Galvez Fernando, Wong Denise, Wood Chris M
Department of Biological, McMaster University, Hamilton, Ontario, Canada.
Am J Physiol Regul Integr Comp Physiol. 2006 Jul;291(1):R170-6. doi: 10.1152/ajpregu.00217.2005. Epub 2006 Feb 9.
A novel cell isolation technique was used to characterize cadmium and calcium uptake in distinct populations of gill cells from the adult rainbow trout (Oncorhynchus mykiss). A specific population of mitochondria-rich (MR) cell, termed the PNA+ MR cell (PNA is peanut lectin agglutinin), was found to accumulate over threefold more 109Cd than did PNA- MR cells, pavement cells (PV cells), and mucous cells during a 1-h in vivo exposure at 2.4 microg/l 109Cd. In vitro 109Cd exposures, performed in standard PBS and Cl- -free PBS, at concentrations from 1 to 16 microg/l 109Cd, were also carried out to further characterize Cd2+ uptake kinetics. As observed during in vivo experiments, PNA+ MR cells accumulated significantly more 109Cd than did other cell types when exposures were performed by an in vitro procedure in PBS. Under such conditions, Cd2+ accumulation kinetics in all cell types could be described with Michaelis-Menten relationships, with Km values of approximately 3.0 microg/l Cd (27 nM) for both MR cell subtypes and 8.6 microg/l Cd (77 nM) for PV cells. In similar experiments performed in Cl- -free conditions, a significant reduction in 109Cd accumulation in PNA+ MR cells was seen but not in PNA- MR or in PV cells. In vitro 45Ca fluxes were also performed to determine the cellular localization of Ca2+ transport in these functionally distinct populations of gill cells. 45Ca uptake was most pronounced in PNA+ MR cells, with levels over threefold higher than those found in either PNA(-) MR or in PV cells. Results from the present study suggest that the PNA+ MR cell type is a high-affinity and high-capacity site for apical entry of Cd2+ and Ca2+ in the gill epithelium of rainbow trout.
一种新型细胞分离技术被用于表征成年虹鳟(Oncorhynchus mykiss)鳃细胞不同群体中镉和钙的摄取情况。发现一种特定的富含线粒体(MR)的细胞群体,称为PNA + MR细胞(PNA是花生凝集素),在2.4微克/升109Cd的体内暴露1小时期间,其积累的109Cd比PNA - MR细胞、扁平上皮细胞(PV细胞)和黏液细胞多三倍以上。还在标准PBS和无Cl-的PBS中进行了体外109Cd暴露实验,暴露浓度为1至16微克/升109Cd,以进一步表征Cd2+摄取动力学。如体内实验所观察到的,当在PBS中通过体外程序进行暴露时,PNA + MR细胞积累的109Cd明显多于其他细胞类型。在这种条件下,所有细胞类型中Cd2+积累动力学可用米氏关系描述,两种MR细胞亚型的Km值约为3.0微克/升Cd(27纳摩尔),PV细胞的Km值为8.6微克/升Cd(77纳摩尔)。在无Cl-条件下进行的类似实验中,PNA + MR细胞中109Cd积累显著减少,但PNA - MR细胞或PV细胞中未出现这种情况。还进行了体外45Ca通量实验,以确定这些功能不同的鳃细胞群体中Ca2+转运的细胞定位。45Ca摄取在PNA + MR细胞中最为明显,其水平比PNA(-)MR细胞或PV细胞中高出三倍以上。本研究结果表明,PNA + MR细胞类型是虹鳟鳃上皮中Cd2+和Ca2+顶端进入的高亲和力和高容量位点。