CHROMIUM DISTRIBUTION AND TOXICITY IN MAMMALIAN CELLS

我们将通过以下方式来检查铬的分布和遗传毒性 体内铬毒性的系统形态计量学分析 在染铬小鼠的肝、肺、肾组织中。至 到目前为止，体内研究主要着眼于在 给药部位并具有铬的特征- 在整个器官水平上诱发恶性肿瘤。我们的 研究体内铬效应的研究方法将是 使用创新的计算机辅助显微镜(CAM) 量化单个细胞中的变化的系统，由 细胞化学、组织学和放射自显影分析。我们的内部 体外研究结果让我们假设，对铬敏感的 单元格是多个单元格中的一个(OCAM)，以及 量化单个细胞的能力是至关重要的。使用 计算机的精度、速度和内存，我们可以 通过实验确定和量化铬对单个细胞的影响 (特别是细胞核)。 铬在体内的遗传毒性将被系统地研究。 使用以下研究方法：1)研究急性和 对肝、肺、肾(与肝脏)的慢性毒性作用 作为原型器官)在给铬后，使用 CAM增强形态计量学分析。急性毒性研究 将为长期研究提供基础。2)测量铬 放射自显影和组织化学方法在组织中的分布 由CAM系统增强的技术，以量化铬- 结合在单个细胞的核和细胞质中。3)至 开发和验证其他增强的CAM技术，以 描述单个细胞的核或基因组变化：a) 使用小鼠cdna探针的原位杂交；b)质地分析 从不同的细胞状态获得的核的特征 细胞死亡(细胞凋亡和坏死)；c)电泳/高 染色质组分的高效液相色谱分析 描述细胞死亡的特征。4)我们研究的最终目的是 将铬器官毒性的研究结果与 分布研究。启用CAM存储的数据库 铬诱导的变化，我们将生成细胞原型 肝、肾、肺，已按治疗分类 间隔和剂量。

We will examine chromium distribution and genotoxicity by doing a systematic morphometric analysis of in vivo chromium toxicity in the liver, lung, kidney of mice injected with chromium. To date, in vivo studies have looked mainly for tumor induction at the sites of administration and have characterized chromium- induced malignancies at the level of the whole organ. Our research approach to the study of in vivo Cr effects will be the use of an innovative CAM (Computer-Assisted Microscopy) system to quantify changes in individual cells, as analyzed by cytochemical, histological, and radioautographic assays. Our in vitro findings led us to hypothesize that the chromium-sensitive cells are on the order of One Cell Among Many (OCAM), and that the ability to quantify individual cells was critical. With the computer's precision, speed, and memory, we can experimentally define and quantify Cr effects on individual cells (particularly, nuclei). Chromium genotoxicity, in vivo, will be studied systematically using the following research approaches: 1) To study acute and chronic toxic effects on the liver, lung, and kidney (with liver being the prototype organ) after chromium administration, using CAM-enhanced morphometric analysis. Acute toxicity studies will provide the basis for chronic studies. 2) To measure Cr distribution in tissues, using radioautographic and histochemical techniques augmented by the CAM system, to quantify Cr- binding in the nucleus and cytoplasm of individual cells. 3) To develop and verify other CAM-enhanced techniques to characterize nuclear or genomic changes in single cells: a) in situ hybridization, using mouse cDNA probes; b) textural analysis of nuclei obtained from different cellular states to characterize cell death (apoptosis and necrosis); c) electrophoresis/high performance liquid chromatography of chromatin components to characterize cell death. 4) The ultimate goal of our research is to correlate the findings of the chromium organ toxicity and distribution studies. With the CAM-stored database on chromium-induced changes, we will generate cell archetypes of liver, kidney and lungs, which have been sorted by treatment interval and dosage.