Concentration of elastic system fibers in the corpus cavernosum, corpus spongiosum, and tunica albuginea in the rabbit penis.

The corpus cavernosum (CC) extracellular matrix is essential for normal penile erection and is implicated in erectile dysfunction. Although investigations of these issues have used the rabbit CC, organization of its components is not well known to date. We characterized and quantified the volumetric density (Vv) of the elastic system fibers in the corpus spongiosum (CS), CC and tunica albuginea (TA) of the rabbit penis. Adult New Zealand rabbits (n = 10) were used. The penile mid-shaft fragments were fixed with 4% phosphate-buffered formalin solution and/or Bouin's liquid for 24-48 h, and processed using standard histological techniques. The sections were stained with Weigert's Fucsin-Resorcin with previous oxidation. The elastic system fibers Vv (%) was determined in 25 random fields of each fragment, using the M-42 test grid. The histochemical methods detected elastic system fibers in CS, CC and TA of all animals. The Vv of elastic fibers average was 25.03+/-2.0% for CC, 32.23+/-1.41% for CS and 22.38+/-3.61% for TA. Results for CC and CS were not significantly different. The great amount of elastic fibers distribution beneath the endothelium suggest that these fibers may have an important role in the erection process in rabbits. The present data should therefore provide important information for devising experiments and interpreting results when using the rabbit penis as a model for penile dysfunctions, especially when making comparisons with humans.

Immunohistochemical analysis of smooth muscle cells and volumetric density of the elastic system fibers of wild boar (Sus scrofa) penis.

The purpose of the present study was to verify the smooth muscle cell distribution and elastic system fibers volumetric density (Vv) in the corpus spongiosum and corpus cavernosum of the wild boar penis. Adult wild boars (n=13) were used. The penile mid shaft fragments were fixed with 4% phosphate buffered formalin solution and/or Bouin's liquid during 24-48 h, and processed using standard histological techniques. The sections were stained with Weigert's Resorcin-Fucsin with previous oxidation. The elastic system fibers Vv was determined in 25 random fields of each fragment using M42 test system. For immunohistochemical analysis, monoclonal anti-alpha actin smooth muscle was used. The histochemical methods detected elastic system fibers in both corpus spongiosum and corpus cavernosum of all animals. The elastic fibers Vv average was 36.6%+/-0.9 for corpus spongiosum and 11.7%+/-0.5 for corpus cavernosum. Through immunocitochemistry, a small quantity of smooth muscle cells was visualized in intimate relationship with blood vessels wall. The great amount of elastic fibers and the smooth muscle cell distribution beneath the endothelium suggest that these fibers may have an important role in penile erection process in the penis of wild boars.

An update of the stereology of the myocyte of the baboon's heart: analysis of the crista terminalis, interatrial and interventricular septa, and atrioventricular bundle.

We used light microscopy to compare the stereology of the myocytes from the following regions of the baboon's heart: a) cardiac walls (crista terminalis, interatrial and interventricular septa) and b) cardiac conduction tissue (atrioventricular bundle). Six hearts were perfused and several pieces of tissue were taken from these regions. A staggered test lattice consisting of cycloids was superimposed on twelve random pairs of photomicrographs in order to make disectors, and on twelve other random fields for each cardiac region in order to estimate certain stereological parameters. Statistically significant differences were found when comparing the number of myocytes from the crista terminalis (highest value) with either the interatrial or interventricular septum. There is also a significant difference between the number of myocytes in the interventricular septum and the atrioventricular bundle (highest value). For the mean profile area of the myocytes, significant differences were found between the myocytes of both the crista terminalis and the interatrial septum, and those of the interventricular septum (highest value). Under the light microscope, myocytes from the two regions of the atrial wall studied were indistinguishable in size, since they constitute a single cell population. It was also impossible to distinguish them from the atrioventricular cells. The myocytes from the interventricular septum differed in both size and number from those of the right atrium, but only in number from those of the atrioventricular bundle.

A rare variation in the high division of the sciatic nerve surrounding the superior gemellus muscle.

