Atlas of Microscopic Anatomy: a functional approach - companion to Histology and Neuroanatomy. Second edition

It presents images and information related to the microscopic anatomy of cells, epithelial tissue, connective tissue, blood, muscular and nervous tissues, integument, cardiovascular, lymphatic, digestive, respiratory, urinary, and female and male reproductive systems, endocrine glands, special senses, and central nervous system.

Molecular determinants of ovarian cancer plasticity.

During development, the formation and remodeling of primary vascular networks occurs by vasculogenesis and angiogenesis. Recently, the term "vasculogenic mimicry" has been used by our laboratory and collaborators to reflect the embryonic-like ability of aggressive, but not nonaggressive, melanoma tumor cells to form a pattern of matrix-rich networks (containing channels) surrounding spheroids of tumor cells in three-dimensional culture, concomitant with their expression of vascular cell markers. Ovarian cancer is usually diagnosed as advanced stage disease in most patients when widespread metastases have already been established within the peritoneal cavity. In this study, we explored whether invasive ovarian carcinoma cells could engage in molecular vasculogenic mimicry reflected by their plasticity, compared with their normal cell counterparts. The data revealed that the invasive ovarian cancer cells, but not normal ovarian surface epithelial cells, formed patterned networks containing solid and hollow matrix channels when grown in three-dimensional cultures containing Matrigel or type I collagen, in the absence of endothelial cells or fibroblasts. Immunohistochemical analysis showed that matrix metalloproteinases (MMP)-1, -2, and -9, and MT1-MMP were discretely localized to these networks, and the formation of the networks was inhibited by treatment with MMP inhibitors. Furthermore, the RNase protection assay revealed the expression of multiple vascular cell-associated markers by the invasive ovarian cancer cells. In patient tumor sections from high-stage, high-grade ovarian cancers, 7 to 10% of channels containing red blood cells were lined by tumor cells. By comparison, all vascular areas in benign tumors and low-stage cancers were endothelial lined. These results may offer new insights and molecular markers for consideration in ovarian cancer diagnosis and treatment strategies based on molecular vascular mimicry by aggressive tumor cells.

A novel immunological model for the study of prostate cancer.

The Dunning R-3327 rat prostatic adenocarcinoma is a widely accepted model for in vivo experimental studies of prostate cancer. We have previously derived phenotypically distinct cell lines from a s.c. tumor resulting from the inoculation of the R-3327-5 subclone into Copenhagen rats. In this study, we report studies using a gelatin sponge model for the delivery of tumor cells and the retrieval of tumor-specific leukocytes responsive to different prostatic cell lines. S.c. preimplanted sponges were inoculated with tumor cells previously selected for differential properties of tumor formation and metastasis and examined for leukocyte content at time points of 1, 3, and 5 weeks after tumor cell inoculation. Cytospin and flow cytometric analyses revealed fewer tumor-associated leukocytes present in sponges inoculated with tumorigenic R-3327-5' and R-3327-5'B lines, with lesser sponge degradation, than in experiments with the nontumorigenic R-3327-5'A line, suggestive of a tumor cell-induced immunomodulatory mechanism. Morphological studies indicate an intermittent tumor growth pattern that gradually disappears in sponges inoculated with the nontumorigenic R-3327-5'A cells but a robust growth pattern in sponges inoculated with the tumorigenic cell lines. Cytokine analyses show the secretion of higher levels of active transforming growth factor-beta by the more invasive and metastatic lines. Total transforming growth factor-beta levels are higher in the epithelial, tumorigenic R-3327-5'B line. Additionally, the more tumorigenic lines secrete interleukin 10, a potent immunosuppressive molecule. In this report, we demonstrate the ability to retrieve viable leukocyte populations from a prostate tumor line bearing sponges, which offers an important model for further in vitro and in vivo manipulations and holds promise for testing adoptive immunotherapeutic strategies.

Fine structural studies of induced tumors arising within the prostatic complex of Lobund-Wistar rats.

