PubMed Health⌕ Search

Biomedical subjects

L Birch

Publications and source records attributed to L Birch.

At least 19 recordsLinked to original sources
PubMed2005

Neurodistribution of androgen receptor immunoreactivity in the male frog, Rana esculenta.

Sexual behavior in vertebrates depends on the cyclic release of steroids and their binding to the brain receptors. Previously, we demonstrated the presence of specific binding of (3)H-testosterone and staining with PG-21 in the brain of the adult male frog, Rana esculenta. Here, we report our further receptor characterization using an anti-androgen receptor antiserum, PG-21, and the androgen site of action in frog brain. Nuclei, which contained cells labeled for the androgen receptor (AR), were mainly identified in the olfactory bulbs, preoptic-septal region, infundibulum, amygdala, thalamus, tectum, torus semicircularis, and medulla. The neuroanatomical AR staining appears similar to that in other lower vertebrates.

Animals↗
PubMed2004

Abnormal morphology of the penis in male rats exposed neonatally to diethylstilbestrol is associated with altered profile of estrogen receptor-alpha protein, but not of androgen receptor protein: a developmental and immunocytochemical study.

Objectives of the study were to determine developmental changes in morphology and expression of androgen receptor (AR) and estrogen receptor (ER)alpha in the body of the rat penis exposed neonatally to diethylstilbestrol (DES). Male pups received DES at a dose of 10 microg per rat on alternate days from Postnatal Day 2 to Postnatal Day 12. Controls received olive oil vehicle only. Tissue samples were collected on Days 18 (prepuberty), 41 (puberty), and 120 (adult) of age. DES-induced abnormalities were evident at 18 days of age and included smaller, lighter, and thinner penis, loss of cavernous spaces and associated smooth muscle cells, and increased deposition of fat cells in the corpora cavernosa penis. Fat cells virtually filled the entire area of the corpora cavernosa at puberty and adulthood. Plasma testosterone (T) was reduced to an undetectable level, while LH was unaltered in all treated groups. AR-positive cells were ubiquitous and their profile (incidence and staining intensity) did not differ between control and treated rats of the respective age groups. Conversely, ERalpha-positive cells were limited to the stroma of corpus spongiosus in all age groups of both control and treated rats, but the expression in treated rats at 18 days was up-regulated in stromal cells of corpora cavernosa, coincident with the presence of morphological abnormalities. Hence, this study reports for the first time DES-induced developmental, morphological abnormalities in the body of the penis and suggests that these abnormalities may have resulted from decreased T and/or overexpression of ERalpha.

Aging↗
PubMed2003

Molecular methods for the assessment of bacterial viability.

A significant number of pathogenic microorganisms can be found in environmental reservoirs (air, water, soil). It is important to assess the viability status of these organisms to determine whether they pose a threat to public health. Classical methods for determining viability are time consuming. Hence, molecular methods have been developed to address this problem. Molecular methods offer speed, sensitivity and specificity. Both DNA and RNA have been analysed using molecular amplification methods such as polymerase chain reaction (PCR), reverse transcriptase PCR (RT-PCR) and nucleic acid sequence-based amplification (NASBA). However, due to the variable persistence of nucleic acids in cells post-death, the correlation between presence of DNA and RNA and viability is not clear-cut. Similarly, the choice of target and sensitivity of the method can significantly affect the validity of the viability assay. This review assesses the molecular methods currently available and evaluates their ability to assess cell viability with emphasis on environmental pathogens.

Bacteria↗
PubMed2001

Estrogen imprinting of the developing prostate gland is mediated through stromal estrogen receptor alpha: studies with alphaERKO and betaERKO mice.

