影响有尾类离体胚胎表皮细胞兴奋性的实验

用常规电生理细胞内记录技术了解了影响蝾螈胚胎非典型表皮细胞兴奋性的一些因素。1.小牛血清、蝾螈胚胎内胚层和肌节中胚层、非洲爪蟾胚胎外胚层及嫁接于受体胚胎都提高非典型表皮显示动作电位样品的百分比。血清琼脂包饺子的作用较培养在含血清溶液明显。内、外胚层的作用更为明显。嫁接于受体胚胎的非典型表皮百分之百地显示兴奋性。2.血清、内、中、外胚层及嫁接于受体胚胎都降低表皮细胞的刺激阈值。各处理组的平均阈值均低于对照组,其巾嫁接于受体胚胎的非典型表皮细胞最低,接近于正常胚胎的表皮细胞。3.血清琼脂块和内胚层包饺子的实验表明,影响兴奋性的因素必须始终与表皮细胞保持接触,才‘能产生作用。一旦脱离接触,其作用就逐渐消失。4.胚胎表皮细胞兴奋性的产生不依赖于外来因素,是一种自主的功能分化。但是外来因素能使培养的表皮细胞的兴奋性增强,对低强度的刺激产生兴奋。这种外来因素可能是一些非专一性的与代谢有关的物质。     

代谢抑制剂对有尾类胚胎表皮细胞兴奋性的影响

离体培养的蝾螈胚胎的非典型表皮,其中的大部分在较高强度的电刺激下细胞不显示兴奋性,少量显示兴奋性的外植块所需的刺激强度也高于正常胚胎表皮。用含有能量物质(碳水化合物、氨基酸、代谢中间产物和ATP)的     

关于有尾类胚胎的表皮及非典型表皮的兴奋性

蝾螈胚胎表皮的兴奋性及传导能力在分期26出现,到分期37末结束,某些部位甚至更晚些消失。为了查明表皮细胞兴奋性的产生是否受到来自其它组织的影响,还是仅依赖于表皮细胞自身的分化状态,我们曾用非典型表皮进行过研究。当时曾报道,离体培养的表皮细胞,即使达到了产生兴奋性的分化状态,用电生理技术在刺激强度为1—2伏时,测试不到动作电位。然而,如果这样的非典型表皮被     

中胚层在有尾类胚胎表皮传导中的作用(简报)

从前我们报道过,从蝾螈的早期原肠胚分离培养的非典型表皮,对电刺激不产生可传播的兴奋,但是嫁接到受体胚胎则成为可能产生可传播的兴奋,并能将冲动传到后者,说明兴奋性的产生不单纯是细胞分化的后果,而是受到其它因素的影响。曾经推断这种因素可能是一种物质。根据非典型表皮与受体胚胎在嫁接中接触的情况,尤其是来自中胚层的物质可能和表皮细胞产生传导能力有密切关系。     

蝾螈胚胎表皮兴奋性在发育中的变化

1.蝾螈胚胎表皮在分期26动作电位刚出现时,同一胚胎不同部位表皮细胞的兴奋性不同,存在一个沿头—尾轴的梯度变化。头部细胞引起动作电位所需的阈值??最低,即细胞兴奋性最强。尾部所需阈值最高,中部在两者之间。这一梯度在分期26末消失。2.胚胎在整个传导期间(分期26至分期37),表皮细胞兴奋性也不是恒定的,呈低一高一低的变化过程。在分期32,阈值线达最低值,即此时表皮细胞最容易兴奋。3.在胚胎表皮动作电位消失过程中表皮细胞兴奋性存在一个与动作电位出现时相反的梯度。不仅头一尾、而且背、腹也存在这种兴奋性梯度。背面头部动作电位最早消失,腹面尾部可维持到分期38中期。4.表皮细胞兴奋性出现时,动作电位有一个发生、发展的过程。离体实验表明,含有头部中胚层的表皮,动作电位的出现早于含有尾部中胚层的表皮,平均早出现2.4小时。这说明表皮兴奋性按时空顺序出现可能受到其下不同部位中胚层的某些影响。5.离体实验还提出了神经嵴对表皮兴奋性的出现也有一定影响,虽然作用似乎比中胚层要弱一些。     

