Paulo A. Abrahamsohn

Collagen remodeling during decidualization in the mouse

SummaryAn ultrastructural study of the features and distribution of collagen fibrils was performed in the endometrium of virgin and pregnant (2nd to 11th day) mice. Collagen-containing structures were observed in the cytoplasm of fibroblasts on the 2nd day of pregnancy. Treatment of tissues with lanthanum nitrate established that these structures were intracytoplasmic. Their association with lysosome-like bodies suggested the occurrence of intracellular digestion of collagen, probably connected with remodeling of the endometrial stroma prior to decidualization. On the 4th day of pregnancy, very few collagen fibrils were present in the intercellular space. From the 6th day of pregnancy onwards, “thick” collagen fibrils were observed between decidual cells. The diameter of these fibrils measured up to 300 nm whereas the fibrils present in the endometrium of virgin mice measured 40–68 nm.

Purification of an adenylyl cyclase-containing plasma membrane fraction from trypanosoma cruzi

A fraction containing plasma membrane fragments has been purified from epimastigote forms of Trypanosoma cruzi. Cells were broken by sonic vibration under well defined conditions and membranes were isolated by differential centrifugation and equilibrium centrifugation in sucrose gradients. The co-purification (approximately 10-fold) of adenylyl cyclase and plasma membrane-bound radioactive iodine is highly suggestive of the localization of this enzyme in the plasma membrane of T. cruzi. Determination of succinate cytochrome c reductase and glucose-6-phosphatase activities, as well as of total amounts of DNA and RNA in the purified fraction, indicates a negligible contamination from other cellular organelles. The co-purification of acid phosphatase activity with bound labeled iodine and adenylyl cyclase was taken as circumstantial evidence that part of this enzyme also belongs to the plasma membrane of T. cruzi. Conventional electron miscroscopy and freeze-fracture images of this fraction are consistent with a highly enriched plasma membrane preparation.

Involution of the antimesometrial decidua in the mouse

SummaryInvolution of the antimesometrial decidua was analysed by electron microscopy on days 9, 10 and 11 of pregnancy in the mouse. During this period, the width of the antimesometrial decidua decreases considerably. Involution begins in the decidual cells situated closest to the embryo (internal decidua) and proceeds towards the myometrium. The cells of the internal decidua showed signs of deterioration characterized by accumulation of clumps of chromatin in the nuclei and dilation of the perinuclear cisterna and endoplasmic reticulum cisternae. Autophagosomes and heterophagosomes accumulated in the cytoplasm of these cells. Cells particularly strongly affected became spherical and were devoid of their plasma membrane. Some cells near the trophoblast as well as the mature decidual cells situated farther from the embryo showed a normal morphology. The trophoblastic cells established close contact with healthy decidual cells and engulfed fragments of disorganized decidual cells. It is suggested that the death of decidual cells is a type of programmed cell death and that it is not due to a direct lytic action by the trophoblast.

Ultrastructural study of the mouse antimesometrial decidua

SummaryThe ultrastructure of mouse antimesometrial decidual cells was analyzed during the development of the decidua between days 5 and 8 of pregnancy. The first decidual cells, appearing on the 5th day, are polygonal with rounded nuclei and prominent nucleoli; free ribosomes predominate in the cytoplasm. On the 6th to the 8th days the cytoplasm of these cells is typically that of cells actively engaged in macromolecular synthesis. Large numbers of granular and agranular endoplasmic reticulum cisternae are present in addition to well-developed Golgi complexes, mitochondria and lysosomes. Many bundles of microfilaments and lipid droplets occur during this period. An intense accumulation of autophagosomes and lysosomes with very heterogeneous content was noted on the 7th and especially the 8th days. The presence of these organelles is an indication that involution of this part of the decidua has begun.

Growth curves, morphology and ultrastructure of ten Paracoccidioides brasiliensis isolates

The yeast phase of ten P. brasiliensis isolates were studied to characterize their growth pattern, morphology and ultrastructure. Growth curves were determined after counts of total and viable fungi units (FU) during 20 days. Three growth patterns were observed: slow, reaching approximately 10–30× 106 FU/tube (Pb 18, Pb 265 and PB 2); intermediate, reaching 60–150×106 FU/tube (IVIC Pb 9, IVIC Pb 267, Pb SN, Pb Vitor and Pb Campo Grande) and fast, reaching 180–370×106 FU/tube (Pb 2052 and Pb 192). The highest percentage of viable cells occurred on the 6th day of culture for Pb 192, Pb Campo Grande, Pb 2052 and IVIC Pb 9; on the 8th day for Pb Vitor, Pb SN, Pb 18 and IVIC Pb 267; on the 10th day for Pb 265 and on the 12th day of culture for Pb 2. Mean generation times varied from approximately 21.2 (Pb 2052) to 102.6 hours (Pb 265). The isolates showed similar morphology, except IVIC Pb 267 which did not present a typical yeast-phase at 35°C and the two fast-growing isolates (Pb 2052 and Pb 192) that presented smaller cell sizes and less tendency to clump. The ultrastructure of the isolates was similar: the cell walls presented a width of 0.1 to 0.2 °; the mitochondria presented few cristae and had equivalent patterns of distribution and morphology; the endoplasmic reticulum was scanty, presenting narrow cisternae; the vacuoles, empty or filled with electrondense material, were numerous and two to five nuclei with pores were constantly observed.