BACKGROUND: Morris Pollard, Phyllis Luckert, and colleagues have reported the occurrence of spontaneously arising tumors of the prostatic complex in aged Lobund-Wistar (L-W) rats, and have also shown that the genesis of such tumors may be accelerated by means of intravenous administration of methylnitrosourea, followed by androgen supplementation. METHODS: Light and electron microscopic investigations of the tumors arising under this regime were conducted, with the objective of documenting morphological changes attending the transformation process; 10 tumor samples were used for the electron microscopic studies. RESULTS: All tumors studied were adenocarcinomas arising within the prostatic complex of induced animals. These tumors varied in size, degree of differentiation, and invasiveness. Foci of prostatic intraepithelial neoplasia were noted in light microscopic studies as well. Consistent fine structural features exhibited by cells of the induced adenocarcinomas included a large nuclear to cytoplasmic ratio; large irregular nuclei with heavily marginated chromatin; conspicuous nucleoli; abundant ribosomes and polysomes and a paucity of rough endoplasmic reticulum; and numerous cytoplasmic vesicles and lipid inclusions. Numerous, short microvilli extended from the cell surface into a copious surrounding extracellular matrix. CONCLUSIONS: Thus, these tumors shared many of the fine structural features characteristic of the Dunning (rat) and human prostatic adenocarcinomas.

Gross and microscopic pathology of induced prostatic complex tumors arising in Lobund-Wistar rats.

The necessity for additional animal models for prostate cancer has recently been stressed. The Pollard model of chemically induced prostate cancer has received attention in this regard although the histiogenetic origin of these tumors has come under question. We independently studied this model for the development of tumors in the prostate region. The tumors, all of which were adenocarcinomas, first became grossly evident 5 months after induction and ultimately occurred in 71% of the animals. Seventy-three % of the tumors involved only the seminal vesicle, 22% involved other portions of the prostatic complex as well as the seminal vesicle, and 5% were located in the coagulating gland (anterior prostate). Although the majority of tumors arose in or involved the seminal vesicle, this may still be a useful model for the study of human prostate cancer because the tumors are adenocarcinomas, occur in the large majority of animals, are hormonally induced, and have the propensity to metastasize.

Peptidergic nerves in the ureter.

Vasoactive intestinal peptide (VIP) and substance P were demonstrated in the pig ureter by immunohistochemical techniques. Nerves containing these materials were related mainly to the smooth muscle layer in the normal and obstructed ureter. In isolated ureteral segments, VIP caused relaxation at doses exceeding 0.18 micrograms/ml, with no significant difference seen in the effect on normal and obstructed ureter. Vasoactive intestinal polypeptide may play a role in the regulation of ureteral smooth muscle tone.

Cell wall deficient bacteria as a cause of idiopathic hematuria.

Idiopathic hematuria in the absence of bacteriuria is a medical challenge. Routine cultures of catheterized bladder and endoscopically obtained ureteral urine specimens from a 22-year-old woman with a 6-week history of hematuria showed no growth after 24 to 48 hours of incubation. However, bacterial variants were grown on enriched media. Colonies were typical cell wall deficient/defective bacteria. Phase and electron microscopy of cystoscopic urine specimens obtained by retrograde ureteral catheterization as well as phase microscopy of the cultures revealed the classic morphology of these organisms. When the variant cultures were subcultured the organisms reverted to their related walled forms, that is Streptococcus agalactiae and Staphylococcus haemolyticus. Because the colonies of these organisms showed various patterns of biochemical reactivity, each phenotype was tested against 15 antimicrobials. Collectively, all biotypes had a common susceptibility to only nitrofurantoin. The patient was treated with nitrofurantoin for 6 weeks. Four days after initiation of therapy she had complete remission of hematuria. During the next 3 years she remained well and free of hematuria.

Human doubled renal and testicular arteries.

Doubled renal and testicular arteries were found in a well-developed 69-year-old caucasian male. The right kidney had two renal arteries, one at its usual midorgan (hilar) position and one inferior polar. One testicular artery arose from the mid-point of the usual renal artery, passed under the renal vein at its junction with the inferior vena cava, and then crossed over the inferior polar renal artery. The second testicular artery arose from the inferior polar renal artery near its origin from the abdominal aorta. The two testicular arteries remained doubled throughout their course and both entered the right testis at separate sites on the organ. The inferior vena cava passed under the inferior polar and over the superior testicular artery and the superior renal artery.

Immunocytochemical localization of metallothionein in the rat prostate gland.

