Koppel Indrek, Aid-Pavlidis Tamara, Jaanson Kaur, Sepp Mari, Palm Kaia, Timmusk Tõnis
Genesis. 2010 Apr;48(4):214-9. doi: 10.1002/dvg.20606.
Brain-derived neurotrophic factor (BDNF), a member of the neurotrophin family of neurotrophic factors, has important functions in the peripheral and central nervous system of vertebrates. We have generated bacterial artificial chromosome (BAC) transgenic mice harboring 207 kb of the rat BDNF (rBDNF) locus containing the gene, 13 kb of genomic sequences upstream of BDNF exon I, and 144 kb downstream of protein encoding exon IX, in which protein coding region was replaced with the lacZ reporter gene. This BDNF-BAC drove transgene expression in the brain, heart, and lung, recapitulating endogenous BDNF expression to a larger extent than shorter rat BDNF transgenes employed previously. Moreover, kainic acid induced the expression of the transgenic BDNF mRNA in the cerebral cortex and hippocampus through preferential activation of promoters I and IV, thus recapitulating neuronal activity-dependent transcription of the endogenous BDNF gene.
脑源性神经营养因子(BDNF)是神经营养因子家族的成员之一,在脊椎动物的外周和中枢神经系统中具有重要功能。我们构建了携带大鼠BDNF(rBDNF)基因座207 kb的细菌人工染色体(BAC)转基因小鼠,该基因座包含该基因、BDNF外显子I上游13 kb的基因组序列以及编码外显子IX下游144 kb的基因组序列,其中蛋白质编码区被lacZ报告基因取代。该BDNF-BAC驱动转基因在脑、心脏和肺中表达,比之前使用的较短的大鼠BDNF转基因在更大程度上重现了内源性BDNF的表达。此外,海藻酸通过优先激活启动子I和IV诱导转基因BDNF mRNA在大脑皮层和海马体中的表达,从而重现了内源性BDNF基因的神经元活性依赖性转录。