Microscopy images of the effects of PDZ-RhoGEF manipulation on dendritic spines and videos of effects on PDZ-RhoGEF on mouse behavioral phenotypes

PDZ-RhoGEF is a RhoA-specific guanine nucleotide exchange factor (GEF) for the RhoA small GTPase. The purpose of these experiments is to determine the effects of PDZ-RhoGEF overexpression and knockdown on mouse behavioral phenotypes. Relevant behaviors including assessments of spatial working memory, recognition memory, social behavior, and stress coping. Herpes simplex viral vectors (HSV) were used to overexpress PDZ-RhoGEF. Knockdown of PDZ-RhoGEF was accomplished via packaging of short hairpin RNA (shRNA) into adeno-associated viral vectors (AAV) against Arhgef11 (the gene that encodes the PDZ-RhoGEF protein). Viral vectors were surgically infused directly into the medial prefrontal cortex (mPFC) of young adult male mice. Using the same HSV approach, the effects of PDZ-RhoGEF overexpression on the density and morphology of dendritic spines was also analyzed. Finally, using structured illumination microscopy (SIM), the spatial location of PDZ-RhoGEF and that of one of its direct inter..., Object in place behavior videos: Mice were infused with HSV-PDZ-RhoGEF-green fluorescence protein (HSV-PRG-GFP) or HSV-GFP into the medial prefrontal cortex of mice. 5 days post-infusion, mice were assessed in an object-in-place recognition task. During trial 1, mice were allowed to freely explore 4 unique, non-identical objects in an open arena. Mice were then returned to their home cages for 7 minutes and then placed back into the open field arena (trial 2) with the same objects as before, however, the location of two of the objects were swapped in location. This results in two objects whose location is fixed between trials and two objects whose location is altered between trials. The amount of time mice spent directly investigating the swapped and non-swapped objects was done manually using a scientific stopwatch with 0.01 second accuracy. For the knockdown of PDZ-RhoGEF, these same procedures were followed except mice were infused with AAV-Arhgef11 shRNA-GFP or AAV-scrambled control..., , # Microscopy images of the effects of PDZ-RhoGEF manipulation on dendritic spines and videos of effects on PDZ-RhoGEF on mouse behavioral phenotypes

[https://doi.org/10.5061/dryad.0k6djhb9p](https://doi.org/10.5061/dryad.0k6djhb9p)

## Description of the data and file structure

The behavioral data involves the use of viral-mediated gene transfer to overexpress or knockdown a particular gene/protein (Arhgef11/PDZ-RhoGEF) in the medial prefrontal cortex of young adult male mice. This same viral approach was used to overexpress PDZ_RhoGEF into the medial prefrontal cortex of young adult mice and assess the effects on the density and morphology of dendritic spines. In addition, in cultured neurons, the spatial localization of PDZ-RhoGEF within dendritic spine neck and head regions was assessed. Finally, the spatial overlap between PDZ-RhoGEF and overexpressed DISC1 protein was also assessed in dendritic spines.

### Files and variables

#### File: AAV-arhgef11\_shrna\_object\_in\_place.zi...

PDZ-RhoGEF是一种针对RhoA小GTP酶（small GTPase）的RhoA特异性鸟苷酸交换因子（guanine nucleotide exchange factor, GEF）。本实验旨在探究PDZ-RhoGEF过表达与敲低对小鼠行为表型的影响。相关行为学评估涵盖空间工作记忆、识别记忆、社交行为及应激应对能力。研究采用单纯疱疹病毒载体（herpes simplex viral vectors, HSV）实现PDZ-RhoGEF的过表达；而PDZ-RhoGEF的敲低则通过将靶向Arhgef11（编码PDZ-RhoGEF蛋白的基因）的短发夹RNA（short hairpin RNA, shRNA）包装至腺相关病毒载体（adeno-associated viral vectors, AAV）中来完成。将上述病毒载体通过外科手术直接注射至年轻成年雄性小鼠的内侧前额叶皮层（medial prefrontal cortex, mPFC）。本研究同时采用相同的HSV载体策略，分析PDZ-RhoGEF过表达对树突棘密度与形态的影响。最后，借助结构照明显微镜（structured illumination microscopy, SIM），探究PDZ-RhoGEF及其一种直接相互作用蛋白的空间定位情况。

物体位置识别行为实验视频：将小鼠分别注射HSV-PDZ-RhoGEF-绿色荧光蛋白（green fluorescence protein, GFP）或HSV-GFP至内侧前额叶皮层。病毒感染5天后，对小鼠开展物体位置识别任务测试。在试验1阶段，允许小鼠在开放场域中自由探索4个独特的非重复物体。随后将小鼠放回饲养笼中7分钟，再将其放回同一开放场域（试验2阶段），此时仍放置与试验1相同的物体，但其中两个物体的位置发生调换。由此场域中存在两类物体：两组物体在两次试验中位置保持不变，另外两组物体的位置发生改变。研究人员使用精度达0.01秒的专业秒表，手动记录小鼠对调换位置与未调换位置物体的直接探索时长。针对PDZ-RhoGEF敲低组的实验，遵循相同流程，仅将病毒载体替换为AAV-Arhgef11 shRNA-GFP或AAV-阴性对照（scrambled control）相关实验……

# PDZ-RhoGEF调控对树突棘的显微镜成像及对小鼠行为表型影响的视频数据集
[https://doi.org/10.5061/dryad.0k6djhb9p](https://doi.org/10.5061/dryad.0k6djhb9p)

## 数据与文件结构说明
本行为学数据集通过病毒介导的基因转移技术，在年轻成年雄性小鼠的内侧前额叶皮层中过表达或敲低特定基因/蛋白（Arhgef11/PDZ-RhoGEF）。研究同样采用该病毒载体策略，在年轻成年小鼠的内侧前额叶皮层中过表达PDZ-RhoGEF，以评估其对树突棘密度与形态的影响。此外，在培养的神经元中，本研究还评估了PDZ-RhoGEF在树突棘颈区与头区的空间定位情况。最终，还探究了PDZ-RhoGEF与过表达的DISC1蛋白在树突棘中的空间共定位情况。

### 文件与变量
#### 文件：AAV-arhgef11_shrna_object_in_place.zi……