Gene pathways and cell cycle-related genes in cultured avian primordial germ cells

Abstract

Primordial germ cells (PGC) from early embryos are applicable to various kinds of research, including the production of transgenic animals. Primordial germ cells eventually migrate and differentiate into germ cells in the gonads, where they settle and rapidly proliferate. However, the proliferation rate of PGC is low in early embryos, and there are many significant pathways that mediate PGC activity. Therefore, in vitro culture of PGC from early embryos with efficient growth factors has been necessary. Recently, we cultured chicken PGC from embryonic d 2.5 with basic fibroblast growth factor and characterized the PGC through analysis of cell morphology, survival, proliferation, and apoptosis. However, large-scale analyses of genes expressed in cultured PGC and the genes involved in associated pathways are limited. The objective of the present investigation was to identify the signaling and metabolic pathways of expressed genes by microarray comparison between PGC and their somatic counterpart, chicken embryonic fibroblasts (CEF). We identified 795 genes that were expressed more predominantly in PGC and 824 genes that were expressed more predominantly in CEF. Among the predominant genes in PGC, 201 were differentially identified in 106 pathways. Among the predominant genes in CEF, 242 were differentially identified in 99 pathways. To further validate the genes involved in at least one candidate pathway, those involved in the cell cycle (12 predominant genes in PGC and 8 predominant genes in CEF) were examined by real-time PCR. To the best of our knowledge, this study is the first to investigate signaling and metabolic pathways in cultured PGC.