Optical Coherence Tomography to Study Effect of Poly - Drug Exposure on Fetal Brain Development

 描述(申请人提供)：本研究的总体目标是开发一种基于光学相干断层扫描(OCT)的高分辨率小鼠胚胎脑成像和分析方法，并将该方法与分子分析相结合，以了解乙醇(Etoh)和尼古丁(NIC)对胚胎脑发育的影响。孕妇接触这些物质与胎儿生长迟缓和神经毒性有关，这些毒素经常在怀孕期间共同滥用。然而，关于NIC和EtoH对胎儿的联合影响的研究非常有限，而且它们对胎儿发育的分子机制，特别是大脑的联合作用，了解得很少。因此，迫切需要通过开发能够对发育中的大脑进行实时纵向分析的高分辨率成像技术来了解无水乙醇和NIC之间的相互作用。有趣的是，我们最近的研究表明，乙醇和NIC对胎儿神经干细胞的发育具有相互拮抗的作用。在这项提议中，我们将通过实施我们开发的创新的更高分辨率的胚胎脑成像和动态定量分析方法来研究这些毒素是否确实对活小鼠胚胎的大脑发育具有拮抗或协同作用。我们开创了基于OCT的方法，用于子宫内活体成像和对小鼠胚胎在子宫内的纵向表型分析。在这里，我们建议进一步发展纵向脑成像和分析的技术和方法。这项研究的重点是相当于第二个三个月的发育期，在这个时期，神经干细胞产生了成年大脑的大部分神经元。这一建议的中心假设是，乙醇和NIC在胎儿大脑发育中具有部分拮抗作用。随着该计划的成功完成，我们将建立一种活的小鼠胚胎脑成像方法，将开发一套方案和详细的评估，以定量描述细胞分辨率的动态胚胎脑发育，并将调查Etoh和NIC是否协同扰乱大脑发育或显示部分拮抗作用。我们还将评估使用尼古丁受体拮抗剂和激动剂预防无水乙醇和NIC单独和联合作用的可行性。因此，从基础生物学和致畸的角度研究这一效应都具有重要意义，因为它可能对逆转畸形学的新颖和创新疗法的开发具有特别重要的影响。

 DESCRIPTION (provided by applicant): The overall objective of this study is to develop an optical coherence tomography (OCT) based high- resolution mouse embryonic brain imaging and analysis approach, and to use this method in correlation with molecular analysis to understand the interplay between ethanol (EtOH) and nicotine (NIC) effects on embryonic brain development. Maternal exposures to these substances are linked to fetal growth retardation and neurotoxicity, and these toxins often co-abused during pregnancy. However, studies on the combined fetal effects of NIC and EtOH are very limited and their combined effects on molecular mechanisms of fetal development, particularly the brain, are poorly understood. Therefore, there is a critical need to understand the interplay between the effects of EtOH and NIC via development of high-resolution imaging technique capable of live longitudinal analysis of developing brain. Intriguingly, our recent studies suggested that EtOH and NIC exert mutually antagonistic effects on fetal neuronal stem cells development. In this proposal, we will investigate if these toxins have indeed antagonistic or synergetic effects on embryonic brain development in live mouse embryos with implementation of an innovative higher-resolution embryonic brain imaging and dynamic quantitative analysis approaches, which we develop. We have pioneered OCT-based methodology for live in utero imaging and longitudinal phenotypic analysis of mouse fetuses in utero. Here we propose to further develop both the technology and the methodology for longitudinal brain imaging and analysis. The study is focused on the second trimester- equivalent period of development, when the neuronal stem cells give rise to most of the neurons of the adult brain. The central hypothesis of this proposal is that EtOH and NIC have partially antagonistic effects in fetal brain development. By successful accomplishment of the proposal, we will establish a live mouse embryonic brain imaging approach, will develop a set of protocols and detailed assessments to quantitatively characterize dynamic embryonic brain development with cellular resolution, and will investigate if EtOH and NIC synergize to disrupt the brain development or exhibit partially antagonistic effects. We will also assess the feasibility of using nicotinic receptor antagonists and agonists t prevent the individual and combined effects of EtOH and NIC. Therefore, studying this effect is highly significant from both fundamental biology and teratogenic points of view since it may have particular significant impact for the development of novel and innovative therapies for reversing teratology.