The sciatic nerve normally leaves the pelvis by passing through the greater sciatic foramen below piriformis. However, it may divide into its common fibular and tibial nerve components within the pelvis and its relationship with piriformis is variable. In this paper, we describe a new anatomical variation in which the common fibular nerve passed superior, and the tibial nerve inferior, to the superior gemellus muscle. Anatomical variations such as these may contribute to piriformis syndrome, coccygodynia and muscle atrophy.

[Anatomic and quantitative study of the human pterygopalatine ganglion. Morphometry and stereology]. / Estudo anatômico e quantitativo do gânglio pterigopalatino humano. Morfometria e estereologia.

The pterygopalatine ganglion is important in the regulation of the intraocular pressure and in the cerebral vasodilatation connected with headache of vascular origin. Four human ganglia were dissected, fixed in formalin and serially sectioned with a 6 microns thickness. The volume of the ganglion was calculated by point-counting and stereological parameters were determined using the test-system M42 with light microscopy. The PG volume was (mean +/- standard error of the mean) 5.6 +/- 0.5 mm3. The volume density of neurons was 51.1% +/- 3.4%, and the unitary volume of the neurons was 41,200.0 +/- 2,250.0 microns. The numerical density was 12,600.0 +/- 677.0 neurons by mm3, therefore approximately 70,560 neurons by ganglion.

[Analysis of prostate growth during human fetal period]. / Analyse de la croissance prostatique au cours de la période foetale humaine.

The objective of this study was to present quantitative data concerning prostatic growth during the foetal period (gestational age: 13 to 36 weeks) and to provide normal curves of the growth of the prostatic volume correlated with foetal age and weight, using the allometric method. This study was performed on 45 non-fixed human male foetuses, in a good state of preservation and not presenting any congenital malformations. The gestational age of the foetuses ranged from 13 to 36 weeks. Analysis of correlations showed that growth curves presented an angle less than 45 degrees, indicating that growth of the foetal prostate is slower than that of the individual as a whole. The authors also found a statistically significant correlation (p < 0.001) between prostatic volume and foetal weight during the foetal period.

Morphology of the fetal bladder during the second trimester: comparing genders.

OBJECTIVES: The aim of the present study was to determine, by histological and stereological analysis, whether there are between-gender structural differences in the bladder in the second gestational trimester in human fetuses. MATERIAL AND METHODS: Forty bladders, which were obtained from 40 human fetuses (20 males and 20 females) ranging in age from 13 to 23 weeks post-conception (WPC), were studied. The fetuses were macroscopically well preserved, without anomalies of the urinary and genital systems; the cases with syndromes were abandoned. The bladders were dissected and embedded in paraffin, from which 5-µm thick sections were obtained and stained with: Masson's trichrome, to quantify connective and smooth muscle tissue; Weigert's resorcin fuchsin, to observe elastic fibers; picrosirius red with polarization, to observe collagen; and anti-beta III tubulin antibody, to observe the bladder nerves. The images were captured with an Olympus BX51 microscope and Olympus DP70 camera. The stereological analysis was performed with the Image Pro and Image J programs, using a grid to determine volumetric densities (Vv). Means were statistically compared using simple linear regression and the paired t-test (P<0.05). RESULTS: The fetuses weighed between 60 and 490 g, and had crown-rump lengths between 9.5 and 20.4 cm. No elastic system fibers were observed in any bladders. Quantitative analysis indicated no differences in the Vv of the smooth muscle cells in the male bladders (26.19-50.16%; mean=35.66%) compared to the female ones (30.60-45.63%; mean=38.73%) (P=0.740) and there were also no differences in the Vv of the connective tissue in females (40.52-60.40%; mean=50.69%) and males (38.84-70.16%; mean=57.04%) (P=0.0506). There were no differences observed in the distribution of the nerves and collagen between the genders. CONCLUSION: The histological analysis of the smooth muscle, collagen, nerves and connective tissue of the developing bladders revealed that there are no gender differences during weeks 13-23 of gestation.

Protective effects of nutritional supplementation with arginine and glutamine on the penis of rats submitted to pelvic radiation.

