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P Tuohimaa

Publications and source records attributed to P Tuohimaa.

At least 55 records · Page 3Linked to original sources
PubMed1997

Expression of the chicken progesterone receptor forms A and B is differentially regulated by estrogen in vivo.

The chicken progesterone receptor (cPR), like its human counterpart (hPR), exists as two isoforms, PR-A and PR-B, displaying different biological activities depending upon cellular and promoter contexts. Here we show that the ratio of PR isoforms observed in the immature chicken oviduct is changed during estrogen-induced differentiation from PR-B dominancy to that of PR-A. This is the first report describing that the expression ratio of PR isoforms is altered by upregulation of PR-A by estrogen action in vivo. This result provides a plausible explanation to the differences in oviduct's response to progesterone depending on hormonal and developmental status of the animal.

Animals↗
PubMed1997

Spermatogenesis in the vitamin A-deficient rat: possible interplay between retinoic acid receptors, androgen receptor and inhibin alpha-subunit.

In order to understand the mechanisms of retinol action on the testis, testicular retinoic acid receptor alpha, beta(RAR alpha and beta), androgen receptor (AR) and inhibin alpha-subunit were studied in normal, vitamin A-deficient (VAD) and vitamin A-supplemented rats by immunohistochemistry and immunoblotting. Compared to the normal testis, expression of 110 K AR was up-regulated by vitamin A withdrawal, whereas 51 K RAR alpha remained unchanged. An additional 55 K RAR alpha signal was observed. Readministration of retinol caused a marked decrease of AR in the VAD testis. By 24 h, AR declined to below the normal level. Although the 51 K RAR alpha signal remained unchanged, the 55 K band was slightly up-regulated at 6 h after retinol administration. A 51 K RAR beta protein was seen in the VAD but in not the normal testis. The intensity of the 51 K RAR beta band remained constant before and after the administration of retinol, but it had a slight up-shift at 6 h after retinol injection, suggesting post-translational modification of the receptor. The inhibin alpha-subunit of 18 K protein was undetectable in the VAD testis and increased to above normal level at 24 h after retinol administration. Immunohistochemically, nuclear AR immunostaining was more intense in the VAD testis than in the normal testis. The intensity of immunostaining declined in all AR-positive cells after the injection of retinol, but the decrease was more evident in Sertoli than in other cells. At 24 h after retinol the immunostaining was undetectable in most Sertoli cells. The regulation of the inhibin alpha-subunit by retinol in the cytoplasm of Sertoli cells detected by immunohistochemistry was correlated to the results in immunoblotting. These results suggest a possible interplay between retinoids, androgen and inhibin signalling systems in Sertoli cells in the regulation of spermatogenesis during retinol action.

Animals↗
PubMed1997

Immunodetection of androgen receptor in human urinary bladder cancer.

We investigated the expression of androgen receptor (AR) protein in transitional cell carcinoma of human urinary bladder in paraffin-embedded sections of tumours obtained from nine patients with urinary bladder cancer treated by radical cystectomy. In addition, immunoblotting of AR was also performed on selected samples. Nuclear immunoreactivity of AR was found in seven of the nine urinary bladder cancers studied. AR showed variable staining intensity within a tumour. In the immunoblots, a 110 kDa AR signal was seen with anti-AR antibody, and faint bands of 90 and 60 kDa were also observed. Immunohistochemistry of p53 and c-erbB-2 was also carried out and compared with the distribution of AR. The high frequency of AR expression suggests a role for androgens in transitional cell carcinoma of human urinary bladder.

Adenocarcinoma↗
PubMed1997

Photoperiod-induced changes in androgen receptor expression in testes and accessory sex glands of the bank vole, Clethrionomys glareolus.

Photoperiodic modulation of androgen levels and androgen receptor (AR) expression in testes and accessory sex glands were studied in a seasonally breeding rodent, the bank vole. Juvenile voles subjected to long photoperiod (20L:4D) for 6-8 wk attained sexual maturity, which was associated with a prominent increase in testicular testosterone (T) levels and weight of testes and accessory sex glands. Pubertal development in short photoperiod-treated (6L:18D for 6-8 wk) juveniles was arrested, and subsequently reproductive regression set in with a marked decrease in testicular T levels and gonadal weight. In sexually active voles, strong AR immunostaining was detected in nuclei of epithelial, smooth muscle, and stromal cells of the epididymis, prostate, and seminal vesicles. In active testes, AR was present in nuclei of Sertoli cells, peritubular cells, Leydig cells, and vascular smooth muscle cells. In juveniles, strong to moderate nuclear immunoreactivity was encountered in epithelial and stromal cells of the epididymis and prostate, whereas a weaker reaction was discerned in seminal vesicles. In juvenile testes, AR was localized to vascular smooth muscle cells, peritubular, and interstitial cells. In sexually regressed animals, nuclear staining was almost absent in accessory sex glands, whereas in testes, moderate immunostaining was retained in all other cell types except the Sertoli cells. Western blots of active and regressed testes indicated a marked photoperiod-induced down-regulation of immunodetectable AR in the regressed gonad.

