Donnelly Callum G, Bellone Rebecca R, Hales Erin N, Nguyen Annee, Katzman Scott A, Dujovne Ghislaine A, Knickelbein Kelly E, Avila Felipe, Kalbfleisch Ted S, Giulotto Elena, Kingsley Nicole B, Tanaka Jocelyn, Esdaile Elizabeth, Peng Sichong, Dahlgren Anna, Fuller Anna, Mienaltowski Michael J, Raudsepp Terje, Affolter Verena K, Petersen Jessica L, Finno Carrie J
Department of Population Health and Reproduction, School of Veterinary Medicine, University of California, Davis, Davis, CA, United States.
Veterinary Genetics Laboratory, School of Veterinary Medicine, University of California, Davis, Davis, CA, United States.
Front Genet. 2021 Mar 8;12:650305. doi: 10.3389/fgene.2021.650305. eCollection 2021.
Following the successful creation of a biobank from two adult Thoroughbred mares, this study aimed to recapitulate sample collection in two adult Thoroughbred stallions as part of the Functional Annotation of the Animal Genome (FAANG) initiative. Both stallions underwent thorough physical, lameness, neurologic, and ophthalmic (including electroretinography) examinations prior to humane euthanasia. Epididymal sperm was recovered from both stallions immediately postmortem and cryopreserved. Aseptically collected full thickness skin biopsies were used to isolate, culture and cryopreserve dermal fibroblasts. Serum, plasma, cerebrospinal fluid, urine, and gastrointestinal content from various locations were collected and cryopreserved. Under guidance of a board-certified veterinary anatomic pathologist, 102 representative tissue samples were collected from both horses. Whole tissue samples were flash-frozen and prioritized tissues had nuclei isolated and cryopreserved. Spatially contemporaneous samples of each tissue were submitted for histologic examination. Antemortem and gross pathologic examination revealed mild abnormalities in both stallions. One stallion (ECA_UCD_AH3) had unilateral thoracic limb lameness and bilateral chorioretinal scars. The second stallion (ECA_UCD_AH4) had subtle symmetrical pelvic limb ataxia, symmetrical prostatomegally, and moderate gastrointestinal nematodiasis. DNA from each was whole-genome sequenced and genotyped using the GGP Equine 70K SNP array. The genomic resources and banked biological samples from these animals augments the existing resource available to the equine genomics community. Importantly we may now improve the resolution of tissue-specific gene regulation as affected by sex, as well as add sex-specific tissues and gametes.
在成功从两匹成年纯种母马创建生物样本库之后，本研究旨在作为动物基因组功能注释（FAANG）计划的一部分，在两匹成年纯种种马中重复样本采集过程。在人道安乐死之前，两匹种马均接受了全面的身体、跛行、神经和眼科（包括视网膜电图）检查。在死后立即从两匹种马身上采集附睾精子并进行冷冻保存。无菌采集的全层皮肤活检样本用于分离、培养和冷冻保存真皮成纤维细胞。收集来自不同部位的血清、血浆、脑脊液、尿液和胃肠道内容物并进行冷冻保存。在一名具有专业认证的兽医解剖病理学家的指导下，从两匹马身上采集了102个代表性组织样本。全组织样本进行速冻，优先处理的组织分离出细胞核并进行冷冻保存。每个组织的空间同步样本被送去进行组织学检查。生前和大体病理学检查显示两匹种马均有轻度异常。一匹种马（ECA_UCD_AH3）有单侧胸肢跛行和双侧脉络膜视网膜瘢痕。第二匹种马（ECA_UCD_AH4）有轻微的对称性骨盆肢共济失调、对称性前列腺肿大和中度胃肠道线虫病。对每匹马的DNA进行全基因组测序，并使用GGP马70K SNP阵列进行基因分型。这些动物的基因组资源和保存的生物样本增加了马基因组学界现有的资源。重要的是，我们现在可以提高受性别影响的组织特异性基因调控的分辨率，以及增加性别特异性组织和配子。
