Expression and distribution of THRβ mRNA and protein in HPO axis of Ganja Tibetan sheep during estrus cycle and anestrus

doi:10.3969/j.issn.1004-1524.2022.03.08

Acta Agriculturae Zhejiangensis ›› 2022, Vol. 34 ›› Issue (3): 480-488.DOI: 10.3969/j.issn.1004-1524.2022.03.08

• Animal Science • Previous Articles Next Articles

Expression and distribution of THRβ mRNA and protein in HPO axis of Ganja Tibetan sheep during estrus cycle and anestrus

LIU Lili¹(), GAO Hexuan², YANG Yawen¹, BAO Yingying¹, HE Yuqin¹^,^*(), YANG Zhijie¹, CHEN Weigang¹, GE Wenbo³

1. College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China
2. Network Finance Department of Lanzhou Bank, Lanzhou 730070, China
3. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China

Received:2021-05-06 Online:2022-03-25 Published:2022-03-30
Contact: HE Yuqin

Abstract

Abstract:

Thyroid hormone (TH) exerted its biological effects by specifically binding to its target cell receptor. In order to clarify the regulation of thyroid hormone receptor β (THRβ) gene and its protein in the pineal gland and Hypothalamic-pituitary-ovary (HPO) axis of the reproductive activity of seasonal breeding animals, Ganjia Tibetan sheep (Ovis aries). In this study, the healthy Ganjia Tibetan sheep during the estrus cycle and anestrus were used as the research object. RT-qPCR, Western blot and immunohistochemical staining were used to detect and analyze the expression and distribution of THRβ mRNA and its protein in the pineal gland and HPOA. The results showed that THRβ mRNA and its protein were expressed and distributed in the pineal gland and HPOA tissues. The expression of THRβ mRNA and its protein in the pineal gland in the anestrus were significantly (P<0.05) higher than that in the estrus cycle. In hypothalamus, the expression of THRβ mRNA in the metestrus were significantly (P <0.05) higher than that in other stages while the expression of its protein in the disestrus were significantly (P<0.05) higher than that in other stages; In pituitary gland, the expression of THRβ mRNA and its protein in the diestrus were significantly (P<0.05) higher than that in other stages. In ovary, the expression of THRβ mRNA and its protein in the metestrus were significantly (P<0.05) higher than that in other stages. The immunohistochemical results showed that THRβ was mainly distributed in the eosinophilic cytoplasm of the pineal, glial cells, large neuron cell bodies and vertebral cell cytoplasm of the hypothalamic, eosinophilic and basophilic cytoplasm of adenohypophysis, granulosa cell cytoplasm, follicular inner membrane cells and luteal cells of the ovary. THRβ mRNA and its protein in the pineal gland and HPOA showed different changes during the estrus and anestrus, which indicated that THRβ gene and its protein were involved in the regulation of the reproductive activities of the Ganjia Tibetan sheep.

Key words: Ganga Tibetan sheep, hypothalamus-pituitary-ovary axis, estrus cycle, anestrus

CLC Number:

S826

LIU Lili, GAO Hexuan, YANG Yawen, BAO Yingying, HE Yuqin, YANG Zhijie, CHEN Weigang, GE Wenbo. Expression and distribution of THRβ mRNA and protein in HPO axis of Ganja Tibetan sheep during estrus cycle and anestrus[J]. Acta Agriculturae Zhejiangensis, 2022, 34(3): 480-488.

Figures/Tables 4

Table 1 Primers sequences of THRβ and β-actin gene

基因 Gene	引物序列 Primer sequences(5'-3')	产物大小 Product size/bp	退火温度 Annealing temperature/℃	登录号 Login ID
THRβ	F:ACCGCCATCTCCCCGTTCAG R:TTGTGAGCTGCCATGTGAAGACC	129	60	NM001190391.1
β-actin	F:CCATTGAGCACGGCATTGT R: GCAGGGGTGTTGAAGGTCTC	184	58	HM067830.1

Fig.1 Relative expression of THRβ mRNA and protein in Pineal gland and HPO axis during estrus cycle and anestrus of Ganjia Tibetan sheep A,Pineal gland;B,Hypothalamus;C,Pituitary;D, Ovary. Bars marked without the same letters on the same series indicated significant differences in the relative expression of mRNA or protein at different periods at P<0.05.

Fig.2 Content of THRβprotein in pineal gland and HPO axis during estrus cycle and anestrus of Ganjia Tibetan sheep PE, Proestrus; E, Estrus; ME, Metestrus; DE, Diestrus; AE, Anestrus.

Fig.3 Location of THRβ immunoreactive products of in pineal and HPO axis of Ganjia Tibetan sheep during the estrus cycle and anestrus A, F, K, P, Proestrus; B, G, L, Q, Estrus; C, H, M, R, Metestrus; D, I, N, S, Diestrus; E, J, O, T, Anestrusl. P, Pineal cell; BV, Blood vessel; GC, Glial cell; NC, Nerve cell; Nu, Neuronuclei; AC, Acidophilic cell; BC, Basophilic cell; CC, Chromophobe cell; FA, Follicular antrum; GV, Germinal vesicle; LC, Luteal cell; TF, Theca folliculi; SG, Stratum granulosum.

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Expression and distribution of THRβ mRNA and protein in HPO axis of Ganja Tibetan sheep during estrus cycle and anestrus

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[1]	BAO Yingying, CHEN Weigang, GE Wenbo, HE Yuqin, YANG Zhijie, ZHOU Kairen, YANG Dapeng, YANG Yawen, LIU Lili. Expression of ERβ in hypothalamic-pituitary-ovarian axis of Ganjia Tibetan sheep in estrus cycle [J]. Acta Agriculturae Zhejiangensis, 2022, 34(1): 10-16.
[2]	CHEN Weigang, BAO Yingying, HE Yuqin, GE Wenbo, ZHANG Xia, HUADAO Cairang, YANG Dapeng, ZHOU Kairen. Expression and distribution of KISS-1/GPR54 on hypothalamic-pituitary-ovary axis in estrous cycle of Ganjia Tibetan sheep [J]. , 2020, 32(6): 971-977.