Cloning and Tissue Expression of the TTR Gene During the Initiation of Puberty in Dolang Sheep
Cloning and Tissue Expression of the TTR Gene During the Initiation of Puberty in Dolang Sheep
Zhishuai Zhang, Zhiyuan Sui, Jihu Zhang, Qingjin Li, Yongjie Zhang, Chenguang Wang, Xiaojun Li and Feng Xing*
ABSTRACT
To study the expression level and function of the TTR gene in different tissues of Dolang sheep at the periods of prepuberty, puberty and postpuberty, we collected hypothalamic, pituitary, ovarian, oviduct, and uterine tissues. The TTR gene was amplified, cloned, and sequenced using RT-PCR in Dolang sheep. Its expression in the five tissue types was analyzed. The amino acid sequences of the TTR gene were compared to those of other species and a phylogenetic tree was produced. qPCR was used to detect the expression levels of the TTR gene in the tissues. TTR gene was transfected into granulosa cells of the ovaries of Dolang sheep and was detected by fluorescence quantification. The cDNA sequence of the TTR gene of Dolang sheep was 444 bp and encoded 147 amino acids. Amino acid sequence identity of TTR from Dolang sheep was 99.32%, 99.32%, and 93.20% with sheep, goat, and cattle, respectively, and 89.33%, 89.12%, and 86.39% with pigs, horses, and humans, respectively. The TTR gene was expressed in all five tissue types, with higher expression in the pituitary and the hypothalamus (P < 0.05). TTR gene expression in the hypothalamic tissues was significantly higher at the periods of puberty than prepuberty and postpuberty (P < 0.05). The expression of TTR genes in the pituitary and uterus increased significantly from puberty to postpuberty (P < 0.05). Changes of TTR gene expression in the hypothalamus, pituitary, and uterus at different pubertal stages suggest that TTR genes may be involved in the initiation of puberty in sheep. The TTR gene has a repressive effect on the expression of the GnRH gene and may indirectly regulate the initiation of puberty in Dolang sheep. Our results provide a new theoretical basis for studying the initiation of puberty in sheep.
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