Abstract
Bovine spongiform encephalopathy (BSE), which has been documented in 190,000 cases of BSE-infected cattle, to our knowledge, has not been reported in buffalo. Prion protein (PrP) is critical to susceptibility and development of BSE. Moreover, a new PrP-like protein, Shadoo (Sho), has been shown to have overlapping expression patterns and shared functions with PrP. Therefore, we hypothesize that differences in expression at the transcriptional level and/or the post-transcriptional level of the two genes may be associated with these specific differences between cattle and buffalo. We compared the relative mRNA expression of the prion protein gene (PRNP) and the Shadoo gene (SPRN) in 396 RNA samples using real-time PCR. We also analyzed PrP/Sho protein from 96 samples by Western blot. Our results demonstrated significantly lower PrP expression in the cerebellum, obex, mesenteric lymph node, and bronchial lymph node tissues, but higher relative expression of Sho in the cerebrum and spleen in buffalos compared with cattles. Although these results support our primary assumption, Sho and PrP expressions did not correlate with corresponding mRNA expression, suggesting that the biological modulations of both PrP and Sho proteins are at post-translational levels. Moreover, positive correlations between PRNP and SPRN were found in cattle and buffalo cerebrum as well as in buffalo obex. Conversely, negative correlations between PrP and Sho were detected in buffalo cerebellum and obex tissues. These findings suggest that additional post-transcriptional studies are warranted to elucidate mechanisms behind prion diseases.
摘要
全球有超过 19 万头黄牛被感染疯牛病, 却没有水牛患病的报道。 本研究通过分析与疯牛病关联的PRNP 和SPRN 基因转录水平及其蛋白 (PrP 和 Sho)水平的表达, 揭示了水牛小脑、脑闩、肠系膜淋巴结和支气管淋巴结 PrP 的表达量显著低于黄牛, 而水牛大脑和脾 Sho 表达量显著高于黄牛。然而 PrP 和 Sho 表达与它们相应的 mRNA 表达没有呈现出相同 趋势, 说明这两个蛋白的表达受到了转录后的调控作用。另外, 在水牛和黄牛的大脑和脑闩中, PRNP 和 SPRN mRNA 的表达呈正相关, 而水牛的小脑和脑闩组织中 PrP 和 Sho 蛋白的表达呈负相关, 这些结果提示转录后表达水平的研究在朊病毒疾病研究中的重要性。
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Acknowledgments
This work was supported by the National Natural Science Foundation of China (31060302, 31260032 and 31460038), and the Program for Innovative Research Team (in Science and Technology) in the University of Yunnan Province.
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Hui Zhao and Si-Qi Wang contributed equally to this work.
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11434_2016_1130_MOESM1_ESM.pdf
Figure S1. Relative expression of PRNP in cattle and buffalo. RNA was isolated from cerebrum (a), cerebellum (b), obex (c), mesenteric lymph node (d), bronchial lymph node (e), and spleen (f) of 34 cattle and 32 buffalo. Transcript quantities were analyzed by qPCR. The vertical ordinates show the relative expression of PRNP normalized to ACTB. The mean values of each species are indicated by horizontal lines. * Indicates significant differences for PRNP expression between cattle and buffalo. (PDF 27 kb)
11434_2016_1130_MOESM2_ESM.pdf
Figure S2. Relative expression of SPRN in cattle and buffalo. RNA was isolated from cerebrum (a), cerebellum (b), obex (c), mesenteric lymph node (d), bronchial lymph node (e), and spleen (f) of 34 cattles and 32 buffaloes. The vertical ordinates depict relative expression of SPRN normalized to ACTB. The mean values of each species are indicated by horizontal lines. * Indicates significant differences for SPRN expression between cattles and buffalos. (PDF 23 kb)
11434_2016_1130_MOESM3_ESM.png
Figure S3. The expression levels of PrP and Shadoo proteins. Six tissues, including cerebrum (a), cerebellum (b), obex (c), MLN (d), BLN (e) and spleen (f), from 8 cattles and 8 buffaloes were prepared and then immunoblotted for PrP and Sho proteins. (PNG 276 kb)
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Zhao, H., Wang, SQ., Qing, LL. et al. Expression of BSE-associated proteins in the CNS and lymphoreticular tissues of cattle and buffalo. Sci. Bull. 61, 1377–1383 (2016). https://doi.org/10.1007/s11434-016-1130-y
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DOI: https://doi.org/10.1007/s11434-016-1130-y