Androgen metabolism and actions in rat ventral prostate epithelial and stromal cell cultures

1986 ◽  
Vol 64 (6) ◽  
pp. 583-593 ◽  
Author(s):  
J. Orlowski ◽  
A. F. Clark
Keyword(s):  
Epithelial Cells ◽  
Cell Cultures ◽  
Stromal Cells ◽  
Stromal Cell ◽  
Cultured Cells ◽  
Cell Types ◽  
Ventral Prostate ◽  
Oxidoreductase Activity ◽  
Androgen Metabolism ◽  
Rat Ventral Prostate

The rat ventral prostate requires androgens for normal development, growth, and function. To investigate the relationship between androgen metabolism and its effects in the prostate and to examine differences between the epithelial and stromal cells, we have established a system of primary cell cultures of immature rat ventral prostate cells. Cultures of both cell types after reaching confluency (6–7 days) actively metabolized 3H-labelled testosterone (T), 5α-dihydrotestosterone (5α-DHT), 5α-androstane-3α,17β-diol, and 5α-androstane-3β,17β-diol. The epithelial cells actively reduced T to 5α-DHT and formed significant amounts of 5α-androstane-3,17-dione from T, 5α-DHT, and 5α-androstane-3α,17β-diol. All substrates were converted to significant amounts of C19O3metabolites. The stromal cells also metabolized all substrates, but very little 5α-androstane-3,17-dione was formed. The metabolism studies indicate that both cell types have Δ4-5α-reductase, 3α- and 3β-hydroxysteroid oxidoreductase and hydroxylase activities. The epithelial cells have significant 17β-hydroxysteroid oxidoreductase activity. The epithelial cells cultures grown in the presence of T have higher acid phosphatase (AP) contents (demonstrated histochemically and by biochemical assay). Tartrate inhibition studies indicate that the epithelial cells grown in the presence of T are making secretory AP. Stromal cell AP is not influenced by T. The results indicate that the cultured cells maintain differentiated prostatic functions: ability to metabolize androgens and, in the case of the epithelial cells, synthesize secretory AP.

scholarly journals Hyaluronan content and distribution in the rat ventral prostate after castration

2018 ◽  
Vol 96 (5) ◽  
pp. 556-563
Author(s):  
Heloisa H.M. Della-Colleta ◽  
Hernandes F. Carvalho
Keyword(s):  
Epithelial Cells ◽  
Tumor Growth ◽  
Reverse Transcription ◽  
Stromal Cells ◽  
Total Content ◽  
Ventral Prostate ◽  
Rt Pcr ◽  
Qrt Pcr ◽  
Hyaluronan Synthases ◽  
Rat Ventral Prostate

Hyaluronan (HA) has been implicated in tissue remodeling, healing, and tumor growth. This study investigated the variation in hyaluronan content, distribution, and metabolism in the rat ventral prostate (VP) in response to androgen deprivation after castration. The mRNA abundance of hyaluronan synthases (Has1–3) and hyaluronidases (Hyal 1–3) were assessed by reverse transcription (RT)–PCR and immunohistochemistry, respectively. The results demonstrated an increased concentration, but an overall reduction in HA content. HA was located in both epithelium and stroma of the prostate of both the noncastrated and castrated animals. Quantitative RT–PCR (qRT–PCR) showed that Has1 and Has2 are major synthases, and that Hyal 1 was the predominant hydrolase expressed in the VP. qRT–PCR also showed that Has1 and Has2 mRNA increased transiently after castration, whereas Has3 mRNA declined markedly. While Hyal 1 mRNA increased slowly up to day 21 after castration, Hyal 2 and Hyal 3 mRNA dropped significantly. CD44 was found in the epithelial cells and in some stromal cells in both hormonal conditions. In conclusion, castration results in increased abundance of Has1 and Has2 mRNA, but is associated with a decrease in the total content of HA, with an increased concentration, and a predominance of short-chain HA molecules.

