Endometrial expression of progesterone-induced blocking factor and galectins-1, -3, -9, and -3 binding protein in the luteal phase and early pregnancy in cattle

Abstract

Forde N. Endometrial expression of progesterone-induced blocking factor and galectins-1, -3, -9, and -3 binding protein in the luteal phase and early pregnancy in cattle. Physiol Genomics 43: 903-910, 2011. First published May 24, 2011 doi:10.1152/physiolgenomics.00251.2010.-Progesterone-induced blocking factor (PIBF) and galectins modulate the maternal immune response during pregnancy. We hypothesized that the relative transcript abundance of the above genes would be different during the luteal phase/early pregnancy and would be affected by progesterone supplementation. To further test this, hypothesis protein expression analyses were carried out to evaluate the abundance and localization of LGALS9 and PIBF. Following estrus synchronization, heifers were inseminated (n ϭ 140) or not (n ϭ 70). Half the heifers in each status (cyclic or potentially pregnant) were randomly assigned to receive a progesterone-releasing intravaginal device (PRID) on day 3 after estrus, which elevated progesterone concentrations from day 3.5 to 8 (P Ͻ 0.05), resulting in four treatment groups: cyclic and pregnant heifers, each with normal and high progesterone. After confirmation of pregnancy status in inseminated animals, uterine tissue was collected on days 5, 7, 13, or 16 of the luteal phase of the cycle/ pregnancy. Gene and protein expression was determined using Q-RT-PCR and IHC, respectively, on 5 heifers per treatment per time point (i.e., 80 in total). Progesterone concentrations did not affect expression of any of the genes (P Ͼ 0.05).LGALS9 and LGALS3BP were expressed at low levels in both cyclic and pregnant endometria until day 13. On day 16, expression increased only in the pregnant heifers (P Ͻ 0.0001).LGALS1 and LGALS3 decreased on day 7 (P Ͻ 0.0001) and remained low until day 16. Pregnancy had no effect on the expression of LGALS1, LGALS3, and PIBF. Additionally, LGALS9 and PIBF proteins were expressed in distinct uterine cell types. These results indicate that the galectins may be involved in uterine receptivity and/or implantation in heifers.bovine; uterus; early pregnancy; estrous cycle; progesterone DURING THE ESTROUS CYCLE, the uterus undergoes remodeling under the influence of estrogen and progesterone, to create an optimum environment for embryo survival and successful implantation. Numerous studies have identified a linear and quadratic relationship between progesterone concentrations during early pregnancy and probability of embryo survival/ pregnancy outcome in both lactating dairy cows and nonlactating heifers (12,35,52). Elevated concentrations of circulating progesterone in the immediate postconception period have been associated with an advancement of conceptus elongation (8,20,46), an increase in interferon-tau (IFNT) production (32, 33) and higher pregnancy rates in cattle and sheep (35,52

Temporal regulation of fibroblast growth factors and their receptors in the endometrium and conceptus during the pre-implantation period of pregnancy in cattle

“…Gene expression analysis was carried out using qPCR analysis and was performed as described previously (Okumu et al 2011). Briefly, 100 mg samples of endometrium were homogenised and total RNA was extracted using TRIzol reagent (Invitrogen), according to the manufacturer's instructions.…”

“…The procedure for IHC was carried out as described previously (Okumu et al 2011). In summary, 4 mm sections of paraffin wax-embedded tissues were de-waxed in xylene and rehydrated through a series of graded alcohol steps.…”

“…Using a digital camera, four images were captured per tissue section (two images showing the luminal epithelium (LE), superficial glands (SG) and stroma (STR), and two images showing the deep glands (DG) and myometrium (MYO)). Intensity of staining for all regions was determined using Image-Pro Plus Software (version 6.2, MediaCybernetics, Bethesda, MD, USA) as described previously (Okumu et al 2011).…”

Investigation of bovine conceptus galectins and the effect of bovine conceptus secretory proteins on the endometrial epithelial transcriptome

“…ISGs are hypothesized to influence proliferation, migration, attachment, and adhesion of the trophectoderm to support elongation of the conceptus [169]. The gene, LGALS3BP, has been previously reported to be abundant in both pregnant and cyclic d16 uterine luminal fluid and endometrium of heifers in comparison to d13 [138,168,178]. Galectin-9 was upregulated in epithelial and fibroblast cells in response to IFNT treatment, while LGALS3BP was found to only be up-regulated in epithelial cells (Table 3.2).…”

“…Elongating bovine conceptuses express galectin-1 (LGALS1) which has also been associated with the viability of d7 bovine blastocysts. Expression of LGALS1 was found to be elevated in in-vivo produced blastocysts compared to less viable in-vitro produced d7 bovine blastocysts [138]. Further, the endometrium of sheep produces LGALS15 after d10 of gestation to aid in trophectoderm migration and attachment.…”