Characterization of cDNA of lycopene beta-cyclase responsible for a high level of beta-carotene accumulation in Dunaliella salina.

Lycopene beta-cyclase (Lyc-B) is the key enzyme in the catalysis of linear lycopene to form cyclic beta-carotene, an indispensable part of the photosynthetic apparatus and an important source of vitamin A in human and animal nutrition. Studies showing that the microalga Dunaliella salina can accumulate a high level of beta-carotene are lacking. We hypothesize that D. salina is closely involved with the catalytic mechanism of Lyc-B and the molecular regulation of its gene. In this study, we used RT-PCR and RACE-PCR to isolate a 2475 bp cDNA with a 1824 bp open reading frame, encoding a putative Lyc-B, from D. salina. Homology studies showed that the deduced amino acid sequence had a significant overall similarity with sequences of other green algae and higher plants, and that it shared the highest sequence identity, up to 64%, with Lyc-B of Chlamydomonas reinhardtii. Codon analysis showed that synonymous codon usage in the enzyme has a strong bias towards codons ending with adenosine. Two motifs were found in the Lyc-B sequence, one at the N terminus, for binding the hypothetical cofactor FAD, and the other was a substrate carrier motif in oxygenic organisms shared by an earlier carotenogenesis enzyme, phytoene desaturase, and Lyc-B. A tertiary structure prediction suggested that the catalytic or binding site structure within LycB from D. salina is superior to that of both H. pluvialis and C. reinhardtii. The LycB protein from D. salina was quite removed from that of H. pluvialis and C. reinhardtii in the phylogenetic tree. Taken as a whole, this information provides insight into the regulatatory mechanism of Lyc-B at the molecular level and the high level of beta-carotene accumulation in the microalga D. salina.     

Abrogation of the capacity of delayed-type hypersensitivity responses to alloantigens by intravenous injection of neuraminidase-treated allogeneic cells

BALB/c or C3H/He mice were inoculated i.v. with allogeneic spleen cells untreated or treated with neuraminidase. Appreciable or potent anti-allo-delayed-type hypersensitivity (DTH) responses were observed when mice were inoculated i.v. with untreated allogeneic cells or inoculated i.v. with those cells followed by s.c. immunization with untreated allogeneic cells. In contrast, i.v. inoculation of neuraminidase-treated allogeneic cells (presensitization) not only failed to induce any significant anti-allo-DTH responses but also abolished the capability of the animals to develop DTH responses after s.c. immunization, indicating the tolerance induction. This tolerance was alloantigen-specific, and rapidly inducible and long lasting. The induction of suppressor cell activity was demonstrated in tolerant mice. However, this activity was associated only with the tolerant state around 4 to 7 days after the i.v. presensitization, but was no longer detected in mice more than 14 days after the presensitization, although these mice exhibited complete tolerant state. When spleen cells from such tolerant mice were transferred i.v. into 600 R x-irradiated syngeneic recipient mice alone or together with normal syngeneic spleen cells, these tolerant spleen cells themselves failed to induce DTH responses but did not exhibit suppressive effect on the generation of DTH responses induced by normal spleen cells co-transferred. These results indicate that i.v. administration of neuraminidase-treated allogeneic cells results in the induction of alloantigen-specific tolerance which is not always associated with the induction of suppressor cell activity but rather with the elimination or functional impairment of alloantigen-specific clones.     

电刺激大鼠尾核头部对丘脑腹后外侧核痛相关神经元电活动的影响

丘脑腹后外侧核的神经元对伤害性刺激的反应形式主要为以下三种类型:痛兴奋神经元、痛抑制神经元和痛无关神经元。痛相关神经元的放电活动可被吗啡类药物所抑制。电刺激尾核头部对痛相关神经元主要产生抑制作用,尾核头部背侧的作用强于腹侧,且受不同刺激参数的影响。本文就以上结果在镇痛中的意义进行了讨论。     

雄性水狼蛛生殖球顶部的扫描电镜研究

本文对我国已知的十一种水狼蛛(蜘蛛目:狼蛛科)生殖球顶部用扫描电镜进行了研究。结果表明,水狼蛛属蜘蛛插入器细长,其后端位于顶突上一突起的凹槽内,顶突作为功能引导器保护着插入器,而无其他狼蛛属具有的次生真正引导器;顶突的形状和其上的突起、顶突与插入器的相互关系对于种的分类与鉴定,特别是对于近似种的区分有较大的参考价值,但仅根据插入器从顶部升起的位置来作为分新属的主要标准,笔者认为还值得商榷和进一步探讨。     

