Morphometric and stereological analysis of the effects of 17 beta-estradiol on the glandular epithelium of the castrated guinea pig lateral prostate.

Upon administration of pharmacological doses of estradiol to castrated guinea pigs, the secretory cells of the lateral prostate underwent hypertrophy which resulted from significant increases in nuclear and cytoplasmic volume. There were quantitative increases in the small highly electron-dense granules and multivesicular bodies when compared with the castrated control. The dramatic increase in the number of highly electron-dense granules probably occurred at the expense of the low electron-dense granules. The average size of the condensing granules and mitochondria decreased significantly after estradiol administration. However, significant increase in the number of mitochondria was detected when compared with the castrated control. Ultrastructural data revealed no significant changes in the absolute dimensions of granular endoplasmic reticulum or of the Golgi complex, suggesting that estradiol exerted no significant stimulatory effects on these organelles. Pharmacological doses of estrogen appear to regulate the expression of secretory granules and multivesicular bodies in the lateral prostate of castrated guinea pigs.     

Estrogen-induced morphological and immunohistochemical changes in stroma and epithelium of rat ventral prostate.

Prostatic smooth muscle cells have been regarded to play a major pathogenetic role during the development of benign prostatic hyperplasia (BPH) in elderly men. Altered hormonal signals (increased estrogen) have been made responsible for the "metabolic" transformation of prostatic smooth muscle cells, which were thought to produce increased amounts of connective tissue fibers observed in BPH. In order to find out the role of metabolically "activated" smooth muscle cells, hormone stimulation experiments were performed in male rats. The effects of androgen deprivation and estrogen stimulation were recorded by semiquantitative analysis of intermediate and myofilament proteins in stromal smooth muscle cells. In castrated or estrogen-treated or estrogen-treated and castrated animals, the reduction of the glandular lumen is the most obvious morphological alteration, accompanied by an increase in connective tissue. Regressive changes occurred most rapidly in castrated animals (already within the first week), slower in castrated estrogen-treated animals and still slower in normal estrogen-treated animals. Regression of the epithelium was accompanied by a marked decrease in immunoreactivity for prostatic binding protein (PBP) in castrated animals, while PBP immunoreactivity in estrogenized animals was retained for up to 6 weeks. Smooth muscle cells became atrophic in castrated animals. This effect was attenuated in estrogen-treated animals. There was no indication for enhanced collagen synthesis by smooth muscle cells. Actin and desmin-immunoreactivity were only slightly altered in experimental animals and showed a changed distribution pattern. Prostatic smooth muscle cells respond less markedly to hormonal alterations than do the fibroblasts.     

Cell proliferation studies in the rat prostate: II. The effects of castration and androgen-induced regeneration upon basal and secretory cell proliferation

The changes over short and prolonged periods (up to three months) after castration on the proliferative activity of basal and secretory epithelial cells in the rat prostate were studied. Although castration induced widespread apoptosis of the secretory cells, no compensatory hyperplasia of the basal cells in response to this was noted. Instead, observations of the cell kinetics and ultrastructure suggested that both the basal and secretory cells entered a quiescent state as a result of castration. The proliferative potential of secretory cells was not diminished up to three months after castration. During androgen-induced regeneration of the prostate the pattern of basal and secretory cell proliferation was found to be similar to that observed during normal growth, although it was more rapid and of shorter duration.     

Effects of cis-4-hydroxy-L-proline on the androgen-lnduced growth of the prostate of the prepubertally castrated guinea pig

