Ohno Yukihiro, Shimizu Saki, Tatara Ayaka, Imaoku Takuji, Ishii Takahiro, Sasa Masashi, Serikawa Tadao, Kuramoto Takashi
Laboratory of Pharmacology, Osaka University of Pharmaceutical Sciences, Takatsuki, 569-1094, Japan.
Department of Physiology and Neurobiology, Graduate School of Medicine, Kyoto University, Kyoto, 606-8501, Japan.
PLoS One. 2015 May 13;10(5):e0123529. doi: 10.1371/journal.pone.0123529. eCollection 2015.
Genetic factors are thought to play a major role in the etiology of essential tremor (ET); however, few genetic changes that induce ET have been identified to date. In the present study, to find genes responsible for the development of ET, we employed a rat model system consisting of a tremulous mutant strain, TRM/Kyo (TRM), and its substrain TRMR/Kyo (TRMR). The TRM rat is homozygous for the tremor (tm) mutation and shows spontaneous tremors resembling human ET. The TRMR rat also carries a homozygous tm mutation but shows no tremor, leading us to hypothesize that TRM rats carry one or more genes implicated in the development of ET in addition to the tm mutation. We used a positional cloning approach and found a missense mutation (c. 1061 C>T, p. A354V) in the hyperpolarization-activated cyclic nucleotide-gated 1 channel (Hcn1) gene. The A354V HCN1 failed to conduct hyperpolarization-activated currents in vitro, implicating it as a loss-of-function mutation. Blocking HCN1 channels with ZD7288 in vivo evoked kinetic tremors in nontremulous TRMR rats. We also found neuronal activation of the inferior olive (IO) in both ZD7288-treated TRMR and non-treated TRM rats and a reduced incidence of tremor in the IO-lesioned TRM rats, suggesting a critical role of the IO in tremorgenesis. A rat strain carrying the A354V mutation alone on a genetic background identical to that of the TRM rats showed no tremor. Together, these data indicate that body tremors emerge when the two mutant loci, tm and Hcn1A354V, are combined in a rat model of ET. In this model, HCN1 channels play an important role in the tremorgenesis of ET. We propose that oligogenic, most probably digenic, inheritance is responsible for the genetic heterogeneity of ET.
遗传因素被认为在特发性震颤(ET)的病因中起主要作用;然而,迄今为止,很少有能诱发ET的基因变化被确定。在本研究中,为了找到与ET发生相关的基因,我们采用了一个大鼠模型系统,该系统由一个震颤突变品系TRM/Kyo(TRM)及其亚系TRMR/Kyo(TRMR)组成。TRM大鼠为震颤(tm)突变的纯合子,表现出类似于人类ET的自发性震颤。TRMR大鼠也携带纯合的tm突变,但不表现出震颤,这使我们推测,除了tm突变外,TRM大鼠还携带一个或多个与ET发生相关的基因。我们使用定位克隆方法,在超极化激活环核苷酸门控1通道(Hcn1)基因中发现了一个错义突变(c. 1061 C>T,p. A354V)。体外实验中,A354V HCN1无法传导超极化激活电流,这表明它是一个功能丧失突变。在体内用ZD7288阻断HCN1通道会在非震颤的TRMR大鼠中诱发运动性震颤。我们还发现,经ZD7288处理的TRMR大鼠和未处理的TRM大鼠的下橄榄核(IO)均有神经元激活,而IO损伤的TRM大鼠震颤发生率降低,这表明IO在震颤发生中起关键作用。在与TRM大鼠相同的遗传背景下,仅携带A354V突变的大鼠品系不表现出震颤。总之,这些数据表明,在ET大鼠模型中,当两个突变位点tm和Hcn1A354V结合时会出现身体震颤。在这个模型中,HCN1通道在ET的震颤发生中起重要作用。我们提出,寡基因遗传,很可能是双基因遗传,是ET遗传异质性的原因。