Cloned pig fetuses exhibit fatty acid deficiency from impaired placental transport

Ao, Zheng; Wu, Xuehong; Zhou, Jun; Gu, Tao; Wang, Xingwang; Shi, Junsong; Zhao, Chengfa; Cai, Guangyan; Zheng, Enqin; Liu, Dewu; Wu, Zhenfang; Li, Zicong

doi:10.1002/mrd.23242

Cited by 9 publications

(7 citation statements)

References 49 publications

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“…These abnormalities are thought to be associated with the upregulation of nutrient transport-related genes such as SFMBT2 and SLC38A4 in SCNT mice [40]. In contrast, the placentas of cloned piglets possessed low villus density and exhibited severe trophoblast hypoplasia [41][42][43]. Therefore, we expected a downregulation of these genes in the TSC NT lines of the cloned pigs with placental hypoplasia.…”

Section: Discussionmentioning

confidence: 92%

Establishment and characterization of porcin trophoblast stem cell lines from somatic cell nuclear-transferred blastocysts

Kim

Cai

Choi

et al. 2023

Preprint

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Background: Genetically modified pigs are considered ideal models for studying human diseases and potential sources for xenotransplantation research. However, the somatic cell nuclear transfer (SCNT) technique utilized to generate these cloned pig models has low efficiency and fetuses exhibit limited developmental potential for abnormalities in the placenta. Methods: Herein, we report, for the first time, the establishment of porcine trophoblast stem cell (TSC) lines using SCNT and in vitro fertilized (IVF) blastocysts by activation of Wing-less/Integrated (Wnt) and epidermal growth factor (EGF) pathways, inhibition of transforming growth factor-β (TGFβ) and Rho-associated protein kinase (ROCK) pathways, and supplementation with ascorbic acid. We also compared the transcripts of TSC lines originating from SCNT and IVF embryos and their differentiated lineages. Results: A total of 19 porcine TSC lines were established from SCNT and IVF blastocysts (TSCNT and TSCIVF lines) showing typical TSC characteristics. Compared with the TSCIVF lines, TSCNT lines showed distinct expression patterns suggesting unique TSCNT characteristics, including decreased mRNA expression levels of genes related to apposition, steroid hormone biosynthesis, angiopoiesis, and RNA stability. Conclusions: This study provides valuable information and a powerful model for studying the abnormal development and dysfunction of trophoblasts and placentas in cloned pigs.

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Section: Discussionmentioning

confidence: 92%

Establishment and characterization of porcin trophoblast stem cell lines from somatic cell nuclear-transferred blastocysts

Kim

Cai

Choi

et al. 2023

Preprint

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“…The FAs of the placenta tissue samples (50 mg) were extracted with a chloroform–methanol (2:1 v / v ) mixture in accordance with the method as described by a previous study [ 17 ]. The extracted FAs were transesterified with a 2 mL 1% methanol–sulfuric acid mixture for esterification for 30 min at 80 °C.…”

Section: Methodsmentioning

confidence: 99%

“…Western blot analyses were carried out as described previously [ 17 ]. The total protein of the placenta samples was extracted using a Tissue Protein Extraction Kit (CWBIO, Jiangsu, China), and the concentration of the extracted total protein was determined by a bicinchoninic acid assay (CWBIO, Jiangsu, China).…”

Section: Methodsmentioning

confidence: 99%

IGF2 May Enhance Placental Fatty Acid Metabolism by Regulating Expression of Fatty Acid Carriers in the Growth of Fetus and Placenta during Late Pregnancy in Pigs

