Effect of chicken leptin recptor short hairpin RNA on expression of JAK2, STAT3, SOCS3 and CPT1 genes in chicken preadipocytes

Anim Sci J. 2017 Mar;88(3):559-564. doi: 10.1111/asj.12722. Epub 2016 Nov 21.

Abstract

In this study, we detect depressive effect on leptin receptor (LEPR) by LEPR-specific short hairpin RNA (shRNA) expression plasmids in chicken preadipocytes, and effect on messenger RNA (mRNA) expression levels of genes related to signal transduction, including JAK2, STAT3, SOCS3 as well as CPT1, which is associated with fatty acid metabolism. shRNA expression vectors targeting LEPR were constructed and transfected into chicken preadipocytes. The transfection efficiency was evaluated by fluorescence microscopy. Real-time PCR was used to detect its effect on mRNA expression levels of JAK2, STAT3, SOCS3 and CPT1. Results showed that LEPR mRNA was knocked down by 99% (P < 0.01) after transfection for 72 h. In the knockdown preadipocytes, the mRNA levels of JAK2 and CPT1 were down-regulated by 47.56% (P < 0.01) and 42.26% (P < 0.05), respectively; while expression of STAT3 and SOCS3 increased 7.72-fold (P < 0.01), 1.71-fold (P < 0.01), respectively. It is concluded that knockdown of LEPR influences mRNA expression of its down-stream genes, suggesting that chicken LEPR play a certain role in regulating genes in the complicated gene network of preadipocytes.

Keywords: LEPR; RNAi; chicken; preadipocytes.

MeSH terms

  • Adipocytes / metabolism*
  • Animals
  • Carnitine O-Palmitoyltransferase / genetics*
  • Carnitine O-Palmitoyltransferase / metabolism*
  • Cells, Cultured
  • Chickens / genetics*
  • Chickens / metabolism*
  • Fatty Acids / metabolism
  • Gene Expression*
  • Janus Kinase 2 / genetics*
  • Janus Kinase 2 / metabolism*
  • Plasmids
  • Polymerase Chain Reaction
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / genetics*
  • RNA, Small Interfering / physiology*
  • Receptors, Leptin / physiology*
  • STAT3 Transcription Factor / genetics*
  • STAT3 Transcription Factor / metabolism*
  • Signal Transduction / genetics
  • Suppressor of Cytokine Signaling 3 Protein / genetics*
  • Suppressor of Cytokine Signaling 3 Protein / metabolism*
  • Transfection

Substances

  • Fatty Acids
  • RNA, Messenger
  • RNA, Small Interfering
  • Receptors, Leptin
  • STAT3 Transcription Factor
  • Suppressor of Cytokine Signaling 3 Protein
  • Carnitine O-Palmitoyltransferase
  • Janus Kinase 2