The present study examined the effects of cis-4-hydroxy-L-proline (CHP), a proline analog, on the androgen-induced growth of the lateral prostate of prepubertally castrated guinea pigs. Prepubertal male guinea pigs were castrated at the age of 3 weeks and allowed to recover completely before subjection to an experimental regime to alter the stromal collagen synthesis by CHP. The animals were kept on a special proline-deficient diet (PDD) for a week before the subcutaneous injection of CHP (200 mg/kg/day) for 3 days, followed by a combined injection of CHP and dihydrotestosterone (DHT) (10 mg/kg/day) for 10 days. Control animals were injected with saline and DHT only. At the end of the experiment, the lateral prostate was removed and examined by 1) conventional transmission electron microscopy (TEM), 2) staining of proteoglycans (PGs) by Cuprolinic Blue (CB) using the critical electrolyte concentration (CEC) method, 3) carbohydrate and lectin histochemistry, and 4) electron microscopy (EM) lectin-gold labelling. The results showed that the wet weight of prostate from CHP-treated animals was significantly lower than the control and recovery groups. The epithelium was low columnar with an obvious increase in intercellular spaces and number of basal cells. The glandular cells showed little secretory activity with a decrease in number of granular endoplasmic reticulum (GER) profiles, secretory granules, and a small Golgi apparatus. The stroma was composed of stromal cells separated by large intercellular spaces with very sparse collagen fibrils, and a decrease in stromal PGs especially those PGs normally associated with collagen fibrils. CHP treatment also caused perturbation and disorganization in the epithelial basement membrane. The results suggested that stromal collagen is essential in mediating the response of glandular cells to DHT stimulation. Defective stromal collagens hamper the responsiveness of prostatic gland to androgen. © 1993 Wiley-Liss, Inc.     

Comparative characterization of the canine normal prostate in intact and castrated animals

BACKGROUND: Prostate diseases in the dog are generally regarded as representative for their human counterparts. We characterized the normal canine prostate in comparison to the normal human prostate. METHODS: Prostates of dogs were examined histomorphologically and by immunohistochemical detection of the markers CK14, HMWCK, CK5, CK18, CK7, UPIII, PSA, and PSMA. RESULTS: Histomorphologically, the canine prostate lacks the human zonal differentiation, has much more prominent acini, while comprising less stromal tissue. In general, the canine prostate epithelium displayed a highly differentiated character, with no cells expressing CK14, minimal amounts of cells expressing HMWCK/CK5 and the vast majority of cells expressing CK18 and PSA. After castration, the prostate epithelium regressed, and the remaining tubules were largely populated by cells showing a ductal phenotype (HMWCK+/CK5+/CK18+/CK7+). CONCLUSIONS: The human and canine prostate are histologically differently organized. The general scheme of cellular differentiation of the prostate epithelium may however be applicable to both species.     

Cytochemical characterization of cuprolinic blue-stained proteoglycans in the epithelial-stromal interface of the guinea pig lateral prostate

Three types (T1, T2, T3) of proteoglycan (PG) filaments, as demonstrated by cuprolinic blue (CB) under critical electrolyte concentration method in the epithelial-stromal interface of the guinea pig lateral prostate, were characterized cytochemically by using a number of glycosaminoglycan(GAG)-degrading enzymes and nitrous acid. The results showed that T1 filaments located in basement membranes of the epithelium, endothelium, and smooth muscle cells, were removed by nitrous acid, heparitinase, and pronase but resistant to chondroitinase (Ch)-ABC and Ch-AC, heparinase, neuraminidase, and Streptomyces (S) hyaluronidase. The T1 filaments, therefore, contain heparan sulfate. The T2 filaments closely linked to collagen fibrils were removed by Ch-ABC, Ch-ABC plus S-hyaluronidase, and pronase but were resistant to nitrous acid, heparitinase, heparinase, neuraminidase, and S-hyaluronidase. These show that T2 filaments are rich in dermatan sulfate. The T3 filaments in the interstitial spaces and on the surface of fibroblasts were removed by Ch-ABC, Ch-AC, and pronase but were resistant to heparitinase, heparinase, hyaluronidase, neuraminidase, and nitrous acid. They are, therefore, rich in chondroitin sulfate.     

