IFN‐τ Acts in a Dose‐Dependent Manner on Prostaglandin Production by Buffalo Endometrial Stromal Cells Cultured in vitro

Abstract

Interferon-τ (IFN-τ) has been recognized as the primary embryonic signal responsible for maternal recognition of pregnancy. Uterine endometrium produces both prostaglandin F2α (PGF2α ) and prostaglandin E2 (PGE2 ). PGF2α is responsible for the luteolysis; however, PGE2 favours establishment of pregnancy by its luteoprotective action. In this study, the dose-response effect of recombinant bovine IFN-τ (rbIFN-τ) on prostaglandin (PG) production by buffalo endometrial stromal cells cultured in vitro was studied. Buffalo endometrial stromal cells were isolated by double enzymatic digestion, initially with trypsin III followed by a cocktail of trypsin III, collagenase type II and DNase I and subsequently cultured till confluence. Further, cells were treated with different doses of rbIFN-τ (0.001, 0.01, 0.1, 1.0 and 10 μg/ml) and keeping a separate set of control. Culture supernatant was collected after 6, 12 and 24 h of treatment. PG levels in the culture supernatant were measured by enzyme immune assay (EIA) and total cellular protein estimated by Bradford method. Results indicated that buffalo endometrial stromal cells following rbIFN-τ treatment enhanced the secretion of both PGE2 and PGF2α , and also its ratio in a strict dose-dependent manner with a significant increase (p < 0.01) in PGE2 production at 1 μg/ml dose of rbIFN-τ and maximal stimulation for both PG was observed at 10 μg/ml. Further, both PG production and its ratio were increased significantly (p < 0.01) in a time-dependent fashion in all the groups at 6, 12 and 24 h post-treatment with highest level achieved at 24 h as compared with control. Absolute levels of PGE2 remained higher than PGF2α indicating PGE2 as the major PG produced by endometrial stromal cells. The dose-dependent response of rbIFN-τ signifies the importance of optimum concentration of IFN-τ for the embryonic development especially during the critical period to establish successful pregnancy.

Prospecting major genes in dairy buffaloes

“…For age at first calving (AFC), the gene coding for interferon-Tau, IFN-TAU , and other interferon genes were identified. Embryonic production of IFN-TAU is the primary signal for maternal recognition of pregnancy in buffaloes [ 23 ]. Another gene associated with AFC was LOC100299005 ( SELP ), a gene with up-regulated expression during inflammatory processes related to follicular atresia in cattle [ 24 ].…”

Investigation of PAG2 mRNA Expression in Water Buffalo Peripheral Blood Mononuclear Cells and Polymorphonuclear Leukocytes from Maternal Blood at the Peri-Implantation Period

“…Recently, various works have demonstrated the presence of a viable conceptus by measuring levels of ISG in maternal peripheral blood leukocytes 15 days after insemination in sheep [44], 18 days after AI in dairy cattle [45] and between 15 and 20 days after AI in beef cattle [46]. The interferon tau (IFN-τ) is the major paracrine factor involved in pregnancy recognition in ruminant species [47,48,49]. Both PAG-2 and IFN-τ are Day 7/Day 14 and Day 14/Day 21 up-regulated genes in bovine conceptus, respectively [50].…”

Immunomodulatory effect of curcumin on lipopolysaccharide‐ and/or flagellin‐induced production of prostaglandin E2 and relative expression of proinflammatory cytokines in the primary bubaline endometrial stromal cells

“…The primary culture of stromal cell was done as reported elsewhere (Chethan et al, ) with minor modifications. Briefly, endometrial strips were digested in 0.3% trypsin III for 2 hr.…”