Browsing by Author "Velasquez, LA"

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    A segment and epithelium specific messenger ribonucleic acid fragment up-regulated by estradiol in the rat oviduct
    (2001) Rios, M; Ojeda, S; Velasquez, LA; Maisey, K; Croxatto, HB
    Estradiol accelerates oviductal embryo transport in the rat through changes of genomic expression in oviductal cells. However, the genes involved are unknown. We used a differential display by reverse transcription-polymerase chain reaction to detect estradiol (E-2)-dependent genes in the rat oviduct. Rats on day 2 of pregnancy were untreated or treated with 10 mug of E-2 and the oviducts were extracted at 30, 180 and 360 min later and used to isolate RNA. products of reverse transcriptase-PCR, made with pairs of arbitrary and oligo-deoxythymidine primers, were separated on denaturing polyacrylamide gels and candidate bands were excised and reamplified. Truly positive cDNA fragments determined by a single strand conformation polymorphism assay were cloned and sequenced. A ribonuclease protection assay confirmed that clone 25 is up-regulated by E-2 in the oviduct at 30, 180 and 360 min. This clone exhibited no homology with known genes and in situ hybridization showed it is only expressed in the epithelial cells of the isthmic segment. Clone 25 is likely to represent a new gene. which is upregulated by E-2 in the epithelium of the isthmic segment of the rat oviduct. Its rime frame of response is compatible with a mediator or the effect of E-2 on oviductal embryo transport.
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    Expression of platelet-activating factor receptor in the hamster oviduct: Localization to the endosalpinx
    (1997) Velasquez, LA; Ojeda, SR; Croxatto, HB
    Platelet-activating factor (PAF) is a lipid mediator that has a range of biological effects on various cells and tissues. PAF-like activity has been detected in the spent media of two-cell to morula stage hamster embryos, leading to the suggestion that PAF may be the embryonic signal that hastens embryo transport to the uterus in this species. The present study was undertaken to examine whether the PAF receptor (PAFr) gene is expressed in hamster oviduct, and to identify the cell types in which the gene is expressed. DNA fragments complementary to the coding region of mRNA encoding hamster PAFr were cloned by reverse transcription-polymerase chain reaction (RT-PCR), identified by sequencing and used to prepare hamster specific cRNA probes. The presence of mRNA transcripts encoding the PAFr receptor in the oviduct was investigated by subjecting oviduct mRNA to RT-PCR. Southern blot analysis of the RT-PCR products verified the identity of the presumptive PAFr cDNAs. The cloned cDNA fragment of hamster PAFr was found to be highly conserved with respect to the receptor of other species, having 94.3% sequence similarity to the rat PAFr receptor. Hybridization histochemistry demonstrated that PAFr is expressed in the subepithelial cells and occasionally in the epithelium. In conclusion, expression of PAFr in the hamster oviduct is compatible with the proposed paracrine role of early embryo-derived PAF.
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    PAF receptor and PAF acetylhydrolase expression in the endosalpinx of the human Fallopian tube: possible role of embryo-derived PAF in the control of embryo transport to the uterus
    (2001) Velasquez, LA; Maisey, K; Fernandez, R; Valdes, D; Cardenas, H; Imarai, M; Delgado, J; Aguilera, J
    BACKGROUND: Prostaglandin-E-2 and platelet-activating factor (PAF) are embryonic-derived signals that time embryo passage into the uterus in the mare and hamster respectively. PAF-like activity is detectable in the spent media of preimplantation human embryos and it has been suggested that PAF may be the embryonic signal that controls embryo transport to the uterus in our species. The actions of PAF are regulated at the level of its synthesis and degradation as well as the expression of a specific cell surface receptor (PAFr). The enzyme PAF acetylhydrolase (PAF-AH) degrades PAR This study was undertaken to examine whether or not PAFr and PAF-AH are expressed in the human Fallopian tube and to identify the cell types in which they are expressed. METHODS: The presence of PAFr mRNA in tissue extracts was investigated using reverse transcription-polymerase chain reaction. We amplified the predicted amplicon for PAFr mRNA from RNA samples extracted from Fallopian tubes. The expression of PAF-AH was detected by Western blot and the localization of PAFr and PAF-AH proteins was detected by immunohistochemistry. RESULTS: Utilizing antibodies against PAFr and PAF-AH, co-localization of the two proteins in the epithelium and stromal cells were demonstrated. CONCLUSIONS: These observations show that the human Fallopian tube expresses PAFr and PAF-AH at a location compatible with the proposed paracrine role of early embryo-derived PAF.