柯尔莫哥洛夫检验分析了北美Chihuahuan荒漠啮齿动物群落6个生态学变量的频次分布,并用移动平均和指数平滑方法拟合了这些变量的动态变化。结果显示：1)结合种群密度、生物量、物种均匀性和生物量均匀性服从正态分布;2)物种数和物种多样性的频次为向右偏斜的分布,无法用常见的理论分布来近似表示;3)单移动平均和双移动平均分别较好地描述了物种多样性和物种均匀性;4)其余4个变量可用单指数平滑来较好的描述．     

人工饲养条件下高原鼠兔生长和发育的初步研究

本文研究了人工饲养条件下高原鼠兔的生长发育情况,并和其他种鼠兔的生长作了比较。室内高原鼠兔比野外的生长快,人工饲养的阿富汗鼠兔和北美鼠兔生长期短,成熟早。     

Sulfite inhibits the F1F0-ATP synthase and activates the F1F0-ATPase of Paracoccus denitrificans

The F₁F₀ complex of Paracoccus denitrificans (PdF₁F₀) is the fastest ATP synthase but the slowest ATPase. Sulfite exerts maximal activation of the PdF₁F₀-ATPase (Pacheco-Moisés, F., García, J. J., Rodríguez-Zavala, J. S., and Moreno-Sánchez, R. (2000). Eur. J. Biochem. 267, 993–1000) but its effect on the PdF₁F₀-ATP synthase activity remains unknown. Therefore, we studied the effect of sulfite on ATP synthesis and ³²Pi ATP exchange reactions of inside-out membrane vesicles of P. denitrificans. Sulfite inhibited both reactions under conditions of maximal pH and normal sensitivity to dicyclohexylcarbodiimide. Sulfite increased by 10- and 5-fold the K _0.5 for Mg²⁺-ADP and Pi during ATP synthesis, respectively, and by 4-fold the IC₅₀ of Mg²⁺-ADP for inhibition of the PdF₁F₀-ATPase activity. Thus, sulfite exerts opposite effects on the forward and reverse functioning of the PdF₁F₀ complex. These effects are not due to membrane or PdF₁F₀ uncoupling. Kinetic and structural modifications that could account for these results are discussed.     

Identification of F0 subunits in the rat liver mitochondrial F0F1-ATP synthase.

In order to identify the subunits constituting the rat liver F0F1-ATP synthase, the complex prepared by selective extraction from the mitochondrial membranes with a detergent followed by purification on a sucrose gradient has been compared to that obtained by immunoprecipitation with an anti-F1 serum. The subunits present in both preparations that are assumed to be authentic components of the complex have been identified. The results show that the total rat liver F0F1-ATP synthase contains at least 13 different proteins, seven of which can be attributed to F0. The following F0 subunits have been identified: the subunit b (migrating as a 24 kDa band in SDS-PAGE), the oligomycin-sensitivity-conferring protein (20 kDa), and F6 (9 kDa) that have N-terminal sequences homologous to the beef-heart ones; the mtDNA encoded subunits 6 (20 kDa) and 8 (less than 7 kDa) that can be synthesized in isolated mitochondria; an additional 20 kDa protein that could be equivalent to the beef heart subunit d.     

重组人肝再生增强因子在体外能刺激HTC肝癌细胞DNA合成

在成功克隆人肝再生增强因子（ｈＡＬＲ）ｃＤＮＡ的基础上,本研究将人ＡＬＲ编码区ｃＤＮＡ亚克隆于真核瞬间表达质粒ｐＣＤＮＡ Ｉ,构建了真核表达质粒ｐＣＤＮＡ Ｉ－ｈＡＬＲ,转染ｃｏｓ－７细胞后,表达产物生物活性检测表明：真核表达的ｈＡＬＲ在体外能刺激ＨＴＣ肝癌细胞增殖,这一体外活性的确定为重组人ＡＬＲ的纯化提供了简洁、可靠的检测方法。     

Structure of Dimeric F1F0-ATP Synthase

The structure of the dimeric ATP synthase from yeast mitochondria was analyzed by transmission electron microscopy and single particle image analysis. In addition to the previously reported side views of the dimer, top view and intermediate projections served to resolve the arrangement of the rotary c₁₀ ring and the other stator subunits at the F₀-F₀ dimeric interface. A three-dimensional reconstruction of the complex was calculated from a data set of 9960 molecular images at a resolution of 27 Å. The structural model of the dimeric ATP synthase shows the two monomers arranged at an angle of ∼45°, consistent with our earlier analysis of the ATP synthase from bovine heart mitochondria (Minauro-Sanmiguel, F., Wilkens, S., and Garcia, J. J. (2005) Proc. Natl. Acad. Sci. U.S.A. 102, 12356–12358). In the ATP synthase dimer, the two peripheral stalks are located near the F₁-F₁ interface but are turned away from each other so that they are not in contact. Based on the three-dimensional reconstruction, a model of how dimeric ATP synthase assembles to form the higher order oligomeric structures that are required for mitochondrial cristae biogenesis is discussed.     

cAMP在急性胃粘膜损害中作用的研究

本研究应用ＲＩＡ法,观察了大鼠胃组织ｃＡＭＰ含量与急性胃粘膜损害之间的关系。结果表明：（１）由消炎痛或失血性休克引起的急性胃粘膜损害时,胃组织ｃＡＭＰ含量明显降低;事先使用异搏定（１０ｍｇ／ｋｇ）可使胃组织ｃＡＭＰ含量明显增加,并使由消炎痛引起的急性胃粘膜损害相应减轻。（２）在正常情况下,胃窦、胃体组织间ｃＡＭＰ含量并非相等,胃窦部组织ｃＡＭＰ含量高于胃体部组织,与此相应的是急性胃粘膜损害主要局限在胃体部。上述引起急性胃粘膜损害的诸因素均可使这种差别加大。５和１０ｍｇ／ｋｇ剂量的异搏定可使消炎痛所致急性胃粘膜损害的大鼠胃体部组织的ｃＡＭＰ含量明显增加,与胃窦部组织ｃＡＭＰ含量的差值变小,结果是胃体部粘膜的损害明显减轻。提示胃组织ｃＡＭＰ含量变化在急性胃粘膜损害中有一定的作用。