Differential Gene Expression in the Bovine Corpus Luteum During Transition from Early Phase to Midphase and Its Potential Role in Acquisition of Luteolytic Sensitivity to Prostaglandin F2 Alpha1

Goravanahally, Madhusudan P.; Salem, Mohamed; Yao, Jianbo; Inskeep, E. K.; Flores, Jorge A.

doi:10.1095/biolreprod.108.069518

Cited by 41 publications

(30 citation statements)

References 55 publications

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“…The use of GAPDH as a housekeeping gene has been previously validated [36], and we further validated the consistency of steady-state concentrations for this gene in the CL in preliminary experiments (unpublished results). This preliminary experiment indicated similar expression of GAPDH after multiple biopsies of the same animal and similar concentrations in the different treatment groups and different collection times in the present experiment.…”

Section: Discussionmentioning

confidence: 61%

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Patterns of Gene Expression in the Bovine Corpus Luteum Following Repeated Intrauterine Infusions of Low Doses of Prostaglandin F2alpha1

Atlı

Bender

Mehta

et al. 2012

Biology of Reproduction

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Natural luteolysis involves multiple pulses of prostaglandin F2alpha (PGF) released by the nonpregnant uterus. This study investigated expression of 18 genes from five distinct pathways, following multiple low-dose pulses of PGF. Cows on Day 9 of the estrous cycle received four intrauterine infusions of 0.25 ml of phosphate-buffered saline (PBS) or PGF (0.5 mg of PGF in 0.25 ml of PBS) at 6-h intervals. A luteal biopsy sample was collected 30 min after each PBS or PGF infusion. There were four treatment groups: Control (n = 5; 4 PBS infusions), 4XPGF (4 PGF infusions; n = 5), 2XPGF-non-regressed (2 PGF infusions; n = 5; PGF-PBS-PGF-PBS; no regression after treatments), and 2XPGF-regressed (PGF-PBS-PGF-PBS; regression after treatments; n = 5). As expected, the first PGF pulse increased mRNA for the immediate early genes JUN, FOS, NR4A1, and EGR1 but unexpectedly also increased mRNA for steroidogenic (STAR) and angiogenic (VEGFA) pathways. The second PGF pulse induced immediate early genes and genes related to immune system activation (IL1B, FAS, FASLG, IL8). However, mRNA for VEGFA and STAR were decreased by the second PGF infusion. After the third and fourth PGF pulses, a distinctly luteolytic pattern of gene expression was evident, with inhibition of steroidogenic and angiogenic pathways, whereas, there was induction of pathways for immune system activation and production of PGF. The pattern of PGF-induced gene expression was similar in corpus luteum not destined for luteolysis (2X-non-regressed) after the first PGF pulse but was very distinct after the second PGF pulse. Thus, although the initial PGF pulse induced mRNA for many pathways, the second and later pulses of PGF appear to have set the distinct pattern of gene expression that result in luteolysis.

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Section: Discussionmentioning

confidence: 61%

“…Sequences for primers were designed using Primer3 from the NCBI (http:// www.ncbi.nlm.nih.gov/) gene database or were derived from published studies [16,22,[35][36][37]. Primer pair sequences and product sizes are shown in supplemental Table S1 (available online at www.biolreprod.org).…”

Section: Atli Et Almentioning

confidence: 99%

Patterns of Gene Expression in the Bovine Corpus Luteum Following Repeated Intrauterine Infusions of Low Doses of Prostaglandin F2alpha1

Atlı

Bender

Mehta

et al. 2012

Biology of Reproduction

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“…EGR-1 and c-Fos have been shown as important key players of luteolytic acquisition in pig and cows (Chen et al 2001, Hou et al 2008, Atli et al 2012, Diaz et al 2013. PGF 2a -FP interaction increases intracellular Ca 2C which activates multiple cell signaling pathways and mediates the acquisition of luteolytic sensitivity to PGF 2a in the bovine luteal cells (Goravanahally et al 2009, Wright et al 2014. Interactions among PGF 2a , Ca 2C , PKC, ERK1/2, c-Fos, and c-JUN have been shown in luteal cells in cows (Chen et al 2001, Yadav et al 2005.…”

Section: Survival and Apoptosis Proteins In The CLmentioning

confidence: 99%

Early pregnancy modulates survival and apoptosis pathways in the corpus luteum in sheep