Neonatal exposure of rodents to high doses of estrogen permanently imprints the growth and function of the prostate and predisposes this gland to hyperplasia and severe dysplasia analogous to prostatic intraepithelial neoplasia with aging. Because the rodent prostate gland expresses estrogen receptor (ER)-alpha within a subpopulation of stromal cells and ERbeta within epithelial cells, the present study was undertaken to determine the specific ER(s) involved in mediating prostatic developmental estrogenization. Wild-type (WT) mice, homozygous mutant ER (ERKO) alpha -/- mice, and betaERKO -/- mice were injected with 2 microg of diethylstilbestrol (DES) or oil (controls) on days 1, 3, and 5 of life. Reproductive tracts were excised on days 5 or 10 (prepubertal), day 30 (pubertal), day 90 (young adult), or with aging at 6, 12, and 18 months of age. Prostate complexes were microdissected and examined histologically for prostatic lesions and markers of estrogenization. Immunocytochemistry was used to examine expression of androgen receptor, ERalpha, ERbeta, cytokeratin 14 (basal cells), cytokeratin 18 (luminal cells), and dorsolateral protein over time in the treated mice. In WT-DES mice, developmental estrogenization of the prostate was observed at all of the time points as compared with WT-oil mice. These prostatic imprints included transient up-regulation of ERalpha, down-regulation of androgen receptor, decreased ERbeta levels in adult prostate epithelium, lack of DLP secretory protein, and a continuous layer of basal cells lining the ducts. With aging, epithelial dysplasia and inflammatory cell infiltrate were observed in the ventral and dorsolateral prostate lobes. In contrast, the prostates of alphaERKO mice exhibited no response to neonatal DES either immediately after exposure or throughout life up to 18 months of age. Furthermore, neonatal DES treatment of betaERKO mice resulted in a prostatic response similar to that observed in WT animals. The present findings indicate that ERalpha is the dominant ER form mediating the developmental estrogenization of the prostate gland. If epithelial ERbeta is involved in some component of estrogen imprinting, its role would be considered minor and would require the presence of ERalpha expression in the prostatic stromal cells.

Animals↗
PubMed2001

An investigation of how acute muscle pain modulates performance during computer work with digitizer and puck.

The purpose was to investigate the influence of muscle pain on work performance during computer work with digitizer and puck. Muscle pain was induced by infusion of hypertonic saline in the trapezius and the extensor carpi ulnaris (ECU) muscles on two separate days. Twelve healthy subjects participated. A computer task was performed in three 6 min sessions: baseline, pain, after pain. The computer task comprised production of drawings at maximal work pace. One drawing was defined as a work cycle. Work cycle time, number of puck button clicks, and screen pixels the cursor had moved per cycle were assessed. Shoulder pain did not influence these variables. Cycle time decreased from 13.8 (SD 2.2) to 13.0 s (SD 1.9) compared to baseline (p < 0.05) during ECU muscle pain. The increased or unchanged performance suggests that acute moderate muscle pain has minor influence on performance during computer work with digitizer and puck.

Adult↗
PubMed2001

Motor unit recruitment in the trapezius muscle with special reference to coarse arm movements.

Chronic shoulder pain is common in a variety of occupations. The "Cinderella hypothesis" suggests that the pain originates from damaged type I muscle fibres driven into degenerative processes as a result of too long activation and too short recovery time. The main purpose of this study was to investigate if the same motor units are active during all phases of coarse arm movements. Eight healthy volunteers participated in the study. Intramuscular electromyographic signals were picked up with a four-lead fine wire electrode, during a unilateral straight arm movement. The movement started with either (part 1) an abduction or a flexion, then (2) a movement in the horizontal plane from the sagittal to the frontal plane or vice versa, and finally (3) an adduction or an extension to the start position. The movement cycle was performed in three different speeds, slow, medium, and high, with one, two or five cycles per 20 s, respectively. On an average, the motor unit action potentials (MUAPs) of 6 motor units (range, 1-15) were identified per trail. In total 94% of the MUAP trains that were identified showed firings in all 3 parts of the movements. The findings support the Cinderella hypothesis, although there is a need to further investigate the temporal pattern of long-term motor unit activity.

Action Potentials↗
PubMed2001

Food and activity preferences in children of lean and obese parents.