Radiotherapy is widely used to treat pelvic malignancies, but normal tissues near the target tumour are often affected. Our aims were thus to determine whether the structural organization of the rat penis is altered by radiation, and whether supplementation with L-arginine (ARG) or L-glutamine (GLN) would have protective effects against these alterations. Groups of rats were treated with: no intervention (CONTR); pelvic radiation, followed by sacrifice 7 (RAD7) or 15 (RAD15) days later; and pelvic radiation, daily supplementation with ARG or GLN, followed by sacrifice 7 (RAD7+ARG, RAD7+GLN) or 15 (RAD15+ARG, RAD15+GLN) days after radiation. Structural components in the corpus cavernosum (CC), tunica albuginea of the corpus spongiosum (TACS) and urethral epithelium (UE) were analysed using stereological and immunohistochemical methods. The results showed that in the CC, connective tissue was increased by 18% in RAD15 (p < 0.04), but this change was partially prevented in RAD15+GLN (p < 0.05) and RAD15+ARG (p < 0.04). The fibrous matrix of the CC trabeculae stained evenly for collagen type I. In RAD15, the intensity of the labelling was increased, whereas in RAD15+GLN and RAD15+ARG the staining was similar to that of CONTR. No staining changes were seen in the groups that were sacrificed 7 days after radiation. Cavernosal elastic fibre content in RAD15 was increased by 61% (p < 0.004), and this was prevented in RAD15+ARG (p < 0.004) but not in RAD15+GLN. In TACS, the amino acids protected (p < 0.02) against the radiation-induced 92% increase in elastic fibre content, but only in RAD15. Cell density in the UE, as well as UE thickness, were reduced by 30% in RAD15 (p < 0.004), and there were protective effects of both amino acids. In conclusion, radiation-induced alterations in penile structures tend to be more pronounced 15 days after radiation session. Both ARG and GLN have protective effects against these changes, with the former being slightly more effective.

Fine structure and quantitative study in the otic ganglia of the rat.

Observations were concentrated on the ultrastructure of perikarya and satellite cells of the otic ganglion of the adult rat. Characteristics of both cell types were morphologically analysed as well as their relationship. Quantitative data concerning the volumetric density were calculated for the following elements of the ganglion: glial cells together (28.5%), unmyelinated fibres (13.4%), myelinated fibres (2.7%), connective tissue (13.6%). This electron microscopic study is a sequence of our previous light microscopic study which determined neuronal densities (COSTA, MANDARIM-DE-LACERDA and BAUER, in press).

Stereological analysis of the otic ganglia in adult rat: light microscopic study.

Quantifications in parasympathetic ganglia are not available in literature. The otic ganglia of 9 Wistar rats were studied (thick and semi-thin sections) in order to determine the following quantitative data: ganglionic volume (GV), neuronal volume density (Vv), neuronal cellular surface per cell volume (Sv), and neuronal numerical density (Nv). The results showed (mean +/- SD): GV = 0.31 +/- 0.05 mm3, Vv = 39.62 +/- 12.25%, Sv = 0.169 +/- 0.056 microns-1 and Nv = 3.84 x 10(-5) +/- 1.69 x 10(-5) microns-3. These determinations provide a basis for comparisons of parasympathetic with sympathetic ganglia.

Collagen and elastic fibers in the penis of human fetuses at 28 weeks postconception.

OBJECTIVES: The extracellular matrix is a key element in penile function and pathology, yet little is known of its development. Herein we investigated the morphological organization of collagen and elastin in the corpora cavernosa and tunica albuginea of human fetuses. MATERIAL AND METHODS: The penises from 5 fresh human fetuses at 28 weeks postconception (WPC) were routinely fixed and embedded, and all staining procedures were carried out on paraffin sections. Collagen was evidenced by staining with: (1) Gomori's trichrome; (2) sirius red, followed by observation under polarized light, and (3) an antihuman collagen type-III antibody. Elastin and the whole elastic system were revealed using an antihuman elastin antibody and Weigert's resorcin fucsin, respectively. RESULTS: At this stage of fetal development, the albuginea is formed predominantly by dense bundles of collagen. Near the corpora cavernosa, the presence of type-III collagen was also observed. Weigert staining showed numerous fibers of the elastic system in the albuginea. Type-III collagen was found to be strongly positive in the cavernous trabeculae and in the connective sheath surrounding the central artery. Using Weigert staining and an immunolabeling method with primary antibody against human elastin, we found an important quantity of elastic system fibers in the trabeculae of the corpora cavernosa. CONCLUSION: In fetuses at 28 WPC the albuginea is formed predominantly by dense bundles of collagen. The trabecular structures of the corpora cavernosa present a significant quantity of type-III collagen and elastic system fibers.

