Transcriptional analysis of buffalo (Bubalus bubalis) oocytes during in vitro maturation using bovine cDNA microarray.
The need for improving in vitro production of buffalo embryos necessitates a better understanding of the molecular mechanisms regulating early development including oocyte maturation. Here, we used bovine cDNA microarray platform to investigate mRNA abundance of buffalo oocytes before and after in vitro maturation. For this, a total of six pools each contains 50 immature or in vitro matured buffalo oocytes were used for mRNA isolation and subsequent cDNA synthesis. The BlueChip bovine cDNA microarray (with approximately 2000 clones) was used to analyse gene expression profiles between immature and matured oocytes. Statistical analysis of microarray data revealed a total of 104 transcripts to be differentially expressed between the two oocyte groups. Among these, transcription factors (ZFP91), M-phase mitotic cell cycle (MPHOSPH9), growth factor (BMP15) and DNA binding (HMGN2) were found to be up-regulated in immature oocytes. Similarly, matured oocytes were found to be enriched with genes involved in cytoskeleton (ACTB), hydrogen ion transporting (ATP6V1C2) and structural constituent of ribosome (RPS27A). Quantitative real-time polymerase chain reaction validated the expression profile of some selected transcripts during array analysis. In conclusion, to our knowledge, this is the first large-scale expression study to identify candidate genes differentially abundant and with potential role during buffalo oocyte maturation.
Kandil
O M
OM
Department of Animal Reproduction and A.I., National Research Centre, Dokki, Giza, Egypt.
Ghanem
N
N
Abdoon
A S S
AS
Hölker
M
M
Phatsara
C
C
Schellander
K
K
Tesfaye
D
D
D000818:Animals; D002020:Buffaloes / Q000235:genetics*; D002417:Cattle / Q000235:genetics; D002478:Cells, Cultured; D005260:Female; D005307:Fertilization in Vitro; D020869:Gene Expression Profiling / Q000662:veterinary*; D005786:Gene Expression Regulation; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis / Q000662:veterinary*; D009865:Oocytes / Q000737:chemistry* / Q000254:growth & development*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012333:RNA, Messenger / Q000032:analysis*
D012333:RNA, Messenger