Transcripts from a human primordial follicle cDNA library.

Abstract

BACKGROUND: Human primordial follicles (PFs) or the oocyte-pre-granulosa complex, constitute the earliest and most immature stage of human oogenesis. The factors, signalling networks and the precise role of the oocyte and the pre-granulosa cells in initiating growth and recruitment from this finite resting pool remain largely unknown at present. METHODS: To obtain a gene resource of this oogenesis stage and thereby determine a molecular blueprint of the human PF, a cDNA library was constructed from 50 isolated human PFs using the phagemid vector pTriplEx2. RESULTS: Sequence analysis showed that 46.67% of these clones corresponded to known genes while 29.48% were uncharacterized genes that included hypothetical proteins, human cDNA clones and novel genes. Bioinformatics analysis revealed a preponderance of mitochondrial genes and repeat elements followed by ribosomal proteins, transcription and translation genes. Transcripts for heat shock proteins, cell cycle, embryogenesis genes and apoptosis genes were identified. Members of the ubiquitin-proteasome pathway, MAPK, p38/JNK, GPCR, Wnt, NF-kappaB and notch signalling pathways were identified. A mitochondrial pathway and a transcription factor pathway in the human PF were generated. The gene networks in the transcription factor pathway provided a first glimpse of the balance between proliferation and cell death/apoptosis in this earliest stage of oogenesis. CONCLUSIONS: The abundance and diversity of retroviral elements and transcriptional repressor genes in the human PF suggest these could contribute to the maintainance of this oogenesis stage. The role of these genes in initial recruitment and in subsequent oogenesis stages will be greatly facilitated and elucidated by printing a human PF cDNA array of the sequenced clones and using it for gene profiling.