scholarly journals Differential distribution of oestrogen receptor-alpha and -beta mRNAs in the female reproductive organ of rats as revealed by in situ hybridization

2000 ◽  
Vol 165 (1) ◽  
pp. 59-66 ◽  
Author(s):  
CN Mowa ◽  
T Iwanaga
Keyword(s):  
In Situ Hybridization ◽  
Stromal Cells ◽  
Oestrogen Receptor ◽  
Reproductive Tract ◽  
Reproductive Organ ◽  
Differential Distribution ◽  
Oestrogen Receptor Alpha ◽  
Female Reproductive Organ ◽  
Significant Expression

The cellular distribution of two oestrogen receptor (ER) subtypes, ERalpha and ERbeta mRNAs, was studied in the entire female reproductive organ of the rat using in situ hybridization. Expression of ERalpha and ERbeta mRNAs was predominant in the reproductive tract and ovary respectively. ERalpha mRNA had the most pronounced expression in epithelial cells and subepithelial stromal cells from the oviduct to the vagina, while in the ovary it was moderately detected in only the theca folliculi and interstitial glands. The oviduct showed a region-dependent expression of ERalpha mRNA: the isthmus had the most intense signals while the infundibulum revealed a low intensity of expression. Signals for ERbeta mRNA in the ovary were most intense in the granulosa cells of healthy follicles, whereas degenerating follicles lacked any significant expression. Less intense signals for ERbeta mRNA were localized in the theca folliculi and corpus luteum. Detectable levels of ERbeta mRNA were observed in the subepithelial stromal cells from the oviduct to the vagina. This study shows that the two ER subtypes are differentially expressed in cells and compartments of the reproductive organ, suggesting that the mediation of oestrogen action in these tissues may be accomplished through the respective predominant receptor.

scholarly journals Developmental changes of the oestrogen receptor-alpha and -beta mRNAs in the female reproductive organ of the rat--an analysis by in situ hybridization

2000 ◽  
Vol 167 (3) ◽  
pp. 363-369 ◽  
Author(s):  
CN Mowa ◽  
T Iwanaga
Keyword(s):  
In Situ Hybridization ◽  
Oestrogen Receptor ◽  
Reproductive Tract ◽  
Reproductive Organ ◽  
Female Reproductive Tract ◽  
Prenatal Period ◽  
Cellular Expression ◽  
Female Reproductive Organ ◽  
Er Alpha

This study employed an in situ hybridization technique to compare the cellular expression of oestrogen receptor (ER) subtypes, ER alpha and beta, in the female reproductive organ of the rat during prenatal and postnatal periods. Diffuse signals of ER alpha and beta mRNAs were co-expressed in the foetal ovary; they were weak and inconsistent before onset of gonadal differentiation, but increased in intensity with age. ER beta mRNA signals in the ovary sharply increased in intensity to adult levels by postnatal days 6-7, whereas those of ER alpha mRNA remained unchanged after birth. ER alpha was the sole subtype expressed during the prenatal period from the oviduct to the vagina, being localized mainly to the sub-epithelial stromal cells, and remained predominant thereafter. Signals for ER alpha mRNA in the epithelia were confined to the oviduct during prenatal and early postnatal periods; those in uterine and vaginal epithelia first appeared by postnatal days 4-5 and 6 respectively. Expressions of ER beta mRNA in the reproductive tract were absent during the prenatal period, and were weakly expressed during the postnatal period. Thus, oestrogen action in the developing ovary may be co-mediated by both ER alpha and beta, whereas ER alpha may be the primary mediator in the differentiation and growth of the female reproductive tract.

scholarly journals Up-regulation of Per1 expression by estradiol and progesterone in the rat uterus

2007 ◽  
Vol 194 (3) ◽  
pp. 511-519 ◽  
Author(s):  
Pei-Jian He ◽  
Masami Hirata ◽  
Nobuhiko Yamauchi ◽  
Masa-aki Hattori
Keyword(s):  
Stromal Cells ◽  
Clock Genes ◽  
Staining Intensity ◽  
Reproductive Organ ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Rat Uterus ◽  
Ovarian Steroids ◽  
Direct Regulation

It has been established that estrogen can alter circadian rhythms in behavior and endocrine physiology in rodents. The uterus is a reproductive organ that is critically dependent on regulation by ovarian steroids. Here, we examined the expression of Per1 in different compartments of the uterus, and explored whether the ovarian steroids could regulate Per1 expression employing ovariectomized rat uterus. RT-PCR analysis showed that Per1 was cyclically expressed in the uterus. As revealed by in situ hybridization, the staining intensity of Per1 mRNA was stronger at ZT 8 than at ZT 0 in the uterine luminal epithelium (LE), stroma (S), and myometrium (M) compartments, but was not changed in the glandular epithelium (GE). Both in situ hybridization and immunofluorescence analyses revealed that estradiol (E2) administration induced high expression of Per1 in the LE, GE, and M, and less expression in the S compartment. Progesterone (P4) treatment resulted in an obvious enhancement of Per1 expression in the LE, GE, and S, but unchanged in the M compartment. Furthermore, the E2- and P4-activated Per1 expression was significantly repressed by their respective antagonists, ICI182 780 and RU486. These findings were further supported by RT-PCR analysis of Per1 expression in cultured uterine stromal cells. Collectively, the present data indicate that E2 and P4 might be involved in modification of circadian rhythm via direct regulation of the expression of clock genes.