大鼠下丘脑薄片视上核神经元的细胞内生物电记录

从成年Ｓｐｒａｇｕｅ－Ｄａｗｌｅｙ大鼠用振动切片机制备含有视上核的下丘脑冠状薄片（５００μｍ厚）,并对视上核神经元（ｎ＝１７）进行常规细胞内生物电记录。测得静息电位－６０±８ｍＶ,膜输入电阻１７３±５８ＭΩ,时间常数１０．２±３．９ｍｓ,动作电位幅度６５±１２ｍＶ,阈电位－４４±７ｍＶ,由Ｉ─Ｖ曲线测得斜率电阻１５８±６２ＭΩ,大部分细胞还显示内向整流特性。在静息电位状态下,５７％细胞保持静息,４３％有自发锋电位发放。细胞的锋电位发放模式７８％为时相型,２２％为持续型。外源性去甲肾上腺素（ｎ＝７）或谷氨酸钠（ｎ＝５）可引起伴有膜电阻减小的去极化反应,并可导致锋电位的串发放。     

蝾螈胚胎表皮兴奋性和传导能力的出现和消失

两栖纲中有尾目和无尾目胚胎表皮在一定发育阶段都具有兴奋性并能传导兴奋的问题,经过一番争论之后终于被肯定了下来。有尾目中蝾螈的胚胎在发育分期26阶段出现前肾隆起时,表皮出现兴奋性和传导能力;但这时胚胎的肌肉还处于不具有收缩能力的时期。胚胎表皮的兴奋性和传导能力一直维持到分期37。至于     

关于Rohon-Beard细胞的功能发育及其与胚胎早期行为的关系

在前一篇报道里我们断定了R.-B.细胞在神经胚中的定位,叙述了它们被割除干净后,早期胚胎显示不出应有的、对刺激的反应。虽然已经有人从它们的纤维的分布,从膜的兴奋性判断它们的神经元的性质,我们用实验胚胎学的方法证明了它们确实是初级感觉神经元(Primary sensory neuron),在早期胚胎的传导通路中,负责把兴奋传递到脊     

背景氯离子通道研究进展

综述了目前了解得最为充分的一类电压门控氯通道——背景氯通道,内容涉及选择性、门控和药理学以及通道蛋白的克隆和分子结构.氯通道广泛存在于细胞膜和细胞器膜,作为“总管家”参与细胞pH,体积,静息膜电位和兴奋性等多种细胞过程的调节.由于种种原因,对氯通道的研究起步较晚.目前应用膜片钳和分子生物学技术对氯通道结构功能的研究已经成为一个热点.     

缺氧对游离的颈动脉体球细胞和细胞簇内球细胞的不同效应

用细胞内记录法测定了２０１个培养的大鼠颈动脉体球细胞的膜电位及输入电阻。在连二硫酸钠（Ｎａ２Ｓ２Ｏ４）造成缺氧（Ｐｏ２,１．３－８．０ｋＰａ）条件下,细胞簇内球细胞与游离球细胞对缺氧的反应不同。８２％以上的细胞簇内球细胞发生去极化和输入电阻降低,而６０％的游离球细胞产生超极化和输入电阻增加。细胞簇内球细胞与游离球细胞反应不同的主要原因可能取决于支持细胞存在与否。细胞簇内球细胞常被支持细胞的突起所包绕,而游离的球细胞则无支持细胞的包绕。也可能存在不同功能的球细胞。     

代谢抑制剂对有尾类胚胎表皮细胞兴奋性的影响

It has been reported that the cells in atypical epidermis, which developed from the in vitro cultured ectoderm isolated at early gastrula, showed very low excitability or were even non-excitable at 6 V when examined electro-physiologically. If non-excitable explants were treated with energy supplying substances, such as glucose, the action potential (AP) appeared quickly. It indicates that, the excitability of epidermis cells is related to their energy metabolism. In order to verify the above proposition the effects of metabolic inhibitors on the excitability of the epidermis cells were examined using electrophysiological technique. Two kinds of explants were used: explants which developed from the epidermis underlaid with mesoderm isolated at early neurula (epidermis vesicle) and explant which developed from the ectoderm isolated at early gastrula (atypical epidermis). In all experiments explants were stimulated extracellularly and APs were recorded intracellularly. The specimens were stimulated with electric stimulus at 6 V first, and, if they displayed AP, the strength was lowered to determine the stimulus threshold to evoke AP. The duration of stimulus was fixed at 1 ms. The ratio of the resting potential value during treatment to the original value was taken as index of change level of the resting potential (RP). During treatment of epidermis vesicle with 1 mM NaN3 or 1 mM NaCN or 0.1 mM 2,4-dinitrophenol (DNP), the excitability of the epidermis cells was reduced: the stimulus threshold gradually increased and the cells in most explants lost the excitability. The cells became excitable after the drugs were washed out.(ABSTRACT TRUNCATED AT 250 WORDS)     

