David Brandes

The Fine Structure and Histochemistry of Prostatic Glands in Relation to Sex Hormones

Publisher Summary This chapter reviews some of the histochemical and ultrastructural properties of prostatic tissue and how these are affected by various alterations of the hormonal environment of the host. From the various studies on the fine structure of prostatic tissues, it is apparent that differences exist not only between the prostates of the various species, but also between the various prostatic lobes of a given species. However, a prototype prostatic acinus can be described which may reflect the structural patterns prevailing in most cases. Acid phosphatase has been shown at the ultrastructural level in rat prostatic lysosomes. Observations in the dog prostate have revealed the presence of acid phosphatase activity not only in typical lysosomes, but also in the secretory granules. In older rats acid phosphatase activity has been detected in the supranuclear lysosomes, which make their appearance when the rats have reached a weight of 250 gm. Acid phosphatase activity has been also demonstrated in the lysosomes and cytolysomes which appear in rat prostatic epithelial cells after castration. Preliminary studies on the fine structural localization of acid phosphatase in the human prostate have also revealed the enzymic activity in the lysosomes of the secretory cells. Various nucleoside phosphatases have been shown to be localized in the Golgi saccules of rat ventral prostate and human prostate. These and many other nucleoside phosphatases have been demonstrated in the various membranes of numerous cell types. The chapter also discusses the possible functional significance of differences in the various prostatic cells, influence of sex hormones on the ultrastructure and histochemical reactions, and interpretation of hormone-induced changes on the basis of structure-function relationships

Role of lysosomes in cellular lytic processes: II. Cell death during holocrine secretion in sebaceous glands

Abstract The mechanism of holocrine secretion was investigated in rat sebaceous glands by means of combined electron microscopical and histochemical methods. Numerous lysosomes, apparently derived from the Golgi apparatus, were seen in the cells undergoing maturation. During the process of disintegration of the mature cells, the lysosomes seemed to release their hydrolases into the general cytoplasm. Numerous other examples can be found in the current literature which tend to indicate that lysosomal rupture may be a fairly general feature of cells undergoing autolysis.

Observation on the apparent mode of formation of “pure” lysosomes

The intracellular site where lysosomal enzymes are revealed by electron histochemical methods appears to be related to the prevailing type of endoplasmic reticulum, i.e., rough- or smooth-surfaced. In rat prostatic and seminal vesicle epithelial cells the rough-surfaced endoplasmic reticulum predominates. Formation of lysosomes and synthesis of at least some of their enzymes appear to be connected with the ribosome-coated membranes of the endoplasmic reticulum. In sebaceous secretory cells and Euglena cells the endoplasmic reticulum is almost exclusively of the smooth-surface variety. In these cells, formation of lysosomes and synthesis of their enzymes appears to be mediated chiefly through the Golgi apparatus. Lysosomes such as described in the various cell types in this work seem to represent “pure lysosomes,” in the sense that they do not appear to be actively engaged in intracellular lytic processes. That they may do so under various physiological and pathological conditions has been indicated in previous reports (2, 3).

The effects of retinoids on proliferative capacities and macromolecular synthesis in human breast cancer MCF-7 cells.

The effects of various retinoids on the proliferative capacities and on the synthesis of DNA, RNA, and protein have been investigated in MCF‐7 mammary carcinoma cells in culture of the various retinoids tested, retinoic acid revealed maximum activity in inhibiting cell proliferation and thymidine incorporation. The degree of inhibition of cell proliferation by the various retinoids paralleled their capacity to inhibit thymidine incorporation, suggesting suppression of DNA synthesis as a primary cause of restriction of cell growth by these compounds. Two nonepithelial human cell lines were tested for sensitivity to retinoids, and showed diminished responses compared with MCF‐7 cells. This suggests a correlation between the ability of retinoids to exert control of differentiation and cell proliferation for a given cell type. Reversibility of the effect of retinoid treatment, high cell viability, and lack of retinoid‐induced lysosomal enzyme release, as shown in our studies, indicate that cytotoxicity may be excluded as a cause of decreased cell proliferation and inhibition of thymidine incorporation by retinoids.

Ultrastructural and cytochemical changes in cultured human lung cells

Human diploid cells (WI-38) in culture show marked ultrastructural and cytochemical differences from the various mesenchymal cells present in the fetal lung. The cultured cells show hypertrophic RER cisternae studded with helical polyribosomes. A complex system of microtubules and filaments develops in the cultured cells, which also show an abundance of microvesicles formed at both the cell surface and in the Golgi. The vesicles at the cell surface are actively engaged in pinocytosis of macromolecules in the medium, and those in the Golgi represent primary lysosomes. Microtubules and filaments are very labile structures, and appear to play a role in the maintenance of cell form and in intracytoplasmic flow. Secondary lysosomes, residual bodies, and autophagic vacuoles increased with passage level. It is not clear, however, whether this change is associated with cellular degradative processes or whether it pertains to the aging process in vitro .

