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Testing of the course of neurogenesis and gliogenesis in the germinative zones of the CNS of embryonal and early postnatal rats by means of the gel reaction for the histochemical demonstration of the thiamine-pyrophosphatase. Histochemical and autoradiographical study.

Using the histochemical reaction for the demonstration of TPPase the shape and distribution of the Golgi apparatus (GA) of the ventricular zones of the CNS of rats was studied during embryonal and postnatal development. In the cells of the ventricular zone of the spinal cord, the Hypothalamus, the Thalamus, the N. caudatus and the Cerebral cortex two forms of GA can be distinguished: Form 1 GA has the shape of small rods or grains placed in the ventricular process of the cell at various distances from the nucleus. In the spinal cord and the Hypothalamus the 1st form correlates with the period of neuroblastic production, and the same correlation probably exists in other regions of the CNS also. Form 2 GA is in sequence with form 1; it is characterized by a large beam shape, situated supranuclearly near to the nucleus. The relation of form 2 to gliogenesis is discussed. In the alar region of the spinal cord only form 1 GA occurs. The GA of cells of the subventricular zone has on average a lower enzymatic activity than that of the ventricular zone. By day 18 intra uterinam the subventricular cells have the shape of very small grains (1 micron) or are not visible. From day 18 intra uterinam cells with a large GA begin to appear and steadily grow in number until postnatally they form the majority of the subventricular layer. Autoradiographical investigation and a comparison with published data showed that cells without GA and with discrete GA are stem cells and those with a large GA are glioblasts. Glioblasts are arranged in the subventricular zone with the GA pointing in the direction of migration. According to the ratio of glioblasts and stem cells topographical regions can be divided into early, transitional and permanent phases of glioproduction. The gel method of demonstrating TPPase is highly suitable for study of the differentiation of the CNS and for observing the progress of glioproduction.

Animals↗

[Electron microscope localization of thiamine pyrophosphatase and nucleoside diphosphatase in lymphocytes].

In ductus thoracicus lymphocytes of the rat the localization of specific phosphatases was studied by means of ultrahistochemistry. Thiaminepyrophosphatase was found in the outer lamellae of the Golgi-field as well as in lysosomes, whereas nucleosiddiphosphatases could be localized on the plasma membrane at low pH. These findings are correlated with the phaenotype of B- and T-lymphocytes. Their functional significance is discussed.

Animals↗

Inorganic pyrophosphatase activity of purified bovine pulp alkaline phosphatase at physiological pH.

At physiological pH, the hydrolytic activity of purified bovine pulp alkaline phosphatase toward phosphorus compounds was observed to be in the order of inorganic pyrophosphate greater than beta-glycerophosphate greater than phosphorylcholine greater than p-nitrophenylphosphate greater than glucose-6-phosphate. Optimum pH of the enzyme toward inorganic pyrophosphate was shown to be 8.5, with around 60% of the activity at pH 7.5. The activity was increased by the addition of Mg2+, but a different pattern of activation was observed between pH 7.5 and 8.5.

Alkaline Phosphatase↗

Cytochemical localization of beta-NADPase in rat hepatocytes and Kupffer cells. Comparison with thiamine pyrophosphatase (TPPase).

The intracellular localization of beta-NADPase in rat hepatocytes and Kupffer cells has been studied and compared with the pattern of TPPase in these cells. The reaction product for beta-NADPase is present in some but not all hepatocytes in two cisternae on the trans aspect of the Golgi apparatus. It is absent from the trans-most lamella and the GERL of hepatocytes. TPPase, on the other hand, is limited to the first Golgi cisterna on the trans aspect with sprinkles of reaction product in the second lamella. Considering that TPPase is a marker of the trans Golgi lamella and hepatocyte Golgi stacks contain usually 2-4 lamellae, our observations suggest that beta-NADPase is localized in the trans as well as in the intermediate Golgi lamellae of liver parenchymal cells. In Kupffer cells, the reaction product for both beta-NADPase and TPPase was found in some but not in all cells. The enzyme beta-NADPase was localized in the rigid lamella and the tubulovacuolar system of GERL. This pattern differed significantly from that for TPPase, which was found in 2-3 cisternae at the trans aspect of the Golgi complex in Kupffer cells. These observations demonstrate the difference in the localization of beta-NADPase in hepatocytes and Kupffer cells. Such differences should be taken into consideration in studies of Golgi fractions, when phosphatase reactions are used as specific markers of Golgi components.

Animals↗