BACKGROUND: Children of obese parents have a substantially higher risk of adult obesity than children of lean parents. Adoption and twin studies have shown that this risk is largely genetic but the proximal mechanisms of the genetic risk are not known. Comparisons of energy intake or expenditure in children of obese and lean parents have produced mixed, but generally negative results. An alternative hypothesis is that the early expression of obesity risk is through food and activity preferences, which provides a basis for later weight gain. The aim of this study was therefore to compare food and activity preferences in a large sample of young children from obese and lean families using parental obesity as a marker of the obesity-risk phenotype. Because the children from the families with obese parents were not yet overweight, differences observed in the two types of families are more likely to be causes than effects of obesity. METHODS: A total of 428 children aged 4-5 y, whose parents were either obese/overweight or normal-weight/lean were selected from a population sample of families with twin births. Food and activity preferences were assessed with a combination of food intake and taste tasks, and questionnaires completed by the mother during a home visit. FINDINGS: Children from the obese/overweight families had a higher preference for fatty foods in a taste test, a lower liking for vegetables, and a more 'overeating-type' eating style. They also had a stronger preference for sedentary activities, and spent more time in sedentary pastimes. There were no differences in speed of eating or reported frequency of intake of high-fat foods. CONCLUSION: Part of the process whereby a genetic risk of obesity is transmitted to the next generation could be through differences in diet and activity preferences, which would place susceptible individuals at risk of positive energy balance in the permissive nutritional environment of industrialised countries today.

Child, Preschool↗
PubMed2001

A comparison of nucleic acid amplification techniques for the assessment of bacterial viability.

AIMS: The ability to determine the presence and viability status of bacteria by molecular methods could offer significant advantages to the food, environmental and health sectors, in terms of improved speed and sensitivity of detection. METHODS AND RESULTS: In this study, we have assessed three amplification techniques, PCR, RT-PCR and NASBA, for their ability to detect nucleic acid persistence in an E. coli strain following heat-killing. NASBA offered the greatest sensitivity of the three methods tested. The presence of residual DNA and mRNA could be detected by PCR and NASBA, respectively, for up to 30 h postdeath, by which time cell death had been confirmed by culture methods. Thus a single quantitative measurement based on nucleic acid amplification did not permit unequivocal determination of cell viability. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: The correlation between cell viability and persistence of nucleic acids must be well characterized for a particular analytical situation before molecular techniques can be substituted for traditional culture methods.

Bacteria↗
PubMed2001

Influence of neonatal estrogens on rat prostate development.

Brief exposure of rodents to estrogens during early development alters prostate branching morphogenesis and cellular differentiation in a dose-dependant manner. If estrogenic exposures are high, these disturbances lead to permanent imprints of the prostate, which include reduced growth, differentiation defects of the epithelial cells, altered secretory function and reduced responsiveness to androgens in adulthood. This process, referred to as neonatal imprinting or developmental estrogenization, is associated with an increased incidence of prostatic lesions with aging, which include hyperplasia, inflammation and dysplasia. To better understand how early estrogenic exposures can permanently alter prostate growth and function and predispose the gland to neoplasia, the effects of estrogens on prostatic steroid receptors, cell-cell communication molecules and key developmental genes were examined. Transient and permanent alterations in the expression of prostatic androgen receptors, estrogen receptors alpha (ERalpha) and beta, and retinoic acid receptors are observed. It is proposed that the estrogen-induced alterations in these critical transcription factors play a fundamental role in initiating prostatic growth and differentiation defects. Down-stream effects of the altered steroid receptor expression include disruption of TGFbeta paracrine communication, altered expression of gap junction connexin molecules and loss of epithelial cadherin on epithelial cells. Additionally, specific disruptions in the expression of prostatic developmental genes are observed in response to neonatal estrogen. An extended developmental period of hoxa-13 expression, a lack of hoxd-13 increase with maturation, and an immediate and sustained suppression of hoxb-13 was noted within prostatic tissue. A transient decrease in Nkx3.1 expression in the developing prostate was also observed. Thus subtle and overt alterations in Hox-13 and Nkx3.1 genes may be involved in the altered prostate phenotype in response to neonatal estrogen exposure. In summary, estrogen imprinting of the prostate gland is mediated through up-regulated levels of stromal ERalpha, which initiates alterations in steroid receptor expression within the developing gland. Rather than being an androgen-dominated process, as occurs normally, prostatic development is regulated by alternate steroids, including estrogens and retinoids, in the estrogenized animal. This, in turn, leads to disruptions in the coordinated expression of critical developmental genes including TGFbeta, Hox-13 genes and Nkx3.1. Since a precise temporal expression pattern of these and other molecules is normally required for appropriate differentiation of the prostatic epithelium and stroma, the estrogen-initiated disruption in this pattern would lead to permanent differentiation defects of the prostate gland. It is hypothesized that these molecular and cellular changes initiated early in life predispose the prostate to the neoplastic state upon aging.

