Evaluation of ovarian POMC mRNA through quantitative RT-PCR analysis in Rana esculenta.
The evaluation of changes in the expression of specific genes requires accurate measurement of the corresponding mRNA concentration, especially when the gene is expressed at a very low level. We previously showed that the proopiomelanocortin (POMC) gene is expressed in the ovary of the frog Rana esculenta, and, to evaluate its mRNA content in frog ovary, we have now developed a sensitive quantitative RT-PCR method. This study provides evidence for the validation of this method and for the effects of captivity and hypophysectomy on POMC gene expression in the ovary of this anuran. Our data indicate that ovarian POMC gene is involved in short-term captivity stress response and seems not influenced by pituitary. These results are discussed taking into account the knowledge of the role played by opioids in stress response; moreover, a local control of POMC gene expression is also suggested.
Nabissi
M
M
Dipartimento di Scienze Morfologiche e Biochimiche Comparate, Universita' degli Studi di Camerino, 62032 Camerino, Italia.
Soverchia
L
L
Lihrmann
I
I
Vaudry
H
H
Mosconi
G
G
Polzonetti-Magni
A M
AM
D000818:Animals; D001483:Base Sequence; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D066298:In Vitro Techniques; D007986:Luteinizing Hormone / Q000494:pharmacology; D008969:Molecular Sequence Data; D010053:Ovary / Q000378:metabolism*; D010902:Pituitary Gland; D016133:Polymerase Chain Reaction / Q000379:methods; D011333:Pro-Opiomelanocortin / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics*; D011892:Rana catesbeiana; D011893:Rana esculenta; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014020:Tissue Extracts / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects
D012333:RNA, Messenger; D014020:Tissue Extracts; D011333:Pro-Opiomelanocortin; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone