Use of reverse transcription-polymerase chain reaction to detect embryonic interleukin-1 system messenger RNA in individual preimplantation mouse embryos co-cultured with Vero cells.
In this study, we report a total of 292 mouse embryos cultured on Vero cell monolayers and 77 embryos cultured in medium alone at different preimplantation stages examined individually for embryonic mRNA of beta-actin, interleukin-1beta (IL-1beta), interleukin-1 receptor antagonist (icIL-1ra) and interleukin-1 receptor type I (IL-1RtI) using reverse transcription and two-step polymerase chain reaction (RT-PCR). The rates of blastocyst formation and blastocyst hatching were both significantly higher in embryos co-cultured with Vero cells in comparison with the embryos cultured in control medium (81.2 +/- 2.6 versus 42.2 +/- 3.7%, P < 0.001; 75.6 +/- 2.7 versus 19.2 +/- 6.2%, P < 0.001 respectively). We have identified a similar pattern of interleukin-1 family embryonic mRNA transcripts expressed from the compact morula stage through to hatching blastocyst in both control and Vero cell cultured embryos with significantly increased icIL-1ra transcript at hatching blastocyst stage (P < 0.05, P < 0.001 respectively). There was a significant increase in IL-1beta mRNA transcripts of embryos at hatching blastocyst stage compared to compact morula stage in Vero cell cultured embryos (P < 0.05). These findings support the hypothesis that the IL-1 system is an important factor in embryo-maternal molecular communication during the implantation process.
Huang
H Y
HY
Department of Gynecology and Obstetrics, Stanford University Medical Center, California, USA.
Krüssel
J S
JS
Wen
Y
Y
Polan
M L
ML
D000199:Actins / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D001755:Blastocyst / Q000737:chemistry / Q000502:physiology; D002522:Chlorocebus aethiops; D018920:Coculture Techniques; D004622:Embryo, Mammalian / Q000737:chemistry*; D047108:Embryonic Development; D005260:Female; D015870:Gene Expression; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1 / Q000235:genetics*; D051379:Mice; D009028:Morula / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis*; D012194:RNA-Directed DNA Polymerase; D017472:Receptors, Interleukin-1 / Q000235:genetics; D012795:Sialoglycoproteins / Q000235:genetics; D014709:Vero Cells
D000199:Actins; C507764:Il1rn protein, mouse; D053590:Interleukin 1 Receptor Antagonist Protein; D007375:Interleukin-1; D012333:RNA, Messenger; D017472:Receptors, Interleukin-1; D012795:Sialoglycoproteins; D012194:RNA-Directed DNA Polymerase