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United States Department of Agriculture

Agricultural Research Service

Title: Microarray Analysis of Gene Expression in Anterior Pituitary Glands from Anestrous and Cycling Postpartum Beef Cows

Authors
item Roberts, Andrew
item Mclean, D - WSU PULLMAN

Submitted to: Western Section of Animal Science Proceedings
Publication Type: Proceedings
Publication Acceptance Date: May 15, 2006
Publication Date: June 15, 2006
Citation: Roberts, A.J., Mclean, D.J. 2006. Microarray analysis of gene expression in anterior pituitary glands from anestrous and cycling postpartum beef cows. Western Section of Animal Science Proceedings 57:276-279.

Interpretive Summary: Oligionucleotide microarrays (GeneChip Bovine Genome Arrays, Affymetrix Inc., Santa Clara, CA) were used to evaluate gene expression profiles in anterior pituitary glands collected from four anestrous and four cycling postpartum beef cows. Anestrous cows were harvested 40 to 61d after calving at ~2 yr of age. Cycling cows were harvested 7 to 13 d after estrus, at 54 to 77 d after calving. Anterior pituitary tissue was collected and snap-frozen in liquid nitrogen within 22 to 37 min after exsanguination. Total RNA was isolated from each sample and was reverse transcribed into double stranded cDNA and subsequently transcribed in the presence of biotinylated UTP to generate target for hybridization on the Bovine Genome GeneChip . Each sample was hybridized to an individual Genechip containing 24,027 total probe sets including 23,080 bovine transcripts representing 19,000 unigene clusters as of March 2004. Hybridization signal were normalized across arrays using GeneChip Operating Software (GCOS; Affymetrix), and average intensities of each probe set were compared between groups by t-test. Expression of 25 transcripts were greater (P < 0.01) in pituitaries from cycling cows than anestrous cows; including: gastrin-releasing peptide, Ig heavy chain variable region, claudin 1, IGFBP-3, peroxisome proliferative activated receptor gamma, coactivator 1; 11 transcripts that share some homology (41 to 99%) to human proteins, and 9 uncharacterized transcripts. Transcripts expressed at decreased levels in cycling than anestrous cows included: signal transducer and activator of transcription 3, versican), alpha 1 acid glycoprotein, type 1 inositol 1, 4, 5-triphosphate receptor, calmodulin-dependent phospodiesterase 1B, protein S alpha; 9 transcripts that share some homology (56 to 99%) to human proteins and 1 uncharacterized transcript.. Although further study is required to confirm the role of these genes in the transition from anestrous to cycling status, results demonstrate potential of this methodology for identifying novel mechanisms regulating reproductive function..

Technical Abstract: Oligionucleotide microarrays (GeneChip Bovine Genome Arrays, Affymetrix Inc., Santa Clara, CA) were used to evaluate gene expression profiles in anterior pituitary glands collected from four anestrous and four cycling postpartum beef cows. Anestrous cows were harvested 40 to 61d after calving at ~2 yr of age. Cycling cows were harvested 7 to 13 d after estrus, at 54 to 77 d after calving. Anterior pituitary tissue was collected and snap-frozen in liquid nitrogen within 22 to 37 min after exsanguination. Total RNA was isolated from each sample and was reverse transcribed into double stranded cDNA and subsequently transcribed in the presence of biotinylated UTP to generate target for hybridization on the Bovine Genome GeneChip . Each sample was hybridized to an individual Genechip containing 24,027 total probe sets including 23,080 bovine transcripts representing 19,000 unigene clusters as of March 2004. Hybridization signal were normalized across arrays using GeneChip Operating Software (GCOS; Affymetrix), and average intensities of each probe set were compared between groups by t-test. Expression of 25 transcripts were greater (P < 0.01) in pituitaries from cycling cows than anestrous cows; including: gastrin-releasing peptide, Ig heavy chain variable region, claudin 1, IGFBP-3, peroxisome proliferative activated receptor gamma, coactivator 1; 11 transcripts that share some homology (41 to 99%) to human proteins, and 9 uncharacterized transcripts. Transcripts expressed at decreased levels in cycling than anestrous cows included: signal transducer and activator of transcription 3, versican), alpha 1 acid glycoprotein, type 1 inositol 1, 4, 5-triphosphate receptor, calmodulin-dependent phospodiesterase 1B, protein S alpha; 9 transcripts that share some homology (56 to 99%) to human proteins and 1 uncharacterized transcript.. Although further study is required to confirm the role of these genes in the transition from anestrous to cycling status, results demonstrate potential of this methodology for identifying novel mechanisms regulating reproductive function..

Last Modified: 11/23/2014
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