Molecular histochemical analysis of estrogen receptor alpha and beta expressions in the mouse ovary: in situ hybridization and Southwestern histochemistry.
It is well known that estrogen plays important roles in the female reproductive organs, including the ovary, through estrogen receptors (ERalpha and ERbeta). It is suggested that selective effects of estrogen on a discrete set of genes may reside in promoters that are differentially responsive to ERalpha and ERbeta, while the relative biological significance of ERalpha and ERbeta is unclear. Therefore, for a better understanding of the physiological and pathological states of ovarian cells, it seems to be essential to analyze the cellular expression of ERalpha and ERbeta, at both the protein and mRNA levels. For this purpose, we have optimized the protocols for immunohistochemistry and nonradioactive in situ hybridization, respectively, using the mouse ovary. These techniques have been established as useful histochemical methods for the localization of specific protein and/or mRNA expression at the cellular level. Moreover, we also attempted to evaluate ERs as binding proteins to estrogen responsive elements, using Southwestern histochemistry, which permits us to analyze the localization in situ of transcription regulatory proteins. By employing these methods, more precise understanding of specific gene expression and evaluation of its biological significance in the ovary would be possible.
Hishikawa
Yoshitaka
Y
Department of Histology and Cell Biology, Nagasaki University School of Medicine, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan.
Damavandi
Elia
E
Izumi
Shin-ichi
S
Koji
Takehiko
T
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D026501:Blotting, Southwestern; D015342:DNA Probes / Q000235:genetics; D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D005260:Female; D015870:Gene Expression; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D051379:Mice; D010053:Ovary / Q000378:metabolism*; D011960:Receptors, Estrogen / Q000235:genetics* / Q000378:metabolism*
D015342:DNA Probes; D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D011960:Receptors, Estrogen