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Changes of messenger RNAs encoding vascular endothelial growth factor and its receptors during the development and maintenance of caprine corpora lutea
Author(s) -
Kawate Noritoshi,
Tsuji Makoto,
Tamada Hiromichi,
Inaba Toshio,
Sawada Tsutomu
Publication year - 2003
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.10244
Subject(s) - biology , vascular endothelial growth factor , medicine , endocrinology , messenger rna , receptor , ovulation , luteinizing hormone , andrology , angiogenesis , hormone , vegf receptors , gene , biochemistry , genetics , cancer research
The present study was conducted to examine changes of mRNAs encoding vascular endothelial growth factor (VEGF) and its receptors (KDR/Flk‐1 and Flt‐1), and CD34, which is known to be a specific marker for endothelial cells, during the development and maintenance of the caprine corpora lutea (CL). Effects of a potent GnRH antagonist (GA), which was previously shown to suppress release of luteinizing hormone (LH), on expressions of those mRNAs during the CL development were also investigated. Goats were divided into control (n = 12) and GA‐treated groups (n = 6). The goats were treated with saline or GA (50 μg/kg, sc) on days 0 (day of ovulation), 4, and 8 (control only), and CL collected on a subset of goats (n = 3 for each day) on days 0 (no saline), 4, 8, or 14 (control only). Ribonuclease protection assay was performed to quantitate the mRNAs in the CL using specific cRNA probes generated by RT‐PCR and in vitro transcription. Level of CD34 mRNA significantly increased from day 0 to 8 (CL development) in the control group ( P < 0.05). Long and short forms were detected in the caprine CL by RT‐PCR for VEGF mRNA and analyses of their sequences showed that they correspond to mRNAs encoding VEGF 165 and VEGF 121 , respectively. Level of VEGF 165 mRNA significantly increased from day 4 to 8 and day 8 to 14 (CL maintenance) in the control group ( P < 0.05) while VEGF 121 mRNA did not change during the whole period. Level of KDR/Flk‐1 mRNA significantly increased from day 0 to 8 ( P < 0.05) while Flt‐1 mRNA significantly increased from day 8 to 14 ( P < 0.005) in the control group. In the GA‐treated group, levels of all of the mRNAs did not alter remarkably as compared with those in the control group. These results suggest that rise of KDR/Flk‐1 and VEGF 165 mRNAs during the caprine CL development may be associated with enhanced angiogenesis and that increment of VEGF 165 and Flt‐1 mRNAs during the CL maintenance may play nonangiogenic roles. The present study also indicates that the changes of VEGF 165 and KDR/Flk‐1 mRNAs during the CL development are probably not regulated by LH. Mol. Reprod. Dev. 64: 166–171, 2003. © 2003 Wiley‐Liss, Inc.