Organization and relative content of smooth muscle cells, collagen and elastic fibers in the corpus cavernosum of rat penis.

PURPOSE: The corpus cavernosum smooth muscle and extracellular matrix are essential for normal penile erection and are implicated in erectile dysfunction. Although investigations of these issues have used the rat corpus cavernosum, organization of its components is to date not well known. We characterized and quantified the smooth muscle cells and the main extracellular matrix components of the rat corpus cavernosum. MATERIALS AND METHODS: Collagen, elastic fibers and smooth muscle cells were stained on paraffin sections of rat penises using sirius red and Gomori's reticulin, Weigert's resorcin-fuchsin and an anti-smooth muscle cells alpha-actin antibody, respectively. Stained components were then quantified by computer aided morphometry. RESULTS: Smooth muscle cells were restricted to the subendothelial space of corpus cavernosum and had a volumetric density of 9.1%. Collagen was thick, usually in transversely oriented bundles and was the most abundant component of the trabeculae with a volumetric density of 62.7%. Gomori's reticulin disclosed a meshwork of fibrils also in the subendothelial space but did not stain the thicker bundles. Volumetric density of elastic fibers was 4.9%, and at the periphery of the corpus cavernosum the fibers were parallel to the long axis of the penis, while in deeper regions most of them were transversely oriented and at different directions from those of collagen. CONCLUSIONS: Rat corpus cavernosum differs from that of humans by lesser amounts of smooth muscle cells, greater amounts of collagen and the presence of fibrillar collagen and smooth muscle cell subendothelial layers. Therefore, these differences should be considered when using the rat penis for studies on erection.

Prostatic epithelial and luminal area in the transition zone acini: morphometric analysis in normal and hyperplastic human prostate.

OBJECTIVE: To analyse quantitatively the acini and changes in the acinar epithelium and lumen in the transitional zone of normal and hyperplastic human prostates. PATIENTS AND METHODS: Tissue samples of the transitional zone were taken from prostates with benign prostatic hyperplasia (BPH) obtained from 20 patients with clinical symptoms of bladder outlet obstruction who underwent open prostatectomy. The control tissue comprised 20 transitional zones of prostates obtained during autopsy of adults aged < 30 years (killed in accidents). The following variables were measured; the number of acini, total acinar area, area of the lumen, epithelial area, and the median (range) epithelial height, using computerized histomorphometric techniques. RESULTS: The total area of the acini and the luminal area was statistically significantly greater in BPH. In normal and hyperplastic prostates, respectively, the total mean (sd) area (mm2) of the acini was 0.041 (0.007) and 0.056 (0.016), of the lumen was 0.016 (0.003) and 0.036 (0.013), and of the epithelium was 0.025 (0.004) and 0.019 (0.003) (all P < 0.001). There was no statistically significant difference in the number of acini between controls and BPH. The area and the height of the acinar epithelium was statistically significantly greater in BPH; for epithelial height ( micro m) in normal and BPH tissue, respectively the minimum height was 9.92 (1.67) and 6.45 (1.14), the maximum 54.38 (4.09) and 41.52 (4.51) and the median 27.89 (2.48) and 19.96 (2.20) (all P < 0.001). CONCLUSIONS: There was no significant difference in the number of acini between control and BPH tissue, but the area and the height of the acinar epithelium was significantly lower in BPH.

Proximal insertion of gubernaculum testis in normal human fetuses and in boys with cryptorchidism.

