Donnelly D F
Department of Pediatrics, Yale University School of Medicine, New Haven, CT 06510.
Brain Res. 1993 Dec 10;630(1-2):157-68. doi: 10.1016/0006-8993(93)90653-5.
Cells belonging to glomoids of mature rat carotid bodies were studied using the whole-cell patch clamp technique following acute dissociation. The recorded population encompassed two subtypes: one type (n = 202), termed G(out), was characterized by a small voltage-dependent inward current (43 +/- 9 pA, mean +/- S.E.M.), large outward current (671 +/- 31 pA @ +40 mV), high membrane resistance (1910 +/- 110 M omega) and low capacitance (5.1 +/- 0.1 pF). A second subtype (n = 56), termed G(in), had significantly lower membrane resistance (177 +/- 35 M omega), higher membrane capacitance (15.0 +/- 1.0 pF) and little voltage-dependent current. Neither subtype supported generation of multiple action potentials during depolarization in the current clamp mode. Intracellular staining of the recorded cells by Lucifer yellow showed co-localization of both subtypes to clusters of cells which stained positively for catecholamines. Somal diameter was slightly, but significantly, larger for G(in) cells (8.7 +/- 0.4 microM, n = 7) compared to G(out) cells (7.8 +/- 0.2 microM, n = 31) and all cells had fine cytoplasmic processes extending around neighboring cells. During recordings using the perforated patch technique, histotoxic hypoxia significantly decreased a voltage-dependent outward current in G(out) cells by 113 +/- 60 pA (n = 13), and decreased the holding current by 10 +/- 4 pA (n = 13) from a control value of -32 +/- 6 pA. In G(in) cells, cyanide significant decreased membrane resistance and decreased holding current by 55 +/- 28 pA from a control value of +120 +/- 42 pA (n = 7), but caused no significant change in outward current. These results show that glomoids of mature rat carotid bodies contain at least two types of cells which differ in their morphologic and electrophysiologic characteristics. The subtypes rapidly respond to histotoxic hypoxia and thus may mediate separate roles in the organ response to chemostimuli.
运用全细胞膜片钳技术,对急性分离的成年大鼠颈动脉体球样小体中的细胞进行了研究。记录的细胞群体包含两种亚型:一种类型(n = 202),称为G(out),其特征为小的电压依赖性内向电流(43±9 pA,平均值±标准误)、大的外向电流(在+40 mV时为671±31 pA)、高膜电阻(1910±110 MΩ)和低电容(5.1±0.1 pF)。第二种亚型(n = 56),称为G(in),其膜电阻显著更低(177±35 MΩ)、膜电容更高(15.0±1.0 pF)且电压依赖性电流很小。在电流钳模式下,两种亚型在去极化过程中均不支持多个动作电位的产生。用鲁米诺黄对记录的细胞进行细胞内染色显示,两种亚型均共定位于对儿茶酚胺呈阳性染色的细胞簇中。与G(out)细胞(7.8±0.2 μm,n = 31)相比,G(in)细胞(8.7±0.4 μm,n = 7)的胞体直径略大但具有显著性差异,并且所有细胞都有围绕相邻细胞延伸的精细细胞质突起。在使用穿孔膜片技术进行记录时,组织中毒性缺氧使G(out)细胞中电压依赖性外向电流显著降低113±60 pA(n = 13),并使钳制电流从-32±6 pA的对照值降低10±4 pA(n = 13)。在G(in)细胞中,氰化物使膜电阻显著降低,并使钳制电流从+120±42 pA的对照值降低55±28 pA(n = 7),但对外向电流无显著影响。这些结果表明,成年大鼠颈动脉体球样小体包含至少两种细胞类型,它们在形态和电生理特征上有所不同。这些亚型对组织中毒性缺氧迅速做出反应,因此可能在该器官对化学刺激的反应中发挥不同作用。