Effect of p-Nitrophenyl-β-D-xylopyranoside (β-D-xyloside) on the androgen-induced growth of the lateral prostate of the prepubertally castrated guinea pig

The aim of this study was to examine the effects of β-D-xyloside (XYL), a compound which interferes with stromal proteoglycan (PG) synthesis, on androgen induced growth of the lateral prostate (LP). Young male guinea pigs were castrated at 3 weeks of age and divided into three groups 6 weeks after castration. In group one, the animals were injected subcutaneously daily with 80 mg/kg of XYL, followed 3 days later by a daily dose of 10 mg/kg of dihydrotestosterone (DHT) for 2 more weeks. The second group served as control and received DHT only. In the third group, animals were treated first with XYL, like those in group one, and then followed by DHT alone for 2 weeks to check reversibility of the XYL effect. At the end of the experiment, the lateral prostate was removed and processed for morphological and cytochemical examination. The results showed that XYL inhibited the DHT stimulated growth of the lateral prostate. The fibroblasts showed a dilated granular endoplasmic reticulum filled with granular substances. In the interstitial spaces, there was a drastic increase in Cuprolinic Blue (CB) positive filaments and polygonal granules believed to be PGs or glycosaminoglycans (GAGs). Their number was much greater than the control. The distribution and density of the collagen fibers appeared similar to the control. The secretory alveoli were lined by epithelium with few secretory granules of low electron density and a larger number of clear vesicles. There was a slight reduction in glycoconjugate reactivities in the epithelial cells. The lectin binding patterns and the structural features were comparable between the control and recovery groups, indicating the XYL effects were reversible. The results suggest that stromal PG biosynthesis may play a role in epithelial function and an altered stromal matrix would hamper the effects of DHT on the target organ. © 1993 Wiley-Liss, Inc.     

Ultrastructural localization of proteoglycans by cationic dyes in the epithelial-stromal interface of the guinea pig lateral prostate

Proteoglycans (PGs) in the epithelial-stromal interface of the guinea pig lateral prostate were localized at ultrastructural level, using cuprolinic Blue (CB), alcian Blue (AB), and ruthenium red (RR). After staining with CB or AB according to the critical electrolyte concentration method (CEC), PGs appeared as short electron-dense filaments. According to their sizes and location, three type (T1, T2, T3) of CB-stained filaments were identified. T1 filaments were short (25 nm) and were found on both sides of the lamina densa of the basal laminae of the prostatic epithelium, smooth muscle cells, and capillary endothelial cells. They were regularly spaced with an interval of 60 nm. T1 filaments were more randomly distributed in the lamina densa. T2 CB filaments were approximately 30-40 nm long and closely associated with the collagen fibrils. They were usually arranged perpendicular to the long axis of collagen fibrils also at intervals of 60 nm. T3 filaments were found in different regions of the lamina propria, including: 1) reticular layer (pars fibroreticularis) below the basal lamina; 2) interstitial spaces; 3) closely associated with the cell surfaces of fibroblasts; and 4) around the collagen fibrils. Their sizes were variable (60-100 nm) and more densely stained. AB revealed similar patterns of PG distribution, except that the three types of PG filaments were longer but thinner. When the tissues were stained with RR, or RR-AB combined, PGs appeared as dense granules of various sizes, instead of filaments. Their locations and distributions were similar to those of the CB filaments, except that in the case of combined RR-AB treatment the PG granules were linked by a fine filamentous network, suggesting the interconnecting nature of the PGs and other extracellular components.     

Androgen and estrogen effect on guinea pig seminal vesicle muscle: a combined stereological and biochemical study

A combined electron microscopic stereological and biochemical study of the smooth muscle cells of guinea pig seminal vesicles was performed in intact, castrated, castrated and dihydrotestosterone- or estradiol-treated adult animals. Castration led to cell atrophy as determined stereologically by a decreased single cell volume and biochemically by no change in DNA content coupled with an increase in the DNA concentration. Treatment of castrates with dihydrotestosterone restored both the stereological and biochemical parameters of the cell size to slightly supranormal levels. The estrogen-induced increase in muscle weight and DNA content appeared to be due only to hyperplasia of muscle cells and not to a proliferation of fibroblasts or to infiltration by inflammatory cells. In all treatment groups, including the estrogen-treated castrates, more than 95% of the cells in the tissue were smooth muscle cells, and there was no evidence that polyploidy contributed to changes in DNA levels. In addition, in the estrogen-treated muscles, DNA concentration remained high, and the stereologically determined cell size remained low. Therefore, both morphological and biochemical evidence indicate that androgen induces hypertrophy, whereas estrogen induces hyperplasia of muscle cells. The correction of stereological and biochemical data validates the application of stereological cell size determination for smooth muscle cells in organs that hardly can be separated into stromal and epithelial components; eg, the prostate.     

