Abstract

Insects, as poikilotherms, have adaptations to deal with wide ranges in temperature fluctuation. Allelic variations in the foraging gene that encodes a cGMP dependent protein kinase, were discovered to have effects on behavior in Drosophila by Dr. Marla Sokolowski in 1980. This single gene has many pleiotropic effects and influences feeding behavior, metabolic storage, learning and memory and has been shown to affect stress tolerance. PKG regulation affects motoneuronal thermotolerance in Drosophila larvae as well as adults. While the focus of thermotolerance studies has been on the modulation of neuronal function, other cell types have been overlooked. Because glia are vital to neuronal function and survival, we wanted to determine if glia play a role in thermotolerance as well. In our investigation, we discovered a novel calcium wave at the larval NMJ and set out to characterize the wave’s dynamics and the potential mechanism underlying the wave prior to determining what effect, if any, PKG modulation has on the thermotolerance of glia cells. Using pharmacology, we determined that calcium buffering mechanisms of the mitochondria and endoplasmic reticulum play a role in the propagation of our novel glial calcium wave. By coupling pharmacology with genetic manipulation using RNA interference (RNAi), we found that PKG modulation in glia alters thermoprotection of function as well as glial calcium wave dynamics.

摘要

作为变温动物，昆虫具有适应大范围温度波动的能力。1980 年，Marla Sokolowski 博士发现觅食基因中编码 cGMP 依赖性蛋白激酶的等位基因变异对果蝇的行为有影响。这个单一基因具有许多多效性，并影响摄食行为、代谢储存、学习和记忆以及已被证明会影响压力耐受性。PKG 调节影响果蝇的运动神经元耐热性幼虫和成虫一样。虽然耐热性研究的重点是调节神经元功能，但其他细胞类型却被忽视了。因为神经胶质对神经元功能和生存至关重要，我们想确定神经胶质是否也在耐热性中起作用。在我们的调查中，我们在幼虫 NMJ 发现了一种新的钙波，并在确定 PKG 调制对神经胶质细胞的耐热性有什么影响（如果有的话）之前，开始描述该波的动力学和该波背后的潜在机制。使用药理学，我们确定线粒体和内质网的钙缓冲机制在我们新的神经胶质钙波的传播中起作用。通过使用 RNA 干扰 (RNAi) 将药理学与遗传操作相结合，