Scientia Agricultura Sinica ›› 2021, Vol. 54 ›› Issue (4): 845-854.doi: 10.3864/j.issn.0578-1752.2021.04.015

• ANIMAL SCIENCE·VETERINARY SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

Expression Patterns of TETs and Programmed Cell Death Related Genes in Oviduct and Uterus of Early Pregnancy Goats

ZHAO Le(),YANG HaiLi,LI JiaLu,YANG YongHeng,ZHANG Rong,CHENG WenQiang,CHENG Lei,ZHAO YongJu()   

  1. College of Animal Science and Technology, Southwest University/Chongqing Key Laboratory of Forage & Herbivore/Chongqing Engineering Research Center for Herbivores Resource Protection and Utilization, Chongqing 400715
  • Received:2020-03-12 Accepted:2020-07-10 Online:2021-02-16 Published:2021-02-16
  • Contact: YongJu ZHAO E-mail:zhaole0228@163.com;zyongju@163.com

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Abstract:

【Objective】The ten-eleven translocation (TET) protein family play crucial roles in embryo and placental development. The balance of cell death between oviduct and endometrial would affect the development of gametes and early embryos. In this study, the expression patterns and their potential regulatory role of TETs, programmed cell death related genes and blastocyst implantation-related genes in oviduct and uterus of goats in early pregnancy were mainly explored, so as to provide a basis for studying development of gametes and early embryos.【Method】In this study, the uterus and oviduct of Hechuan white goats during non-pregnant 19 days (C19) and pregnant 19 days (P19) were collected, and then the morphology uterine horn was showed by HE staining. Then, the genes, including blastocyst implantation-related genes (Wnt5a, OPN, VEGFA), TET family genes (TET1, TET2, TET3), programmed cell death-related genes (BAX, BCL2, Caspase9, GSDMD, NLRP3 and Caspase1) and inflammation-related genes (NF-kB、TNF-α) were detected by qRT-PCR. Finally, the correlation between TETs and programmed cell death-related genes expression were analyzed.【Result】The results showed that the gland number increased, and its morphology varied in uterine horn in P19 group. Compared with C19 group, OPN gene expression and Bax/BCL2 in uterine horn tissue were significantly increased in P19 group (P＜0.01), however, there was no significant difference between Wnt5a and VEGFA. TET family genes were expressed in both oviduct and uterus. The expression of TET1 and TET2 was significantly higher than TET3 in uterine horn (P＜0.01). TET2 (P＜0.01), NF-kB (P＜0.01), Caspase1 (P＜0.01), and GSDMD (P＜0.05) gene were significantly higher in P19 group of oviduct tissue. Correlation analysis showed that the relationship between TET2 and TET3 (P＜0.01), NF-kB (P＜0.01) and Caspase9 (P＜0.01) in oviduct tissue were positive correlation. The TET3 were negative correlated with VEGFA (P＜0.05) and WNT5a (P＜0.01). Blastocyst implantation-related genes OPN was positively correlated with Caspase1 (P＜0.01), NLRP3 (P＜0.05) and GSDMD (P＜0.05) in uterine horn. The GSDMD was positively correlated with Bax (P＜0.05), Caspase1 (P＜0.01) and NLRP3 (P＜0.05) in oviduct and uterus horn.【Conclusion】It was indicated that there was correlation between TETs and programmed cell death related genes. So, it was speculated that TET family genes and programmed cell death related genes played a key role in regulating the development of oviduct, zygote, early embryo and uterus. The key genes for embryo implantation of OPN were significantly correlated with pyroptosis related genes, suggesting that proptosis had a certain influence on the attachment of early embryos. These finding provided heretical references for studying the effect of TETs family genes and programmed cell death related genes on oviduct and uterine horn of early pregnancy.

Key words: goat, uterine horn, oviduct, programmed cell death, gene expression

Table 1

The information of primers used in qRT-PCR"

基因名称
Gene name		 引物序列（5'→3'）
Primer sequence（5'→3'）		 产物长度
Product size (bp)
β-actin F：TGATATTGCTGCGCTCGTGGT
R：GTCAGGATGCCTCTCTTGCTC		 189
WNT5a F：GCAAGGGCAATGTCTTCCA
R：TTCATACCTAGCGACCACCAA		 104
OPN F: GTTAAACCGACCAGTTCTGGCA
R: GTTGTCATCAGTTTCCTCAGAGG		 148
NF-kB F: GTGTAAAGAAGCGGGACTTGG
R: GCGGTTGTCAAAGATGGGATG		 203
Caspase9 F: CAATAGCAACCCTGAGACGGA
R: AAAGCCTGGGAAGGTGGAGTA		 130
GSDMD F: TTATTGGCTCTGACTGGGACG
R: GGCAAAGCTGAAGCACGAAC		 128
BCL2 F: GAAACCCCGCCACGAATTA
R: CCAGCCAACAGTACGGAACAA		 96
BAX F: TCTCCCCGAGAGGTCTTTTT
R: TGATGGTCCTGATCAACTCG		 151
NLRP3 F: CCGTCTGGGTGAGAGCGTGAA
R: TCCTGTTGGCTCCTGTGTTCCT		 78
Caspase1 F: GGATACAATAAATGGCTTGCTGG
R: CTCGGGCTTTATCCATAGTTGT		 110
TNF-α F: CCACGTTGTAGCCAACATCAG
R: AGATGAGGTAAAGCCCGTCAG		 134
VEGFA F: GTGACCCAGCACAGTTCCTCTT
R: TTCCGGGCTCGGTGATTTAG		 74
TET1 F: CATCACTGTCCGTCTTTGGA
R: ACTTTCCTGTGCCTTTGTGG		 126
TET2 F: GATACCATCTCCGTCTCCCATT
R: CTTGCTGTCTCCATTCACTTCC		 109
TET3 F: CCTTCTCCTTCGGTTGTTCCTG
R: CACTTCTTCCTCTTTGGGGTTGTC		 113

