Histological Studies of the Reactions of Cells and Intercellular Substances of Loose Connective Tissue to the Spreading Factor of Testicular Extracts

Abstract

As a basis for the interpretation of the experimental findings to be presented, it may be well to review the histological changes which occur in the development, differentiation, and maturation of the loose or areolar connective tissue from mesenchyme.Mesenchyme, from which all the connective tissues are derived, is composed of undifferentiated mesenchymal cells separated by amorphous intercellular substance with only a hint of a fibrillar appearance, which has been accentuated by fixation. In the umbilical cord, the intercellular substance is of a soft jelly-like consistency, and only a few tiny fibrils have appeared.As the mesenchymal cells differentiate into fibroblasts within such a matrix, fine fibers, ensheathed by a somewhat modified amorphous substance, first appear. Some of these are later cemented together into bundles as collagen fibers.FIGURE 1 is a phase-contrast photomicrograph of a fresh preparation of adult-mouse subcutaneous tissue lightly stained with toluidine blue. Here are fully clifferentiated cells such as the fibroblast and several mast cells. The intercellular substances, both amorphous and fibrous, i.e., collagen fibers of various sizes, reticular fibers, and elastic fibers (just out of focus), have been derived from the optically homogeneous amorphous substance of the mesenchyme.Material and Methods. The following experiments were undertaken in an attempt to determine the relation of connective tissue cells to the intercellular substances and the specific effect on both of the spreading factor of testicular extracts.Testicular extracts were prepared in the laboratory and compared with extracts kindly supplied by the Schering Corporation. No attempts were made to determine the potency of the extract other than the rate and degree of spread of India ink injected with the extract subcutaneously. Though the results varied in degree, they were similar in kind in all the series. 0.1 cm3 of testicular extract was injected subcutaneously under the shaved skin of each animal of the series. Tissues were removed from a control mouse, from an injected mouse within 5 minutes after the injection, and subsequently from each succeeding mouse in the series at daily intervals following the day of injection. In each case, fresh tissue studies were made by the "bulh" method and by dried spreads of the connective tissue stained metachromatically with toluidine blue. Pieces of skin were removed from the site of injection, fixed in formol-saline, embedded in paraffin, sectioned, and stained with a variety of stains.The two routine stains most useful for the fixed tissue were found to be hematoxylin and eosin for tissue relationships and a neutral dye of pina- 983Others become coated to form elastic fibers.Several series of mice and rats were used.

Histochemical Demonstration of Hyaluronic Acid in Human Dermal Mast Cells

“…Velican and Velican (1959), after studying the effect of hyaluronidase on the metachromatic staining of mast cells in autopsy specimens of human bronchial lymph nodes, concluded that the metachromatic substance in some of the granules consists exclusively of hyaluronic acid. Similar results were observed by Bensley (1950), and others. On the other hand, West (1958) pointed out that hyaluronidase did not destroy the metachromasia of mast cell granules, and that sometimes there was poor correlation between the mast cell count and the hyaluronic acid content of a tissue.…”

The mast cells

“…However, they and many later authors agree that in acute inflammation the mast cells temporarily disappear (Neumann, 1890;Neuberger, 1894;Maximow, 1904;Ernst, 1926;Stockinger, 1927;Brodersen, 1928;Quensel, 1933;Larsson and Sylven, 1947;Drennan, 1951; Gustafsson and Cronberg, 1957;McGovern, 1957). Later, as the acute inflammation subsides and as spindle-shaped fibroblasts elaborate their fibrils, there occurs a progressive increase in the number of mast cells in the affected area; and as long as this proliferative phase persists the local mast-cell population continues to increase (Staemmler, 1921;Bensley, 1950). Enormous numbers of mast cells are thus found in the hypertrophied connective tissue of elephantiasis (Baumer, 1896;Ehrich et al, 1949).…”

Antigenic evidence for host origin of exudative fluids in lesions of Treponema pallidum-infected rabbits

“…As mucopolysaccharides are synthesized intracellularly and secreted into the extracellular matrix (8), an increased cellular metabolism would cause increased biosynthesis and secretion of mucopolysaccharides. Such a series of events is supported by reports that the local injection of hyaluronidase causes the depolymerization of ground substance, tissue cell proliferation, and increased cell mitosis (6,7,42). Much of the reported data can be explained by this proposed mechanism.…”