Localization of the human oestrogen receptor gene to chromosome 6q24→q27 by in situ hybridization

1986 ◽  
Vol 43 (3-4) ◽  
pp. 218-220 ◽  
Author(s):  
J.R. Gosden ◽  
P.G. Middleton ◽  
D. Rout
Keyword(s):  
In Situ Hybridization ◽  
Oestrogen Receptor ◽  
Receptor Gene ◽  
Chromosome 6Q24

Chromosome preparations from protoplasts: in situ hybridization banding pattern of a dispersed DNA sequence in rye (Secale cereale L.)

Genome ◽  
1991 ◽  
Vol 34 (4) ◽  
pp. 524-527 ◽  
Author(s):  
N. Jouve ◽  
C. L. McIntyre ◽  
J. P. Gustafson
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Banding Pattern ◽  
Banding Technique ◽  
Differential Distribution ◽  
Cellular Debris ◽  
Biotin Labeling ◽  
Secale Cereale L

The utilization of genome-specific DNA sequences coupled with in situ hybridization for chromosome karyotyping in wheat, rye, and triticale has been of limited value because of the presence of cellular and cytoplasmic debris. The use of protoplasts, thus eliminating cellular debris, has been shown to improve the level of detection of low-copy and unique DNA sequences in cereals. Therefore, the use of protoplasts could represent an appropriate tool to improve the results of karyotyping cereal chromosomes with genome-specific DNA sequences. This paper describes the results on the comparative application of protoplasts and squash preparations in the analysis of physical mapping of a dispersed DNA sequence (pSc119.1) to rye chromosomes by in situ hybridization. Individual chromosomes of rye were not distinguishable by their hybridization patterns to pSc119.1 when squash preparations were used. These showed an undefined distribution of the DNA probe that covered apparently the entire length of each rye chromosome. However, considerable improvement was observed for the differential distribution of the pSc119.1 DNA sequence in protoplast preparations. The karyotypic banding pattern of pSc119.1 showed a better banding pattern than can be observed using the C-banding technique. Therefore, the use of protoplasts hybridized with dispersed DNA markers could be of more value in monitoring chromosome karyotypes than existing cytological techniques.Key words: biotin labeling, dispersed sequences, rye.

scholarly journals Detection of phosphoenolpyruvate and ribulose 1,5-bisphosphate carboxylase transcripts in maize leaves by in situ hybridization with sulfonated cDNA probes.

1989 ◽  
Vol 37 (4) ◽  
pp. 423-428 ◽  
Author(s):  
C Perrot-Rechenmann ◽  
M Joannes ◽  
D Squalli ◽  
P Lebacq
Keyword(s):  
In Situ Hybridization ◽  
Cell Types ◽  
Small Subunit ◽  
Bundle Sheath ◽  
Differential Distribution ◽  
Mesophyll Cells ◽  
Bisphosphate Carboxylase ◽  
Maize Leaves ◽  
Specific Mrnas

This report outlines an efficient in situ hybridization method for locating specific mRNAs in tissue cryosections using sulfonated cDNA probes. The method involves chemical modification of DNA probes by insertion of a sulfone radical on cytosine residues, which generates a specific epitope. Sulfonated DNA is then detected by using indirect immunochemical procedure. Alternatively, antibodies conjugated to fluorescein or to alkaline phosphatase were used for mRNA detection. In situ hybridization was developed to study aspects of mesophyll and bundle sheath cell differentiation in maize leaves. Our results indicate that phosphoenolpyruvate carboxylase (PEP C) mRNA is restricted to mesophyll cells, and the nucleus-encoded mRNA of the small subunit (SSU) ribulose 1,5-bisphosphate carboxylase (RuBP C) is limited to the cytosol of bundle sheath cells. Thus, using in situ hybridization, we have demonstrated that the differential distribution of PEP C and RuBP C proteins in the two cell types also reflects the location of their mRNAs. These data imply either a tissue-specific transcriptional regulation or a selective mRNA degradation.