Table 2

qRT-PCR Set-up"

组成成分
Components		 10 μL反应体系
10 μL reaction
Nuclease-Free Water 3.4 μL
cDNA 1 μL
Forward primer 0.3 μL
Reverse primer 0.3 μL
EvaGreen2x qPCR MasterMix 5 μL
Total 10 μL

Fig. 1

The morphology of goat uterine horn A: C19 goat uterine horn (4×); B: C19 goat uterine horn (10×);C: P19 goat uterine horn (4×); D: P19 goat uterine horn (10×)"

Fig. 2

The expression of blastocyst implantation-related genes in goat uterine horn * P0.05;** P0.01。下同 The same as below"

Fig. 3

The TETs family genes expression in goat oviduct and uterine horn"

Fig. 4

The apoptosis-related genes expression in goat oviduct and uterine horn"

Fig. 5

The pyroptosis-related genes expression in goat oviduct and uterine horn"

Fig. 6

The inflammation-related genes expression in goat oviduct and uterine horn"

Table 3

Correlation of TETs and programmed cell death related genes in oviduct of early pregnancy goat"

TET2 TET3 NFkB TNFa BAX Bcl2 Caspase9 Caspase1 NLRP3 GSDMD
TET1 r 0.657 0.667 0.657 0.829* -0.086 0.829* 0.371 0.314 0.257 0.429
p 0.156 0.148 0.156 P＜0.05 0.872 P＜0.05 0.468 0.544 0.623 0.397
TET2 0.928** 1.000** 0.714 0.486 0.657 0.829* 0.714 0.771 0.771
P＜0.01 P＜0.01 0.111 0.329 0.156 P＜0.05 0.111 0.072 0.072
TET3 0.928** 0.696 0.522 0.667 0.696 0.696 0.638 0.754
P＜0.01 0.125 0.288 0.148 0.125 0.125 0.173 0.084
NF-kB 0.714 0.486 0.657 0.829* 0.714 0.771 0.771
0.111 0.329 0.156 P＜0.05 0.111 0.072 0.072
TNFa 0.086 0.886* 0.543 0.657 0.6 0.6
0.872 P＜0.05 0.266 0.156 0.208 0.208
Bax 0.314 0.771 0.771 0.714 0.829*
0.544 0.072 0.072 0.111 P＜0.05
Bcl2 0.657 0.714 0.6 0.771
0.156 0.111 0.208 0.072
Caspase9 0.886* 0.943** 0.943**
P＜0.05 P＜0.01 P＜0.01
Caspase1 0.943** 0.943**
P＜0.01 P＜0.01
NLRP3 0.886*
P＜0.05

Table 4

Correlation of TETs, programmed cell death genes and implantation-related genes in uterine horn of early pregnancy goat"

TET2 TET3 NFkB TNFa BAX Bcl2 Caspase9 Caspase1 NLRP3 GSDMD OPN WNT5a VEGFA
TET1 r -0.771 0.486 -0.257 0.086 0.029 0.143 -0.143 0.371 0.429 0.257 0.257 -0.429 -0.086
p 0.072 0.329 0.623 0.872 0.957 0.787 0.787 0.468 0.397 0.623 0.623 0.397 0.872
TET2 -0.200 0.371 0.371 0.314 -0.086 0.600 -0.257 -0.429 0.029 -0.200 0.086 -0.029
0.704 0.468 0.468 0.544 0.872 0.208 0.623 0.397 0.957 0.704 0.872 0.957
TET3 0.314 0.314 0.143 0.143 0.600 0.029 -0.086 0.086 0.200 -0.943** -0.886*
0.544 0.544 0.787 0.787 0.208 0.957 0.872 0.872 0.704 P＜0.01 P＜0.05
NFkB 0.714 0.600 0.600 0.714 -0.086 -0.029 0.086 -0.029 -0.543 -0.314
0.111 0.208 0.208 0.111 0.872 0.957 0.872 0.957 0.266 0.544
TNFa 0.943** 0.257 0.771 0.429 0.371 0.657 0.371 -0.486 -0.143
P＜0.01 0.623 0.072 0.397 0.468 0.156 0.468 0.329 0.787
BAX 0.029 0.600 0.657 0.600 0.829* 0.600 -0.257 -0.029
0.957 0.208 0.156 0.208 P＜0.05 0.208 0.623 0.957
Bcl2 0.200 -0.486 -0.257 -0.429 -0.600 -0.429 0.086
0.704 0.329 0.623 0.397 0.208 0.397 0.872
Caspase9 -0.029 -0.200 0.257 0.086 -0.714 -0.600
0.957 0.704 0.623 0.872 0.111 0.208
Caspase1 0.943** 0.943** 0.943** 0.086 0.086
P＜0.01 P＜0.01 P＜0.01 0.872 0.872
NLRP3 0.829* 0.829* 0.143 0.257
P＜0.01 P＜0.05 0.787 0.623
GSDMD 0.886* -0.029 0.029
P＜0.05 0.957 0.957
OPN -0.029 -0.200
0.957 0.704
WNT5a 0.771
0.072
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