| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Regular Article |
a Department of Physiological Sciences,
b College of Veterinary Medicine, Departments of Statistics
c Animal Science, Division of Agricultural Science and Natural Resources, Oklahoma State University, Stillwater, Oklahoma 74078-2006
Successful embryonic development is dependant on time and location-specific expression of appropriate genes. Unfortunately, information on stage-specific gene expression during early embryonic development in the bovine is lacking. In the present study, we compared gene expression between in vitro-produced Day 7–8 intact blastocysts (driver) and Day 9–10 hatched blastocysts (tester) using suppression-subtractive hybridization. Pools of 30 embryos for both driver and tester were used in the RNA extraction process. From limited amounts of starting material (
400 ng of total RNA), a reverse transcription-polymerase chain reaction (PCR) procedure was used to amplify the mRNA and generate sufficient cDNA to conduct suppression-subtractive hybridization. The subtracted cDNA products were cloned, and 126 cDNAs representing expressed mRNAs were isolated, sized, single-pass sequenced, and compared to known sequences in GenBank. Ninety-two clones provided sequence information for further analysis. Among these, 31 exhibited high homology to known genes. Three, 26S proteasomal ATPase (PSMC3), casein kinase 2 
subunit (CK2), and phosphoglycerate kinase (PGK) were selected and further characterized using real-time quantitative PCR to assess their differential expression in hatched blastocysts. Overall, a 1.3-, 1.6-, and 1.5-fold increase in expression level was observed in hatched blastocysts compared with intact blastocyst for PSMC3, CK2, and PGK, respectively. These results show that construction of subtracted cDNA libraries from small numbers of embryos is feasible and can provide information on gene expression patterns during preattachment embryogenesis.
1 This study was supported by the Oklahoma Agricultural Experiment Station. We thank the Lew Wentz Foundation, Wentz Project Scholarship program for support for S.R.
2 Correspondence: J.R. Malayer, Department of Physiological Sciences, Oklahoma State University, 264 McElroy Hall, Stillwater, OK 74078-2006. FAX: 405 744 7110; malayer{at}okstate.edu
This article has been cited by other articles:
|
|
 |
|
  A. El-Sayed, M. Hoelker, F. Rings, D. Salilew, D. Jennen, E. Tholen, M.-A. Sirard, K. Schellander, and D. Tesfaye Large-scale transcriptional analysis of bovine embryo biopsies in relation to pregnancy success after transfer to recipients Physiol Genomics, December 13, 2006; 28(1): 84 - 96. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P Froment, F Gizard, D Defever, B Staels, J Dupont, and P Monget Peroxisome proliferator-activated receptors in reproductive tissues: from gametogenesis to parturition. J. Endocrinol., May 1, 2006; 189(2): 199 - 209. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A.G.B. Hurst, D.W. Goad, M. Mohan, and J.R. Malayer Independent Downstream Gene Expression Profiles in the Presence of Estrogen Receptor {alpha} or {beta} Biol Reprod, October 1, 2004; 71(4): 1252 - 1261. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Yao, X. Ren, J. J. Ireland, P. M. Coussens, T. P. L. Smith, and G. W. Smith Generation of a bovine oocyte cDNA library and microarray: resources for identification of genes important for follicular development and early embryogenesis Physiol Genomics, September 16, 2004; 19(1): 84 - 92. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