Aging↗
PubMed1996

Measurement of activin B in human saliva and localization of activin subunits in rat salivary glands.

The present paper is the first report to demonstrate that measurable amounts of activin B are secreted into the saliva. The results show wide fluctuations in activin B concentrations during the menstrual cycle with peak values detected at the follicular phase. Estrogen replacement therapy was found to increase salivary activin B levels in post-menopausal subjects. The concentration in males was negligible. These data suggest that activin B concentrations in the human saliva may be under hormonal regulation and propose that salivary activin B measurements may prove useful in investigating the as yet less well defined local and/or physiological roles of activin B. The present paper reports, in addition, the immunohistochemical localization of activin/inhibin subunits in the duct systems of the rat submandibular, sublingual and parotid salivary glands.

Activins↗
PubMed1996

Androgen receptor in rat Harderian and submandibular glands.

Androgens regulate the development and sexual dimorphism of rodent Harderian and submandibular glands. This effect is believed to be mediated by the androgen receptor. Immunohistochemistry and immunoblotting were carried out to study the receptor in normal, castrated and dihydrotestosterone-supplemented rat Harderian and submandibular glands. Immunohistochemically, the most intense nuclear staining was observed in the acinar cells of the submandibular glands, followed by intercalated duct cells. The granular convoluted tubules showed weak immunostaining and the striated ducts were negative. In the Harderian gland, nuclear staining was seen in both type I and II secretory cells. Castration and treatment had no effect on the expression of the androgen receptor protein in either gland. A 110 K androgen receptor signal was detected by immunoblotting in the Harderian gland but not in the submandibular gland. An experiment was designed to explore the possible effect of proteinases on the receptor protein in the homogenate of submandibular gland. Our results demonstrate the cell-specific location of the receptor in Harderian and submandibular glands, and show that the expression of the receptor protein is androgen-independent.

Animals↗
PubMed1996

Immunochemical and in situ hybridization analyses of retinoic acid receptor alpha, beta, and gamma in murine Harderian and submandibular glands.

Retinoic acid (RA), through its cognate receptors (retinoic acid receptors, RARs), plays an important role in the ontogenesis and maintenance of the normal function of murine Harderian and submandibular glands. In the present study, autoradiography was used to study RA binding to these glands. Both glands showed high radioactive labelling after [14C]-RA administration in normal and partially vitamin A-deficient (VAD) mice. The peak uptake was at 6 h after [14C]-RA administration in normal mice and at 0.5 h in VAD mice. At 24 h, RA binding remained high in normal mice, while it decreased significantly in VAD mice. In western blots with an antibody recognizing all forms of RARs, a band of molecular weight 51 kDa was seen in homogenates of both glands. Immunohistochemically, RAR staining was found in the nuclei of the glandular cells. The Harderian gland exhibited more intense staining than the submandibular gland. In the latter, the most intense staining was seen in the acinar cells, followed by the intercalated duct cells. The granular convoluted tubule showed weak immunostaining and the striated duct was negative. In the Harderian gland, RAR immunostaining was observed in both type I and II cells, but only part of them stained with RAR antibody. The expression of RAR alpha, beta, and gamma transcripts was studied by in situ hybridization using specific oligonucleotide probes. The cell-specific expression of RAR alpha mRNA in the submandibular gland corresponded to the RAR proteins detected by immunohistochemistry, while the RAR beta transcript was mainly seen in the striated duct. The transcripts of RAR alpha and beta were evenly distributed in type I and II glandular cells of the Harderian gland. RAR gamma labelling was below detectable levels in both glands. This result suggests that RA and RARs regulate the functions of Harderian and submandibular glands in a cell-specific manner.

Animals↗
PubMed1996

Mechanisms of action of sex steroid hormones: basic concepts and clinical correlations.