Regional and cellular distribution within the rat prostate of two mRNA species undergoing opposite regulation by androgens

1992 ◽  
Vol 132 (3) ◽  
pp. 361-NP ◽  
Author(s):  
S. Bettuzzi ◽  
M. Zoli ◽  
F. Ferraguti ◽  
M. C. Ingletti ◽  
L. F. Agnati ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cell Types ◽  
High Rate ◽  
Ventral Prostate ◽  
Photographic Emulsion ◽  
Cell Multiplication ◽  
Rat Ventral Prostate ◽  
A Cell ◽  
Rat Prostate ◽  
Different Cell Types

ABSTRACT We have investigated, by in-situ hybridization histochemistry, the distribution within the rat ventral prostate of the mRNAs for sulphated glycoprotein-2 (SGP-2) and ornithine decarboxylase (ODC; EC 4.1.1.17), two proteins that appear to be inversely regulated by androgens in this organ, in that the level of SGP-2 mRNA is lowered, while the activity of ODC is enhanced by the latter hormones. Low-magnification autoradiograms of whole ventral prostate sections showed that, in intact animals, the SGP-2 transcript was only detectable in restricted areas and not diffused evenly throughout the section. Reciprocally, the ODC transcript was not detectable in areas where the SGP-2 transcript was detected, but appeared distributed uniformly in the remaining parts of the section. The effect of castration on the levels of the two mRNAs was evaluated by a semiquantitative analysis of autoradiograms from whole ventral prostate sections using an autoimmune image analyser. Four days after castration, SGP-2 mRNA increased by about 11-fold, while ODC mRNA decreased by fivefold. The distribution of the two mRNAs among the different cell types, studied by treating the slides with photographic emulsion and counterstaining, showed that both were expressed exclusively in the epithelial luminal cells of the ducts. Furthermore, each of the two mRNAs preferentially accumulated in a cell population which was morphologically distinct while accumulation of the other was negligible. SGP-2 mRNA was mostly found in cuboidal epithelial cells and ODC mRNA in columnar epithelial cells. Castration caused a dramatic accumulation of SGP-2 and a decrease in ODC mRNAs in the cells of the columnar epithelium 4 days later. These data suggest that, in the ventral prostate of intact animals, SGP-2 is expressed in the proximal ducts whose luminal epithelium consists of cuboidal cells undergoing apoptotic phenomena and not in the intermediate and distal ducts characterized by columnar epithelia with active protein synthesis and cell multiplication. In the intermediate and distal parts of the ductal system the ODC gene would be expressed at a high rate while being turned off in the proximal ducts. Castration, resulting in apoptosis of the epithelial cells of the intermediate and distal ducts, caused SGP-2 to be actively expressed and ODC to be repressed in the latter segments. Journal of Endocrinology (1992) 132, 361–367

Androgen 5α-reductase and 3α-hydroxysteroid dehydrogenase activities in ventral prostate epithelial and stromal cells from immature and mature rats

1983 ◽  
Vol 99 (1) ◽  
pp. 131-139 ◽  
Author(s):  
J. Orlowski ◽  
C. E. Bird ◽  
A. F. Clark
Keyword(s):  
Epithelial Cells ◽  
Stromal Cells ◽  
Reductase Activity ◽  
Age Groups ◽  
Cell Types ◽  
Hydroxysteroid Dehydrogenase ◽  
Ventral Prostate ◽  
Testosterone Metabolism ◽  
Immature Rats

To study androgen-mediated differentiation in the rat ventral prostate, we separated the two principal cell types (epithelial and stromal) derived from prostates of immature and mature rats on two continuous Percoll gradients. Cells were immediately placed in culture medium. Testosterone metabolism by the two prostatic cell types was evaluated using [3H]testosterone and quantifying the formation of 5α-[3H]dihydrotestosterone (5α-DHT) and 5α-[3H]androstane-(3α or 3β), 17β-diols. In epithelial cells from both immature and mature rat prostates the major testosterone metabolites were 5α-DHT and 5α-androstane-3α, 17β-diol. Stromal cells metabolized less testosterone than did the epithelial cells. Differences in the relative levels of the various metabolites were observed for the two age groups. To examine in more detail the changes in testosterone metabolism observed in vitro both types of cells and unfractionated cells from immature and mature rat prostates were assayed for testosterone 5α-reductase (using testosterone as substrate) and 3α-hydroxysteroid dehydrogenase (using 5α-DHT as substrate) activities (expressed as pmol substrate reduced/min per 106 cells). In immature rats both 5α-reductase and 3α-hydroxysteroid dehydrogenase activities were localized in the epithelial cell fraction (17 and 52 respectively); stromal cells showed lower 5α-reductase and 3α-hydroxysteroid dehydrogenase activity (4 and 4). Relative to epithelial cells from immature rats epithelial cells from mature rats showed a decrease in 5α-reductase (7) and an increase in 3α-hydroxysteroid dehydrogenase (160) activity while stromal 5α-reductase showed little change (3) and 3α-hydroxysteroid dehydrogenase increased to 22. Because there are more epithelial than stromal cells in the rat prostate, the former can be considered important sites for 5α-reductase and 3α-hydroxysteroid dehydrogenase activities. This contrasts with the human prostate where there is more 5α-reductase activity in the stroma than in the epithelium.

