Retrieval of estradiol receptor in paraffin sections of resting porcine uteri by microwave treatment. Immunostaining patterns obtained with different primary antibodies

Sierralta, Walter; Thole, Hubert

doi:10.1007/bf01463656

Cited by 15 publications

(8 citation statements)

References 28 publications

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“…For the receptor isoforms, porcine uterus was used as a positive control. Tissue sections of 5 μ m were mounted onto Super Frost‐plus glass slides (Menzel Glaeser, Braunschweig, Germany) and mouse monoclonal primary antibodies raised against a peptide mapping at the carboxy terminus of the porcine ER α (clone HT277) (Sierralta and Thole 1996; Qualmann et al. 2000) and against the highly conserved carboxy terminus of the ER β 1 of human origin (clone PPG5/10; Serotec, Düsseldorf, Germany) were used for localization of ER α and ER β , respectively, while mouse monoclonal antibody against a conserved epitope within human P450 aromatase (cone SM1671P; Acris, Hiddenhausen, Germany) was used to detect P450 aromatase.…”

Section: Methodsmentioning

confidence: 99%

Expression of Oestrogen Receptors α and β and of Aromatase in the Testis of Immature and Mature Boars

Mutembei

Pesch

Schuler

et al. 2005

Reprod Domestic Animals

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The boar testis secretes high amounts of oestrogens. In order to test for a likely local significance, we investigated the expression of oestrogen receptors (ER) in immature and mature boar testes using immunohistochemistry (IHC), in vitro and in situ reverse transcription-polymerase chain reaction (RT-PCR). Samples were from 25 boars castrated at ages of 50, 100, 150, 200 and 250 days. Mouse monoclonal primary antibodies against porcine ERalpha (clone HT227), human ERbeta1 (clone PPG5/10) and human P450 aromatase (clone SM1671P) were used. Expression of the mRNA was tested utilizing primers specific for the respective porcine mRNA sequences. ER immunoreactivity was exclusively localized to the nuclei. In immature boars, 90.6 +/- 1.2% of prespermatogonia and 71.0 +/- 2.6% of the Leydig cells showed a strong staining for ERalpha; 95.5 +/- 3.5% of the prespermatogonia but none of the Leydig and Sertoli cells were ERbeta-positive. In mature boars a strong staining for the ERbeta was observed in virtually all Sertoli, Leydig and germ cells, except for the elongating/ed spermatids, which were clearly negative; for the ERalpha, strong immunoreaction signals were restricted to spermatogonia and primary spermatocytes with 93.6 +/- 2.7% of these cells being positive; distinctly less intensive signals were observed in 51.4 +/- 0.27% of the secondary spermatocytes, round spermatids and Leydig cells. In vitro RT-PCR was positive for both receptors and results of in situ RT-PCR matched those obtained by IHC. P450 aromatase immunoreaction was restricted to the cytoplasm of Leydig cells. These findings suggest that testicular ER may be important factors contributing to onset and maintenance of spermatogenesis in the boar.

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Section: Methodsmentioning

confidence: 99%

Expression of Oestrogen Receptors α and β and of Aromatase in the Testis of Immature and Mature Boars

Mutembei

Pesch

Schuler

et al. 2005

Reprod Domestic Animals

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“…ERα cytoplasmic distribution has also been described in other tissues [50,51]. It was shown that different accessibility of the ER to the reagents depending upon the functional activity of the estradiol-controlled cells could interfere with subcellular immunolocalization of the receptor [50,52]. In agreement with this finding, ERα and ERβ were more frequently localized to the cytoplasm of resting uterine cells [51] and human breast cells [45].…”

Section: Discussionmentioning

confidence: 56%

The antiestrogen ICI 182,780 decreases the expression of estrogen receptor-alpha but has no effect on estrogen receptor-beta and androgen receptor in rat efferent ductules

