Pla Laura, Kühne Britta Anna, Guardia-Escote Laia, Vázquez-Aristizabal Paula, Loreiro Carla, Flick Burkhard, Gratacós Eduard, Barenys Marta, Illa Miriam
BCNatal-Barcelona Center for Maternal-Fetal and Neonatal Medicine (Hospital Clínic and Hospital Sant Joan de Déu), Fetal i+D Fetal Medicine Research Center, IDIBAPS, University of Barcelona, Center for Biomedical Research on Rare Diseases (CIBER-ER), Barcelona, Spain.
GRET, INSA-UB and Toxicology Unit, Pharmacology, Toxicology and Therapeutical Chemistry Department, Faculty of Pharmacy, University of Barcelona, Barcelona, Spain.
Front Toxicol. 2022 Jul 22;4:918520. doi: 10.3389/ftox.2022.918520. eCollection 2022.
The rabbit model is gaining importance in the field of neurodevelopmental evaluation due to its higher similarity to humans in terms of brain development and maturation than rodents. In this publication, we detailed 14 protocols covering toxicological relevant endpoints for the assessment of neurodevelopmental adverse effects in the rabbit species. These protocols include both and techniques, which also cover different evaluation time-points, the neonatal period, and long-term examinations at postnatal days (PNDs) 50-70. Specifically, the protocols (P) included are as follows: neurosphere preparation (GD30/PND0; P2) and neurosphere assay (P3), behavioral ontogeny (PND1; P4), brain obtaining and brain weight measurement at two different ages: PND1 (P5) and PND70 (P12), neurohistopathological evaluations after immersion fixation for neurons, astrocytes, oligodendrocytes and microglia (PND1; P6-9) or perfusion fixation (PND70; P12), motor activity (P11, open field), memory and sensory function (P11, object recognition test), learning (P10, Skinner box), and histological evaluation of plasticity (P13 and P14) through dendritic spines and perineuronal nets. The expected control values and their variabilities are presented together with the information on how to troubleshoot the most common issues related to each protocol. To sum up, this publication offers a comprehensive compilation of reliable protocols adapted to the rabbit model for neurodevelopmental assessment in toxicology.
由于兔子在大脑发育和成熟方面比啮齿动物与人类有更高的相似性，兔子模型在神经发育评估领域正变得越来越重要。在本出版物中，我们详细介绍了14种方案，涵盖了与毒理学相关的终点，用于评估兔种的神经发育不良影响。这些方案包括体内和体外技术，还涵盖了不同的评估时间点、新生儿期以及出生后第50 - 70天的长期检查。具体而言，所包括的方案（P）如下：神经球制备（妊娠第30天/出生后第0天；P2）和神经球测定（P3）、行为个体发育（出生后第1天；P4）、在两个不同年龄获取大脑并测量脑重量：出生后第1天（P5）和出生后第70天（P12）、对神经元、星形胶质细胞、少突胶质细胞和小胶质细胞进行浸入固定（出生后第1天；P6 - 9）或灌注固定（出生后第70天；P12）后的神经组织病理学评估、运动活动（P11，旷场试验）、记忆和感觉功能（P11，物体识别试验）、学习（P10，斯金纳箱）以及通过树突棘和神经元周围网进行可塑性的组织学评估（P13和P14）。文中给出了预期的对照值及其变异性，以及有关如何解决与每个方案相关的最常见问题的信息。总之，本出版物提供了一套全面的可靠方案汇编，适用于兔模型在毒理学中的神经发育评估。