刺激杏仁基底外侧核对外侧缰核神经元单位放电的影响

用玻璃微电极细胞外记录大鼠外侧缰核(LHN)神经元的单位放电。共记录了110个神经元。其中痛兴奋神经元(LHPE)75个;痛抑制神经元(LHPI)11个;广动力型神经元2个;无反应神经元17个;此外还有5个对躯体与内脏伤害性刺激反应不同的神经元。电刺激杏仁基底外侧核(以下简称杏仁核,AMG)对LHPE和LHPI的自发放电主要产生抑制作用,分别占总数的81.1％和72.7％,并抑制其对伤害性刺激的反应;对无反应神经元和广动力型神经元无明显影响。AMG内微量注射吗啡能抑制LHPE的伤害性刺激反应,但对其自发放电无明显影响。微量注射纳洛酮则可增加LHPE的自发放电频率,并加强其对伤害性刺激的反应。注射纳洛酮还可以取消电针对LHPE的伤害性刺激反应的抑制作用。     

冷冻或切除小脑蚓部山顶(Culmen)对豚鼠“踏步自动作用”(Stepping automatism)的影响

本研究探讨部分冷冻或切除小脑蚓部(vermis)对整体豚鼠“踏步自动作用”(steppingautomatism)的影响。“踏步自动作用”由我们近年来发现的诱发踏步物质(SIS)(4-R-2,2,5,5-四(三氟甲基)-咪唑啉)所引起。结果表明部分冷冻或切除小脑蚓部的山顶(culmen,Ⅴ和Ⅳ叶)和中央叶(Centralis,Ⅲa,b)明显增强豚鼠的“踏步自动作用”。冷冻小脑不能触发,但仅能调控“踏步自动作用”。这种调控作用对自动化程度差的弱“踏步自动作用”特别显著。蚓部山顶(Ⅴ叶为主)同时调控左右前肢踏步,而一侧蚓部山顶及其半球则主要调控同侧前肢踏步。此外,本研究的结果表明当介面温度(冷冻头和小脑幕间)致冷至5℃—0℃左右,冷冻小脑便可基本模拟部分切除小脑效应。     

辣椒素引起脊髓P物质释放及其对血压的影响

为进一步研究脊髓 P 物质(SP)在调节心血管活动中的作用,在大鼠脊髓蛛网膜下腔注射(ith)辣淑素(cap),以刺激脊髓 SP 能神经末梢释放 SP,结果引起血浆去甲肾上腺素(NA)和肾上腺素(AD)含量增高,及具有剂量依赖性的动脉血压上升,心率升高。ith 具有高度特异性的 SP 受体拮抗剂或 SP 抗血清均可阻断 cap 引起的升压效应,免疫组化测定也观察到注入的cap 剂量越大,脊髓胸段 SP 样免疫阳性反应物的致密度越低,这些观察结果支持 cap 可以引起脊髓内 SP 的释放的说法。在第一颈段(C_1)横断脊髓后 ith cap 所引起的升压效应与完整动物 ith cap 的升压效应无显著差异。以上结果提示脊髓 SP 能神经末梢释放的 SP 可以通过交感肾上腺髓质系统引起心血管兴奋效应,SP 可能是引起交感节前神经元兴奋的神经递质。     

大鼠隔—海马通路损伤对海马内递质含量及酶活力的影响

单侧切断大鼠海马缴和部分穹窿可使海马部分去神经。损伤后7d,海马内胆碱能系统中乙酰胆碱(ACh)含量下降72.5%,胆碱乙酰基转移酶(ChAT)活力下降45.7%,胆碱酯酶活力下降52.2%,在单胺能系统中,去甲肾上腺素(NA)含量下降16.3%,多巴胺(DA)含量下降31.3%,5-羟色胺含量下降30.3%。在损伤过程中,海马内氨基酸含量没有改变。实验结果表明,海马缴和穹窿是脑内胆碱能和单胺能传入神经到达海马靶区的部分共同通路。