Suppression Subtractive Hybridization Identifies Genes Expressed in Oviduct during Mouse Preimplantation Period

Lee, Ckf; Kwok, Ka-Leung; Yeung, William S.B.

doi:10.1006/bbrc.2000.3736

Cited by 20 publications

(12 citation statements)

References 42 publications

(34 reference statements)

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“…Thirty clones were expressed at a same or higher level in the 48-h PMSG (preovulatory) ovarian mRNA relative to the post-hCG (ovulatory) mRNA, giving a false-positive rate of 41%. This rate is within the accepted range of the reported false-positive rate for the SSH technique, as it varies very much depending on experimental circumstances (Lee et al 2000, Tanaka et al 2003, Fayad et al 2004.…”

Section: Discussionsupporting

confidence: 75%

“…We chose to use the SSH technique, since the relative advantages of SSH include the fact that it does not rely on an existing cDNA library and therefore is not limited by its quality. Other advantages are the normalization of the representation of high and low abundance transcripts, and the elimination of the physical subtraction step in the isolation of target cDNAs (Lee et al 2000, Levesque et al 2003, Fayad et al 2004, Rebrikov et al 2004. Moreover, the successful use of this PCR-based method has previously been reported in the context of constructing testis-specific library (Diatchenko et al 1996) and by our laboratory in constructing an ovary-specific library (Tanaka et al 2003).…”

Section: Discussionmentioning

confidence: 97%

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Ovulation-selective genes: the generation and characterization of an ovulatory-selective cDNA library

Hourvitz

Gershon

Hennebold³

et al. 2006

Journal of Endocrinology

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Ovulation-selective/specific genes, that is, genes preferentially or exclusively expressed during the ovulatory process, have been the subject of growing interest. We report herein studies on the use of suppression subtractive hybridization (SSH) to construct a 'forward' ovulationselective/specific cDNA library. In toto, 485 clones were sequenced and analyzed for homology to known genes with the basic local alignment tool (BLAST). Of those, 252 were determined to be nonredundant. Of these 252 nonredundant clones, 98 were analyzed by probing mouse preovulatory and postovulatory ovarian cDNA. Twentyfive clones (26%) failed to show any signal, and 43 cDNAs tested thus far display a true ovulation-selective/specific expression pattern. In this communication, we focus on one such ovulation-selective gene, the fatty acid elongase 1 (FAE-1) homolog, found to be localized to the inner periantral granulosa and to the cumulus granulosa cells of antral follicles. The FAE-1 gene is a -ketoacyl-CoA synthase belonging to the fatty acid elongase (ELO) family, which catalyzes the initial step of very long-chain fatty acid synthesis. All in all, the present study accomplished systematic identification of those hormonally regulated genes that are expressed in the ovary in an ovulation-selective/specific manner. These ovulationselective/specific genes may have significant implications for the understanding of ovarian function in molecular terms and for the development of innovative strategies for both the promotion of fertility and its control.

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Section: Discussionsupporting

confidence: 75%

Section: Discussionmentioning

confidence: 97%

Ovulation-selective genes: the generation and characterization of an ovulatory-selective cDNA library

Hourvitz

Gershon

Hennebold³

et al. 2006

Journal of Endocrinology

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“…PCR was carried out at 948C for 10 min, then 30 cycles of 948C for 30 s, 608C for 30 s, and 728C for 60 s. The PCR products (50 ng) were purified and dotted onto Hybond N þ membranes (Amersham Pharmacia Biotech.) as described previously [Lee et al, 2000. Non-subtracted cDNA probes from testers and drivers were 32 P-labeled using PCR-select Subtraction Hybridization Screening Kit (Clontech, Inc.) in the presence of [aÀ 32 P]-dCTP (Dupont NEN, Boston, MA).…”

