Brzustowicz L M, Gardner J P, Hopp L, Jeanclos E, Ott J, Yang X Y, Fekete Z, Aviv A
Hypertension Research Program, University of Medicine and Dentistry of New Jersey, New Jersey Medical School, Newark 07103-2714, USA.
Hypertension. 1997 Jan;29(1 Pt 2):158-64. doi: 10.1161/01.hyp.29.1.158.
Epstein-Barr virus-transformed lymphoblasts from patients with essential hypertension demonstrate enhanced G protein-mediated cytosolic free calcium ([Ca2+]i) response to platelet-activating factor (PAF). To map genes responsible for variation in G protein-coupled signaling, we used this cellular phenotype for a linkage study of transformed cell lines from the Centre d'Etude du Polymorphisme Humain (CEPH) reference pedigrees. The PAF-evoked change in [Ca2+]i ranged from 20 to 392 mmol/L and was highly reproducible within each cell line. PAF-elicited [Ca2+]i responses were obtained in lymphoblastic cell lines from five densely mapped pedigrees of the CEPH collection. Using PAF-evoked [Ca2+]i responses as a quantitative trait, two-point sibpair linkage analyses were conducted using 5150 markers from the Collaborative Human Linkage Center (CHLC) database. Nine loci, located on chromosomes 1, 4, 10, 11, 13, 16, and 17, were suggestive of linkage, with values of P < 7.4 x 10(-4). Multipoint linkage analysis produced a significant linkage finding (P = 2.1 x 10(-5) in one family at D16S151, with suggestive linkage results for seven additional markers spanning a 40-cM interval of chromosome 16. Multipoint analysis produced suggestive findings of linkage to eight loci from two distinct regions of chromosome 11 in another family. These results indicate that loci involved in the control of G protein-mediated mechanisms, suggested to be involved in the pathophysiology of essential hypertension, can be identified using cell lines from general pedigrees selected without any knowledge of the blood pressure status of the donors. This strategy represents an approach to rapidly and inexpensively mapping loci related to common, complex disorders, using phenotypes that are stable in immortalized lymphoblasts together with existing reference pedigree cell lines and genotype databases.
原发性高血压患者的爱泼斯坦-巴尔病毒转化淋巴细胞对血小板活化因子（PAF）表现出增强的G蛋白介导的胞质游离钙（[Ca2+]i）反应。为了定位负责G蛋白偶联信号传导变异的基因，我们利用这种细胞表型对来自人类多态性研究中心（CEPH）参考家系的转化细胞系进行连锁研究。PAF诱发的[Ca2+]i变化范围为20至392 mmol/L，并且在每个细胞系中具有高度可重复性。在CEPH收集的五个高密度定位家系的淋巴细胞系中获得了PAF诱发的[Ca2+]i反应。使用PAF诱发的[Ca2+]i反应作为数量性状，使用来自协作人类连锁中心（CHLC）数据库的5150个标记进行两点同胞对连锁分析。位于1、4、10、11、13、16和17号染色体上的9个位点提示存在连锁，P值<7.4×10(-4)。多点连锁分析产生了一个显著的连锁发现（在一个家系中，D16S151处P = 2.1×10(-5)），另外七个标记在16号染色体的40 cM区间内提示连锁结果。多点分析在另一个家系中产生了与11号染色体两个不同区域的八个位点连锁的提示性发现。这些结果表明，参与控制G蛋白介导机制的位点，被认为与原发性高血压的病理生理学有关，可以使用从普通家系中选择的细胞系来鉴定，而无需了解供体的血压状况。这种策略代表了一种利用永生化淋巴细胞中稳定的表型以及现有的参考家系细胞系和基因型数据库，快速且廉价地定位与常见复杂疾病相关位点的方法。