Metallothionein (MT), which binds zinc and other metals, was localized within the rat prostatic complex at light and electron microscopic levels utilizing immunocytochemistry. Two groups of mature, male Sprague-Dawley rats were injected with cadmium chloride subcutaneously (2 mg/kg body weight) to induce the synthesis of MT, and were sacrificed 48 h postinjection. From the first group, prostatic tissue from the three lobes was prepared for light microscopy. The indirect peroxidase antiperoxidase procedure was used for MT localization using rabbit anti-MT as the primary antibody. The three lobes of the rat prostate demonstrated a positive reaction for MT, but among the lobes variations in the intensity and localization of the reaction product occurred. Tissues from the second group of animals were prepared for electron microscopy. Using the indirect immunogold labeling technique with anti-MT, MT localization was confirmed in the rough endoplasmic reticulum, secretory vesicles, secretory products, and subepithelial connective tissue. These localizations suggest that MT binds zinc both intra- and extracellularly, where it may function in zinc storage and metabolism.

Stereological study of Leydig cell density in the guinea pig testis.

Stereological analysis of Leydig cell and macrophage volume density along the long axis of the guinea pig testis was assessed quantitatively by histometric point counting. Morphological identification of Leydig cells was accomplished by staining for 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), whereas macrophages were identified by vital staining (trypan blue). The average percentage of Leydig cell density constitutes about 10.17 +/- 0.23% at the cranial level (level 1) and about 8.8 +/- 0.21% at the caudal level (level 4). Leydig cell density through four testicular levels showed no significant difference among levels 1, 2, and 3, whereas level 4 was significantly different. The percentage of macrophage density, on the other hand, was approximately 0.4% +/- SEM per cross-sectional profile. It may thus be concluded that the macrophage density within the guinea pig testis does not bias histometric studies of the Leydig cell population to any significant degree, but that regional differences in Leydig cell volume density could influence validity of sampling if tissue procurement is from different testicular levels in successive experiments.

Characterization of the heterogeneity of R3327 rat prostatic tumors derived from single-cell clones.

Prostatic adenocarcinoma is characterized by cellular diversity, which is well demonstrated in the Dunning R3327 rat prostatic adenocarcinoma. This heterogeneity may arise from epigenetic influences, ie, cellular adaptation or selection, and/or from genetic changes. To investigate the question of genetic instability, four tissue culture cell lines were derived from single cells isolated from the uncloned late (UCL) passage of the Dunning R3327H prostate cell culture. Each of these clonally derived tissue cultures was injected into castrated and intact young adult male rats for tumor production. Uncloned early (UCE) and UCL passage tissue cultures were also propagated as solid tumors. Tumors and the cultures from which they were derived were examined for evidence of phenotypic and genetic changes using morphological and cytometric methods. Transmission and scanning electron microscopy revealed only slight differences among the cell cultures. A single population of diploid cells was demonstrated in each of the cell cultures by propidium iodide staining and subsequent flow cytometric measurement of DNA content/nucleus. Tumors of unicellular as well as multicellular origin exhibited extreme heterogeneity of histological features, both among animals as well as within a single tumor. Tumors were surveyed and tissue types were characterized and cataloged. Clone 3 was generally better differentiated than the others; tumors from castrated animals were better differentiated than those from intact animals. Flow cytometry revealed multiple hyperdiploid cell populations that were variable from one sample to another. We concluded that changes in genotype as well as phenotype occurred in the tumors derived from single cells. Some of these changes may have occurred in the cells while still in culture.

Androgen receptor binding characteristics in the cytosol of the rat dorsolateral prostate gland and the Dunning R-3327 prostatic adenocarcinoma.