PURPOSE: We determine how the proximal gubernaculum testis is attached to the testis and epididymis in human fetuses, and compare these data with findings in boys who had undergone surgery for cryptorchidism. MATERIALS AND METHODS: We analyzed 280 testes and epididymides with the gubernacula of 140 well preserved, fresh human fetuses ranging from 10 to 35 weeks after conception with no detectable congenital malformations and 36 undescended testes of 28 boys 2 to 15 years old (mean age 6.8) who had undergone surgery for cryptorchidism. In both groups the different conformations of the relationship among the proximal gubernaculum, testis and epididymis were classified according to a system used for patients with cryptorchidism. In group A the gubernaculum is attached to the testis and epididymis, in group B the gubernaculum is attached only to the testis with a tail disjunction epididymal anomaly, in group C the gubernaculum is attached only to the testis with total disjunction of the epididymis, in group D the gubernaculum is attached only to the epididymal tail and in group E there are no attachments among gubernaculum, testis and epididymis. RESULTS: Of the 280 fetal testes studied 194 (69.2%) were in the abdomen, 38 (13. 57%) in the inguinal canal and 48 (17.14%) in the scrotum. There were 277 cases (98.9%) in group A and 3 (1.1%) in group B. Of the 36 undescended testes analyzed 2 (5.6%) were abdominal and 34 (94.4%) were inguinal. There were 26 cases (72.2%) in group A, 8 (22.2%) in group B and 2 in group D. CONCLUSIONS: In fetuses without congenital malformations or epididymal alterations, such as tail disjunction or elongated epididymis, the proximal portion of the gubernaculum was attached to the testis and epididymis in all cases. In undescended testes there was an increased incidence of paratesticular structure malformations accompanied by gubernacular attachment anomalies compared to the testes in normal fetuses.

Stereology of the pterygopalatine ganglion of the rat.

The right pterygopalatine ganglia (PG) of 9 male Wistar-strain rats were dissected, embedded in Epon (3 specimens) or paraffin (6 specimens), and prepared for stereological examination under light microscopy. The perikarya were quantitatively characterized, and the ganglionic volume was determined. Stereology is an efficient method for the quantitative evaluation of the perikarya of the PG. The results(expressed as mean +/- standard deviation) were: a) areal fraction occupied by the perikarya = 53.8 +/- 7.4%; b) the perikaryal surface area per volume = 0.101 +/- 0.013 microns-1; c) the number of perikarya per volume x 10(-5) = 5.26 +/- 0.99 microns-3; d) the mean profile area of the perikarya (apk) = 505.93 +/- 78.29 microns 2; e) the mean perikaryal volume (vpk) = 9,179.33 +/- 1,533.52 microns 3; and f) the ganglionic volume = 0.210 +/- 0.127 mm3. The low coefficient of variation the apk and vpk values suggests the presence of only one population of neurons in the PG of the rat. The number of perikarya in the PG is about 11,046 per ganglion. As compared to analogous data in the otic ganglion of the rat, the PG did not show statistically significant stereological differences, but the relatively higher number of neurons found in the PG is probably associated with the higher functional activity of this ganglion.

Stromal and acinar components of the transition zone in normal and hyperplastic human prostate.

OBJECTIVE: To analyse the histological components of the transition zone in both normal human prostate and benign prostatic hyperplasia (BPH), and to determine the volumetric density (Vv) of the different elements (connective tissue, smooth muscle cells, acini and total stroma). MATERIALS AND METHODS: Samples of BPH tissue from the transition zone were obtained from 16 patients with clinical symptoms of bladder outlet obstruction who underwent open prostatectomy. The control samples comprised 16 transition zone samples from prostates obtained during necropsy of adults aged < 30 years (killed in accidents). The Vv of these components was determined by stereological methods. RESULTS: The mean (sd) Vv in the controls and BPH samples were, respectively: connective tissue 40.47 (5.16) and 46.71 (9.91)%; smooth muscle cells 24.86 (2.74) and 31.56 (5.49)%; acini 28.73 (6.25) and 17.78 (10.87)%; all differences were statistically significant. CONCLUSIONS: These results confirm the hypothesis that in BPH there is an increase in the stromal component, both smooth muscle fibres and connective tissue.