Epidermal growth factor (EGF)-induced morphological changes in the basement membrane of chick embryonic skin

Summary The effect of epidermal growth factor (EGF) on the basement membrane structure of chick embryonic skin cultured in a chemically defined medium (BGJb) containing 20 mM hydrocortisone, and EGF at 10, 50, or 100 ng/ml supplemented with 5% delipidized fetal calf serum, was examined by electron microscopy. During development of the epidermis in vitro, EGF (100 ng/ml) caused striking changes to occur in the basement membrane structure and in the keratinization process. The basement membrane frequently became discontinuous with many gaps apparent in section, and occasionally became folded following detachment from the basal surface of the epidermis and protruded into the underlying dermis. In the basal and intermediate cells of EGF-treated epidermis, tonofilament bundles were decreased in number, while desmosomes and hemidesmosomes revealed no significant changes in morphology.     

Use of aminopterin in selecting electrically active neuroblastoma cells

A marked increase in electrical excitability and process formation occurs in the N-18 clone of mouse neuroblastoma as these cells go from the logarithmic phase of growth to the stationary state in confluent cultures. Even more excitable cells can be selected by growth in culture medium containing 10⁻⁵ M aminopterin which kills about 90% of the cells. Clone 1A-103 does not develop significant processes or exhibit marked electrical excitability under any of the culture conditions studied. Thus, our results show that one or more of the steps required for generation of the action potential is sensitive to regulation in cultured cells. Methods are presented for obtaining populations of either electrically passive cells or electrically excitable cells which can easily be maintained for several weeks. Clones differ markedly in their capacity to extend processes and their ability to generate action potentials.     

日本血吸虫卵黄培养细胞超微结构动态的研究

从超微结构水平研究日本血吸虫卵黄细胞培养的动态。在体外培养过程中,卵黄细胞对外界环境的改变比日本血吸虫的其它体细胞更加敏感。随着培养时间的延长,卵黄细胞发生不同程度的变性,成熟卵黄细胞比未成熟卵黄细胞发生变性。变性主要表现在核和胞质的电子密度降低;卵黄球相互融合,或卵黄球与膜之间的空隙逐步增大,最终卵黄球从中释放出来,变成裸露体;脂滴数目增多,体积增大;粗面内质网扩大和囊泡变,其上的核糖体脱颗粒等     

花背蟾蜍蝌蚪发生类坏死的皮肤在恢复过程中的超微结构

本实验以花背蟾蜍后肢芽期的蝌蚪为材料,用0.001 mol/L氨水(氨水组)和0.01mol/L醋酸(醋酸组)处理皮肤使之发生类坏死。以细胞核、细胞表面、细胞质及细胞间隙等的超微结构变化为指标,研究了各组皮肤发生类坏死后恢复过程中的可逆变化。结果观察到:两组均能引起核染色质浓缩,粘液分泌,液泡增多,线粒体及内质网膨大变形,细胞间隙变窄;醋酸还引起张力原纤维的凝集及紊乱等。恢复过程中,细胞核内染色质的分布趋于均匀,粘液的分泌渐正常,液泡减少,线粒体及内质网的形态逐步恢复,但细胞间隙一直很??窄;醋酸组中张力原纤维恢复成束且排列较整齐。作者认为氨水及醋酸引起蝌蚪皮肤发生类坏死后,在一定时间内是可以恢复的,而且此可逆变化也反映在超微结构的变化上。     

The influence of plasma membrane ion permeability modulators on superoxide formation and bioelectrical characteristics of wheat root cells

