Contents lists available at ScienceDirect Research in Veterinary Science journal homepage: www.elsevier.com/locate/rvsc Transcriptome analysis of MAPK signaling pathway and associated genes to angiogenesis in chicken erythrocytes on response to thiram-induced tibial lesions Ali Raza Jahejo, Sheng Niu, Ding Zhang, Guan-bao Ning, Afrasyab Khan, Raza Ali Mangi, Muhammad Farhan Qadir, Ajab Khan, Jian-hui Li, Wen-xia Tian ⁎ College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China ARTICLE INFO Keywords: Angiogenesis Erythrocytes MAP kinase Tibial dyschondroplasia Thiram Transcriptome ABSTRACT This study was planned to investigate TD (Tibial dyschondroplasia) on the potential MAPK signaling pathway and angiogenesis related genes. Forty-eight broilers were allotted into control (C) and treatment (T) groups of 2, 6 and 15 days as C1, C2, C3, T1, T2 and T3. The histopathology results revealed that tibiotarsus bone of chickens had more lesions on day 6 (T2 group). The chondrocytes were disordered, and the size, shape and proliferation were affected. Transcriptome results revealed that differentially expressed genes (DEGs) identified were 63, 1026, 623, 130, 141 and 146 in C1 (2 days control vs 6 days control); C2 (2 days control vs 15 days control); C3 (6 days control vs 15 days control); T1 (2 days treatment vs 6 days treatment); T2 (2 days treatment vs 15 days treatment) and T3 (6 days treatment vs 15 days treatment) groups respectively. Whereas, 10 angiogenesis re- lated-genes RHOC, MEIS2, BAIAP2, TGFBI, KLF2, CYR61, PTPN11, PLXNC1, HSPH1 and NRP2 were down- regulated on day 6 in the treatment group. The pathway which was found enriched in the control and treatment groups was MAPK signaling pathway. Therefore selected 10 MAPK signaling pathway-related genes RAC2, MAP3K1, PRKCB, FLNB, IL1R1, PTPN7, RPS6KA, MAP3K6, GNA12 and HSPA8 which were found significantly downregulated in the treatment group on day 6. It is concluded that angiogenesis and MAPK signaling pathway related genes has an essential role in TD, as those top screened genes found downregulated in the thiram fed chickens when TD observed severed on day 6. 1. Introduction Worldwide pesticides are being used in fields to control the agri- cultural pests, weeds, and diseases which can damage crops and reduce productivity. Though, extensive usage of pesticides is threatening to the human population because of its toxicity which carries into living or- ganisms via the food chain, air, water and soil (Osman et al., 2012). Tetramethyl thiuram disulfide (thiram) is a pesticide which is being extensively used in agriculture for treating and storing food grains (Kunkur et al., 2010). It is lipophilic which combines with the cell membrane to cause cytotoxicity, bone formation disorder, cartilage cell damage, immune depressions, membrane damage and inhibit angio- genesis (Beckmann et al., 2014; Zhang et al., 2018b). It has a very deleterious effect on the poultry chickens and causes a growth plate cartilage disease tibial dyschondroplasia (TD) in many avian species, by feeding diets containing thiram (Tian et al., 2013; Zhang et al., 2018a). The incidence of TD in birds is significantly increased because of providing the thiram-contaminated feed (Rath et al., 2005; Niu et al., 2019; Zhang et al., 2018a). TD is one of the most widespread leg disorders, which is associated with fast-growing chickens and characterized by non-vascularized and non-mineralized tibia (Nabi et al., 2016). The growth of longitudinal bone begins with the division of cells in the proliferative zone (Genin et al., 2008). The cartilage cell damaged in TD due to the apoptosis process and abnormal protein secretions in the cartilage cells, causing a decrease in the cartilage extracellular matrix degradation rate, which limits the space for bone deposition (Shim et al., 2012; Velada et al., 2011). Thiram destroyed the regular column of chondrocytes, and the number of cells decreased in the tibial growth plate (TGP) of chickens (Zhang et al., 2018b). The cells degraded, dead and arranged in irre- gular columns in the hypertrophic zone of TD (Iqbal et al., 2018). La- meness in TD is an animal welfare problem for broiler chickens. Thus, any information regarding the mechanisms related to TD is of utmost importance. https://doi.org/10.1016/j.rvsc.2019.10.013 Received 18 April 2019; Received in revised form 28 August 2019; Accepted 21 October 2019 ⁎ Corresponding author. E-mail address: wenxiatian@126.com (W.-x. Tian). Research in Veterinary Science 127 (2019) 65–75 0034-5288/ © 2019 Elsevier Ltd. All rights reserved. T