The review deals with the clinically important aspects of the basic mechanisms of sex steroid hormones. Steroids can act through two basic mechanisms: genomic and non-genomic. The classical genomic action is mediated by specific intracellular receptors, whereas the primary target for the non-genomic one is the cell membrane. Many clinical symptoms seem to be mediated through the non-genomic route. Furthermore, membrane effects of steroid and other factors can interfere with the intranuclear receptor system inducing or repressing steroid-and receptor-specific genomic effects. These signalling pathways may lead to unexpected hormonal or anti-hormonal effects in patients treated with certain drugs. Steroid receptors (SRs) are members of a large family of nuclear transcription factors that regulate gene expression by binding to their cognate steroid ligands, to the specific enhancer sequences of DNA (steroid response elements) and to the basic transcription machinery. SRs are phosphoproteins, which are further phosphorylated after ligand binding. The role of phosphorylation in receptor transaction is complex and may not be uniform to all SRs. However, phosphorylation/dephosphorylation is believed to be a key event regulating the transcriptional activity of steroid receptors. SR activities can be affected by the amount of SR in the cell nuclei, which is modified by the rate of transcription and translation of the SR gene as well as by proteolysis of the SR protein. There is an auto- and heteroregulation of receptor levels. Some of the SRs appear to bind specific protease inhibitors and exhibit protease activity. The physiological significance of this weak proteolytic activity is not clear. Some SRs are expressed as two or more isoforms, which may have different effects on transcription. Receptor isoforms are different translation or transcription products of a single gene. Isoform A of the progesterone receptor is a truncated form of PR isoform B originating from the same gene, but it is able to suppress not only the gene enhancing activity of PR-B but also that of other steroid receptors. From the clinical point of view, it is important to note that the final hormonal effect in a target tissue is dependent on the cross talk between different nuclear steroid receptors and on expression of receptor isoforms.

Animals↗
PubMed1996

Hormone-dependent changes in A and B forms of progesterone receptor.

The influence of different estrogen and/or progesterone treatments on concentrations of A and B forms of progesterone receptor (PR-A and PR-B) in the different cell types of chick oviduct was studied. A semiquantitative immunohistochemical assay for cellular PR concentrations was developed using a computer-assisted image analysis system. The staining intensity of nuclear PR in the basal layer of epithelial cells, glandular, smooth muscle and mesothelial cells was analysed separately using two monoclonal antibodies, PR6 and PR22. The measured concentrations of PR varied between different cell types and from cell to cell. A significant decrease in PR concentration, as noted by a decrease in staining intensity, was observed in all cell types studied 2 or 6 h after a single injection of progesterone with or without simultaneous estrogen administration. The decrease was also verified with immunoblotting and an immunoenzymometric assay (IEMA) for chicken PR. After down-regulation the concentration of PR recovered to the control level within 48 h after progesterone or estrogen administration. Estrogen administration alone was observed to cause changes in the concentration of PR-A only, having little or no effect on PR-B concentration depending on the cell type studied. These findings indicate that estrogen and progesterone cause cell-specific changes not only to the total concentration of PR but also to the cellular ratio of PR-A and PR-B.

Animals↗
PubMed1996

Vitamin D3- and retinoic acid-induced monocytic differentiation: interactions between the endogenous vitamin D3 receptor, retinoic acid receptors, and retinoid X receptors in U-937 cells.

Retinoic acid (RA) and 1,25 alpha-dihydroxycholecalciferol (VitD3) are potent regulators of hematopoletic differentiation. Yet, little is known as to how the RA and VitD3 receptor network operates in hematopoietic cells, and whether receptor interactions can explain the interplay between the RA- and VitD3-signaling pathways during differentiation. Therefore, we analyzed the expression, DNA binding, and transcriptional activity of the endogenous RA and VitD3 receptors [retinoic acid receptors (RARs), retinoid X receptors (RXRs), and VitD3 receptor (VDR)] in the U-937 cell line, in which RA and VitD3 induce distinct monocytic differentiation pathways. VitD3 induction resulted in the formation of VDR/RXR DNA-binding complexes on both VitD3 response elements and RA response elements (RAREs). However, transcriptional activation was only observed from a VitD3 response element-driven reporter construct. Several DNA-binding complexes were detected on RAREs in undifferentiated cells. Stimulation by RA resulted in increased RAR beta/RXR DNA binding, activated RARE-dependent transcription, and increased expression of RAR-beta. Concomitant stimulation by VitD3 inhibited the RA-stimulated formation of RAR beta/RXR heterodimers, favoring VDR/RXR binding to the RARE. Also, VitD3 inhibited the expression of CD23 and CD49f, characteristic markers of retinoid-induced U-937 cell differentiation. In contrast, neither the RA-stimulated, RARE-mediated transcription nor the induced RAR-beta expression was suppressed by VitD3, suggesting that VitD3 selectively inhibited the retinoid-induced differentiation program but not the RARE-mediated signal. These results demonstrate a complex role for VitD3 in modifying the retinoid differentiation pathway and may have implications for differentiation-inducing therapy of hematopoietic tumors.