Changes in superoxide radical formation and bioelectrical characteristics of excised wheat root cells under modification of plasma membrane ion permeability were studied. It was shown that a 2 h treatment of excised roots with valinomycin (Val, 20 microM), N, N'-dicyclohexylcarbodimide (DCCD, 100 microM), gramicidin S (Gr, 20 microM), chlorpromazine (CPZ, 100 microM) caused an increased loss of potassium by cells, lowering of membrane potential (MP) and electrical input resistance (Rin) of the cells. The superoxide formation by excised root cells diminished (under DCCD) or remained at the control level (under Val), which was accompanied by a minor decrease of MP and Rin of the cells, a small increase in potassium loss by excised roots, and in no change of pH of incubation medium. Significant depolarization of plasma membrane, dropping of Rin and essential loss of potassium ions by the cells correlated with a rise in the medium alkalinization and superoxide formation by excised roots (in the presence of Gr, CPZ). Ion channel blocker gadolinium (Gd3+, 200 microM) caused an increase of MP and Rin reduction of potassium loss by cells, and a decrease of pH of the incubation medium, and also enhancement of superoxide formation by excised root cells. It is suggested that upon plasma membrane ion permeability modification the activity of superoxide generating systems depends on the specificity and mechanisms of action of modulators, and is determined by their influence on redox state of plasma membrane as well as by peculiarities of ion transport disturbance.     

Reconstituted skin from murine embryonic stem cells

Embryonic stem (ES) cell lines can be expanded indefinitely in culture while maintaining their potential to differentiate into any cell type. During embryonic development, the skin forms as a result of reciprocal interactions between mesoderm and ectoderm. Here, we report the in vitro differentiation and enrichment of keratinocytes from murine ES cells seeded on extracellular matrix (ECM) in the presence of Bone Morphogenic Protein-4 (BMP-4) or ascorbate. The enriched preparation of keratinocytes was able to form an epidermal equivalent composed of a stratified epithelium when cultured at the air-liquid interface on a collagen-coated acellular substratum. Interestingly, an underlying cellular compartment that belongs to the fibroblast lineage was systematically formed between the reconstituted epidermis and the inert membrane. The resulting tissue displayed morphological patterns similar to normal embryonic skin, as evidenced by light and transmission electron microscopy. Immunohistochemical studies revealed expression patterns of cytokeratins, basement membrane (BM) proteins and late differentiation markers of epidermis, as well as fibroblast markers, similar to native skin. The results demonstrate the capacity of ES cells to reconstitute in vitro a fully differentiated skin. This ES-derived bioengineered skin provides a powerful tool for studying the molecular mechanisms controlling epidermal and dermal commitments.     

杉木雄性不育株与可育株小孢子囊发育的电镜研究

杉木雄性不育属“无花粉型”,败育从无孢原细胞到四分体时期,中层细胞增生,压迫小孢子母细胞,使之养分更加缺乏并引起减数分裂异常。其表皮层和药室内壁细胞中具大量蛋白体,影响了绒毡层对小孢子发育的外源蛋白的供应;表皮层具膜状溶酶体,引起淀粉粒缺乏和酶系统代谢异常;绒毡层发育后期,缺少包被小泡、乌氏体及绒毡层膜,内质同少、短而光滑,严重影响了绒毡层对小孢子母细胞或小孢子养分和合成细胞壁物质的供给;线粒体发育异常,能量代谢减弱,导致小孢子母细胞或小孢子一系列生理活动紊乱及其原生质体解体。     

Electrical properties of motoneurons in the spinal cord of rat embryos

Electrical properties of immature motoneurons were studied in vitro using isolated segments of spinal cords of rat embryos aged 14-21 days of gestation. Stable resting potentials and evoked synaptic potentials were recorded for more than 9 hr, indicating that motoneurons remain viable for many hours. Motoneurons are electrically excitable at 14 days of gestation and from the onset of excitability the action potentials are Na+-dependent but slow rising long-duration Ca2+-dependent action potentials can be evoked if K+ conductance is reduced. Thus, during embryonic development the regenerative potential inward current is Na+-and Ca2+-dependent. During motoneurons' differentiation there are some changes in their electrical properties: resting membrane potential increases, input resistance decreases, input capacitance increases, threshold for action potential decreases, and maximum rate of rise of action potential increases. Afferent motoneuron contacts are formed at 16-18 days of gestation when excitatory synaptic potentials can first be evoked in response to dorsal root stimulation. The changes in input capacitance and threshold for action potential occur at the onset of functional afferent motoneuron contacts, but it is not known whether these changes are autonomous or are influenced by the newly formed sensory inputs.