Intracellular pH shifts capable of uncoupling cultured oligodendrocytes are seen only in low HCO3- solution

Electrical coupling between cultured mouse oligodendrocytes was transiently blocked when pHi was decreased below about 6.5 using the NH4+ prepulse method. This uncoupling could, however, only be achieved if the dominant pHi regulating mechanism in these cells, the Na+/HCO3- cotransporter, was blocked by lowering bath [HCO3-]. Under this condition, an NH4+ prepulse caused pHi to decrease toward the passive distribution for H+ (i.e., about pH 6.2). In the presence of normal bath [HCO3-] an NH4+ prepulse did not decrease pHi below 6.5 even when the second pHi regulating mechanism, the Na+/H+ exchanger, was blocked by amiloride, and consequently oligodendrocytes could not be uncoupled. Increasing CO2, which uncouples glial cells in situ (Connors et al: J. Neurosci. 4:1324-1330, 1984), did not uncouple cultured oligodendrocytes in the presence of normal bath [HCO3-], but did cause uncoupling in low [HCO3-] solution. These results indicate that electrical coupling between cultured oligodendrocytes is sensitive to pHi; in normal bath [HCO3-], however, the pHi regulation of these cells is so effective that standard techniques for intracellular acidification are unable to lower pHi to levels which cause the closure of oligodendrocyte gap junctions.     

Response of rat cerebral cortical astrocytes to freeze- or cobalt-induced injury: an immunocytochemical and gap-FRAP study

Astrocytic response in the immediate vicinity of freeze- and cobalt-induced lesions has been examined at the light and ultrastructural level. However, the temporal and spatial distribution of astrocytic reactivity throughout the rat cerebral cortex, using glial fibrillary acidic protein (GFAP) immunolabeling, has not been examined. The first purpose of this study was to establish the chronological distribution of astrocytic reactivity, as measured by changes in GFAP immunoreactivity, following freeze- or cobalt-induced injury to the rat cerebral cortex. Cobalt metal also has been proposed to have a direct effect on astrocytes and has been shown to stimulate in vitro astrocytes to become reactive. The second purpose of this report was to determine if cobalt had an effect on in vitro astrocytic gap junctional dye coupling as measured by fluorescence recovery after laser-photobleaching (gap-FRAP). Although the chronological development of the increased GFAP immunoreactivity was different for the freeze- and cobalt-induced lesions, astrocytes initially showed an increase in GFAP immunoreactivity in the region surrounding these lesions. This initial response was followed by a spread of increased GFAP immunoreactivity throughout certain regions of the ipsilateral cerebral hemisphere and then by a restriction of the increased immunolabeling to the lesion site. Cobalt also had a direct effect on in vitro astrocytes as demonstrated by the inhibition of astrocytic gap junctional dye coupling. Based on gap-FRAP analysis, cobalt significantly blocked fluorescence recovery (2.5%) as compared to the fluorescence recovery in control astrocytes (26%). It is proposed that the initial increase in GFAP immunoreactivity may be due to decreased gap junctional activity.     

电磁噪声阻断极低频磁场对细胞缝隙连接功能的抑制效应

目的　探讨电磁噪声对 5 0Hz磁场诱导的细胞缝隙连接通讯功能 (GJIC)抑制的干预作用。方法　小鼠成纤维细胞 (NIH 3T3)受 5 0Hz 0 .4mT极低频磁场或磁场加同等强度的电磁噪声联合作用 2 4h后 ,在激光共聚焦显微镜下 ,采用荧光光漂白后恢复技术 (FRAP)测定细胞的GJIC功能。结果　 0 .4mT磁场单独作用明显抑制细胞的GJIC ,其荧光恢复率为 2 7.6 7%± 5 .12 % ,与对照组(45 .5 7%± 9.72 % )相比 ,差异有显著性 (P <0 .0 1) ;而磁场与电磁噪声联合作用 (5 2 .6 1%± 8.30 % )明显拮抗磁场对GJIC的抑制作用 ,与 0 .4mT磁场组的差异有显著性 (P <0 .0 1) ,并与对照组的差异无显著性 (P >0 .0 5 )。结论　电磁噪声对极低频磁场诱导的GJIC的抑制具有阻断作用     