Lee¹,

Banu²,

McCracken³

et al. 2016

Reproduction

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The corpus luteum (CL) is a transient endocrine gland. Functional and structural demise of the CL allows a new estrous cycle. On the other hand, survival of CL and its secretion of progesterone are required for the establishment of pregnancy. Survival or apoptosis of the luteal cells is precisely controlled by interactions between survival and apoptosis pathways. Regulation of these cell signaling components during natural luteolysis and establishment of pregnancy is largely unknown in ruminants. The objective of the present study was to determine the regulation of survival and apoptosis signaling protein machinery in the CL on days 12, 14, and 16 of the estrous cycle and pregnancy in sheep. Results indicate that: i) expressions of p-ERK1/2, p-AKT, b-catenin, NFkB -p65, -p50, -p52, p-Src, p-b -arrestin, p-GSK3b, X-linked inhibitor of apoptosis protein (XIAP), and p-CREB proteins are suppressed during natural luteolysis; in contrast, their expressions are sustained or increased during establishment of pregnancy; ii) expressions of cleaved caspase-3, apoptosis inducing factor (AIF), c-Fos, c-Jun, and EGR-1 proteins are increased during natural luteolysis; in contrast, their expressions are decreased during establishment of pregnancy; and iii) expressions of Bcl-2, Bcl-XL, Bad, and Bax proteins are not modulated during natural luteolysis while expressions of Bcl2 and Bcl-XL proteins are increased during establishment of pregnancy in sheep. These proteomic changes are evident in both large and small luteal cells. These results together indicate that regression of the CL during natural luteolysis or survival of the CL during establishment of pregnancy is precisely controlled by distinct programmed suppression or activation of intraluteal cell survival and apoptosis pathways in sheep/ruminants.

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“…Tissues from Day 4 (n ¼ 4) and Day 10 (n ¼ 4) bovine CL were homogenized individually and total RNA was isolated using Tri-reagent according to the manufacturer's instructions (Molecular Research Center) and as previously described [36]. Briefly, frozen CL tissue immersed in liquid nitrogen was pulverized mechanically using an RNase-free mortar and pestle.…”

Section: Isolation Of Total Rna and Cdna Synthesismentioning

confidence: 99%

“…Tissue processing was performed as previously described [36] with minor modifications. Briefly, tissue samples were fixed for at least 48 h in Bouin fixative.…”

Section: Immunohistochemistrymentioning

confidence: 99%

Mechanisms of Intracellular Calcium Homeostasis in Developing and Mature Bovine Corpora Lutea1

Wright

Bowdridge

McDermott

et al. 2014

Biology of Reproduction

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Although calcium (Ca(2+)) is accepted as an intracellular mediator of prostaglandin F2 alpha (PGF2alpha) actions on luteal cells, studies defining mechanisms of Ca(2+) homeostasis in bovine corpora lutea (CL) are lacking. The increase in intracellular Ca(2+) concentration ([Ca(2+)]i) induced by PGF2alpha in steroidogenic cells from mature CL is greater than in those isolated from developing CL. Our hypothesis is that differences in signal transduction associated with developing and mature CL contribute to the increased efficacy of PGF2alpha to induce a Ca(2+) signal capable of inducing regression in mature CL. To test this hypothesis, major genes participating in Ca(2+) homeostasis in the bovine CL were identified, and expression of mRNA, protein, or activity, in the case of phospholipase Cbeta (PLCbeta), in developing and mature bovine CL was compared. In addition, we examined the contribution of external and internal Ca(2+) to the PGF2alpha stimulated rise in [Ca(2+)]i in LLCs isolated from developing and mature bovine CL. Three differences were identified in mechanisms of calcium homeostasis between developing and mature CL, which could account for the lesser increase in [Ca(2+)]i in response to PGF2alpha in developing than in mature CL. First, there were lower concentrations of inositol 1,4,5-trisphosphate (IP3) after similar PGF2alpha challenge, indicating reduced phospholipase C beta (PLCbeta) activity, in developing than mature CL. Second, there was an increased expression of sorcin (SRI) in developing than in mature CL. This cytoplasmic Ca(2+) binding protein modulates the endoplasmic reticulum (ER) Ca(2+) release channel, ryanodine receptor (RyR), to be in the closed configuration. Third, there was greater expression of ATP2A2 or SERCA, which causes calcium reuptake into the ER, in developing than in mature CL. Developmental differences in expression detected in whole CL were confirmed by Western blots using protein samples from steroidogenic cells isolated from developing and mature CL. Localization of these genes in steroidogenic luteal cells was confirmed by immunohistochemistry. Therefore, it is concluded that the cellular mechanisms that allow PGF2alpha to induce a calcium signal of greater magnitude in mature than in developing CL involve 1) greater PLCbeta activity with enhanced generation of IP3, 2) an enhanced Ca(2+) release from the ER via unrestrained RYR2 due to a decrease in SRI expression, and 3) a reduction in calcium reuptake to the ER due to lower expression of ATP2A2. Accordingly, the increase in [Ca(2+)]i induced by PGF2alpha in mature large steroidogenic cells had less dependency from extracellular calcium than in those isolated from immature CL.

show abstract

Differential Gene Expression in the Bovine Corpus Luteum During Transition from Early Phase to Midphase and Its Potential Role in Acquisition of Luteolytic Sensitivity to Prostaglandin F2 Alpha1

Cited by 41 publications

References 55 publications

Patterns of Gene Expression in the Bovine Corpus Luteum Following Repeated Intrauterine Infusions of Low Doses of Prostaglandin F2alpha1

Patterns of Gene Expression in the Bovine Corpus Luteum Following Repeated Intrauterine Infusions of Low Doses of Prostaglandin F2alpha1

Early pregnancy modulates survival and apoptosis pathways in the corpus luteum in sheep

Mechanisms of Intracellular Calcium Homeostasis in Developing and Mature Bovine Corpora Lutea1

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