Expression of endothelial and inducible nitric oxide synthases is modulated in the endometrium of cyclic and early pregnant mares

Welter, Harald; Bollwein, Heinrich; Weber, Frank; Rohr, Sven Olaf; Einspanier, Ralf

doi:10.1071/rd03103

Cited by 22 publications

(23 citation statements)

References 44 publications

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“…To expose antigenic sites, dewaxed sections were heated four times to 95 C in a 600 W microwave oven in citrate buffer for 5 min. Endogenous peroxidase activity was then eliminated by incubation with 0·5% (v/v) H 2 O 2 solution in absolute methanol for 15 min at 20 C. Non-specific protein binding was eliminated by incubation with 10% normal goat serum in PBS for 1 h at 20 C. Sections were then incubated with either polyclonal rabbit antibody against iNOS (Upstate, Lake Placid, NY, USA) (Sherman et al 1999), which is known to cross-react with bovine iNOS, or monoclonal rabbit antibody against eNOS (Alpha Diagnostic, San Antonio, TX, USA), from which the human peptide sequence is 100% conserved in the bovine (Marsden et al 1992, Welter et al 2004. Each was used at a dilution of 1:200.…”

Section: Immunohistochemistrymentioning

confidence: 99%

Region-specific expression of nitric oxide synthases in the bovine oviduct during the oestrous cycle and in vitro

Ulbrich¹,

Rehfeld²,

Bauersachs³

et al. 2006

Journal of Endocrinology

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Nitric oxide synthases (NOS) account for the endogenous production of nitric oxide (NO), a small and permeable bioreactive molecule. NO is known to act as a paracrine mediator during various processes associated with female reproduction. In the present study, the mRNA expression of the endothelial (eNOS) and inducible (iNOS) NO synthases were examined in bovine oviduct epithelial cells (BOEC) during the oestrous cycle. In addition, eNOS and iNOS mRNA and protein were localised by in situ hybridisation and immunocytochemistry respectively. Furthermore, the effects of exogenously applied oestradiol-17 and progesterone on NOS mRNA regulation were studied in a suspension culture of BOEC. The eNOS mRNA abundance was low around ovulation (day 0) and increased significantly until pro-oestrus (day 18) in the ampulla. Immunoreactive protein of eNOS was detected predominantly in endothelial cells as well as in secretory oviduct epithelial cells at pro-oestrus. The iNOS mRNA concentration was significantly reduced in the isthmus at pro-oestrus (day 18) and oestrus (day 0) compared with persistently high levels in the ampulla. By in situ hybridisation, specific iNOS transcripts were additionally demonstrated in the oviduct epithelium. Immunoreactive iNOS protein was localised in secretory epithelial cells as well as in the lamina muscularis. The in vitro stimulation showed that both NOS were stimulated by progesterone, but not by oestradiol-17 . The region-specific modulated expression of eNOS and iNOS provides evidence for an involvement of endogenously produced NO in the regulation of oviductal functions.

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Section: Immunohistochemistrymentioning

confidence: 99%

Region-specific expression of nitric oxide synthases in the bovine oviduct during the oestrous cycle and in vitro

Ulbrich¹,

Rehfeld²,

Bauersachs³

et al. 2006

Journal of Endocrinology

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“…Endothelial nitric oxide synthases (eNOS) and inducible nitric oxide synthases (iNOS), which are responsible for the production of NO, have been detected in the endometrium in humans, sheep, rodents [18,19], cows [20] and horses [21][22][23]. Since there were changes in the expression of eNOS during the estrous cycle in mares, it was speculated that the NOS system was involved in the regulation of their endometrial cyclicity [21,22].The objective of the present study was to investigate the involvement of steroid hormones and nitric oxide synthases in the regulation of uterine blood flow in cycling mares. …”

mentioning

confidence: 99%

“…Contamination by genomic DNA was minimized by an additional DNase digestion according to the manufacturer's protocol (RQ1, Promega, Madison, WI, USA). Total RNA was purified using NucleoSpin ® RNA II (Macherey-Nagel, Düren, Germany) and quantified spectroscopically (260 nm) using a Biophotometer (Eppendorf, Hamburg, Germany).A two-step quantitative real-time RT-PCR was undertaken, as previously described [21]. In brief, total RNA was reverse transcribed to cDNA, as previously described [27].…”

mentioning

confidence: 99%

Relationships Between Uterine Blood Flow, Peripheral Sex Steroids, Expression of Endometrial Estrogen Receptors and Nitric Oxide Synthases During the Estrous Cycle in Mares

