Linkage analysis using platelet-activating factor Ca2+ response in transformed lymphoblasts.
Linkage analysis using platelet-activating factor Ca2+ response in transformed lymphoblasts.
复制标题
使用转化淋巴母细胞中血小板激活因子 Ca2 反应进行连锁分析。
DOI：
10.1161/01.hyp.29.1.158
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Aviv,A
中科院分区：
文献类型：
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作者：
Brzustowicz,LM;Gardner,JP;Hopp,L;Jeanclos,E;Ott,J;Yang,XY;Fekete,Z;Aviv,A
原发性高血压患者EB病毒转化的淋巴母细胞对血小板活化因子（PAF）的反应性增强。为了绘制G蛋白偶联信号变化的基因，我们使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引起的[Ca~（2+）]变化范围为20~392 mmol/L，在各细胞系内重复性好。PAF引起的[Ca2 +] i反应，从五个密集映射的CEPH收集家系的淋巴母细胞系。使用PAF诱发的[Ca~（2+）] 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。位于第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。多点连锁分析在D16S151的一个家族中产生了显著的连锁发现（P = 2.1 × 10 − 5），并提示了跨越16号染色体40 cM间隔的7个额外标记的连锁结果。多点分析产生了暗示性的结果，连锁的8个位点从两个不同的区域的11号染色体在另一个家庭。这些结果表明，基因座参与控制G蛋白介导的机制，建议参与原发性高血压的病理生理学，可以确定使用细胞系从一般家系选择没有任何知识的血压状态的捐助者。该策略代表了一种快速且廉价地绘制与常见复杂疾病相关的基因座的方法，使用在永生化淋巴母细胞中稳定的表型以及现有的参考谱系细胞系和基因型数据库。
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+]iranged from 20 to 392 mmol/L and was highly reproducible within each cell line. PAF-elicited [Ca2+]iresponses were obtained in lymphoblastic cell lines from five densely mapped pedigrees of the CEPH collection. Using PAF-evoked [Ca2+]iresponses 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 ofP<7.4×10−4. Multipoint linkage analysis produced a significant linkage finding (P=2.1×10−5) 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.