acid-phosphatase and uranyl-acetate

Ultrastructural localization of acid phosphatase activity was investigated in the jejunal absorptive cells of 21 d fasted rats (about 500 g). The enzyme activity was localized on the membrane of microvilli, the lateral cell membrane, the Golgi complex, the lysosomes and the GERL of Novikoff (a part of the smooth surfaced endoplasmic reticulum located in close proximity to the Golgi saccules) of jejunal absorptive cells. Moreover, the lysosomes of various sizes and shapes with acid phosphatase activity was characteristically encountered in the infranuclear cytoplasm. The lysosomes appeared to be autolysosomes .

Topics: Acid Phosphatase; Animals; Cell Membrane; Citrates; Citric Acid; Endoplasmic Reticulum; Fasting; Golgi Apparatus; Intestinal Mucosa; Jejunum; Lysosomes; Male; Microscopy, Electron; Microvilli; Organometallic Compounds; Rats; Staining and Labeling; Uranium

Lead ions at similar concentrations to those used for Gomori type phosphatase localization stain some parts of the vacuolar system, particularly compartments of the Golgi complex (GC) and isolation envelopes (im) in a characteristic way in both vertebrates and invertebrates. After fixation in 2.5% glutaraldehyde, lead citrate in acetate or aspartate buffer (pH 5.5-7.2) leaves the contents of GC cisternal compartments with a fine particulate stippling. In the fat body of Calpodes ethlius and in mouse pancreas the staining is faint but definite without further enhancement of contrast, although it is easily overlooked after section staining. The distribution of lead stain differs from that of the lead phosphate precipitated after Gomori type acid phosphatase reactions. Whereas lead stain may be in all GC and im compartments, acid phosphatase is restricted to the innermost saccules and nearby vacuoles. The compartment specific staining by led also differs from the generalized staining in all compartments given by uranyl. Thus the contents of luminal membrane surfaces of some parts of the vacuolar system can be characterized by their ability to bind lead. In cells where protein synthesis has been blocked by cycloheximide, secretory vesicles are absent and the RER and GC from the generalized staining in all compartments given by uranyl. Thus the contents of luminal membrane surfaces of some parts of the vacuolar system can be characterized by their ability to bind lead. In cells where protein synthesis has been blocked by cycloheximide, secretory vesicles are absent and the RER and GC from the generalized staining in all compartments given by uranyl. Thus the contents of luminal membrane surfaces of some parts of the vacuolar system can be characterized by their ability to bind lead. In cells where protein synthesis has been blocked by cycloheximide, secretory vesicles are absent and the RER and GC cisternae are devoid of uranyl stainable material. However, lead staining and acid phosphatase activity in the GC continue. We presume that they mark the environment within these cisternae rather than the proteins passing through them. This environment is itself not static. Several observations suggest that the function of cisternae that is detectable by lead staining is temporally discontinuous and related to a stage of maturation or development. Only early stage ims stain: the staining ceases by the beginning of autophagy after hydrolytic enzymes are presumed to ha

Topics: Acid Phosphatase; Adipose Tissue; Animals; Aspartic Acid; Cell Compartmentation; Endoplasmic Reticulum; Golgi Apparatus; Lead; Lepidoptera; Mice; Microscopy, Electron; Organometallic Compounds; Pancreas; Protein Biosynthesis; Staining and Labeling; Uranium

Ultrastructural evaluation of eosinophilic leukocytes from a 2-yr-old asymptomatic girl with chronic benign neutropenia (CBN) revealed a variety of morphological abnormalities. All eosinophils obtained from blood and marrow specimens contained multipole microcrystalloids in most of the mature cytoplasmic granules. An increase in crystalloid-free, immature granules in late (bilobed nuclei) eosinophils suggested a delay in granule maturation. The eosinophil granules appeared to be of normal size and demonstrated normal acid phosphatase reactivity. Eosinophilic myelocytes contained abnormal cisternae of rough endoplasmic reticulum (RER) and lacked abundant elongated RER cisternae seen in normal cells. A few eosinophilic myelocytes in specimens of bone marrow from the child contained large intranuclear crystalloids measuring up to 3 mu in length. The intranuclear crystalloid contained as cubic lattice of dense material with a periodicity similar to that described for cytoplasmic crystalloids. The ultrastructural morphology of marrow neutrophils was normal, as described in other cases of CBN. Ultrastructural examination of blood eosinophils from the father demonstrated microcrystalloids in cytoplasmic granules identical to those seen in the child. The father was asymptomatic and had normal leukocyte counts. Thus, anomalous crystalloid granule genesis occurred in the father and daughter and was not necessarily associated with neutropenia or clinical symptomatology. This anomaly is associated with the accumulation of intranuclear crystalloid material in eosinophilic myelocytes, which do not appear to be released from the marrow compartment.

Topics: Acid Phosphatase; Bone Marrow Cells; Cell Nucleus; Child, Preschool; Crystalloid Solutions; Cytoplasmic Granules; Eosinophils; Female; Humans; Isotonic Solutions; Organometallic Compounds; Plasma Substitutes; Uranium