Effect of miR-302b MicroRNA Inhibition on Chicken Primordial Germ Cell Proliferation and Apoptosis Rate

Abstract

The primordial germ cells (PGCs) are the precursors for both the oocytes and spermatogonia. Recently, a novel culture system was established for chicken PGCs, isolated from embryonic blood. The possibility of PGC long-term cultivation issues a new advance in germ cell preservation, biotechnology, and cell biology. We investigated the consequence of gga-miR-302b-5P (5P), gga-miR-302b-3P (3P) and dual inhibition (5P/3P) in two male and two female chicken PGC lines. In treated and control cell cultures, the cell number was calculated every four hours for three days by the XLS Imaging system. Comparing the cell number of control and treated lines on the first day, we found that male lines had a higher proliferation rate independently from the treatments. Compared to the untreated ones, the proliferation rate and the number of apoptotic cells were considerably reduced at gga-miR-302b-5P inhibition in all PGC lines on the third day of the cultivation. The control PGC lines showed a significantly higher proliferation rate than 3P inhibited lines on Day 3 in all PGC lines. Dual inhibition of gga-miR-302b mature miRNAs caused a slight reduction in proliferation rate, but the number of apoptotic cells increased dramatically. The information gathered by examining the factors affecting cell proliferation of PGCs can lead to new data in stem cell biology.

Epigenetic programming of chicken germ cells: a comparative review

“…This miRNA expression pattern differs from those of other miRNAs; for instance, miR-302 is upregulated in E5.5–6.5 epiblasts but is almost absent in mouse PGCs upon their specification ( Parchem et al, 2014 ; Paikari et al, 2017 ). This is in contrast with the miR-302 family in chicken, involved in regulating PGC proliferation ( Rengaraj et al, 2013 ; Lázár et al, 2021 ). gga-miR-17-302b and miR-17-5p affect PGC proliferation by regulating glucose phosphate isomerase ( GPI ) expression ( Rengaraj et al, 2013 ).…”

“…gga-miR-17-302b and miR-17-5p affect PGC proliferation by regulating glucose phosphate isomerase ( GPI ) expression ( Rengaraj et al, 2013 ). Another study showed that the inhibition of gga-miR-302b-5p (5p) and/or gga-miR-302b-3p (3p) differentially influences PGCs proliferation and the rate of apoptosis, implying the complex influence of the miR-302b family on PGCs proliferation ( Lázár et al, 2021 ).…”

A Novel Tumor Glycolysis Inhibitor: 4-Methylumbelliferone

“…When GPI is knocked down, oxygen consumption is restored and glycolytic activity is reduced. Downregulation of GPI expression can reduce cell migration and sensitize tumor cells to drugs (Lázár et al, 2021; Shao et al, 2020). In acute hepatitis, acute myocardial infarction, malignant tumor and other diseases, the serum GPI activity is significantly increased, and it is positively correlated with the severity of the disease.…”

Fate Decisions of Chicken Primordial Germ Cells (PGCs): Development, Integrity, Sex Determination, and Self-Renewal Mechanisms

“…They demonstrated that miR-181a* downregulates the expression of nuclear receptor subfamily 6 group A member 1 ( NR6A1 ), and homeobox A1 ( HOXA1 ), suggesting that miR-181a* regulates inappropriate meiosis and somatic differentiation of chicken PGCs in different pathways [ 110 ]. Other studies have also identified and characterized miRNAs involved in maintaining the integrity of chicken PGCs, such as the regulation of DNA methylation [ 54 ], glucose metabolism [ 111 ], and apoptosis [ 112 ]. In contrast, lncRNAs are non-coding RNAs composed of over 200 nucleotides and possess various functions, such as epigenetic, transcriptional, and translational regulation, via lncRNA–RNA or lncRNA–protein interactions [ 113 ].…”