Honnens¹,

Weisser²,

Welter

et al. 2011

J. Reprod. Dev.

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Abstract. The objective of this study was to investigate the relationships between uterine perfusion and estrogen, progesterone and the uterine nitric oxide synthase (NOS) system in five trotter mares during the estrous cycle. Color Doppler sonography for measurement of uterine blood flow and collection of blood for determination of plasma estrogen and progesterone concentrations were performed on days 0 (= ovulation), 1, 5, 11 and 15 and daily during estrus (days -1 to -4) of one estrous cycle; endometrial biopsy collection for mRNA expression analysis of NOS and estrogen receptors was performed on days 0, 1, 5, 11, 15 and -3. Blood flow in each uterine artery was assessed by calculating the mean time-averaged maximum velocity (TAMV) and the pulsatility index (PI). Plasma concentrations of estrogen and progesterone were determined using specific enzyme immunoassays. The mRNA expressions of endothelial NOS (eNOS), inducible NOS (iNOS) as well as estrogen receptors α (ERα) and β (ERβ) were quantified using reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. The TAMV and PI had a biphasic pattern during the estrous cycle (P<0.05), with maximum and minimum, respectively, values on days 5 and -4. Estrogen receptor mRNA concentrations increased significantly during days 15 (ERα) and -3 (ERβ). Transcript expression of eNOS, but not iNOS, had a biphasic pattern during the cycle (P<0.05) with maximum levels on days 5 and -3 and correlated positively with TAMV (r=0.81, P=0.05). We infer that the uterine NOS system, especially eNOS, plays an important role in the regulation of uterine blood flow during the estrous cycle in mares. Key words: Endometrial messenger RNA expression, Estrous cycle, Nitric oxide synthases, Steroid hormones, Uterine perfusion (J. Reprod. Dev. 57: [43][44][45][46][47][48] 2011) haracteristic uterine blood flow patterns have been reported during the estrous cycle in mares based on color Doppler sonography [1,2]. Uterine perfusion had a wave-shaped profile throughout the cycle, showing maximal blood flow during estrus and the early luteal phase [1][2][3][4]. Variations in uterine blood flow in cycling mares have been attributed to changes in circulating estrogen concentrations. However, due to a weak correlation between the resistance to uterine blood flow and plasma estrogen concentrations, other factors may also regulate uterine blood flow [2].Changes in uterine blood flow during the estrous cycle in many species are mediated mainly by estrogens [5]; although flow was enhanced by high plasma estradiol concentrations [6,7] and decreased during the luteal phase [8,9], the mechanisms involved have not been elucidated. Estrogens appear to act via two pathways: the genomic and non-genomic pathways [10]. Briefly, in the classical genomic pathway, estrogens bind to their nuclear receptors, subsequently regulating the transcription of target genes. For example, administration of estradiol increased estrogen receptors within 48 h in cyclic ewes [11]. Furthermore, a non-genomic pathway wit...

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“…Moreover, Nitric oxide appears, under normal physiologic conditions, to be the most important regulator of blood flow (Klubunde, 2015). Welter et al, (2004) reported that different isoforms of NO synthase were present in the equine endometrium of early pregnant mares. NO is considered a vital mediator concerned in maintenance of pregnancy (Yalampalli et al, 1994).…”

Section: Discussionmentioning

confidence: 99%

Preovulatory Dominant Follicular Size and Serum Nitric Oxide Can Predicate the Conception Rate after Foaling Heat in Straight Egyptian Arabian Mares

Elmetwally¹,

El‐Adl²,

Marghani³

et al. 2017

AJVS

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Key words:Arabian horse, dominant follicle, nitric oxide, alkaline phosphatase Postpartum heat is very important for ability of mare's to conceive. The present study aimed to evaluate ovarian changes and its relation to oxidative stress during early postpartum period in Arabian horses. Further, the predication of conception according to the relationship between sonographic and biochemical changes analyses. Ultrasound imaging of follicular changes and oxidative analyses changes were investigated at day 7-9 postpartum whereas Receiver Operating Characteristic (ROC) curves were used to determine the ability of oxidative stress and ultrasound imaging to predict the conception rate. ROC analyses indicated that nitric oxide predicate the conception rate with sensitivity (79% P<0.003) and dominant follicle size (80-100% P<0.0001). Alkaline phosphatase, catalase, glutathione, glutathione S transferase and total antioxidant capacity (P>0.05) between pregnant and nonpregnant mares. In conclusion, nitric oxide and dominant follicle diameter plays a crucial role in enhancement the conception rate during foaling heat.

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Expression of endothelial and inducible nitric oxide synthases is modulated in the endometrium of cyclic and early pregnant mares

Cited by 22 publications

References 44 publications

Region-specific expression of nitric oxide synthases in the bovine oviduct during the oestrous cycle and in vitro

Region-specific expression of nitric oxide synthases in the bovine oviduct during the oestrous cycle and in vitro

Relationships Between Uterine Blood Flow, Peripheral Sex Steroids, Expression of Endometrial Estrogen Receptors and Nitric Oxide Synthases During the Estrous Cycle in Mares

Preovulatory Dominant Follicular Size and Serum Nitric Oxide Can Predicate the Conception Rate after Foaling Heat in Straight Egyptian Arabian Mares

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