Animals↗
PubMed2001

Maternal milk consumption predicts the tradeoff between milk and soft drinks in young girls' diets.

Milk intake constitutes an important source of dietary calcium for young girls but declines throughout childhood. Recent work shows that the intake of soft drinks may contribute to this decline. Influences on the apparent tradeoff between soft drinks and milk in young girls' diets are not well described. The objective of this research was to test a model depicting maternal beverage choices as predictors of their daughters' milk and soft drink intake. A structural equation model describing maternal influences on daughters' milk, soft drink and calcium intakes was tested using data from 180 non-Hispanic, white families with 5-y-old daughters. Mothers' calcium, milk and soft drink intakes were evaluated as predictors of their daughters' intakes. Mothers' and daughters' soft drink intakes were also examined as predictors of their own milk and calcium intakes. The model provided a good fit to the data, revealing mother-daughter similarities in beverage intake. Mothers who drank milk more frequently had daughters who drank milk more frequently and drank fewer soft drinks. For both mothers and daughters, soft drink consumption was negatively related to both milk and calcium intake. This research provides evidence that mothers' beverage choices influence the tradeoff between milk and soft drinks in their daughters' diets. In particular, mothers' milk and soft drink intakes may affect their daughters' calcium adequacy in early childhood by influencing the frequency with which their daughters consume those beverages.

Adult↗
PubMed2001

Developmental exposure to estrogens alters epithelial cell adhesion and gap junction proteins in the adult rat prostate.

Brief exposure to estrogens during the neonatal period interrupts rat prostatic development by reducing branching morphogenesis and by blocking epithelial cells from entering a normal differentiation pathway. Upon aging, ventral prostates exhibit extensive hyperplasia and dysplasia suggesting that neonatal estrogens may predispose the prostate gland to preneoplastic lesions. To determine whether these prostatic lesions may be manifested through aberrant cell-to-cell communications, the present study examined specific gap junction proteins, Connexins (Cx) 32, and Cx 43, and the cell adhesion molecule, E-cadherin, in the developing, adult and aged rat prostate gland. Male rat pups were given 25 microgram estradiol benzoate or oil on days 1, 3, and 5 of life. Prostates were removed on days 1, 4, 5, 6, 10, 15, 30, or 90 or at 16 months, and frozen sections were immunostained for E-cadherin, Cx 43, and Cx 32. Colocalization studies were performed with immunofluorescence using specific antibodies for cell markers. Gap junctions in undifferentiated epithelial cells at days 1-10 of life were composed of Cx 43, which always colocalized with basal cell cytokeratins (CK 5/15). Cx 32 expression was first observed between days 10-15 and colocalized to differentiated luminal cells (CK 8/18). Cx 43 and Cx 32 never colocalized to the same cell indicating that gap junction intercellular communication differs between basal and luminal prostatic cells. While epithelial connexin expression was not initially altered in the developing prostates following estrogen exposure, adult prostates of neonatally estrogenized rats exhibited a marked decrease in Cx 32 staining and an increased proportion of Cx 43 expressing cells. In the developing prostate, E-cadherin was localized to lateral surfaces of undifferentiated epithelial cells and staining intensity increased as the cells differentiated into luminal cells. By day 30, estrogenized prostates had small foci of epithelial cells that did not immunostain for E-cadherins. In the adult and aged prostates of estrogenized rats, larger foci with differentiation defects and dysplasia were associated with a decrease or loss in E-cadherin staining. The present findings suggest that estrogen-induced changes in the expression of E-cadherin, Cx32 and Cx43 may result in impaired cell-cell adhesion and defective cell-cell communication and may be one of the key mechanisms through which changes toward a dysplastic state are mediated. These findings are significant in light of the data on human prostate cancers where carcinogenesis and progression are associated with loss of E-cadherin and a switch from Cx32 to Cx43 expression in the epithelium.

