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The regulation of nitric oxide synthase isoform expression in mouse and human fallopian tubes: potential insights for ectopic pregnancy.

Hu J, Ma S, Zou S, Li X, Cui P, Weijdegård B, Wu G, Shao R, Billig H, Feng Y - Int J Mol Sci (2014)

Bottom Line: Nitric oxide (NO) is highly unstable and has a half-life of seconds in buffer solutions.It is synthesized by NO-synthase (NOS), which has been found to exist in the following three isoforms: neuro nitric oxide synthase (nNOS), inducible nitric oxide synthase (iNOS), and endothelial nitric oxide synthase (eNOS).Elevated iNOS activity might influence ovulation, cilia beats, contractility, and embryo transportation in such a manner as to increase the risk of ectopic pregnancy.

View Article: PubMed Central - PubMed

Affiliation: Department of Integrative Medicine and Neurobiology, State Key Lab of Medical Neurobiology, Shanghai Medical College and Institute of Acupuncture Research (WHO Collaborating Center for Traditional Medicine), Institute of Brain Science, Fudan University, Shanghai 200032, China. 11211250005@fudan.edu.cn.

ABSTRACT
Nitric oxide (NO) is highly unstable and has a half-life of seconds in buffer solutions. It is synthesized by NO-synthase (NOS), which has been found to exist in the following three isoforms: neuro nitric oxide synthase (nNOS), inducible nitric oxide synthase (iNOS), and endothelial nitric oxide synthase (eNOS). NOS activity is localized in the reproductive tracts of many species, although direct evidence for NOS isoforms in the Fallopian tubes of mice is still lacking. In the present study, we investigated the expression and regulation of NOS isoforms in the mouse and human Fallopian tubes during the estrous and menstrual cycles, respectively. We also measured isoform expression in humans with ectopic pregnancy and in mice treated with lipopolysaccharide (LPS). Our results confirmed the presence of different NOS isoforms in the mouse and human Fallopian tubes during different stages of the estrous and menstrual cycles and showed that iNOS expression increased in the Fallopian tubes of women with ectopic pregnancy and in LPS-treated mice. Elevated iNOS activity might influence ovulation, cilia beats, contractility, and embryo transportation in such a manner as to increase the risk of ectopic pregnancy. This study has provided morphological and molecular evidence that NOS isoforms are present and active in the human and mouse Fallopian tubes and suggests that iNOS might play an important role in both the reproductive cycle and infection-induced ectopic pregnancies.

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The mRNA and protein levels of the NOS isoforms in human Fallopian tubes. (A) Fold changes in nNOS, iNOS, and eNOS mRNA levels at different stages of the menstrual cycle. * p < 0.05, ** p < 0.01; (B,C) iNOS expression in late ovulatory phase and middle secretory phase. Slides were subsequently counterstained with DAPI to visualize the cell nuclei. Sections were immunolabeled for iNOS (red) and DAPI (blue). iNOS was found both in the epithelial cells and muscle cells of the human Fallopian tube and was located in the cytoplasm of the epithelial cells. The amount and density of iNOS expression in the late ovulatory phase was higher than in the mid-secretary phase; and (D) Negative control using goat serum instead of primary antibody resulted in no immunostaining. Epi, epithelial cells; Sm, smooth muscle cells. Scale bar = 100 µm.
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ijms-16-00049-f006: The mRNA and protein levels of the NOS isoforms in human Fallopian tubes. (A) Fold changes in nNOS, iNOS, and eNOS mRNA levels at different stages of the menstrual cycle. * p < 0.05, ** p < 0.01; (B,C) iNOS expression in late ovulatory phase and middle secretory phase. Slides were subsequently counterstained with DAPI to visualize the cell nuclei. Sections were immunolabeled for iNOS (red) and DAPI (blue). iNOS was found both in the epithelial cells and muscle cells of the human Fallopian tube and was located in the cytoplasm of the epithelial cells. The amount and density of iNOS expression in the late ovulatory phase was higher than in the mid-secretary phase; and (D) Negative control using goat serum instead of primary antibody resulted in no immunostaining. Epi, epithelial cells; Sm, smooth muscle cells. Scale bar = 100 µm.

Mentions: mRNAs of all three NOS isoforms were expressed in the human Fallopian tube (Figure 6A). The nNOS mRNA level was increased in the mid-secretory stage compared with the other stages (p < 0.05). The iNOS mRNA level was increased in the late ovulatory phase compared with the early ovulatory phase (p < 0.01) and post-ovulatory phase (p < 0.05), and the iNOS mRNA level in the mid-secretory phase was higher than in the early ovulatory phase (p < 0.05). However, there were no significant changes in eNOS mRNA levels at the different stages. Immunochemical analysis showed that iNOS was found in both the epithelial cells and muscle cells of the human Fallopian tube. It was located only in the cytoplasm of the epithelial cells but in both the cytoplasm and nucleus in muscle cells. Both the quantity and the density of iNOS expression in the late ovulatory phase was much higher than in the mid-secretory phase (Figure 6B,C).


The regulation of nitric oxide synthase isoform expression in mouse and human fallopian tubes: potential insights for ectopic pregnancy.

Hu J, Ma S, Zou S, Li X, Cui P, Weijdegård B, Wu G, Shao R, Billig H, Feng Y - Int J Mol Sci (2014)

The mRNA and protein levels of the NOS isoforms in human Fallopian tubes. (A) Fold changes in nNOS, iNOS, and eNOS mRNA levels at different stages of the menstrual cycle. * p < 0.05, ** p < 0.01; (B,C) iNOS expression in late ovulatory phase and middle secretory phase. Slides were subsequently counterstained with DAPI to visualize the cell nuclei. Sections were immunolabeled for iNOS (red) and DAPI (blue). iNOS was found both in the epithelial cells and muscle cells of the human Fallopian tube and was located in the cytoplasm of the epithelial cells. The amount and density of iNOS expression in the late ovulatory phase was higher than in the mid-secretary phase; and (D) Negative control using goat serum instead of primary antibody resulted in no immunostaining. Epi, epithelial cells; Sm, smooth muscle cells. Scale bar = 100 µm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4307235&req=5

ijms-16-00049-f006: The mRNA and protein levels of the NOS isoforms in human Fallopian tubes. (A) Fold changes in nNOS, iNOS, and eNOS mRNA levels at different stages of the menstrual cycle. * p < 0.05, ** p < 0.01; (B,C) iNOS expression in late ovulatory phase and middle secretory phase. Slides were subsequently counterstained with DAPI to visualize the cell nuclei. Sections were immunolabeled for iNOS (red) and DAPI (blue). iNOS was found both in the epithelial cells and muscle cells of the human Fallopian tube and was located in the cytoplasm of the epithelial cells. The amount and density of iNOS expression in the late ovulatory phase was higher than in the mid-secretary phase; and (D) Negative control using goat serum instead of primary antibody resulted in no immunostaining. Epi, epithelial cells; Sm, smooth muscle cells. Scale bar = 100 µm.
Mentions: mRNAs of all three NOS isoforms were expressed in the human Fallopian tube (Figure 6A). The nNOS mRNA level was increased in the mid-secretory stage compared with the other stages (p < 0.05). The iNOS mRNA level was increased in the late ovulatory phase compared with the early ovulatory phase (p < 0.01) and post-ovulatory phase (p < 0.05), and the iNOS mRNA level in the mid-secretory phase was higher than in the early ovulatory phase (p < 0.05). However, there were no significant changes in eNOS mRNA levels at the different stages. Immunochemical analysis showed that iNOS was found in both the epithelial cells and muscle cells of the human Fallopian tube. It was located only in the cytoplasm of the epithelial cells but in both the cytoplasm and nucleus in muscle cells. Both the quantity and the density of iNOS expression in the late ovulatory phase was much higher than in the mid-secretory phase (Figure 6B,C).

Bottom Line: Nitric oxide (NO) is highly unstable and has a half-life of seconds in buffer solutions.It is synthesized by NO-synthase (NOS), which has been found to exist in the following three isoforms: neuro nitric oxide synthase (nNOS), inducible nitric oxide synthase (iNOS), and endothelial nitric oxide synthase (eNOS).Elevated iNOS activity might influence ovulation, cilia beats, contractility, and embryo transportation in such a manner as to increase the risk of ectopic pregnancy.

View Article: PubMed Central - PubMed

Affiliation: Department of Integrative Medicine and Neurobiology, State Key Lab of Medical Neurobiology, Shanghai Medical College and Institute of Acupuncture Research (WHO Collaborating Center for Traditional Medicine), Institute of Brain Science, Fudan University, Shanghai 200032, China. 11211250005@fudan.edu.cn.

ABSTRACT
Nitric oxide (NO) is highly unstable and has a half-life of seconds in buffer solutions. It is synthesized by NO-synthase (NOS), which has been found to exist in the following three isoforms: neuro nitric oxide synthase (nNOS), inducible nitric oxide synthase (iNOS), and endothelial nitric oxide synthase (eNOS). NOS activity is localized in the reproductive tracts of many species, although direct evidence for NOS isoforms in the Fallopian tubes of mice is still lacking. In the present study, we investigated the expression and regulation of NOS isoforms in the mouse and human Fallopian tubes during the estrous and menstrual cycles, respectively. We also measured isoform expression in humans with ectopic pregnancy and in mice treated with lipopolysaccharide (LPS). Our results confirmed the presence of different NOS isoforms in the mouse and human Fallopian tubes during different stages of the estrous and menstrual cycles and showed that iNOS expression increased in the Fallopian tubes of women with ectopic pregnancy and in LPS-treated mice. Elevated iNOS activity might influence ovulation, cilia beats, contractility, and embryo transportation in such a manner as to increase the risk of ectopic pregnancy. This study has provided morphological and molecular evidence that NOS isoforms are present and active in the human and mouse Fallopian tubes and suggests that iNOS might play an important role in both the reproductive cycle and infection-induced ectopic pregnancies.

Show MeSH
Related in: MedlinePlus