Fetal membrane distention: II. Differentially expressed genes regulated by acute distention in vitro.
OBJECTIVE
This study was undertaken to identify genes with expression up-regulated by acute distention in the human fetal membranes.


METHODS
Fetal membrane explants were distended reproducibly in a novel device in vitro for 4 hours, and suppression subtractive hybridization was used to identify the candidate genes for up-regulation of expression in response to this stimulus. The up-regulation in response to distention was confirmed by quantitative Northern blot analysis both after a 4-hour in vitro distention and after labor in vivo.


RESULTS
Suppression subtractive hybridization identified 3 genes with expression up-regulated by acute distention: an interferon-stimulated gene encoding a 54-kd protein, the gene for huntingtin-interacting protein 2 (a ubiquitin-conjugating enzyme), and a novel transcript. Expression of each of the distention-responsive genes found to be up-regulated in vitro was also up-regulated in fetal membranes in association with labor.


CONCLUSIONS
Suppression subtractive hybridization was successfully applied to a complex tissue, the human fetal membranes, and 3 novel distention-responsive genes were identified. Both acute in vitro distention and labor in vivo up-regulate expression of at least 3 genes in the human fetal membranes.
Nemeth
E
E
Pacific Biomedical Research Center, University of Hawaii, Honolulu 96822, USA.
Millar
L K
LK
Bryant-Greenwood
G
G
D051017:Apoptosis Regulatory Proteins; D001696:Biomechanical Phenomena; D015152:Blotting, Northern; D046508:Culture Techniques; D016207:Cytokines / Q000235:genetics; D004268:DNA-Binding Proteins; D005321:Extraembryonic Membranes / Q000502:physiology*; D005189:False Positive Reactions; D005260:Female; D005786:Gene Expression Regulation; D005865:Gestational Age; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics; D007743:Labor, Obstetric / Q000502:physiology; D008025:Ligases / Q000235:genetics; D008969:Molecular Sequence Data; D050778:NFATC Transcription Factors; D054409:Nicotinamide Phosphoribosyltransferase; D009687:Nuclear Proteins; D009693:Nucleic Acid Hybridization; D011247:Pregnancy; D016601:RNA-Binding Proteins; D014157:Transcription Factors / Q000235:genetics; D044763:Ubiquitin-Conjugating Enzymes
D051017:Apoptosis Regulatory Proteins; D016207:Cytokines; D004268:DNA-Binding Proteins; C484289:IFIT2 protein, human; D016209:Interleukin-8; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; D014157:Transcription Factors; C100879:UBE2K protein, human; D044763:Ubiquitin-Conjugating Enzymes; D054409:Nicotinamide Phosphoribosyltransferase; C085455:nicotinamide phosphoribosyltransferase, human; D008025:Ligases