Lysosomes in mice mammary tumors treated with cyclophosphamide: Distribution related to course of disease

Transplantable carcinomas of mouse mammary gland were treated with cyclophosphamide at various dose levels ranging from 50 to 300 mg/kg body weight. The degree and the length of the period of tumor growth inhibition increased with the dose. Increased activity of the lysosomal enzyme, acid phosphatase, was observed in the tumor cells of treated mice, especially at the dose levels of cyclophosphamide that inhibited tumor growth (200 to 300 mg/kg). Lysosomal acid phosphatase activity remained present during the entire period of growth inhibition but decreased at the time when tumor growth was resumed and disappeared almost completely when the growth rate approximated that of the untreated controls. Combined treatment with the cytotoxic agent and a lysosomal labilizer, vitamin A, increased both the inhibitory effect of cyclophosphamide and the acid phosphatase activity in the tumor cells. In the electron microscopic preparations, the tumor cells of treated mice showed areas of focal degradation and widespread ultrastructural alterations which frequently were associated with lysosomes.

Effects of a choline-deficient diet on the induction of drug- and ethanol-metabolizing enzymes and on the alteration of rates of ethanol degradation by ethanol and phenobarbital

Abstract Phosphatidylcholine has been shown to be an essential component for electron transport to cytochrome P-450 and for hydroxylation of a number of substrates by microsomes in vitro . A choline-deficient diet was fed to rats for 3 weeks in order to study the effect in vivo of alterations of liver phospholipids on the activity of microsomal enzymes, on parameters of ethanol metabolism, and on the adaptive responses of both to ethanol and phenobarbital administration. Choline deficiency resulted in an increase in total liver lipids and triglycerides, but in a decrease in total phospholipids, due mostly to a decrease in phosphatidylcholine. Choline deficiency did not result in changes in microsomal enzymes or parameters of ethanol metabolism. However, it did prevent optimal induction of aniline hydroxylase activity and cytochrome P-450, by both ethanol and phenobarbital, and of microsomal protein concentration and cytochrome b 5 by phenobarbital; it also prevented ethanol-induced increases both in the activity of the microsomal ethanol-oxidizing system and in the rates of ethanol disappearance from the blood. Alcohol dehydrogenase activity remained unchanged. This study demonstrates that dietary choline is required for optimal induction of microsomal enzymes by both ethanol and phenobarbital, and for increases in ethanol metabolism induced by ethanol administration. It is suggested that a decrease in available hepatic phosphatidylcholine, due to choline deficiency, is a cause of inhibition of the optimal induction of microsomal enzymes.

Studies of L-1210 leukemia: IV. Ultrastructural findings after in vitro treatment with cyclophosphamide and Vitamin A

Abstract The influence of normal liver and Vitamin A on the activity of cyclophosphamide on the L-1210 leukemia was investigated in short-term organ cultures. The experimental data presented here support the hypothesis that some principle in the liver is instrumental in the biologic activation of cyclophosphamide. It is also suggested that Vitamin A may alter the permeability of lipoprotein membranes and favor the release and diffusion of lysosomal or microsomal enzymes, or both, and that such enzymes may play a role in the activation process and in tumor-cell damage. Vitamin A also appeared to induce changes in the leukemic cells, reflected in a pseudoepithelization of the lymphoid elements. Increased phagocytosis of red cells by leukemic cells and macrophages, and characteristic changes in the phagocytized corpuscles is also attributed to the action of Vitamin A.

Role of lysosomes in cellular lytic processes. IV. Ultrastructural and histochemical changes in lymphoid tissue of thymectomized mice with wasting disease.

Lymphoid tissues obtained from mice with wasting disease resulting from neonatal thymectomy were examined by histochemical and electron microscopical techniques. The lymphoid organs showed loss of architecture, depletion of lymphocytes, and marked infiltration of macrophages. Plasma cells, however, were numerous. Macrophages were frequently engaged in the phagocytosis of altered lymphocytes, and their degradation within the phagocytic vacuoles appeared to be due to hydrolytic enzymes derived from macrophage lysosomes. Stages in the maturation of myeloid cells, including the development of lysosomes in the Golgi apparatus, were readily detected in the spleen. Neither bacteria nor viruses were detected in the lymphoid tissues, and a possible role of lymphocytic phagocytosis in the depletion of lymphocytes is suggested.