Digital multiplexed mRNA analysis of functionally important genes in single human oocytes and correlation of changes in transcript levels with oocyte protein expression.
Objective: To investigate functionally important transcripts in single human oocytes with the use of NanoString technology and determine whether observed differences are biologically meaningful.Design: Analysis of human oocytes with the use of NanoString and immunoblotting.Setting: University-affili...
| Publicado en: | Fertility & Sterility Vol. 101; no. 3; pp. 857 - 865 |
|---|---|
| Autores principales: | , , |
| Formato: | research Journal Article |
| Publicado: |
Elsevier B.V.
Mar2014
|
| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=104031654&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 104031654 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 00150282 1GO jtl: Fertility & Sterility issn: 00150282 maglogo: N pubinfo: dt: Mar2014 vid: 101 iid: 3 pid: 467 pub: Elsevier B.V. place: New York, New York artinfo: ui: 104031654 NLM24444598 2012486127 10.1016/j.fertnstert.2013.11.125 NLM24444598 PMC3969224 104031654 ppf: 857 ppct: 8 formats: tig: atl: Digital multiplexed mRNA analysis of functionally important genes in single human oocytes and correlation of changes in transcript levels with oocyte protein expression. aug: au: Riris, Solon Webster, Philippa Homer, Hayden affil: Reproductive Medicine Unit, Elizabeth Garrett Anderson Wing, University College London Hospitals, London, United Kingdom. sug: subj: Genes Physiology Nanotechnology Methods Ovum Physiology RNA RNA Physiology Adult Female Human Adult: 19-44 years Female ab: Objective: To investigate functionally important transcripts in single human oocytes with the use of NanoString technology and determine whether observed differences are biologically meaningful.Design: Analysis of human oocytes with the use of NanoString and immunoblotting.Setting: University-affiliated reproductive medicine unit.Patients: Women undergoing in vitro fertilization.Intervention: Human oocytes were analyzed with the use of NanoString or immunoblotting.Main Outcome Measures: The abundance of transcripts for ten functionally important genes-AURKA, AURKC, BUB1, BUB1B (encoding BubR1), CDK1, CHEK1, FYN, MOS, MAP2K1, and WEE2-and six functionally dispensable genes were analyzed with the use of NanoString. BubR1 protein levels in oocytes from younger and older women were compared with the use of immunoblotting.Result(s): All ten functional genes but none of the six dispensable genes were detectable with the use of NanoString in single oocytes. There was 3- to 5-fold variation in BUB1, BUB1B, and CDK1 transcript abundance among individual oocytes from a single patient. Transcripts for these three genes-all players within the spindle assembly checkpoint surveillance mechanism for preventing aneuploidy-were reduced in the same oocyte from an older patient. Mean BUB1B transcripts were reduced by 1.5-fold with aging and associated with marked reductions in BubR1 protein levels.Conclusion(s): The abundance of functionally important transcripts exhibit marked oocyte-to-oocyte heterogeneity to a degree that is sufficient to affect protein expression. Observed variations in transcript abundance are therefore likely to be biologically meaningful, especially if multiple genes within the same pathway are simultaneously affected. pubtype: Academic Journal doctype: research Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
|---|