Determination of serotonin and 5-hydroxyindoleacetic acid in guinea pig and human prostate using HPLC

The finding of significant numbers of endocrine-paracrine (EP) cells in the prostate glands of guinea pigs and man suggests that these cells may be important in the regulation or modulation of prostatic function. Serotonin is a biogenic amine common to most prostatic EP cells. In order to extend current knowledge relating to these cells, an assay was developed using high-performance liquid chromatography to quantitate serotonin and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) in guinea pig and human prostatic tissue extracts. Levels of serotonin and 5-HIAA in the guinea pig whole-gland preparation were 105.4 +/- 70.6 ng/g and 48.4 +/- 95.7 ng/g, respectively. Normal human prostatic tissue contained 1423.9 +/- 750.8 ng/g serotonin and 66.7 +/- 92.8 ng/g 5-HIAA. Recoveries ranged between 60 and 100%. The detection limits were 24 pg/injection for serotonin and 12 pg/injection for 5-HIAA. This assay provides an expeditious, specific and highly sensitive method for the simultaneous determination of monoamines in guinea pig and human prostatic tissue.     

Cell proliferation studies in rat prostate. I. The proliferative role of basal and secretory epithelial cells during normal growth

In the prostates of rats ranging from 10 days to 8 months of age, the proliferative activity of basal and secretory epithelial cells was studied. No clear evidence was found that basal cells alone represented the proliferative compartment, or that they were responsible for the replacement of secretory cells during normal turnover. Cell kinetic and morphological evidence indicated that basal and secretory cells were self-replicating cell types with discrete functions.     

Spontaneous slow wave and contractile activity of the guinea pig prostate

PURPOSE: We characterized the electrical events underlying spontaneous contractions of the stroma of the guinea pig prostate. MATERIALS AND METHODS: Membrane potential of the stroma was recorded using standard electrophysiological recording techniques. The structure of the prostate was viewed using confocal or electron microscopy. RESULTS: In stromal cells spontaneous depolarizing membrane transients (12 mV. in amplitude) occurred at 5 minutes-1 and triggered 1 or more spikes. The membrane potential, and frequency and duration of the potential transients were not affected by the calcium channel blocker nifedipine (1 microM. for greater than 5 minutes), or blockers of neuronal propagation (tetrodotoxin), and the effects of cholinergic (atropine), adrenergic (guanethidine or prazosin) and sensory blockers (capsaicin) of neurotransmission. However, the amplitude of the superimposed spikes was significantly reduced by nifedipine. A network of c-Kit immunoreactive cells was evident in the interstitial layer between the stroma and glandular lined lumen. These prostatic interstitial cells contained many morphological features distinguishing interstitial cells of Cajal, the pacemaker cells of the gastrointestinal tract. Prostatic interstitial cells formed close appositions with each other, with neighboring smooth muscle cells and with varicose axon bundles. CONCLUSIONS: Movement of the contents of the peripheral prostatic acini into the minor and major prostatic ducts is likely to occur via spontaneous contractions triggered by myogenic slow wave activity in the stromal wall. By analogy with the intestine and urethra prostatic interstitial cells may well act as the pacemaker for prostatic slow waves as well as form an intercellular communication network interacting with the intrinsic nerves and stromal cells.     