Atrioventricular nodal reverse facilitation in connexin40-deficient mice

BACKGROUND: Facilitation is an important physiologic property of the atrioventricular (AV) node. Previous studies demonstrated abnormal AV conduction in connexin (Cx)40-deficient mice. OBJECTIVES: We hypothesize that Cx40-deficient mice display altered patterns of AV nodal facilitation compared with wild-type mice. METHODS: Sixteen 36-week-old mice (eight Cx40(-/-) mice and eight Cx40(+/+) controls) underwent in vivo closed chest electrophysiologic study. A 2Fr octapolar catheter was advanced into the right ventricle to record a His-bundle electrogram. A special facilitation stimulation protocol was performed in each mouse to evaluate facilitation. Following atrial drive pacing (S1S1) at 150 ms, a facilitating beat S2 was delivered prior to the test beat S3. S3H3 was measured for varying S1S2 values at fixed H2S3 intervals. RESULTS: Progressive shortening of S1S2 (from 150 ms to 130, 110, and 90 ms) resulted in gradual prolongation of S2H2. The prolongation was more pronounced in Cx40(-/-) mice for each S1S2 compared with wild-type mice (P <.001). In each wild-type mouse, for a given H2S3 interval, this gradual increase in S2H2 produced progressive shortening of S3H3, so-called AV nodal facilitation phenomenon. However, in each Cx40(-/-) mouse, facilitation was seen only at S1S2 of 130 ms (P <.001 vs S1S2 of 150 ms). Evidence of reverse facilitation was documented at S1S2 of 110 and 90 ms. CONCLUSION: Facilitation is observed in wild-type mice. With similar S1S2 intervals in Cx40-deficient mice, facilitation is seen only at longer S1S2 intervals, whereas reverse facilitation is seen at shorter S1S2 intervals, suggesting that Cx40 is involved in the generation of AV nodal facilitation.     

The differentiation of visually guided and anticipatory saccades in gap and overlap paradigms

Summary Saslow and others have shown that the latency of foveating saccades can be altered by changing the offset time of the current fixation point relative to the onset of the peripheral target. Whether anticipatory saccades contributed to these results was not known. By the criteria of direction error and amplitude error the minimum latency for visually guided saccades is 110–130 ms for three subjects and 160 ms for a longer latency subject. Excluding anticipatory responses did not eliminate offset-onset effects. The genesis of express saccades and the role of higher neural levels is discussed.     

Age-related changes in junctional and non-junctional conductances in two electrically coupled peptidergic neurons of the mollusc Lymnaea stagnalis

Age-related changes in electrotonic coupling ratio of two identified neurons in Lymnaea stagnalis were studied together with the underlying changes in the steady-state conductance properties of the network. Two phases were distinguished in the development of coupling ratio across lifespan. During the first phase (age of 3-13 months), coupling ratio decreased from decreased from 60% to 30%. The second phase (age 13-20 months) was characterized by an increase in coupling ratio. Values of up to 60% were reached again in the oldest animals. Voltage clamp measurements showed that the biphasic trend of the age-related changes in coupling ratio is paralleled by changes in conductance properties of the junction between VD1 and RPD2. During the first phase junctional conductance decreased, whereas during the second phase junctional conductance increased. In addition to the decrease in junctional conductance, a growth-related increase in non-junctional conductance of VD1 and RPD2 contributed to the decrease in coupling ratio observed during the first phase. Thus our results indicate that in Lymnaea junctional connections between neurons may undergo considerable and discontinuous changes after sexual maturation. In addition to these changes in steady-state electrical properties, indications were obtained that age-related changes of kinetically slower conductance(s) may occur in the non-junctional membrane of VD1 and RPD2.     

高血压大鼠心脏中缝管连接蛋白的表达及连接结构观察

目的　为临床高血压心脏病的相关研究提供形态学依据。方法　 Dahl盐敏感高血压大鼠于生后 5w开始喂 8%高盐饲料 ,分别于 9、1 1、1 3周龄处死 ,采用透射电镜及免疫组化方法对大鼠心肌细胞的连接结构及缝管连接蛋白 Cx43、Cx40的含量进行研究。结果　心肌细胞的连接结构闰盘似山峰样 ,台阶消失 ,缝管连接位于突起之间 ;实验组 Cx43含量较相应对照组显著减少 (9周龄 P<0 .0 5;1 1、1 3周龄 P<0 .0 1 ) ,分布紊乱 ;Cx40含量在早期 (9周龄 )时显著减少 (P<0 .0 1 ) ,随后显著增加 (P<0 .0 1 )。结论　缝管连接分布的紊乱及 Cx43含量的下降可能在高血压心脏病心律失常的发生上起了重要作用 ,而 Cx40含量增加可能是增加传导速度的一个补偿机制。     

微泡在肿瘤进展的作用及其临床应用

微泡(microvesicles)能运载多种特异性蛋白、微小RNA及DNA片段,其为细胞间信号交流提供了新的途径。肿瘤细胞分泌微泡(tumor-derived membrane microvesicles,TMV),而微泡参与肿瘤进展的多个方面,TMV向周围的普通细胞传递肿瘤特异性蛋白改变细胞功能,传递microRNA改变细胞表型,增加逆转录干扰基因的稳定性,从而建立肿瘤微环境;同时,其可促进血管新生、破坏细胞基质,从而增强肿瘤细胞的侵袭性;其还可通过强化抑制性免疫T细胞的功能和诱导抗肿瘤T细胞的凋亡,参与肿瘤对机体免疫监视的逃避。另外,微泡作为肿瘤抗原递呈载体,可扩大机体的抗肿瘤免疫,而由此研制的新型肿瘤疫苗,目前已处于临床试用早期阶段。循环系统中微泡运载microRNA及DNA片段的发现,为研究无创性肿瘤标志物提供新思路。然而,微泡的分离技术有待提高,这对微泡的鉴别和亚型分类有重要意义。