Mediators of interferon gamma-initiated signaling in bovine luteal cells.
Interferon gamma (IFNgamma) has been implicated as a mediator of luteal steroidogenesis and cell fate. IFNgamma-initiated signaling events, although implied by studies in cell lines, have yet to be described in primary luteal cells. The objective of these studies was to begin to characterize IFNgamma-initiated signaling within luteal cells. Dispersed bovine luteal cell cultures were challenged with increasing levels of bovine recombinant IFNgamma (0-1000 U) or IFNgamma (200 U) in the presence or absence of tumor necrosis factor alpha (TNFalpha, 10 ng/ml) over time (short term, 0-60 min; long term, 0, 24, 48 h). Fractionated or total cell lysates were evaluated by the Western blotting technique to determine the changes in the levels of signal transducers and activators of transcription (STAT), interferon regulatory factor 1 (IRF-1), and I kappa B alpha (IkappaB-alpha). Utilizing antibodies that recognize the nonphosphorylated forms of STAT-1 and STAT-3, it was determined that levels of STAT-1 and STAT-3 in total cell lysates were constitutively expressed and did not change in response to treatment with IFNgamma or TNFalpha. In contrast, nuclear levels of STAT-1 and phosphorylated STAT-3 were elevated in a time-dependent manner in response to IFNgamma treatment. Furthermore, IFNgamma and TNFalpha treatment elevated levels of IRF-1 within 2 h. TNFalpha-induced increases in the levels of IRF-1 were transient, whereas the levels of IRF-1 in response to IFNgamma treatment remained elevated at 48 h. These data suggest that IFNgamma treatment can activate members of the STAT pathway, resulting in increased levels of IRF-1. TNFalpha treatment induced a rapid decrease in the levels of IkappaB-alpha. IFNgamma treatment did not alter the levels of IkappaB-alpha and failed to inhibit the TNFalpha-initiated decrease in the levels of IkappaB-alpha. The present experiment demonstrates that the steroidogenic cells of the corpus luteum have the capacity to respond to IFNgamma via activation of STAT and IRF-1, providing further evidence that IFNgamma may be involved in the luteolytic process. These data also suggest that IFNgamma does not signal through the nuclear factor kappa B cell survival signaling pathway.
Suter
J
J
The Women's Research Institute, Wichita, Kansas 67214-3199, USA.
Hendry
I R
IR
Ndjountche
L
L
Obholz
K
K
Pru
J K
JK
Davis
J S
JS
Rueda
B R
BR
D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002478:Cells, Cultured; D003338:Corpus Luteum / Q000166:cytology / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005260:Female; D020672:I-kappa B Proteins; D050856:Interferon Regulatory Factor-1; D007371:Interferon-gamma / Q000008:administration & dosage / Q000494:pharmacology*; D000072000:NF-KappaB Inhibitor alpha; D010750:Phosphoproteins / Q000378:metabolism; D011247:Pregnancy; D011374:Progesterone / Q000378:metabolism; D011994:Recombinant Proteins; D050794:STAT1 Transcription Factor; D050796:STAT3 Transcription Factor; D015398:Signal Transduction; D015534:Trans-Activators / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D004268:DNA-Binding Proteins; D020672:I-kappa B Proteins; D050856:Interferon Regulatory Factor-1; D010750:Phosphoproteins; D011994:Recombinant Proteins; D050794:STAT1 Transcription Factor; D050796:STAT3 Transcription Factor; D015534:Trans-Activators; D014409:Tumor Necrosis Factor-alpha; D000072000:NF-KappaB Inhibitor alpha; D011374:Progesterone; D007371:Interferon-gamma