四环素调控A30P突变α-synuclein转基因小鼠的构建

Establishment of Tetracycline-controlled A30P Mutation α-synuclein Transgenic Mice

为了构建四环素调控的人A30P突变α-synuclein转基因小鼠模型,将外源基因pTRE2-syn和pBC-rtTA同时显微注射到FVB小鼠(Mus muscculus)受精卵的雄原核中,将注射后存活的受精卵移植到同期发情的假孕受体鼠输卵管中,出生个体经PCR检测,获得rtTA和A30P突变α-synuclein双阳性转基因雌鼠1只,A30P单基因阳性雄鼠13只并传代。强力霉素诱导后双阳性后代脑区各部分A30P突变α-synucleinmRNA均有表达,而在诱导满4周后,脑干α-synuclein蛋白表达明显增加,8周后增加更明显。结果表明,通过强力霉素诱导后,可在小鼠小脑、脑干、海马、皮层检测到A30P mRNA表达,脑干α-synuclein表达量显著增加。

To establish tetracycline-controlled A30P mutation α-synuclein transgenic mice, pTRE2-syn and pBC-rtTA dual transgenic mice were generated by pro-nuclear microinjection and embryo transplantation, Positive founder mice were identified through PCR. An rtTA and A30P dual positive transgenic founder and thirteen A30P single transgenic founders were obtained. The expression of A30P mRNA was detected by RT-PCR in the brain of the dual transgenic mice after administration of tetracycline. In addition, the expression of α-synuclein protein in the brainstem was also significandy increased in double-transgenic mice after 4-week continuous treatment with doxycycline.