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Title: REGULATION OF ENDOMETRIAL GRANULOCYTE MACROPHAGE COLONY STIMULATING FACTOR (GM-CSF) PRODUCTION IN THE EWE.

Author
item TAMURA, KAZUHIRO - WRI, UNIV KS SCH MED
item HARBISON, LISA - WRI, UNIV KS SCH MED
item Christenson, Ronald
item IMAKAWA, KAZUHIKO - WRI, UNIV KS SCH MED

Submitted to: Society for the Study of Reproduction Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 5/1/1995
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Maternal GM-CSF is involved in conceptus production of oIFN-tau during pre- and peri-implantation periods but the physiological mechanisms that regulate endometrial GM-CSF expression remain obscure. To determine the effects of estradiol-17-beta (E2) and progesterone (P4) on the production of GM-CSF, 13 cyclic crossbred ewes were ovariectomized 30 days before assignment on day 0 to one of four subcutaneous silastic implant treatments: 1) control (n=3), 2) E2 (n=3), 3) P4 (n=3), or 4) a combination of E2 and P4 (n=4). On day 14, all ewes were bled, hysterectomized, and endometrium was dissected from the uterus. Endometrial tissue was fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned for immunohistochemical and in situ hybridization studies. Endometrium (300 mg) was minced and cultured for 24 hr at 37 C in 10 ml Eagle's MEM and 70 uCi 3H-leucine. After culture, the medium samples were dialyzed, concentrated and subjected to western blot analysis using human GM-CSF antibody. Concentrations of GM-CSF in the culture media were quantitated by a proliferation bioassay using GM-CSF sensitive TF-1 cells. Amounts of GM-CSF mRNA in RNA extracted from cultured tissues were determined by quantitative RT-PCR. GM-CSF mRNA levels were 15+/-3, 46+/-8, 16+/-4, and 43+/-5 pg/ug RNA for control, E2, P4, and E2/P4 treatments, respectively. Concentrations of GM-CSF, as determined by bioassay, were 53+/-11, 55+/-2, 58+/-9, and 190+/-19 pg/ml, respectively. Western blot analysis also revealed that immunoreactive GM-CSF was present only in the culture media obtained from the E2/P4 treatment group. These data indicate that E2 is effective in enhancing levels of GM-CSF mRNA, but translation of GM-CSF mRNA into GM-CSF polypeptide requires both E2 and P4.