Dendroaspis natriuretic peptide and its functions in pig ovarian granulosa cells.
Dendroaspis natriuretic peptide (DNP), a 38-amino acid peptide, was isolated from the venom of green mamba. It has structural and functional similarities to the other members of the natriuretic peptide family. The purpose of this study was to determine whether DNP is present in pig ovarian granulosa cells and to define its biological functions. The serial dilution curves of extracts of granulosa cells and follicular fluid were parallel to the standard curve of DNP, and a major peak of molecular profile of both extracts by HPLC was synthetic DNP. The concentration of DNP was 7.51+/-1.46 pg/10(7) cells and 24.81+/-2.38 pg/ml in granulosa cells and follicular fluid, respectively. Natriuretic peptides increased cGMP production in the purified membrane of granulosa cells with a rank order of potency of C-type natriuretic peptide (CNP)>atrial natriuretic peptide (ANP)=DNP. mRNAs for natriuretic peptide receptor-A (NPR-A), NPR-B and NPR-C were detected by RT-PCR. The binding site of (125)I-DNP was also observed in granulosa cell layer by in vitro autoradiography. Synthetic DNP inhibited the secretion of ANP from granulosa cells in a concentration-dependent manner and the potency was similar to CNP. The concentration of DNP and CNP, which inhibited the secretion of ANP by 50%, was about 1 nM. Increases in production of cGMP in granulosa cells were observed by DNP or CNP. Therefore, these results show the existence of DNP system and the cross-talk between natriuretic peptides in pig ovarian granulosa cells.
Department of Physiology, Medical School, Institute for Medical Sciences, Chonbuk National University, 2-20 Keum-Am-Dong-San, Jeonju 560-181, South Korea.
Feng Lian
FL
Piao
Sung Hun
SH
Park
Jeong Hee
JH
Han
Chunhua
C
Cao
Sung Zoo
SZ
Kim
Suhn Hee
SH
Kim
D000818:Animals; D009320:Atrial Natriuretic Factor / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology; D001345:Autoradiography; D001667:Binding, Competitive; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D048429:Cell Size; D002851:Chromatography, High Pressure Liquid; D006152:Cyclic GMP / Q000378:metabolism; D004546:Elapid Venoms / Q000302:isolation & purification / Q000494:pharmacology; D005260:Female; D015870:Gene Expression; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D006162:Guanylate Cyclase / Q000235:genetics; D036341:Intercellular Signaling Peptides and Proteins; D020098:Natriuretic Peptide, C-Type / Q000494:pharmacology / Q000502:physiology; D045265:Natriuretic Peptides / Q000378:metabolism / Q000494:pharmacology / Q000502:physiology*; D010053:Ovary / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D010455:Peptides / Q000302:isolation & purification / Q000494:pharmacology / Q000502:physiology*; D011863:Radioimmunoassay; D017461:Receptors, Atrial Natriuretic Factor / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine; D000099242:Atrial Natriuretic Factor Receptor B
D009320:Atrial Natriuretic Factor; D006152:Cyclic GMP; D004546:Elapid Venoms; D006162:Guanylate Cyclase; D036341:Intercellular Signaling Peptides and Proteins; D020098:Natriuretic Peptide, C-Type; D045265:Natriuretic Peptides; D010455:Peptides; D017461:Receptors, Atrial Natriuretic Factor; D000099242:Atrial Natriuretic Factor Receptor B; C075597:Dendroaspis natriuretic peptide; C091715:atrial natriuretic factor receptor A; C073616:atrial natriuretic factor receptor C