Zhang

et al. 2023

Genes

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Fatty acids (FAs) are essential substances for the growth and development of the fetus and placenta. The growing fetus and placenta must obtain adequate FAs received from the maternal circulation and facilitated by various placental FA carriers, including FA transport proteins (FATPs), FA translocase (FAT/CD36), and cytoplasmic FA binding proteins (FABPs). Placental nutrition transport was regulated by imprinted genes H19 and insulin-like growth factor 2 (IGF2). Nevertheless, the relationship between the expression patterns of H19/IGF2 and placental fatty acid metabolism throughout pig pregnancy remains poorly studied and unclear. We investigated the placental fatty acid profile, expression patterns of FA carriers, and H19/IGF2 in the placentae on Days 40 (D40), 65 (D65), and 95 (D95) of pregnancy. The results showed that the width of the placental folds and the number of trophoblast cells of D65 placentae were significantly increased than those of D40 placentae. Several important long-chain FAs (LCFAs), including oleic acid, linoleic acid, arachidonatic acid, eicosapentaenoic acid, and docosatetraenoic acid, in the pig placenta showed dramatically increased levels throughout pregnancy. The pig placenta possessed higher expression levels of CD36, FATP4, and FABP5 compared with other FA carriers, and their expression levels had significantly upregulated 2.8-, 5.6-, and 12.0-fold from D40 to D95, respectively. The transcription level of IGF2 was dramatically upregulated and there were corresponding lower DNA methylation levels in the IGF2 DMR2 in D95 placentae relative to D65 placentae. Moreover, in vitro experimentation revealed that the overexpression of IGF2 resulted in a significant increase in fatty acid uptake and expression levels of CD36, FATP4, and FABP5 in PTr2 cells. In conclusion, our results indicate that CD36, FATP4, and FABP5 may be important regulators that enhance the transport of LCFAs in the pig placenta and that IGF2 may be involved in FA metabolism by affecting the FA carriers expression to support the growth of the fetus and placenta during late pregnancy in pigs.

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“…qRT-PCR was performed on QuantStudio™ 7 Flex Real-Time PCR System (Thermo Fisher Scientific) by following the parameters recommended by the manufacturer. The reactions were run and performed in triplicate as previously described [ 55 ]. GAPDH was used as the house-keeping gene, and the relative mRNA expression was calculated using the 2-ΔΔCt method.…”

Section: Methodsmentioning

confidence: 99%

Interleukin 17D Enhances the Developmental Competence of Cloned Pig Embryos by Inhibiting Apoptosis and Promoting Embryonic Genome Activation

Zhao

Lai

et al. 2021

Animals

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Cloned animals generated by the somatic cell nuclear transfer (SCNT) approach are valuable for the farm animal industry and biomedical science. Nevertheless, the extremely low developmental efficiency of cloned embryos hinders the application of SCNT. Low developmental competence is related to the higher apoptosis level in cloned embryos than in fertilization-derived counterparts. Interleukin 17D (IL17D) expression is up-regulated during early mouse embryo development and is required for normal development of mouse embryos by inhibiting apoptosis. This study aimed to investigate whether IL17D plays roles in regulating pig SCNT embryo development. Supplementation of IL17D to culture medium improved the developmental competence and decreased the cell apoptosis level in cloned porcine embryos. The transcriptome data indicated that IL17D activated apoptosis-associated pathways and promoted global gene expression at embryonic genome activation (EGA) stage in treated pig SCNT embryos. Treating pig SCNT embryos with IL17D up-regulated expression of GADD45B, which is functional in inhibiting apoptosis and promoting EGA. Overexpression of GADD45B enhanced the developmental efficiency of cloned pig embryos. These results suggested that IL17D treatment enhanced the developmental ability of cloned pig embryos by suppressing apoptosis and promoting EGA, which was related to the up-regulation of GADD45B expression. This study demonstrated the roles of IL17D in early development of porcine SCNT embryos and provided a new approach to improve the developmental efficiency of cloned porcine embryos.

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Cloned pig fetuses exhibit fatty acid deficiency from impaired placental transport

Cited by 9 publications

References 49 publications

Establishment and characterization of porcin trophoblast stem cell lines from somatic cell nuclear-transferred blastocysts

Establishment and characterization of porcin trophoblast stem cell lines from somatic cell nuclear-transferred blastocysts

IGF2 May Enhance Placental Fatty Acid Metabolism by Regulating Expression of Fatty Acid Carriers in the Growth of Fetus and Placenta during Late Pregnancy in Pigs

Interleukin 17D Enhances the Developmental Competence of Cloned Pig Embryos by Inhibiting Apoptosis and Promoting Embryonic Genome Activation

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