Cell Differentiation↗
PubMed1995

Progesterone receptor does not form oligomeric (8S), non-DNA-binding complex in intact cell nuclei.

We raised a polyclonal antibody, alpha D, against a synthetic peptide (amino acids 522-535) of chicken progesterone receptor (PR). The sequence is located between the DNA-binding domain and the hormone-binding domain in the region within the sequences required for stability of the oligomeric form of PR. In the immunoblot, alpha D reacted with both A and B forms of PR. In the sucrose gradient and dot-blot the antibody did not recognize the so-called 8S form of PR, which is an oligomeric complex of PR and other proteins. When the oligomeric complex was dissociated by salt treatment, the antibody recognized the resulting 4S form of PR. This would suggest that the epitope is masked in the 8S form of PR and exposed in the 4S form. To study whether a similar complex exists in vivo, we used the antibody for immunohistochemistry. Two different fixation techniques were employed, freeze-drying-vapor fixation and liquid fixation. In the animals not treated with progesterone, intensive nuclear staining was detected independent of the fixation technique. When receptor from similarly treated animals was analyzed by sucrose gradient, all of the receptor molecules were in the oligomeric complex (8S). Ligand binding is known to promote a dissociation of this complex. Thus progesterone treatment should lead to an increased immunodetection of the epitope; however, progesterone treatment decreased the intensity of PR immunostaining. These results suggest that the oligomeric complex (8S), present in tissue extracts, does not exist in intact cell nuclei. They also call into question the proposed role of hsp90 in regulating progesterone receptor function.

Amino Acid Sequence↗
PubMed1995

Distribution of all-trans-retinoic acid in normal and vitamin A deficient mice: correlation to retinoic acid receptors in different tissues of normal mice.

The distribution of all-trans-retinoic acid (RA) was investigated using whole-body autoradiography of normal and partial vitamin A deficient (VAD) mice. Retinoic acid receptors (alpha, beta, and gamma) were also studied in normal mice using immunoblotting. Normal and VAD mice were injected with 5 muCi 14C RA. The distribution of RA was quantitatively studied using a computer-assisted image analysis system. 14C RA was incorporated 0.5 hr after RA administration in both normal and VAD mice, while the labelling peak was at 6 hr in most organs in normal and VAD mice. The most intense labeling was found in liver, kidney, intestine, lung, Harderian gland, and salivary gland at all time points. A band of M(r) 51K was found in all mouse tissues by immunoblotting using the polyclonal antibody RAR82 against total RARs or the RAR alpha-specific monoclonal antibody R alpha 13. In some tissue, an additional band of 55-58K was also found. Lung, large intestine, small intestine, testis, seminal vesicle, and spleen contained highest concentration of total RARs, while heart, lung, small intestine, spleen, salivary gland, and preputial gland had the highest concentration of RAR alpha. The uptake of labeled RA correlated well with RAR or RAR alpha concentration in the corresponding tissues.

Animals↗
PubMed1995

Activin beta A- and beta B-subunit expression in the developing chicken bursa of fabricius.

