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Expression and Tissue Localization of Soluble Guanylyl Cyclase in the Human Placenta Using Novel Antibodies Directed against theα2Subunit
- Source :
- The Journal of Clinical Endocrinology & Metabolism. 86:909-912
- Publication Year :
- 2001
- Publisher :
- The Endocrine Society, 2001.
-
Abstract
- The cytoplasmic or soluble forms of guanylyl cyclase (sGC) are heme-containing heterodimeric enzymes that are regulated by nitric oxide (NO) and carbon monoxide (CO). These gaseous messenger molecules are produced in the human placenta and are potential regulators of vasodilation and trophoblast invasion. The alpha(2)-subunit of sGC has only recently been shown to naturally occur in placental extracts. In the present study, two novel antibodies directed against different epitopes of the alpha(2) subunit, were generated. Western Blot analysis confirmed the presence of a 82 kDa protein, identical with alpha(2) protein overexpressed in Sf9 cells. According to RNase protection analysis the alternatively spliced alpha(2i) variant was absent from human placenta. Immunohistochemical analysis showed the presence of alpha(2) protein in syncytiotrophoblast and villous and umbilical blood vessels, which are known sites of NO production. Strong expression was observed in the extravillous (intermediate) trophoblast, where the expression of CO-generating hemeoxygenases has recently been documented. Localization of alpha(2) subunit expression suggests a role for sGC in mediating the actions of both NO and CO. The novel antibodies characterized in the present study will be powerful tools to further elucidate the role of the NO/CO/cGMP signaling pathways in pathologic states such as preeclampsia and intrauterine growth retardation.
- Subjects :
- medicine.medical_specialty
Placenta
Endocrinology, Diabetes and Metabolism
Protein subunit
Blotting, Western
Molecular Sequence Data
Clinical Biochemistry
Spodoptera
Biology
Transfection
Biochemistry
Epitope
Cell Line
Epitopes
Cytosol
Endocrinology
Syncytiotrophoblast
Western blot
Antibody Specificity
Pregnancy
Internal medicine
medicine
Placental Extracts
Animals
Humans
Amino Acid Sequence
medicine.diagnostic_test
Reverse Transcriptase Polymerase Chain Reaction
Biochemistry (medical)
Trophoblast
Immunohistochemistry
Peptide Fragments
Protein Subunits
medicine.anatomical_structure
Guanylate Cyclase
embryonic structures
Female
Soluble guanylyl cyclase
Subjects
Details
- ISSN :
- 19457197 and 0021972X
- Volume :
- 86
- Database :
- OpenAIRE
- Journal :
- The Journal of Clinical Endocrinology & Metabolism
- Accession number :
- edsair.doi.dedup.....7326e185eacb408613fd862d8b152bcb