Stromal cell and human prostatic epithelial cell in-vitro co-coltures: Growth and morphology

1996 ◽  
Vol 63 (1_suppl) ◽  
pp. 65-68
Author(s):  
S. De Angeli ◽  
A. Fandella ◽  
C. Gatto ◽  
S. Buoro ◽  
C. Favretti ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Stromal Cells ◽  
Marked Reduction ◽  
Stromal Cell ◽  
Optical Microscope ◽  
Neutral Red ◽  
Cellular Growth ◽  
Murine Fibroblasts ◽  
Prostatic Epithelial Cell

A study was carried out on the effect of stroma-epithelium interaction on cellular growth and morphology in co-coltures of U285 prostatic epithelial cells with human prostatic and esophageal stromal cells and with murine fibroblasts of the 3T3-J2 line. The proliferation rate was determined by growth tests of neutral red and kenacid blue. Morphological observations were made under optical microscope on the same cultures used for the growth tests. Results highlighted a marked reduction in cellular growth in the co-cultures compared to control cultures, as well as the tendency of the stromal and epithelial cells to re-organise themselves in pseudo-acinous structures.

Ca2+-activated Cl? channel currents in rat ventral prostate epithelial cells

The Prostate ◽  
2003 ◽  
Vol 55 (2) ◽  
pp. 118-127 ◽  
Author(s):  
Sung Joon Kim ◽  
Sun Young Shin ◽  
Ji Eun Lee ◽  
Jun Hee Kim ◽  
Dae-Yong Uhm
Keyword(s):  
Epithelial Cells ◽  
Ventral Prostate ◽  
Cl Channel ◽  
Prostate Epithelial Cells ◽  
Rat Ventral Prostate ◽  
Channel Currents

Contact behaviour during the reassociation of dissociated epithelial cells in primary culture

1987 ◽  
Vol 88 (4) ◽  
pp. 521-526
Author(s):  
R.M. Brown ◽  
C.A. Middleton
Keyword(s):  
Epithelial Cells ◽  
Chick Embryo ◽  
Primary Culture ◽  
Cell Cultures ◽  
Corneal Epithelium ◽  
Time Lapse ◽  
Cell Types ◽  
Epidermal Cells ◽  
Isolated Cells ◽  
Contact Behaviour

The behaviour in culture of dissociated epithelial cells from chick embryo pigmented retina epithelium (PRE), corneal epithelium (CE) and epidermis has been studied using time-lapse cinematography. The analysis concentrated on the contact behaviour of 60 previously isolated cells of each type during a 24 h period starting 3.5 h after the cells were plated out. During the period analysed the number of isolated cells in cultures of all three types gradually decreased as they became incorporated into islands and sheets of cells. However, there were significant differences in behaviour between the cell types during the establishment of these sheets and islands. In PRE cell cultures, islands of cells developed because, throughout the period of analysis, collisions involving previously isolated cells almost invariably resulted in the development of a stable contact. Once having established contact with another cell these cells rarely broke away again to become reisolated. In contrast the contacts formed between colliding CE and epidermal cells were, at least initially, much less stable and cells of both these types were frequently seen to break away and become reisolated after colliding with other cells. Sheets and islands of cells eventually developed in these cultures because the frequency with which isolated cells become reisolated decreased with increasing time in culture. The possible reasons underlying the different behaviour of PRE cells, when compared with that of CE and epidermal cells, are discussed. It is suggested that the decreasing tendency of isolated CE and epidermal cells to become reisolated may be related to the formation of desmosomes.

Cultured epithelial cells derived from human foetal pancreas as a model for the study of cystic fibrosis: further analyses on the origins and nature of the cell types

1988 ◽  
Vol 90 (1) ◽  
pp. 73-77
Author(s):  
A. Harris ◽  
L. Coleman
Keyword(s):  
Cystic Fibrosis ◽  
Tissue Culture ◽  
Epithelial Cells ◽  
Culture System ◽  
Cultured Cells ◽  
Cell Types ◽  
Cultured Epithelial Cells ◽  
Different Cell Types ◽  
Foetal Pancreas

The establishment of a tissue-culture system for epithelial cells derived from human foetal pancreas has recently been reported. Further analyses have now been made on these cells in vitro, together with parallel investigation of the distribution of different cell types within the intact foetal pancreas. Results support the view that the cultured cells are ductal in origin and nature. Pancreatic epithelial cell cultures have also been established from foetuses with cystic fibrosis.

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