Aging↗
PubMed2000

The influence of experimental muscle pain on motor unit activity during low-level contraction.

In the present study we compared motor unit (MU) activity in a painful extensor carpi ulnaris (ECU) muscle to that of a pain-free control. According to the pain adaptation model the activity of the painful ECU muscle may be inhibited and its antagonist activity increased during wrist extension performed as a pre-defined low-force ramp. The pre-defined low force may then be maintained by increased activity in the pain-free synergist muscles such as the extensor carpi radialis (ECR) muscle. Nine females (31-47 years old) participated in the study. Maximal voluntary contraction (MVC) of the wrist extensors was performed. A catheter was inserted into the ECU muscle to allow the injection of hypertonic saline to evoke muscle pain, and a concentric needle was inserted for the recording of MU activity. Surface electromyograms were recorded from a synergist and an antagonist (ECR and flexor carpi radialis) to the painful ECU muscle. A force ramp of isometric wrist extensions up to 10% MVC, with a force increase of 1% MVC x s(-1), were performed followed by 60 s of sustained contraction at 10% MVC. The number of MUs recruited was almost identical for baseline and with pain, and no effect of experimental muscle pain was found on the properties of the MUs (amplitude, area) or their firing characteristics (mean firing rate, firing variability) during low-force ramp contraction. During the sustained 10% MVC, no effect of pain was found for concentric or surface EMG of the forearm muscles. At low force levels no pain-induced modulations were found in MU activity, when the mechanical condition was similar to that of a control situation.

Adult↗
PubMed2000

Experimental muscle pain modulates muscle activity and work performance differently during high and low precision use of a computer mouse.

The aim was to investigate the influence of experimental muscle pain on performance and upper extremity muscle activity during occupational work requiring different levels of precision. Experimental muscle pain was induced by infusing hypertonic saline (0.3 ml, 5% NaCl) into the extensor carpi ulnaris (ECU) muscle. The same amount of isotonic saline was infused on a separate day to act as a control. Tasks requiring use of a computer mouse with high and low levels of precision were performed during the two sessions. Electromyographic (EMG) activity was measured from the ECU, the flexor carpi radialis (FCR) and the trapezius muscles. A group of 13 men participated in the study. Performance measured as work cycle time, cursor movements on the screen, and velocity of cursor movement were unaffected by muscle pain. The ECU muscle pain did not modulate EMG profiles of either the trapezius or FCR muscles either during high or during low precision work. During the low precision work the painful ECU muscle showed lower EMG activity in specific phases of the work cycle (highest activity phases) compared to the control session (P<0.05), whereas during the high precision work, experimental pain had no effect on the activity of the ECU muscle. In conclusion experimental muscle pain seems to modulate motor control differently depending on the precision level of the task. This may be of importance for our understanding of why some tasks lead to chronic musculoskeletal disorders.

Adult↗
PubMed2000

Acute response to precision, time pressure and mental demand during simulated computer work.