The incidence of prostatic cancer is highly correlated with advanced age, and it has been suggested that changes in androgen binding may be important in age-associated alterations in growth regulatory mechanisms of prostatic epithelial cells. In this study the effects of age on androgen binding characteristics in the dorsolateral prostate glands of young and aged Copenhagen rats were determined and the binding properties in the Dunning R3327/130 subline of rat prostatic adenocarcinoma were characterized. Tritium-labeled and nonlabeled methyltrienolone analogs (R1881) were used to study the binding properties of 5 alpha-dihydrotestosterone receptor in the cytosol of tumors and prostate glands. Binding of R1881 was low but specific for the androgen receptor as shown by competition studies in which nonlabeled R1881, 5 alpha-dihydrotestosterone, and testosterone competed successfully with 3H-R1881 for binding sites, but 17 beta-estradiol and low levels of progesterone did not. In Copenhagen dorsolateral prostate, Scatchard analysis suggested a single class of binding sites. In young animals (three to five months) the average binding capacity was 10.36 fmol/mg cytosol protein with a dissociation constant (Kd) of 2.28 nmol/L. The dorsolateral prostate of aged rats (11-16 months) showed no significant difference in specific binding characteristics as compared to the younger age group. Specific binding of 3H-R1881 in R3327/130 tumor was saturable with a single class of high-affinity binding sites having an average binding capacity of 64.77 fmol/mg cytosol protein and a Kd of 2.76 nmol/L. These data show that the tumor had approximately 6.5 times the number of binding sites as did the normal Copenhagen rat dorsolateral prostate gland. However, no age-related changes were detected through 11-16 months of age in the androgen binding characteristics of normal rat dorsolateral prostate gland that could be correlated with the higher concentration of androgen binding sites in the R3327/130 tumor subline.

Hormonal effects on zinc concentration and morphology of rat lateral prostate gland.

Hypophysectomized rats were injected with prolactin and/or testosterone, and luteinizing hormone (LH) and prolactin and/or follicle-stimulating hormone (FSH). Tissue from the lateral prostate was processed for spectrophotometric determination of zinc and for electron microscopy. Changes in zinc concentration, epithelial height, and morphological characteristics were used to assess the effect of hormonal replacement on functional activity. Although testosterone administration resulted in full restoration of all parameters, there was no evidence of a synergistic effect of prolactin and testosterone. Prolactin alone decreased the zinc concentration and increased the epithelial height compared with control animals, and there was evidence of epithelial hyperplasia. Luteinizing hormone, alone or combined with prolactin or prolactin and FSH, resulted in a marked increase of epithelial height and restoration of morphological features found in intact animals; however, only a slight increase of zinc concentration over control values was elicited. Combined LH and FSH administration reduced epithelial height and zinc concentration compared with the other LH regimes, and there was evidence of cellular degeneration.

Electron microscopic and histochemical characterization of intra-arterial cushions of the rat and porcine uterine vascular bed.

Scanning and transmission electron microscopic studies, together with histochemical investigations, were conducted on rat and porcine intra-arterial cushions from the uterine vascular bed. In the rat, the fine structure of these cushions closely resembled that previously described in the rat kidney. The cushions were composed of modified smooth muscle, circularly disposed in an incomplete, raised band surrounding the entrance to arterial branches. These muscle cells projected as attenuated processes throughout the loosely organized, PAS-positive stroma, and established close contact with thin endothelial extensions projecting from the base of the surface endothelial cells. Scanning electron microscopic observations of furrows on the endothelial surface gave rise to the suggestion that such contacts might mediate muscular control of endothelial surface topology. In similar cushions from the pig uterine artery, the smooth muscle of the cushions was much more compactly organized, and was disposed radially, rather than circumferentially, within the cushion structure. The enzyme histochemical profile of porcine cushions did not differ appreciably from that of normal vascular smooth muscle and endothelium, suggesting the maintenance of a metabolic similarity with adjacent tissues. These studies clarify the fine-structural basis for recently reported contraction and relaxation of uterine artery cushions during ischemia and perfusion of the rat uterine vascular bed, and thus, for their functional role in the regulation of uterine vascular flow.

Isolation, in vitro cultivation, and electron microscopy of normal and malignant prostatic epithelial cells from the Copenhagen rat.

Procedures are described for the isolation and cultivation of normal rat prostatic epithelial cells. The techniques, which involve collagenase digestion and Ficoll purification of the epithelial population, are efficient, inexpensive, and produce pure monolayers. Included is a scanning and transmission electron microscopic study comparing cells isolated in vitro to rat prostatic epithelial cells in situ. Further ultrastructural comparisons are made to a malignant cell line, the Dunning R3327H Copenhagen rat prostatic adenocarcinoma.

Effects of estrogen upon the fine structure of epithelium and stroma in the rat ventral prostate gland.