The bursa of Fabricius (BF) is a site for B-lymphocyte maturation in birds. The BF is also known to function as an endocrine organ, with regulatory effects on steroid-secreting glands (testes, ovaries, and adrenal glands) during embryonic development. To study the possible involvement of the growth and differentiation factors, inhibins and activins, in bursal differentiation and function, the expression of inhibin/activin subunits in chicken BF was examined using specific antibodies against inhibin/activin alpha-, beta A-, and beta B-subunits. Bursae from chickens from 11 days of embryonic development until 22 weeks after hatching were studied. Immunoreactive beta A- and beta B-subunits were demonstrated in bursal epithelial cells throughout the time period studied. Immunoreactivities for both subunits were most intense and widespread from day 18 of embryonation until 1 week after hatching. Inhibin alpha-subunits were not detected. Partially different locations were shown for beta A- and beta B-subunits, suggesting different roles for activin-A (beta A beta A homodimer) and activin-B (beta B beta B homodimer) in bursal development and function. As activin beta A-subunit immunoreactivity was predominantly localized in medullary epithelia, it may be assumed that the developing B-cells within follicular medullae might be the target for activin-A action in the chicken BF. The most conspicuous site for activin-B production, on the other hand, was the follicle-associated epithelium, which is able to take up antigens from the cloacal environment and pass them to medullary cells. The data suggest that activin-A and -B may have different roles in modulating bursal microenvironment during B-lymphocyte differentiation. The possible role of bursal activins in the regulation of steroidogenesis in birds is discussed.

Activins↗
PubMed1994

Progesterone receptor and hsp90 are not complexed in intact nuclei.

In hypotonic cell extract (cytosol), unliganded progesterone receptor (PR) is known to form an oligomeric complex with heat shock protein 90 (hsp90), and this complex does not bind to DNA. Since ligand binding has been shown to render the complex less stable in vitro, it has been proposed that ligand binding regulates DNA binding and receptor activity in vivo by altering the stability of the oligomeric complex. However, there is no direct evidence as to whether this oligomeric complex is present in vivo. The present study addressed this problem. First, we used an immunoelectron-microscopic technique and monoclonal antibodies to ascertain the location of PR and hsp90 in chick oviduct cells. Hsp90 was found in the cytoplasm and PR in the nucleus. To study the relative affinities of the PR and hsp90 antibodies, we then constructed a chimeric protein (PR-hsp90), which was expressed in the HeLa cells. Both hsp90 and PR antigens of the chimera were detected in the nuclei with the same intensity, which indicates that the antibodies have equal sensitivities in detecting their antigens. This suggests that if significant amounts of nuclear hsp90 were present in intact cells, it should have been detected by our method. Our results indicate that the PR does not exist in vivo as an oligomeric, nonDNA-binding form in the cell nuclei and that the oligomeric form found in tissue extracts is possibly formed during tissue processing.

Animals↗
PubMed1994

Immunolocalization of retinoic acid receptors in rat, mouse and human ovary and uterus.

We raised an antibody against a synthetic peptide corresponding to amino acids 155-174 of human retinoic acid receptor alpha (RAR-alpha). The sequence is highly homologous in all RARs and their isoforms. When mouse and human RARs (alpha, beta and gamma) expressed in Cos cell were analysed with immunoblot, all receptors gave a specific 51 K signal. Mouse RAR-gamma gave an additional signal corresponding to 58 K. In human teratocarcinoma cells (F9) both 51 and 58K molecule sizes were detected. The RAR expression in F9 cells was slightly down-regulated in charcoal-stripped culture medium and returned to normal level after retinoic acid treatment. The 51 K protein was found in all ovarian and uterine samples, but the quantity of the 58 K protein varied in different species and organs, being highest in the mouse uterus and the rat and human ovary. Using immunohistochemistry the RARs were found in the nuclear compartment. In the rat uterus, positive immunoreaction was found mainly in the nuclei of epithelial, uterine glandular and stromal cells. In the rat ovary, positive reaction was found in the nuclei of germinal epithelial, follicular and stromal cells.

Amino Acid Sequence↗
PubMed1994

Prognostic variables in predicting pregnancy. A prospective follow up study of 907 couples with an infertility problem.

This report describes a prospective follow-up study on 907 infertile men presenting as partners of infertile couples. Couples who conceived were compared to those who remained infertile during a 3-year follow up. A univariate life table analysis showed that the duration of infertility, the age of both spouses, a history of male urethritis, the body mass index, the percentage of motile spermatozoa, the quality of motility, the motile sperm density, the total motile count, the sperm morphology, and the pH of semen were significant predictors of fertility. Another eleven socio-demographic and nine semen variables were not associated with fertility. Cox multiple regression analysis identified the duration of infertility, percentage total motility, the quality of motility combined, a history of male urethritis, the wife's age, and the morphology of the spermatozoa to be independent predictors of the reproductive outcome. A prognostic 'risk' ratio (chance for achieving pregnancy) was calculated as a function of these variables for each couple and a life table analysis was made between three classified subgroups of couples with different risk ratios. These life table curves make it possible to define the probability of an individual couple to achieve pregnancy at any given time.

Demography↗