Gonadotropin-releasing hormone messenger ribonucleic acid and protein expression in Vero cells.
OBJECTIVE
Interleukin-1 (IL-1) is a major regulator of local cellular interactions during embryonic implantation. We hypothesized that gonadotropin-releasing hormone (GnRH) may also play a role in the embryonic/epithelial dialogue during early implantation. To examine this hypothesis, we examined the ability of IL-1 to regulate GnRH mRNA and protein expression in Vero cells.


METHODS
Viable Vero cells (1 x 10(5)/well) were cultured in multiple-well tissue culture plates for in vitro studies and in 4-well chamber slides for immunohistochemical study. Confluent Vero cells were cultured with increasing concentrations of recombinant human IL-1 beta for an additional 24 hr. Vero cell expression of GnRH and GnRH receptor mRNAs was measured with polymerase chain reaction (PCR) and nested PCR, respectively. GnRH protein expression was validated by immunohistochemistry study. The quantitative level of GnRH mRNA expression regulated by IL-1 beta in Vero cells was determined by quantitative competitive PCR (QC PCR) with standard curve methodology.


RESULTS
RT-PCR revealed beta-actin, GnRH, and GnRH receptor mRNA expression in Vero cell cultures. Immunostaining confirmed the presence of GnRH protein in Vero cells. Quantitative PCR demonstrated IL-1 beta up-regulation of Vero cell GnRH mRNA expression (p < 0.05).


CONCLUSIONS
These results suggest that Vero cell mRNA and protein expression of GnRH may play a substantial role in early embryo/epithelial dialogue during embryo coculture, with an embryotrophic effect due to expression of GnRH by Vero cells.
Huang
H Y
HY
Department of Obstetrics and Gynecology, Lin-Kou Medical Center, Chang Gung Memorial Hospital, 5 Fu-Hsing Street, Kwei-Shan, Tao-Yuan, Taiwan.
Raga
F
F
Kruessel
J S
JS
Wen
Y
Y
Soong
Y K
YK
Polan
M L
ML
D000199:Actins / Q000096:biosynthesis; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002522:Chlorocebus aethiops; D018920:Coculture Techniques; D016895:Culture Media, Serum-Free / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel; D010064:Embryo Implantation; D007987:Gonadotropin-Releasing Hormone / Q000096:biosynthesis / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism*; D011966:Receptors, LHRH / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation; D014709:Vero Cells
D000199:Actins; D016895:Culture Media, Serum-Free; D018076:DNA, Complementary; D007375:Interleukin-1; D012333:RNA, Messenger; D011966:Receptors, LHRH; D011994:Recombinant Proteins; D007987:Gonadotropin-Releasing Hormone