Preimplantation embryology: Expression of leukaemia inhibitory factor receptor subunits LIFRβ and gp 130 in human oocytes and preimplantation embryos

Abstract

The expression of both components of the high-affinity leukaemia inhibitory factor receptor, LIFR beta and glycoprotein 130 (gp130), was investigated in human oocytes and individual in-vitro cultured preimplantation embryos by reverse transcription-polymerase chain reaction (RT-PCR). Messenger RNA of both LIFR beta and gp130 was detected in as little as 1/30 and 1/12 sample equivalents of cDNA respectively, in oocytes (n = 4), 4-cell and expanded, blastacyst stage embryos. LIFR beta but not gp130 transcripts were detected at the 2-, 8- and 10-cell stages, and in cavitating and hatched blastocysts. In order to exclude a simian origin of these PCR products resulting from the Vero cell line that was used as a feeder during culture to the blastocyst stage, they were digested with restriction endonucleases Taql (LIFR beta) or Kpnl (gp130). Their human origin was confirmed. The results support an earlier finding of LIFR beta mRNA expression in human blastocysts, and extend these results to earlier stages and oocytes. This is the first report of LIFR beta and gp130 transcription in human oocytes. Taken together these results demonstrate that transcription of LIFR beta and gp130 takes place throughout human preimplantation development, and suggest that functional LIF receptors might be present at these stages. These results further confirm the feasibility of performing mRNA phenotyping of multiple genes with RNA derived from a single preimplantation stage embryo.

Synthesis and regulation of leukaemia inhibitory factor in cultured bovine oviduct cells by hormones

“…However, no mRNA encoding LIF has been observed in preimplantation embryonic stages (Sharkey et al, 1995;Van Eijk et al, 1996). This finding supports the hypothesis that the effects of LIF have to be of maternal origin.…”

“…The samples were transferred into Eppendorf tubes and kept frozen at -70°C until analysed. Extraction of total RNA and analysis of mRNA expression was accomplished by using RT-PCR processing and polyacrylamide gels stained with ethidium bromide as described previously (Van Eijk et al, 1996). Briefly, oligonucleotide primers used for amplification were based on human sequences in the databases, as follows: pLIF1 (5Ј-AACCTCATGAA-CCAGATCAG-3Ј) and pLIF2 (5Ј-TGACAGCCCAGCTTCTTCTT-3Ј), which yield a 431 bp product; actin-1 (5Ј-TGAACCCCAAGGCC-AACCGCG-3Ј) and actin-2 (5Ј-GCTCATAGCTC TTCTCC AGGG-3Ј) which yield a 396 bp product.…”

The clinical role of interleukin-6 and interleukin-6 soluble receptor in human follicular fluids

“…A variety of IL-6-response cells is present in the preovulatory follicles at ovulation. It may be possible for IL-6 to cause oocyte maturation, because gp 130, which is shared by a family of related growth factors that includes IL-6, was present in the oocytes [27]. After ovulation and disintegration of the basal lamina, theca vessels invade the granulosa layer, giving rise to the luteal vascular network.…”

Developmental exposure to environmental endocrine disruptors: Consequences within the ovary and on female reproductive function☆

“…Examples include the neurotrophins, bFGF, leukemia inhibitory factor (LIF), and BMP-4. Neurotrophins and their receptors [36,49]; bFGF and its receptor FGFR2IIIb [50,51]; LIF and its receptors LIFR-beta and gp130 [52,53]; and BMP-4 and its receptors BMPR-IA, -IB, and -II [54] are all expressed in the ovary. The roles of bFGF [50], LIF [53], and BMP-4 [55] as regulators of various aspects of folliculogenesis beyond follicular assembly (i.e., the primordial-to-primary follicle transition) have been shown in vitro using neonatal rat ovary organ culture.…”