Plasma membrane vesicles isolated from epimastigote forms of trypanosoma cruzi

Membrane vesicles can be obtained from epimastigote forms of Trypansoma cruzi by incubating cells with either cross-linking reagents or acid pH. Acetate, phtalate or citrate, at pH 4.0, but not at higher pH values, were able to induce plasma membrane vesiculation. Vesicles have been purified by sucrose density centrifugation and their membrane origin was demonstrated by the following criteria: (a) Vesicles are 5--10 times richer in protein-bound iodine when they are prepared from cells previously labeled with 131I by the lactoperoxidase catalyzed reaction. (b) Electron microscopy of vesiculating cells shows physical continuity between cell plasma membrane and vesicle membrane. (c) Antibodies prepared against purified vesicles are able to agglutinate epimastigote forms of T. cruzi with sera dilutions up to 1 : 256 to 1 : 512. (d) Freeze-fracture studies of the purified vesicles have shown images of faces P and E compatible with known images of the intact cell plasma membrane. Typical preparations of acetate vesicles present the following characteristics: total carbohydrate : protein=1.5--2.0; orcinol : protein-0.07 and absence of diphenylamine reaction. Vesicles contain 0.2--0.5% and 0.3--1.0% of the total homogenate protein and carbohydrate, respectively. The presence of 10 major protein bands and 30--50-fold enrichment of the four sugar-containing macromolecules present in epimastigote forms of T. cruzi have been demonstrated in these preparations.

Ultrastructure of the endometrial blood vessels during implantation of the rat blastocyst

SummaryThe ultrastructure of the blood vessels of the endometrium was analysed during implantation of the blastocyst in rats, at the time of appearance of the Pontamine Blue Reaction. Vessels from implantation sites and from interimplantation sites were compared. In vessels from implantation sites the endothelial cells showed fenestrations covered by diaphragms. In addition, small interruptions (gaps) between the endothelial cells were observed. These features were present in vessels larger than 5 μm in diameter and more than 100 μm away from the uterine epithelium, both in the mesometrial and antimesometrial wall of the endometrium. Vessels from interimplantation sites displayed neither fenestrations nor interruptions. The endothelial cells of the implantation sites displayed morphological signs of metabolic activation. These consisted of increased numbers of polyribosomes, well developed Golgi complexes and prominent nucleoli. The fenestrations and gaps in the vessel wall were interpreted as constituting the morphological basis for the increase in vascular permeability and the consequent edema which characterize the Pontamine Blue Reaction.

Light and Electron Microscopic Analysis of Controlled Injury to Follicular Unit Grafts

BACKGROUND. Careful manipulation of hair units is essential for a good yield of transplanted hair. OBJECTIVE. To analyze the morphology of dissected follicular units submitted to crushing, stretching, bending, and drying. METHODS. Follicular units were either crushed, bent, stretched with forceps, or left drying on surgical gloves for 3 minutes. The specimens were fixed and prepared for observation with light microscopy or transmission and scanning electron microscopy. RESULTS. No alterations were detected in follicular units that had been crushed, bent, or stretched. Major damage occurred in samples that were left to dry on gloves. CONCLUSION. Letting the follicular unit dry appears to be the worst mishandling to which the follicular units may be subjected during routine hair transplantation.

Collagen Distribution in the Mouse Endometrium during Decidualization

The distribution of collagen and reticular fibers was studied in the endometrium of virgin and pregnant mice. The collagen and reticular fibers were examined in picrosirius-stained sections observed in a polarizing microscope and in silver-impregnated sections. Picrosirius-stained sections of animals in estrus, diestrus and on the 2nd day of pregnancy had fine greenish fibers distributed irregularly in the endometrium and thicker red fibers concentrated near the myometrium. Argyrophyl fibers in virgin mice were scarce and irregularly distributed. On the 4th day of pregnancy very few fibers were observed in the endometrium. On the 5th, 6th, and 7th days of pregnancy long greenish fibers were found surrounding decidual cells. A network of argyrophyl fibers was observed in the silver-impregnated decidua. It is suggested that new fibers are produced by decidual cells.

Ultrastructural cytochemical study of proteoglycans in the endometrium of pregnant mice using cationic dyes

Decidualization in rodents is accompanied by remarkable modifications of both fibrillar and non-fibrillar components of the endometrial extracellular matrix. Biochemical studies have shown that the levels of synthesis of hyaluronic acid and sulfated glycosaminoglycans change during decidualization in rodents. As the rodent decidua has regions containing cells in different stages of decidual transformation, we decided to analyse, by an ultrastructural cytochemical technique, the distribution of proteoglycans (PGs) in each region of the decidua of mice on different days of pregnancy. Endometria of mice on days 4, 5 and 7 of pregnancy were processed for electron microscopy in the presence of safranin O, a cationic dye which preserves most of the tissue PGs. The endometrium of non-pregnant mice was used as control. We observed evident differences in the arrangement and distribution of the network of PGs between non-pregnant and 4-day pregnant endometria, as well as between different regions of pregnant endometria. The possible relationship between these modifications and cell transformation that occurs during decidualization is discussed.