OBJECTIVES: The electromyographic (EMG) activity of shoulder and forearm muscles was recorded during a standardized computer task with different combinations of time pressure, precision demands, and mental demands to study the interaction of these factors and their effect on muscular response during simulated computer work. METHODS: The computer task lasted 5 minutes, and it was performed by 14 female computer-aided design (CAD) operators during 8 exposure combinations that differed with respect to time pressure, precision demand, and mental demand. Performance (number of produced drawings, mouse clicks, and errors) were recorded. The EMG activity was recorded from the trapezius, infraspinatus, deltoid, and extensor digitorum muscles. An electrogoniometer was used to measure wrist postures and movements. RESULTS: High time pressure (combined with low precision and low mental demands) resulted in higher EMG activity for all the muscles and in a small increase in the number of produced drawings. High precision demands caused a large reduction in the number of produced drawings, but not always a change in EMG activity. High precision demands and high mental demands led to no change or a reduction in muscle activity because the number of drawings was greatly reduced. CONCLUSIONS: The interaction between work pace and other exposure factors must be taken into account when the effects of changes in exposure demands on muscular response are predicted. Only then can it be predicted whether changing demands will constitute a risk of developing musculoskeletal disorders.

Adult↗
PubMed1999

Neonatal estrogen stimulates proliferation of periductal fibroblasts and alters the extracellular matrix composition in the rat prostate.

The purpose of this study was to examine whether changes in extracellular matrix (ECM) molecules are associated with the growth inhibition and differentiation defects of the prostate gland following neonatal exposure to estradiol. Using immunocytochemistry (ICC), laminin and collagen IV were localized to the basement membrane (BM) as well to the basal lamina of the periductal smooth muscle of the control developing prostates. In contrast, fibronectin and collagen III were localized throughout the stromal ECM. Exposure to neonatal estrogen altered the staining profile for specific ECM molecules. In the estrogenized rats, a thick layer of cells negative for laminin and collagen IV was observed adjacent to the BM. Electron microscopy and ICC for alpha-actin, fibronectin, and vimentin identified this multicellular layer of periductal cells as differentiated fibroblasts. Peripheral to these fibroblasts, actin-positive smooth muscle formed a second layer of periductal stromal cells. PCNA labeling showed that estrogen exposure increased the fibroblast proliferation. Because many periductal fibroblasts were positive for estrogen receptor alpha (ER alpha) in estrogenized rats, a direct effect of estradiol on their proliferation is suggested. Gelatinolytic gels revealed that estrogen exposure did not alter the activity of matrix metalloproteinases associated with tissue remodeling during prostate morphogenesis. However, the periductal fibroblast layer in estrogenized prostates was devoid of urokinase- and tissue-plasminogen activator, which may potentially alter the localized proteolysis involved in matrix remodeling. It is proposed that proliferation of a multicellular layer of periductal fibroblasts in estrogenized prostates results in a physical barrier that constrains branching morphogenesis and blocks paracrine communications between smooth muscle and epithelial cells which normally regulate differentiation.

Animals↗
PubMed1999

Neonatal estrogen exposure alters the transforming growth factor-beta signaling system in the developing rat prostate and blocks the transient p21(cip1/waf1) expression associated with epithelial differentiation.

Exposure of male rats to estrogens during the neonatal period retards prostate branching morphogenesis, blocks epithelial differentiation, and predisposes the adult prostate to hyperplasia and dysplasia. The mechanism of neonatal estrogenization is not well understood. The present study evaluated transforming growth factor-beta (TGFbeta) in the neonatally estrogenized ventral prostate to determine whether this paracrine/autocrine factor may in part mediate the effects ofestrogen on the developing prostate gland. Immunocytochemistry using antibodies against active TGFbeta1 and its latency-associated peptide localized this molecule to the periductal smooth muscle cells in the developing prostate. Although neonatal estrogenization increased the accumulation of total and active TGFbeta1 in the smooth muscle layer as early as day 6 of life, it was physically separated from the epithelial ducts by a proliferating layer of fibroblasts surrounding the basement membrane. RT-PCR demonstrated that alterations in TGFbeta1 levels were not due to alterations in TGFbeta1 transcription. TGFbeta2 and TGFbeta3 were primarily immunolocalized to differentiating epithelial cells in developing prostates, and this was markedly dampened between days 10-30 after neonatal estrogen exposure. Immunocytochemistry for TGFbeta signaling components revealed that neonatal estrogenization transiently reduced TGFbeta type I receptor levels in the prostate epithelium, but not in stroma, between days 6-15, whereas there was no effect on TGFbeta type II receptor. Levels of the intracellular signal Smad2 (52 kDa) were detected in epithelial cells but were not altered after estrogenization. To analyze the functional status of the TGFbeta signaling pathway, immunocytochemistry was performed for p21(cip-1/waf-1), a cyclin-dependent kinase inhibitor that is inducible by TGFbeta1 in the prostate. Transient nuclear localization of p21(cip-1/waf-1) was normally observed in epithelial cells between days 6-15 and was associated with entry of cells into a terminal differentiation pathway. Neonatal estrogenization prevented this transient expression of p21(cip-1/waf-1). The present findings demonstrate that the TGFbeta signaling system is perturbed at several levels in the estrogenized prostate, which may in part account for the epithelial cell differentiation blockade as well as the proliferation of periductal fibroblasts in this model.