Section: Reverse Dot-blot Analysismentioning

confidence: 99%

“…The developing embryos, in turn, altered the gene expression in the mouse oviduct [Lee et al, 2000. By comparing the gene expression in the oocyte-containing oviduct with that of the embryo-containing oviduct, we successfully isolated more than a dozen genes differentially expressed in the embryo-containing oviduct .…”

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confidence: 99%

Phospholipid transfer protein (PLTP) mRNA expression is stimulated by developing embryos in the oviduct

Lee

Kwok

Chung

et al. 2005

J of Cellular Biochemistry

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In mammal, fertilization and early preimplantation embryo development occurs in the oviduct. Evidence is accumulating that the oviductal epithelia secrete various biomolecules to the lumen during the secretory phase of the estrus cycle to enhance embryo development. This secretory activity of the oviduct is under the regulation of steroid hormones. Observations also suggested that the gametes and embryos modulate the physiology and gene-expressing pattern of the oviduct. However, the underlying molecular changes remain elusive. We hypothesize that the developing embryos interact with the surrounding environment and affect the gene expression patterns of the oviduct, thereby modulating the oviductal secretory activity conducive to the preimplantation embryo development. To test this hypothesis, suppression subtractive hybridization (SSH) was used to compare the gene expressions in mouse oviduct containing transferred in vitro cultured preimplantation embryos with that of oviduct containing oocytes during the preimplantation period. We reported here the identification and characterization of phospholipids transfer protein (PLTP), which is highly expressed in the embryo-containing oviduct and localized at the oviductal epithelium by in situ hybridization. PLTP contains signal peptide putative for secretory function. More importantly, PLTP mRNA increases in the oviductal epithelia of pregnant, but not pseudo-pregnant mice when assayed by real-time PCR. Taken together, our data suggested that PLTP may play important role(s) during in vivo preimplantation embryo development. This molecule would be a target to delineate the mechanisms and the roles of oviductal secretory proteins on early embryonic development.

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“…Gene content and cDNA clone availability requirements are especially exigent to satisfy the growing interest in expression profiling of both stem cell populations (Phillips et al 2000;Billia et al 2001;Terskikh et al 2001;Steidl et al 2002;Testa et al 2002) and embryos in early developmental stages (Ko et al 2000;Lee et al 2000;Tanaka et al 2000;Hwang et al 2001;Stanton and Green 2002). As a step toward relevant gene content for microarray platforms, we described a sequence-verified mouse cDNA clone set representing up to 15,000 unique transcripts (Kargul et al 2001) derived primarily from preimplantation embryos.…”

mentioning

confidence: 99%

In Situ-Synthesized Novel Microarray Optimized for Mouse Stem Cell and Early Developmental Expression Profiling

et al. 2003

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Applications of microarray technologies to mouse embryology/genetics have been limited, due to the nonavailability of microarrays containing large numbers of embryonic genes and the gap between microgram quantities of RNA required by typical microarray methods and the miniscule amounts of tissue available to researchers. To overcome these problems, we have developed a microarray platform containing in situ-synthesized 60-mer oligonucleotide probes representing approximately 22,000 unique mouse transcripts, assembled primarily from sequences of stem cell and embryo cDNA libraries. We have optimized RNA labeling protocols and experimental designs to use as little as 2 ng total RNA reliably and reproducibly. At least 98% of the probes contained in the microarray correspond to clones in our publicly available collections, making cDNAs readily available for further experimentation on genes of interest. These characteristics, combined with the ability to profile very small samples, make this system a resource for stem cell and embryogenomics research.

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Suppression Subtractive Hybridization Identifies Genes Expressed in Oviduct during Mouse Preimplantation Period

Cited by 20 publications

References 42 publications

Ovulation-selective genes: the generation and characterization of an ovulatory-selective cDNA library

Ovulation-selective genes: the generation and characterization of an ovulatory-selective cDNA library

Phospholipid transfer protein (PLTP) mRNA expression is stimulated by developing embryos in the oviduct

In Situ-Synthesized Novel Microarray Optimized for Mouse Stem Cell and Early Developmental Expression Profiling

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