Intermediate basal cells of the prostate: in vitro and in vivo characterization

BACKGROUND: Progenitor cells within the prostate basal layer may play important roles in differentiation and carcinogenesis; however, prostate stem cell populations remain uncharacterized. METHODS: Immunohistochemical and immunoblot analyses were used to characterize prostate epithelial cells (PrEC), a commercially available prostate basal cell isolate. RESULTS: Proliferating PrECs exhibited immunophenotypic characteristics most consistent with basal cells, but during senescence PrECs up-regulated androgen receptor (AR) mRNA, p27, and low-molecular-weight cytokeratin (LMWCK) expression, suggestive of partial differentiation. PrECs also stained strongly for involucrin, which marked a subset of intermediate prostate basal cells in vivo. Basal hyperplasia consisting of involucrin-positive cells was prevalent in prostate tissue from androgen-ablated patients, and formed epithelial clusters flanked by involucrin-negative basal and luminal monolayers. Cultivation of PrECs on matrigel together with androgen-treated stromal conditioned media resulted in dense aggregates, with a peripheral rim of basal-like cells expressing p63 and basal cytokeratins. CONCLUSIONS: PrEC represents an epithelial population whose basal characteristics are modified in response to matrigel, stromal factors, and senescence, consistent with a transient amplifying population. These cells may derive from a previously unrecognized, involucrin-positive subset present in vivo.     

Thyrotropin-releasing hormone (TRH) activates the adenylyl cyclase of nonsecretory cells in the rat ventral prostate

Prostatic secretory and basal or stem cells were isolated from rat ventral prostate lobes by collagenase dispersion and density centrifugation in a Percoll gradient. The membrane-bound adenylyl cyclase of secretory cells could be activated in a dose-dependent manner by vasoactive intestinal peptide (VIP ED₅₀ 10^?7M) but not thyrotropin-releasing hormone (TRH). Conversely, only TRH could significantly stimulate the adenylyl cyclase in basal cell membranes (ED₅₀ 5 × 10^?7). In two separate studies enzyme activity was stimulated seven- and 13-fold by this peptide. This action of TRH on prostatic basal cells supports previous reports that high levels of immunologically active TRH have been found in prostate tissue and that TRH stimulates the growth of prostatic cancer cells in vitro.     

豚鼠膀胱Cajal样间质细胞类型及其与钙离子振荡特性的关系

目的 探讨豚鼠膀胱组织中Cajal样间质细胞(ICC)形态学类型特点及其与钙离子振荡特征的关系.方法 取豚鼠膀胱制作冰冻切片,体外培养膀胱ICC.免疫荧光染色激光共聚焦显微镜下观察ICC形态学特点,根据细胞不同形态类型及添加2-氨基乙氧基双苯萘酯(2-APB)干预剂将体外培养膀胱ICC分4组:二聚体ICC组(n=10)、单体ICC组(n=20)、2-APB抑制二聚体ICC组(n=10)、2-APB抑制单体ICC组(n=20).应用Fluo-4AM标记细胞内钙离子浓度含量及在2-APB抑制二聚体ICC组、2-APB抑制单体ICC组加入2-APB,共聚焦显微镜下观察记录钙离子振荡功能.结果 二聚体ICC组钙振荡频率(13.4±0.8)次/min及振幅38.5±8.7明显高于单体ICC组钙振荡频率(7.5±0.6)次/min及振幅10.5±3.9(P＜0.05),加入高浓度2-APB后,2-APB抑制二聚体ICC组钙振荡频率(8.7±1.5)次/min及幅度15.5±8.2明显低于二聚体ICC组(P＜0.05),而2-APB抑制单体ICC组钙振荡频率(8.5±0.5)次/min及幅度9.2±4.6与单体ICC组差异无统计学意义(P＞0.05).结果 豚鼠膀胱中存在单体和二聚体2种不同形态类型的ICC,单体ICC自发性钙振荡波较低,二聚体ICC自发性钙振荡波具有高兴奋性,二聚体ICC的特有结构可能是ICC高兴奋性钙波形成的结构基础.     

Cajal样间质细胞在豚鼠膀胱肌层的分布及意义

目的研究豚鼠膀胱Cajal样间质细胞的分布、超微结构特点并探讨其功能。方法选择健康豚鼠10只,利用激光共聚焦显微镜和透射电镜观察膀胱Cajal样间质细胞在肌层的分布和结构特点。结果豚鼠膀胱肌层Cajal样间质细胞形成网状和平行状两种不同的分布。超微结构核大、具有丰富的细胞器。结论膀胱Cajal样间质细胞分布和超微结构与胃肠道的ICCs相似,具有起搏细胞特征。