Asala Tolulope E, Dasmahapatra Asok K, Myla Anitha, Tchounwou Paul B
RCMI Center for Environmental Health, Jackson State University, 1400 JR Lynch Street, Jackson, Mississippi 39217, USA.
Department of BioMolecular Sciences, Environmental Toxicology Division, University of Mississippi, University, Mississippi 38677, USA.
Data Brief. 2021 Nov 23;39:107625. doi: 10.1016/j.dib.2021.107625. eCollection 2021 Dec.
This article presents the experimental datasets obtained from the histological/histochemical studies of endocrine disrupting effects of graphene oxide (GO) on thyroid follicles and gas gland (GG) cells of Japanese medaka larvae at the onset of maturity. The experiment was conducted on one day-post hatch (dph) starved fries (orange-red variety) immersed in different concentrations of GO (2.5-20.0 mg/L) and no GO (controls) in embryo-rearing medium (ERM) for 96 h under laboratory conditions (25 ± 1 °C; light cycle 16 h light: 8 h dark). After treatment, larvae were maintained in balanced salt solution (BSS) with food and allowed depuration for 6 more weeks in a GO-free environment. On 47 dph, the larvae were anesthetized in MS 222 and their total lengths (mm) and weights (mg) were measured, and they were then cut into three small pieces (head, trunk, and tail). Head and trunk regions were fixed in 4% PFA in 20 mM PBS for 48 h at room temperature and the post-anal tail was preserved in TRI reagent and kept at -20 °C until analysis. Tissues in 4% PFA were used for cutting 5µm thick paraffin sections in a manual rotary microtome. Sections of head regions were evaluated for thyroid follicles after hematoxylin-eosin (HE) or Periodic acid-Schiff (PAS) staining. Trunk sections were used for swim bladder (SB) inflation studies and for phenotypic sex (ovary and testis) of the larvae after HE staining. Genetic sex assessment was made from tail DNA by genotyping Y chromosome-specific male sex-determining gene . Digital images were captured by using either an Olympus B-max 40 microscope attached to a camera with Q-capture Pro 7 software or an Olympus CKX53 microscope with DP22 camera and CellSens software. Images of thyroid follicles and GG cells were analyzed using imagej software. HE stained histological sections of thyroid follicles near the heart and branchial regions were captured and the area (µm) of individual follicles (minimum 3) available in the entire section were measured. The heights of thyrocytes (µm) were determined directly. Manual counting of GG cells was made from the digital images captured in several regions of the SB avoiding blood cells and other cells which have indistinct nucleus and pale cytoplasm; results were expressed as the number of GG cells/mm. Data were analyzed by GraphPad prism version 7.04. For normally distributed data, one-way ANOVA followed by post-hoc Tukey's test or unpaired parametric "t" test including Welch's correction was used. Otherwise, Kruskal-Wallis test followed by nonparametric Mann-Whitney's test as a post hoc test was used. Data were expressed as means ±SEM and the level of significance was set at < 0.05.
本文展示了从组织学/组织化学研究中获得的实验数据集，该研究针对氧化石墨烯（GO）对日本青鳉幼鱼成熟初期甲状腺滤泡和鳔腺（GG）细胞的内分泌干扰效应。实验使用孵化后一天（dph）饥饿的鱼苗（橙红色品种），将其置于胚胎饲养培养基（ERM）中不同浓度的GO（2.5 - 20.0 mg/L）和无GO（对照组）中，在实验室条件（25 ± 1°C；光照周期16小时光照：8小时黑暗）下浸泡96小时。处理后，幼鱼在含有食物的平衡盐溶液（BSS）中饲养，并在无GO的环境中净化6周。在47 dph时，将幼鱼用MS 222麻醉，测量其全长（mm）和体重（mg），然后切成三块（头部、躯干和尾部）。头部和躯干区域在室温下用含20 mM PBS的4%多聚甲醛固定48小时，肛门后的尾巴保存在TRI试剂中并在 - 20°C保存直至分析。用4%多聚甲醛固定的组织用于在手动旋转切片机上切成5μm厚的石蜡切片。头部区域的切片在苏木精 - 伊红（HE）或过碘酸 - 希夫（PAS）染色后评估甲状腺滤泡。躯干切片用于鳔（SB）充气研究以及HE染色后幼鱼的表型性别（卵巢和睾丸）鉴定。通过对尾部DNA进行Y染色体特异性雄性性别决定基因的基因分型来进行遗传性别评估。使用连接有配备Q - capture Pro 7软件的相机的Olympus B - max 40显微镜或配备DP22相机和CellSens软件的Olympus CKX53显微镜拍摄数字图像。使用imagej软件分析甲状腺滤泡和GG细胞的图像。拍摄心脏和鳃区域附近甲状腺滤泡的HE染色组织学切片，并测量整个切片中单个滤泡（至少3个）的面积（μm）。直接测定甲状腺细胞的高度（μm）。从SB几个区域拍摄的数字图像中手动计数GG细胞，避开血细胞和其他细胞核不清晰且细胞质淡染的细胞；结果以GG细胞数/mm表示。数据用GraphPad prism 7.04版本进行分析。对于正态分布的数据，使用单因素方差分析，随后进行事后Tukey检验或包括Welch校正的未配对参数 “t” 检验。否则，使用Kruskal - Wallis检验，随后进行非参数Mann - Whitney检验作为事后检验。数据表示为平均值±SEM，显著性水平设定为<0.05。
