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E-GEOD-10662 - Identification of Differential Gene Expression During Transition of Bovine Corpus Luteum from Early To Mid–Phase

Status
Released on 5 May 2010, last updated on 2 May 2014
Organism
Bos taurus
Samples (2)
Array (1)
Protocols (7)
Description
Prostaglandin F2 alpha (PGF2alpha) brings about regression of the bovine corpus luteum (CL). This luteolytic property of PGF2alpha is used in the beef and dairy cattle to synchronize estrus. A limitation of this protocol is an insensitivity of the early CL to luteolytic actions of PGF2alpha. The mechanisms underlying luteal sensitivity (LS) are poorly understood. The early CL has maximum number of PGF2alpha receptors; therefore differences in signaling events might be responsible for LS. Hence differential gene expression at two developmental stages, days 4 (D-4) and 10 (D-10) post estrus, might account for differences in signal transduction pathways associated with LS. This possibility was examined in these studies. Microarray analysis (n=3 per stage) identified 167 genes that were differentially expressed (p<0.05). These were categorized into genes involved in cell signaling, steroidogenesis and metabolism, protein degradation, transcription regulation and DNA biosynthesis, protein biosynthesis and modification, extracellular matrix and cytoskeletal proteins, antioxidant property, miscellaneous, and unknown functions. Real-time PCR confirmed the differential expression of 9 randomly selected genes, including protein kinase C inhibitor protein-1 (KCIP-1) and regulator of G-protein signaling 2 (RGS2), observed in microarray. Further, the in vivo effect of exogenous PGF2 (n=3 per stage) on selected genes that were found differentially expressed during this developmental transition was examined. PGF2alpha increased the expression of a guanine nucleotide binding protein beta polypeptide 1 (GNB1) in D-4 CL and Ca2+/calmodulin dependent kinase kinase 2 beta (Camkk2) in D-10 CL. Therefore, GNB1, Camkk2, KCIP-1, and RGS2 are candidate genes that might play significant role in acquisition of luteal sensitivity to PGF2alpha. Keywords: Direct comparison between Day-4 and Day-10 CL, Developmental type comparision The present experiment made a direct comparision of gene expression in early (Day-4) vs mid-phase CL (Day-10). Each experiment contained two technical relicates per slide. Total of three biological replicates were used ( 3 animals for Day-4 and 3 animals for Day-10). Two Day-4 samples were labelled Cy5 and one Day-10 with Cy5. Two Day-10 samples were labelled with Cy3 and one Day-4 with Cy3. The Day-4 sample was control for our experiments.
Experiment type
transcription profiling by array 
Contacts
MADHUSUDHAN P GORAVANAHALLY <mgoravan@mix.wvu.edu>, JIANBO YAO, JORGE FLORES, KEITH INSKEEP, MADHUSUDAN P GORAVANAHALLY, MOHAMED SALEM
Citation
MIAME
PlatformsProtocolsVariablesProcessedRaw
Files
Investigation descriptionE-GEOD-10662.idf.txt
Sample and data relationshipE-GEOD-10662.sdrf.txt
Raw data (1)E-GEOD-10662.raw.1.zip
Processed data (1)E-GEOD-10662.processed.1.zip
Array designA-GEOD-3810.adf.txt
Links