Epidermal Growth Factor-Induced Proliferation of Chicken Primordial Germ Cells: Involvement of Calcium/Protein Kinase C and NFKB11

Abstract

Epidermal growth factor (EGF) has been shown to stimulate survival in diverse cells in vitro. In the present study, the effects of EGF and the EGF-related signaling pathway on proliferation of chicken primordial germ cells (PGCs) were investigated. Results showed that EGF (10-100 ng/ml) increased the number and area of PGC colonies in a time- and dose-dependent manner. EGF also activated PKC, a process that was inhibited by AG1478 (an EGFR tyrosine kinase inhibitor) and ethyleneglycol-bis-(beta-aminoethyl ether)-N,N'-tetraacetic acid (EGTA; an intracellular Ca(2+) chelator). In addition, the degradation of NFKBIA and NFKB1 (p65) translocation was observed after EGF treatment, which was significantly blocked by pretreatment with AG1478, EGTA, H(7), or SN50 (NFKB1-specific inhibitor). Furthermore, we found that EGF-induced cell proliferation was significantly attenuated by AG1478, EGTA, H(7), and SN50, respectively. On the other hand, inhibition of EGFR, Ca(2+)/PKC, or NFKB1 abolished the EGF-stimulated increase in the expression of cyclins CCND1 and CCNE1, cyclin-dependent kinase 6 (CDK6), CDK2, and BCL2, and restored the EGF-induced inhibition of BAX expression and caspase 3/9 activity, indicating that EGFR, PKC, and NFKB1 signaling cascades were involved in EGF-stimulated DNA synthesis and antiapoptosis action. In conclusion, EGF stimulated proliferation of chicken PGCs via activation of Ca(2+)/PKC involving NFKB1 signaling pathway. These observations suggest that EGF signaling is important in regulating germ cell proliferation in the chicken embryonic gonad.

Effect of epidermal growth factor on follicle-stimulating hormone-induced proliferation of granulosa cells from chicken prehierarchical follicles

“…It has been shown that there is a paracrine action within the follicle and the oocyte-derived EGF family ligands may mediate mutual communications between oocytes and follicle cells (Tse and Ge, 2010). Our previous studies revealed that EGF stimulated proliferation of chicken primordial germ cells directly (Ge et al, 2009). In this study, PCNA immunocytochemistry and TUNEL assay demonstrated that EGF promoted the proliferation of granulosa cells from chicken SYF in a dosedependent manner, and inhibited cell apoptosis.…”

Induced Multilineage Differentiation of Chicken Embryonic Germ Cells via Embryoid Body Formation

“…In our previous study, the quantity of chicken PGCs was amplifi ed by epidermal growth factor via activation of Ca 2+ / PKC involving NFKB1 signaling pathway in culture [18]. In this study, we derived EG-like cells from chicken PGCs and investigated the potentials of differentiation.…”

“…The cDNA was synthesized from total RNA using a AMV First-Strand cDNA synthesis kit with oligo(dT) 18 primers according to the manufacturer's instructions (Sangon, Shanghai, China). PCR was subsequently carried out on 1 μL cDNA template for 30-40 cycles with primers as listed in Table 1.…”

G Protein-Coupled Receptor 30 Mediates Estrogen-Induced Proliferation of Primordial Germ Cells Via EGFR/Akt/β-Catenin Signaling Pathway

“…We previously established a chicken PGC culture system that allows us to study the effect of endogenous growth factors on PGC proliferation (23). Here, we have investigated the direct effect of E 2 on PGC proliferation and aim to elucidate the possible underlying mechanism.…”