Oliveira

Nie

Carnes

et al. 2003

Reprod Biol Endocrinol

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BackgroundThe antiestrogen ICI 182,780 has been used successfully as an alternative experimental model for the study of estrogen action in the rodent adult male reproductive tract. Although ICI 182,780 causes severe alterations in testicular and efferent ductule morphology and function, the effects on the expression of estrogen and androgen receptors in the male have not been shown.MethodsIn the present study, adult male rats were treated with ICI 182,780 for 7 to 150 days, to evaluate the time-response effects of the treatment on the pattern of ERα, ERβ and AR protein expression in the efferent ductules. The receptors were localized using immunohistochemistry.ResultsERα, ERβ and AR have distinct cellular distribution in the testis and efferent ductules. Staining for ERα is nearly opposite of that for ERβ, as ERα shows an increase in staining intensity from proximal to distal efferent ductules, whereas ERβ shows the reverse. Androgen receptor follows that of ERα. ICI 182,780 caused a gradual but dramatic decrease in ERα expression in the testis and efferent ductules, but no change in ERβ and AR expression.ConclusionsThe differential response of ERα and ERβ proteins to ICI 182,780 indicates that these receptors are regulated by different mechanisms in the male reproductive tract.

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“…The apparent discrepancy to reports that describe the absence of ESR1 expression in epithelial cells during early pregnancy may be the result of different antibodies being used. Subcellular staining patterns of ESR1 can depend on the primary antibody used for localization studies; for instance, a particular antibody can show a preference for cytoplasmic staining versus nuclear staining and vice versa (Schuler et al 2002 ;Sierralta and Thole 1996 ). It should be noted that during early pregnancy in the pig, luminal and glandular epithelial cells maintain ESR1 expression, albeit with immunoreactivity confi ned to nuclei and not the cytoplasm (Geisert et al 1993 ;Knapczyk-Stwora et al 2011 ).…”

Section: Spatial and Temporal Regulation Of Endometrial Receptors Formentioning

confidence: 97%

Pregnancy Recognition and Implantation of the Conceptus in the Mare

Klein

2015

Regulation of Implantation and Establishment of Pregnancy in Mammals

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Few, if any, biological processes are as diverse among domestic species as establishment of early pregnancy, in particular maternal recognition of pregnancy. Following fertilization and initial development in the mare oviduct, selective transport of the embryo through the uterotubal junction driven by embryo-derived PGE2 occurs. Upon arrival in the uterus, an acellular glycoprotein capsule is formed that covers the embryo, blastocyst, and conceptus (embryo and associated extraembryonic membranes) between the second and third weeks of pregnancy. Between Days 9 and 15/16 of pregnancy, the conceptus undergoes an extended phase of mobility. Conceptus mobility is driven by conceptus-derived PGF2α and PGE2 that stimulate uterine contractions which in turn propel migration of the conceptus within the uterine lumen. Cessation of conceptus mobility is referred to as fixation and appears to be attributable to increasing size of the conceptus, preferential thickening of the endometrium near the mesometrial attachment referred to as encroachment, and a reduction in sialic acid content of the capsule. During maternal recognition of pregnancy, endometrial PGF2α release is attenuated, a consequence of reduced expression of key enzymes involved in prostaglandin production. Oxytocin responsiveness is altered during early pregnancy, and reduced expression of the oxytocin receptor appears to be regulated at the posttranscriptional level rather than the transcriptional level. Prostaglandin release is attenuated temporarily only during early pregnancy; during the third week of pregnancy, the endometrium resumes the ability to secrete PGF2α. The equine conceptus initiates steroidogenesis as early as Day 6 and synthesizes estrogens, androgens, and progesterone. Estrogens are metabolized locally, presumably regulating their bioavailability and actions. Results of experiments attempting to prove that conceptus-derived estrogens are responsible for extension of corpus luteum function have been inconclusive. By the fourth week of pregnancy, the chorionic girdle becomes visible on the trophoblast. Subsequent invasion of chorionic girdle cells leads to formation of endometrial cups which secrete equine chorionic gonadotropin. Equine chorionic gonadotropin has luteinizing hormone functions in the mare, causing luteinization of follicles resulting in the formation of secondary corpora lutea essential to production of progesterone and maintenance of pregnancy.

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Retrieval of estradiol receptor in paraffin sections of resting porcine uteri by microwave treatment. Immunostaining patterns obtained with different primary antibodies

Cited by 15 publications

References 28 publications

Expression of Oestrogen Receptors α and β and of Aromatase in the Testis of Immature and Mature Boars

Expression of Oestrogen Receptors α and β and of Aromatase in the Testis of Immature and Mature Boars

The antiestrogen ICI 182,780 decreases the expression of estrogen receptor-alpha but has no effect on estrogen receptor-beta and androgen receptor in rat efferent ductules

Pregnancy Recognition and Implantation of the Conceptus in the Mare

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