Animals↗
PubMed1998

Estrogen receptor-beta messenger ribonucleic acid ontogeny in the prostate of normal and neonatally estrogenized rats.

Neonatal exposure to estrogens permanently alters rat prostate growth and epithelial differentiation leading to prostatic dysplasia on aging. The effects are lobe-specific, with the greatest response observed in the ventral lobe. Recently, a novel estrogen receptor (ER) complementary DNA was cloned from the rat prostate and termed ER-beta (ER beta) due to its high homology with the classical ER alpha. The protein possesses high affinity for 17beta-estradiol, indicating that ER beta is an alternate molecule for mediating estrogenic effects. Importantly, ER beta messenger RNA (mRNA) was localized to rat prostatic epithelial cells, which contrasts with the stromal localization of ER alpha in the rat prostate. The present study was undertaken to determine the ontogeny of ER beta mRNA expression in the rat prostate lobes and to examine the effects of early estrogen exposure on prostatic ER beta expression. Male rat pups were given 25 microg estradiol or oil on days 1, 3, and 5; were killed on day 1, 3 (oils only), 6, 10, 30, or 90; and prostate lobes were frozen. Longitudinal sections were processed for in situ hybridization using an 35S-labeled antisense mRNA probe corresponding to a 400-bp EcoRI-AccI fragment in the 5' untranslated region of rat ER beta complementary DNA. Image analysis was used to quantitate silver grains. In addition, total RNA was isolated from the ventral prostate (VP) and used for semiquantitative RT-PCR. Results from in situ hybridization revealed that at birth, ER beta was equivalently expressed at low levels in both mesenchymal and epithelial cells in oil-treated rats. From day 1 onwards, expression in all stromal cells slowly and significantly declined, so that in the control adult prostate, stromal ER beta mRNA was slightly above background. In the oil-treated control rats, epithelial ER beta mRNA increased to moderate levels between days 6-10 in the VP and days 10-15 in the dorsal and lateral lobes as cells began differentiation and ducts lumenized. A further significant increase in ER beta message was observed at day 30, which indicates that full epithelial ER beta expression may require the completion of functional differentiation. By day 90, expression levels were maximal and similar between the lobes. RT-PCR substantiated this developmental increase in ER beta between days 1-90. Neonatal exposure to estrogens did not have an immediate effect on prostatic ER beta mRNA levels as determined by in situ hybridization and RT-PCR. However, the marked increase in epithelial cell expression at day 30 observed in the control VP was dampened in the VP of animals exposed neonatally to estrogens. By day 90, the VP of estrogenized rats possessed low ER beta message levels compared with the high expression in oil controls. In contrast, the dorsal and lateral lobes of neonatally estrogenized rats possessed high levels of ER beta mRNA at day 90, equivalent to controls. The present data demonstrate that ER beta mRNA expression in the rat prostate is developmentally regulated, and that neonatal estrogen can affect this expression in the adult VP. Because the effect of neonatal estrogens was not immediate, the data imply that early estrogen exposure may not directly autoregulate ER beta expression, and suggests that the adult effects on ER beta mRNA expression may be indirect. The differences in ER beta mRNA imprinting in the separate lobes may account for or reflect the lobe-specific neonatal estrogen imprints previously observed in the rat prostate.

Age Factors↗