Levels of mRNA for bone morphogenetic proteins, their receptors and SMADs in goat ovarian follicles grown in vivo and in vitro.
This study investigated the stability of housekeeping genes (glyceraldehyde-3-phosphate dehydrogenase, β-tubulin, β-actin, phosphoglycerate kinase (PGK), 18S rRNA, ubiquitin and ribosomal protein 19) and the levels of mRNA for bone morphogenetic protein-2 (BMP-2), -4 (BMP-4), -6 (BMP-6), -7 (BMP-7) and -15 (BMP-15), their receptors (BMPR-IA, -IB and -II) and Similar to Mothers Against Decapentaplegic (SMADs) (-1, -5 and -8) in goat follicles of 0.2, 0.5 and 1.0mm, as well as in secondary follicles before and after culture for 18 days. β-tubulin and PGK were the most stable housekeeping genes and the levels of mRNA for BMP-2 in follicles of 0.2mm were higher than in follicles of 0.5 and 1.0mm. For BMP-4, -6 and -7, the highest levels of mRNA were found in follicles of 1.0mm. The expression of BMPR-IB was higher in follicles of 0.2mm, whereas the levels of BMPR-II were higher in follicles of 0.5mm. The levels of mRNA for SMAD-5 were higher in follicles of 0.2mm, whereas SMAD-8 had higher levels in 0.5-mm follicles. After culture, follicles showed increased levels of mRNA for BMP-2 and reduced mRNA for BMP-4, BMP-7, BMPR-IA and SMAD-5. In conclusion, β-tubulin and PGK are the most stable reference genes, and BMPs, their receptors and SMADs have variable levels of mRNA in the follicular size classes analysed.
Biotechnology Nucleus of Sobral - NUBIS, Federal University of Ceara, CEP 62042-280, Sobral, CE, Brazil.
J J N
JJ
Costa
M J
MJ
Passos
C C F
CC
Leitão
G L
GL
Vasconcelos
M V A
MV
Saraiva
J R
JR
Figueiredo
R
R
van den Hurk
J R V
JR
Silva
D000818:Animals; D052004:Bone Morphogenetic Protein Receptors / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D019485:Bone Morphogenetic Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D048429:Cell Size; D002478:Cells, Cultured; D005260:Female; D006041:Goats / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D006080:Ovarian Follicle / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D055550:Protein Stability; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D051785:Smad Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D013997:Time Factors
D019485:Bone Morphogenetic Proteins; D012333:RNA, Messenger; D051785:Smad Proteins; D052004:Bone Morphogenetic Protein Receptors