The transfer of dPNA:GFP from the Bolwig's Organ to Laterna Neurons and two other candidate neuronal clusters

<strong>S1. </strong>Confocal optical sections of a Pdf-RFP1 expressing Lateral Neuron (LN) in the 2n instar larval brain that colocalized dPNA:GFP label (S1', arrowheads). S1" is a merged image of S1 and S1' channels. <strong>S2. </strong>Yet another example of dPNA:GFP (arrowheads) co-localization in Pdf-RFP1 expressing LNs. RFP expression often times inside the LNs appear to be aggregated. <strong>S3. </strong>Confocal image of the left hemisphere of a larval brain expressing dPNA:GFP in Bolwig's Organ terminals (BOt). Arrowhead indicates dPNA:GFP puncta in one LN that colocalizes with the corresponding Pdf-RFP1 expressing LN in S3". The encircled cluster of GFP-label farther away from the BOt and the highly localized dPNA:GFP relatively closer (arrows) represent candidate neurons that are not LNs. This supports the notion that other interneurons, besides the LNs, also contact the BOt. The background RFP label in the brain excluding the Pdf-RFP1 is due to the activity of 3xP3-RFP that is a part of the chromosomal attP landing site. I used the PhiC31 method (Bischoff et al. 2007) to integrate the UAS-dPNA:GFP-attB transgene into the 3rd chromosome. Removal of the 3xP3-RFP transgene from the landing site is currently underway.

S1. 表达Pdf-RFP1的2龄幼虫脑中侧神经元（Lateral Neuron, LN）的共聚焦光学切片，该切片的荧光信号与dPNA:GFP标记存在共定位（S1'，箭头所示）。S1''为S1与S1'通道的合并图像。
S2. 又一例表达Pdf-RFP1的侧神经元中dPNA:GFP（箭头所示）的共定位现象。侧神经元内的RFP表达有时会出现聚集情况。
S3. 博尔维格器官终末（Bolwig's Organ terminals, BOt）表达dPNA:GFP的幼虫脑左半球共聚焦图像。箭头指示单个侧神经元内的dPNA:GFP斑点，该斑点与S3''中表达Pdf-RFP1的对应侧神经元共定位。远离博尔维格器官终末的环状GFP标记簇，以及相对更靠近、局部高度富集的dPNA:GFP（箭头所示），均为非侧神经元的候选神经元。这一结果支持了“除侧神经元外，其他中间神经元也可与博尔维格器官终末建立连接”的观点。脑中除Pdf-RFP1外的背景RFP信号，源自作为染色体attP整合位点组分的3xP3-RFP的表达活性。本研究采用PhiC31重组酶系统（Bischoff等，2007）将UAS-dPNA:GFP-attB转基因整合至3号染色体。目前正在开展从该整合位点移除3xP3-RFP转基因的相关工作。