Regulation of vascular endothelial growth factor production by Leydig cells in vitro: The role of protein kinase A and mitogen-activated protein kinase cascade

被引:29
作者
Anand, RJK [1 ]
Paust, HJ [1 ]
Altenpohl, K [1 ]
Mukhopadhyay, AK [1 ]
机构
[1] Univ Hamburg, Inst Hormone & Fertil Res, D-22529 Hamburg, Germany
关键词
angiogenesis; protein kinase; testes; testicular microcirculation; testosterone; vascular permeability;
D O I
10.1095/biolreprod.102.009795
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
We previously reported the presence of vascular endothelial growth factor (VEGF) in testicular cells, and high concentrations of VEGF have been measured in semen, although its role in male reproduction remains obscure. In the present study we focus on understanding the mechanism of VEGF production by mouse Leydig cells cultured in vitro. Production of VEGF protein in medium by testicular cells was markedly increased by the addition of hCG in a time- and dose-dependent manner. Gonadotropin-stimulated VEGF production was mediated by cAMP-dependent protein kinase A (PKA), as evidenced by the effect of hCG being mimicked by 8Br-cAMP and being abolished in the presence of a PKA-specific inhibitor, H-89. Protein kinase C was not involved, as evidenced by phorbol 12-myristate 13-acetate having no influence on VEGF production by Leydig cells. in addition to hCG, atrial natriuretic peptide was also able to stimulate VEGF production, suggesting that cGMP is able to cross-activate PKA. A specific Src kinase inhibitor, PP2, could completely block the stimulatory effects of both gonadotropin and 8Br-cAMP on VEGF production by Leydig cells, implying an involvement of the Src kinase pathway. Furthermore, addition of U0126, an inhibitor of MEK 1/2, abolished the increase in VEGF production stimulated by both hCG and 8Br-cAMP. A similar inhibitory effect was observed by the addition of SB203580, a p38 mitogen-activated protein kinase inhibitor. Thus, in conclusion, Leydig cells are able to produce VEGF by a process under gonadotropic control, and PKA plays a key role in this process. Downstream of PKA, it appears that both MEK 1/2 and Src kinase-dependent pathways are involved, although further research will be necessary to determine the precise link between PKA and other kinases involved.
引用
收藏
页码:1663 / 1673
页数:11
相关论文
共 61 条
[1]   Vascular endothelial growth factor gene expression in human fetal lung in vitro [J].
Acarregui, MJ ;
Penisten, ST ;
Goss, KL ;
Ramirez, K ;
Snyder, JM .
AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY, 1999, 20 (01) :14-23
[2]   Relaxin-like factor expression as a marker of differentiation in the mouse testis and ovary [J].
Balvers, M ;
Spiess, AN ;
Domagalski, R ;
Hunt, N ;
Kilic, E ;
Mukhopadhyay, AK ;
Hanks, E ;
Charlton, HM ;
Ivell, R .
ENDOCRINOLOGY, 1998, 139 (06) :2960-2970
[3]  
CHEN YH, 1994, J BIOL CHEM, V269, P27372
[4]  
Cheng T, 1998, INVEST OPHTH VIS SCI, V39, P581
[5]  
CLAFFEY KP, 1992, J BIOL CHEM, V267, P16317
[6]   Progestins, progesterone receptor modulators, and progesterone antagonists change VEGF release of endometrial cells in culture [J].
Classen-Linke, I ;
Alfer, J ;
Krusche, CA ;
Chwalisz, K ;
Rath, W ;
Beier, HM .
STEROIDS, 2000, 65 (10-11) :763-771
[7]   AMINO-ACID AND CDNA SEQUENCES OF A VASCULAR ENDOTHELIAL-CELL MITOGEN THAT IS HOMOLOGOUS TO PLATELET-DERIVED GROWTH-FACTOR [J].
CONN, G ;
BAYNE, ML ;
SODERMAN, DD ;
KWOK, PW ;
SULLIVAN, KA ;
PALISI, TM ;
HOPE, DA ;
THOMAS, KA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (07) :2628-2632
[8]   TUMOR VASCULAR-PERMEABILITY FACTOR STIMULATES ENDOTHELIAL-CELL GROWTH AND ANGIOGENESIS [J].
CONNOLLY, DT ;
HEUVELMAN, DM ;
NELSON, R ;
OLANDER, JV ;
EPPLEY, BL ;
DELFINO, JJ ;
SIEGEL, NR ;
LEIMGRUBER, RM ;
FEDER, J .
JOURNAL OF CLINICAL INVESTIGATION, 1989, 84 (05) :1470-1478
[9]   Requirement for ERK1/2 activation in the regulation of progesterone production in human granulosa-lutein cells is stimulus specific [J].
Dewi, DA ;
Abayasekara, DRE ;
Wheeler-Jones, CPD .
ENDOCRINOLOGY, 2002, 143 (03) :877-888
[10]   Vascular endothelial growth factor and its receptors in normal human testicular tissue [J].
Ergun, S ;
Kilic, N ;
Fiedler, W ;
Mukhopadhyay, AK .
MOLECULAR AND CELLULAR ENDOCRINOLOGY, 1997, 131 (01) :9-20