Expression of mRNAs for interleukin-4, interleukin-6 and their receptors in porcine corpus luteum during the estrous cycle

Sakumoto, Ryosuke; Komatsu, Tokushi; Kasuya, Etsuko; Okuda, Kiyoshi

doi:10.1016/j.domaniend.2005.11.001

Cited by 67 publications

(50 citation statements)

References 27 publications

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“…Using iScript RT Supermix for RT-qPCR (170-8841; Bio-Rad Laboratories), 1 μg of each total RNA was reverse transcribed. Quantifications of mRNA expressions were determined by Quantitative RT-PCR using MyiQ (Bio-Rad Laboratories) and SooAdvanced SYBR Green Supermix (1725261B10; Bio-Rad Laboratories) starting with 4 ng reverse-transcribed total RNA as described previously (Sakumoto et al 2006). All primers were designed to amplify specific products for NOS2 (forward: 5′-TAC CCT CAG TTC TGC GCT TT-3′; reverse: 5′-GGG ATC TCA ATG TGG TGC TT-3′), NOS3 (forward: 5′-AGG CTC TCA CCT TCT TCC TG-3′; reverse: 5′-AAC CAC TTC CAC TCC TCG TA-3′), ESR1 (forward: 5′-CAG GCA CAT GAG CAA CAA AG-3′; reverse: 5′-TCC AGC AGC AGG TCG TAG AG-3′), and ESR2 (forward: 5′-CTG AAG CAT GAA CTC CAG CAC-3′; reverse: 5′-CAG GAA GGA CCA CAT AGC AGA-3′).…”

Section: Total Rna Extraction and Quantitative Rt-pcrmentioning

confidence: 99%

Regulation of bovine oviductal NO synthesis by follicular steroids and prostaglandins

et al. 2016

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Nitric oxide (NO) is a regulator of sperm motility, oocyte/embryo survival, and waves of contraction/relaxation in mammalian oviducts. As follicles control oviductal functions by two routes at least, (1) a systemic way via blood vessels before ovulation, (2) a direct way by entering of follicular fluid through fimbria at ovulation, we hypothesized that NO synthesis in the bovine oviduct is regulated by follicular steroids and prostaglandins (PGs). Quantification of mRNA expressions in the ampullary tissues showed that inducible NO synthase (NOS2) mRNA expression was highest on the day of ovulation (day 0). By contrast, NOS2 mRNA expression in the isthmus was highest on days 5-6 and lowest on days 19-21. Endothelial NOS (NOS3) mRNA expressions in either the ampulla or the isthmus did not change during the estrous cycle. PGE2 and PGF2α increased NOS2 mRNA expressions in cultured ampullary oviductal epithelial cells after 1-h incubation. These increases were suppressed by an antagonist of E-prostanoid receptor type 2, one of the PGE2 receptor. Estradiol-17β decreased the expression of NOS2 mRNA expression in cultured isthmic epithelial cells 24 h after treatment. This effect was suppressed by an antagonist of estrogen receptor α (ESR1). Expression of ESR1 was highest on days 19-21 in the isthmic tissues. The overall findings indicate region-specific difference of NO synthesis in the oviduct. PGs flowed from ruptured follicle may up-regulate NO synthesis in the oviductal epithelium, whereas circulating E2 seems to inhibit NO synthesis via ESR1 in the isthmus at the follicular stage.Reproduction (2016) 151 577-587

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Section: Total Rna Extraction and Quantitative Rt-pcrmentioning

confidence: 99%

Regulation of bovine oviductal NO synthesis by follicular steroids and prostaglandins

et al. 2016

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“…PCR: Total RNA isolation and subsequent RT-and realtime PCR steps were carried out as previously described [20]. The following primers encoding the porcine sequences were designed and synthesized: 5'-GCCGCCTA-CAACTATTTCCA-3', forward, and 5'-CCAGCCAATG-CAGATAAACA-3 (132 bp), reverse, for CRH-R1…”

Section: Collection Of Porcine Corpora Luteamentioning

confidence: 99%

“…Luteal cell culture: Porcine luteal cells were prepared and cultured as described previously with some modifications [18,20]. ) for up to 48 hr in a humidified atmosphere of 5% CO 2 in air at 37.5C.…”

Section: Collection Of Porcine Corpora Luteamentioning

confidence: 99%

“…Each pig was euthanized by electric shock as is usually done in local abattoirs. The luteal stage was classified as early (days 3-5 after ovulation), mid (days 8-11), late (days [14][15][16] or regressed (days [19][20][21] by macroscopic observation of the ovaries [1] and confirmed by measuring P concentrations in CLs (n=4/each stage). For gene expression studies, the CLs were immediately separated from the ovaries, frozen rapidly in liquid nitrogen and stored at -80C until use.…”

Section: Collection Of Porcine Corpora Luteamentioning

confidence: 99%

“…Ovaries with mid-stage CLs were collected, submerged in ice-cold physiological saline and transported to the laboratory. Ten to sixteen CLs were pooled from each pig (n=4).PCR: Total RNA isolation and subsequent RT-and realtime PCR steps were carried out as previously described [20]. The following primers encoding the porcine sequences were designed and synthesized: 5'-GCCGCCTA-CAACTATTTCCA-3', forward, and 5'-CCAGCCAATG-CAGATAAACA-3 (132 bp), reverse, for CRH-R1…”

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Possible Actions of Corticotropin-Releasing Hormone in Regulating Porcine Corpus Luteum Function

