Isolation of differentially expressed genes in conceptuses and endometrial tissue of sows in early gestation.
The implantation period is a critical time for embryonic survival in pigs. During this period, numerous growth factors are secreted by the conceptuses and the uterine endometrium in order to establish pregnancy and to provide a proper environment for embryonic development. It is well known that the Chinese Meishan sows have a larger litter size when compared with occidental sows mainly because of a superior embryonic survival rate. As a further step toward understanding the mechanisms involved in embryonic survival, we used a suppression subtractive hybridization technique to identify genes that were differentially expressed in Meishan-Landrace conceptuses and endometrial tissue at Day 15 of gestation when compared with conventional Landrace sows. Of the 1000 subtractive clones isolated from each library, 137 endometrial and 166 conceptus-enriched cDNAs were single-pass sequenced and examined by BLAST analysis for identification. Sixty-two percent of the clones found in the endometrial library and 78% of the clones found in the conceptus library showed homology with known genes. Among these genes, the 20 most relevant to embryonic survival based on the available literature were validated through real-time reverse-transcriptase polymerase chain reaction (RT-PCR) analysis. Our results show that suppression subtractive hybridization is a powerful method applicable in identifying putative candidate genes that might be used for selection of high litter-size breeds.
Vallée
Maud
M
Département de Biologie, Université de Sherbrooke, Sherbrooke, Québec, Canada J1K 2R1.
Beaudry
Daniele
D
Roberge
Claude
C
Matte
J Jacques
JJ
Blouin
Richard
R
Palin
Marie-France
MF
D000818:Animals; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D010064:Embryo Implantation / Q000502:physiology; D004622:Embryo, Mammalian / Q000378:metabolism*; D004717:Endometrium / Q000378:metabolism*; D030762:Estrous Cycle / Q000502:physiology; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D015723:Gene Library; D017403:In Situ Hybridization; D011247:Pregnancy; D011270:Pregnancy, Animal / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013045:Species Specificity; D013552:Swine
D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger