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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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Related in: MedlinePlus

Immunochemical detection of the cellular distribution of iNOS expression in the mouse Fallopian tube at different stages of the estrous cycle (A–D) with diaminobenzidine (DAB). All slides were counterstained with hematoxylin. Enlarged views of one part are shown in the lower right corner in each picture. Epi: epithelial cells; Sm: smooth muscle cells. (A) diestrus; (B) proestrus; (C) estrus; (D) metestrus. Scale bar = 100 µm in the main picture and 50 µm in the enlarged images.
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ijms-16-00049-f003: Immunochemical detection of the cellular distribution of iNOS expression in the mouse Fallopian tube at different stages of the estrous cycle (A–D) with diaminobenzidine (DAB). All slides were counterstained with hematoxylin. Enlarged views of one part are shown in the lower right corner in each picture. Epi: epithelial cells; Sm: smooth muscle cells. (A) diestrus; (B) proestrus; (C) estrus; (D) metestrus. Scale bar = 100 µm in the main picture and 50 µm in the enlarged images.

Mentions: Immunochemistry showed that both iNOS (Figure 3A–D) and eNOS (Figure 4A–D) were expressed in the mouse Fallopian tube at all four stages of the estrous cycle, and there were clear differences in immunoreactivity density and quantity at the different stages of the estrous cycle. The intensity of iNOS was highest in the estrus stage (Figure 3C) while eNOS was most abundant in the proestrus stage (Figure 4B). Both of them showed the weakest signal in the metestrus stage (Figure 3D and Figure 4D). iNOS and eNOS protein were mostly located in the epithelium of mouse Fallopian tubes and were found in both the cytoplasm and in the nucleus. However, they were not detectable in every epithelial cell. iNOS was expressed in about half of the epithelial cells, and eNOS was only detectable in about 1/3 of the epithelial cells. Similar to the mRNA results, there were no nNOS-positive cells in any of the Fallopian tubes. As a negative control, normal goat serum replaced the primary antibody (Figure 4F).


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)

Immunochemical detection of the cellular distribution of iNOS expression in the mouse Fallopian tube at different stages of the estrous cycle (A–D) with diaminobenzidine (DAB). All slides were counterstained with hematoxylin. Enlarged views of one part are shown in the lower right corner in each picture. Epi: epithelial cells; Sm: smooth muscle cells. (A) diestrus; (B) proestrus; (C) estrus; (D) metestrus. Scale bar = 100 µm in the main picture and 50 µm in the enlarged images.
© Copyright Policy
Related In: Results  -  Collection

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

ijms-16-00049-f003: Immunochemical detection of the cellular distribution of iNOS expression in the mouse Fallopian tube at different stages of the estrous cycle (A–D) with diaminobenzidine (DAB). All slides were counterstained with hematoxylin. Enlarged views of one part are shown in the lower right corner in each picture. Epi: epithelial cells; Sm: smooth muscle cells. (A) diestrus; (B) proestrus; (C) estrus; (D) metestrus. Scale bar = 100 µm in the main picture and 50 µm in the enlarged images.
Mentions: Immunochemistry showed that both iNOS (Figure 3A–D) and eNOS (Figure 4A–D) were expressed in the mouse Fallopian tube at all four stages of the estrous cycle, and there were clear differences in immunoreactivity density and quantity at the different stages of the estrous cycle. The intensity of iNOS was highest in the estrus stage (Figure 3C) while eNOS was most abundant in the proestrus stage (Figure 4B). Both of them showed the weakest signal in the metestrus stage (Figure 3D and Figure 4D). iNOS and eNOS protein were mostly located in the epithelium of mouse Fallopian tubes and were found in both the cytoplasm and in the nucleus. However, they were not detectable in every epithelial cell. iNOS was expressed in about half of the epithelial cells, and eNOS was only detectable in about 1/3 of the epithelial cells. Similar to the mRNA results, there were no nNOS-positive cells in any of the Fallopian tubes. As a negative control, normal goat serum replaced the primary antibody (Figure 4F).

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