We studied the effects of estradiol upon the fine structure of the rat ventral prostate, with special emphasis on the interstitial tissue. Intact and castrated rats were injected with estradiol or testosterone in combination, or with peanut oil (control). Estrogen exerted an antiandrogenic effect on the epithelium of intact animals resulting in lower epithelial height and reduction in the number of cell organelles and secretory bodies; however, the prostate gland did not atrophy to the same degree as that in castrated controls. In castrates, estradiol had no discernible effect upon the glandular epithelium. Although testosterone alone was able to restore glandular morphology to normal in these animals, estrogen in combination prevented regeneration of rough endoplasmic reticulum and elicited the formation of large lipid-like inclusions and the accumulation of secretory bodies in the apical zone of many cells. Intact animals treated with these hormones in combination exhibited many of the same effects except that a more normal configuration of rough endoplasmic reticulum was present. In castrates, the prostatic stroma became thickened, with a large increase in fibrous material between and surrounding each acinus, although smooth muscle cells retained their normal cytology. With estradiol treatment, singly or in combination with testosterone, smooth muscle cells increased in size and number and organelles decreased in number, cytoplasm became more electron dense, and nuclei became more heterochromatic. Surface vesicles were profuse in smooth muscle cells in animals treated with estradiol alone; larger phagocytic-like vacuoles were characteristic of the glands of animals treated with estrogen and testosterone in combination. In these latter treatment groups, testosterone prevented the overall hypertrophy and hyperplasia of the interstitial tissue; to a degree, endogenous testosterone levels in intact animals treated with estradiol alone prevented these effects as well.

Spermatic granuloma of vas deferens after vasectomy in rhesus monkeys and men: light and electron microscopic study.

Spermatic granuloma of the vas deferens is a common complication of vasectomy which has received scant morphologic study. This study investigated the light and electron microscopic structure of such granulomas detected in rhesus monkeys (Macaca mulatta) and man after various modes of vasectomy and postoperative periods. Unilateral experimental vasectomy in monkeys was performed by either silk ligation or clasp occlusion; in 4 of 13 ligated animals and 5 of 5 clasp vasectomized animals granulomas developed at the site of fasectomy. In man, portions of the vas deferens were excised adjacent to the site of vasectomy preparatory to vasovasostomy. Of 5 patients studied, unilateral spermatic granulomas developed in 3. Such granulomas in both monkey and man were characterized by (1) masses of sperm surrounded by epithelioid cells and connective tissue, and (2) multiple epithelial-lined channels which often contained sperm and spermiophages. In both species, fine structural characteristics of the epithelium lining such channels closely resembled those of the principal cells of the normal vas. Spermiophagic cells included macrophages, epithelioid cells, and, in the monkey only, neutrophils. Lymphocytic invasion was a common feature of the human granulomas but was found only occasionally in the monkey granulomas. As a greater number of granulomas are studied in humans and monkeys, it is hoped that the processes underlying granuloma formation and the role of such granulomas in the development of complications after vasectomy will be clarified.

Fine structural studies of the rat vas deferens.

A fine structural study of the normal rat vas deferens was undertaken utilizing perfusion fixation. Morphological features not previously appreciated were revealed using this technique of fixation, and included the following. The rat vas deferens exhibited a gross morphological and microscopic differentiation along its length: A proximal segment was characterized by a thin muscular wall, an epithelium of low height (comparable to that of the cauda epididymidis) and a distended lumen typically filled with an accumulation of sperm; a distal segment exhibited a thick muscular wall, a convoluted mucosa, and a pseudostratified columnar epithelium with long stereocilia extending into the lumen. The transition from the morphology typical of the proximal segment to that of the distal segment was gradual and progressive, marked by an increase in the mass of the muscular wall and in the height and ultrastructural complexity of the epithelium. Clear or "foamy" cells, characteristic of the cauda epididymidis, were observed in the initial centimeter of the vas deferens. Also, a cell type designated as "mitochondrion-rich" was observed in the distal vas segment. The structure of the small mitochondria in such cells, however, did not conform to the description of mitochondria in similar cells found in the human (Hoffer, '76). Intraepithelial macrophages containing residual accumulations which often resembled spermatozoan remnants in advanced stages of dissolution were present in all segments of the rat vas deferens, confirming in this species a spermiophagic role for such cells.