Effects on differentiation of reproductive organs and sexual behaviour in Japanese quail by excessive embryonic ERα activation

2010 ◽  
Vol 22 (2) ◽  
pp. 416 ◽  
Author(s):  
Anna Mattsson ◽  
Björn Brunström
Keyword(s):  
Japanese Quail ◽  
Oestrogen Receptor ◽  
Reproductive Tract ◽  
Reproductive Organs ◽  
Positive Control ◽  
Reproductive Organ ◽  
Testis Size ◽  
Left Testis ◽  
The Right ◽  
Copulatory Behaviour

Exposure of Japanese quail (Coturnix japonica) embryos to oestrogenic substances disrupts sexual differentiation of the reproductive tract of both sexes and impairs the copulatory behaviour of the adult male. To examine whether these effects can be induced by selective activation of oestrogen receptor α (ERα), Japanese quail eggs were injected with various doses of the selective ERα agonist 16α-lactone-oestradiol (16α-LE2). The natural oestrogen 17β-oestradiol (E2) was used as a positive control. Both 16α-LE2 and E2 induced formation of an ovary-like cortex in the left testis (ovotestis) and reduced the size of the right testis in male embryos. The asymmetry in testis size remained in sexually mature males. Both substances induced retention and malformation of the Müllerian ducts in embryos of both sexes and malformed oviducts in juveniles. Male copulatory behaviour was suppressed by embryonic exposure to E2 and the highest dose of 16α-LE2. However, the lower dose of 16α-LE2, which markedly affected development of the reproductive organs, was without effects on behaviour. It can therefore not be excluded that the behavioural demasculinisation at the 100-fold higher dose involved cross-activation of oestrogen receptor β (ERβ). In conclusion, our results suggest that oestrogen-induced disruption of reproductive organ development in Japanese quail can be mediated via ERα, whereas the role of ERα in demasculinisation of copulatory behaviour remains to be clarified.

scholarly journals Localization of oestrogen receptor alpha, oestrogen receptor beta and androgen receptors in the rat reproductive organs

2000 ◽  
Vol 165 (2) ◽  
pp. 359-370 ◽  
Author(s):  
G Pelletier ◽  
C Labrie ◽  
F Labrie
Keyword(s):  
In Situ Hybridization ◽  
Epithelial Cells ◽  
Leydig Cells ◽  
Oestrogen Receptor ◽  
Muscle Cells ◽  
Seminal Vesicles ◽  
Secretory Cells ◽  
Cell Nuclei ◽  
Sites Of Action

There is now evidence that oestrogens and androgens can influence male and female reproductive systems. In order to accurately identify the sites of action of oestrogens and androgens, we have proceeded to the histological localization of the two oestrogen receptor (ER) subtypes, ERalpha and ERbeta, and the androgen receptor (AR) in the reproductive tissues of adult rats of both sexes. AR was detected by immunocytochemistry, while ERalpha and ERbeta were localized by both immunocytochemistry and in situ hybridization. In the pituitary gland of animals of both sexes, ERalpha was found in the majority of nuclei of secretory cells in the anterior pituitary. The intermediate and posterior lobes did not show any staining. ERbeta was not found to be expressed in any of the pituitary lobes. Using AR antibodies, nuclear staining was detected in about 50% of secretory cells of the anterior lobe, the intermediate and posterior lobes being completely unstained. In the testis, ERalpha was localized in nuclei of Leydig cells as well as in round spermatocytes and spermatids, while ERbeta could only be detected in Sertoli cell nuclei. AR immunoreactivity was found in nuclei of Sertoli, peritubular myoid and Leydig cells. In the prostate, ERbeta was observed in epithelial cells of tubulo-alveoli, while the stroma was unlabelled. ERalpha was not found to be expressed in any prostate cells. In the prostate, AR was detected in nuclei of epithelial, stromal and endothelial cells. In seminal vesicles, staining of ERalpha was found in nuclei of epithelial and stromal cells. Similar findings were observed using AR antibodies. While ERbeta mRNA could not be detected by in situ hybridization, weak staining for ERbeta was localized in epithelial cells of seminal vesicles. In the ovary, both ERalpha and ERbeta were found to be expressed. ERbeta mRNA was found in granulosa cells of growing follicles, while ERalpha was present in theca cells, interstitial gland cells and germinal epithelium. AR immunoreactivity was detected in granulosa cell nuclei in growing follicles and also in scattered interstitial cells. In the oviduct and uterus, ERalpha was observed in nuclei of epithelial cells as well as of stromal and muscle cells. Similarly, AR immunoreactivity was present in nuclei of epithelial cells, stromal and muscle cells in both the oviduct and uterus. ERbeta was not detected in the oviduct and uterus. The present findings indicate a cell-specific localization of ERalpha, ERbeta and AR in reproductive tissues in rats of both sexes. By establishing the precise sites of action of oestrogens and androgens they contribute to a better understanding of the respective role of these steroids in reproduction function.

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