Histochemical Features of the Extracellular Matrix in Rattus rattus norvegicus Otic Ganglion / Características Histoquímicas de la Matriz Extracelular en el Ganglio Ótico de Rattus rattus norvegicus

The otic ganglion is a cranial component of the parasympathetic division of the autonomic nervous system (ANS). Similar to other parasympathetic ganglia, otic ganglion presents multipolar neurons that are completely surrounded by satellite cells and intercellular substance as well, which allow us to use this ganglion as a good experimental model for studying the relationship neuron / extracellular matrix. We have studied rat otic ganglion in 10 animals through light microscopy. After routine histological methods, 5 µm sections were obtained and coloured by Gomori`s trichrome, periodic acid-Schiff (PAS), Alcian Blue pH 2.5 and pH1.0, acetylation + PAS, acetylation + deacetylation + PAS, acid hydrolysis + PAS, phenylhydrazine + PAS and thiosemicarbazide + PAS. The presence of neutral glycoproteins was demonstrated by PAS reactivity. PAS inhibition following Alcian Blue staining in pH 2.5 and 1.0 showed the presence of a small quantity of acid glycoprotein. The extracellular matrix analysis showed the presence of neutral and acid glycoconjugates. These findings suggests a mutual interaction and a complex role in ganglionic physiology.

El ganglio ótico es un componente craneal de la división parasimpática del sistema nervioso autónomo (SNA). Similar al otro ganglio parasimpático, el ganglio ótico presenta neuronas multipolares que están rodeadas totalmente por las células satélites y la sustancia intercelular, lo cual permite la utilización de este ganglio como un buen modelo experimental para estudiar las relaciones matriz extracelular/neurona. Examinamos, con microscopio de luz, el ganglio ótico 10 ratones. Con los métodos histológicos rutinarios fueron obtenidas 5 secciones y coloreadas con tricrómico de Gomori, PAS, Azul de Alcián pH 2.5 y pH1.0, acetilación + PAS, acetilación + desacetilación + PAS, hidrólisis de ácido + PAS, fenilhidrazina + PAS y tiosemicarbacida + PAS. La presencia de glicoproteínas neutras fue demostrada por la reactividad de PAS. La inhibición de PAS y la posterior tinción con Azul de Alcian en pH 2.5 y 1.0, demostró la presencia de una cantidad pequeña de glicoproteínas ácidas. El análisis extracelular de la matriz demostró la presencia de glicoconjugados neutros y ácidos. Estos resultados sugieren una interacción mutua y un papel complejo en la fisiología ganglionar.

Morphometrical and stereological analysis of the superior cervical ganglion of rattus norvegicus submitted to chronic treatment with cortisol

The present work investigates the corticoid action on the growth of the superior cervical ganglion of the rat and describes the cortisol effect during early stages of development. The study was based on morphometric and stereological analysis of the perikarya. Eight rats were treated intraperitoneally with cortisol (1mg/Kg/day) during 36 days. Treatment was initiated in the 8th day after birth and was withdrawn one day before the sacrifice. There was a significant difference (úP0,05) for the neural mean diameter between the control group (16.78 ± 1.11mm) and treated animals (15.84 ± 0.99mm). The decrease of perikarya neuronal diameter was also demonstrated by stereological methods. Morphometrical findings may suggest alterations in superior cervical ganglion neuronal activity in rats treated for long term with cortisol.

Estudo anatômico e quantitativo do gânglio pterigopalatino humano. Morfometria e estereologia / Anatomic and quantitative study of the human pterygopalatine ganglion. Morphometry and stereology

The pterygopalatine ganglion is important in the regulation of the intraocular pressure and in the cerebral vasodilatation connected with headache of vascular origin. Four human ganglia were dissected, fixed in formalin and serially sectioned with a 6 microns thickness. The volume of the ganglion was calculated by point-counting and stereological parameters were determined using the test-system M42 with light microscopy. The PG volume was (mean +/- standard error of the mean) 5.6 +/- 0.5 mm3. The volume density of neurons was 51.1% +/- 3.4%, and the unitary volume of the neurons was 41,200.0 +/- 2,250.0 microns. The numerical density was 12,600.0 +/- 677.0 neurons by mm3, therefore approximately 70,560 neurons by ganglion.