Sakumoto

Ito

Komatsu

2010

The Journal of Veterinary Medical Science

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ABSTRACT. The objective of the present study was to determine whether corticotropin-releasing hormone (CRH) influences porcine corpus luteum function. The gene expressions of CRH receptors (CRH-R) were determined in the CLs of Chinese Meishan pigs during the estrous cycle. The effects of CRH on progesterone (P), estradiol-17 (E) and prostaglandin (PG) F 2 secretion by cultured luteal cells were also investigated. Messenger RNAs of the CRH-R were clearly expressed in the CL throughout the estrous cycle, and the mRNA level was higher at the regressed stage than at the other stages (P<0.05). When the cultured luteal cells obtained from the mid-luteal stage CL (days 8-11) were exposed to CRH (50-5000 ng/ml), P secretion by the cells was significantly reduced at the highest dose (P<0.05), whereas CRH had no effect on E and PGF 2 secretion by the cells. The overall results suggest that CRH inhibits local P production from luteal cells via its specific CRH-R, implying that CRH plays some roles in regulating porcine CL function during the estrous cycle, especially in the period of luteolysis.KEY WORDS: corpus luteum, corticotropin-releasing hormone, receptor, swine.J. Vet. Med. Sci. 72(9): 1173-1177, 2010 Corticotropin-releasing hormone (CRH) is known as a stress-induced peptide that is secreted from the paraventricular nucleus of the hypothalamus [22]. Hypothalamic CRH induces adrenocorticotropic hormone (ACTH) secretion by the pituitary via its specific receptors (CRH-R), and the ACTH stimulates glucocorticoid (GC) production from the adrenal gland; this is considered to be the hypothalamicpituitary-adrenal (HPA) axis [22]. GC is produced in the adrenal cortex during periods of stress and is widely recognized as an immunosuppressive and anti-inflammatory agent [2]. It has been shown to inhibit expression of cytokines, adhesion molecules and enzymes involved in the inflammatory process [4]. Our recent study demonstrated the presence of GC and its converting enzymes in the porcine corpus luteum (CL) and that GC inhibits steroid production from cultured luteal cells [21], suggesting that GC may regulate CL function locally in pigs.Recent studies indicate that a CRH/CRH-R system also exists outside the central nervous system. CRH and CRH-R have been identified in the human placenta [8,17], endometrium [11,12] and ovaries [3,14]. In monkeys, the genes and proteins for CRH and CRH-R are expressed in the CL during the menstrual cycle [24]. Moreover, CRH inhibits progesterone (P) and estradiol-17 (E) secretion by cultured human granulosa-lutein cells [7]. Since the E production from the CL in pigs is a specific feature that is similar to those of the CLs in humans and monkeys [23], the previous studies lead us to hypothesize that CRH directly regulates CL function in pigs as well as in primates.In the present study, therefore, we measured expression of CRH-R in the porcine CL from different stages of the estrous cycle using reverse transcription (RT) polymerase chain reaction (PCR) and real-time PCR analyses. The possible...

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Hyperthermia alters interleukin‐6 production in response to lipopolysaccharide via endoplasmic reticulum stress in bovine endometrial cells

Sakai

Inoue

Tanaka

et al. 2021

Journal Cellular Physiology

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In the postpartum period, cows experience the uterine bacterial infection and develop the endometritis. To eliminate bacteria and recover from endometritis, endometrial epithelial and stromal cells secrete the cytokine and chemokine, such as interleukin 6 (IL-6), IL-8, and monocyte chemotactic protein 1 (MCP1), to recruit immune cells. Moreover, the symptom of endometritis is prolonged in summer and we have recently indicated that hyperthermia suppresses and enhances the IL-6 production in response to lipopolysaccharide (LPS) challenge in endometrial epithelial and stromal cells, respectively. However, the mechanisms for the opposite reaction of IL-6 secretion in response to LPS challenge in both types of endometrial cells under hyperthermia conditions were still unclear. To reveal these mechanisms, both types of endometrial cells were cultured with LPS under the control (38.5°C) or hyperthermia (40.5°C) conditions and comprehensively analyzed differential gene expressions of them by RNA-seq. In addition, based on these results, we examined the effect of endoplasmic reticulum (ER) stress on the IL-6 production in both types of endometrial cells cultured with LPS under hyperthermia conditions. In comprehensive analysis, hyperthermia induced the ER stress in the endometrial stromal cells but not in the endometrial epithelial cells. Actually, we confirmed that hyperthermia increased the gene expression of BiP, ATF4, and sXBP1 and protein expression of BiP and phosphorylated inositol requiring 1, ER stress marker, in the endometrial stromal cells but not in the endometrial epithelial cells. Moreover, in the endometrial stromal cells exposed to LPS, activation and inhibition of ER stress enhanced the IL-6 production under control conditions and suppressed it under hyperthermia conditions, respectively. In this study, we could uncover the one of causes for the disruption of IL-6 production in response to LPS challenge in the endometrial cells under hyperthermia conditions. This finding might be a clue for the improvement of the symptom of endometritis in cows during summer.

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Expression of mRNAs for interleukin-4, interleukin-6 and their receptors in porcine corpus luteum during the estrous cycle

Cited by 67 publications

References 27 publications

Regulation of bovine oviductal NO synthesis by follicular steroids and prostaglandins

Regulation of bovine oviductal NO synthesis by follicular steroids and prostaglandins

Possible Actions of Corticotropin-Releasing Hormone in Regulating Porcine Corpus Luteum Function

Hyperthermia alters interleukin‐6 production in response to lipopolysaccharide via endoplasmic reticulum stress in bovine endometrial cells

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