scholarly journals ELECTRON MICROSCOPIC LOCALIZATION OF ACID PHOSPHATASE AND THIAMINE PYROPHOSPHATASE ACTIVITY IN HYPOTHALAMIC NEUROSECRETORY CELLS OF THE RAT

1964 ◽  
Vol 21 (1) ◽  
pp. 35-47 ◽  
Author(s):  
Joseph Osinchak

Supraoptic nuclei in the hypothalamus of rats were fixed for the electron microscope by vascular perfusion with solutions of glutaraldehyde followed by post fixation with osmium tetroxide. Cytochemical methods for detection of acid phosphatase and thiamine pyrophosphatase activity have been applied to glutaraldehyde-fixed frozen sections containing the neurosecretory cells. The enzyme activities have been localized to certain Golgi cisternae. Acid phosphatase activity is present in the large (0.4 µ to 1.0 µ) granules or dense bodies which are surrounded by a single limiting membrane; both features characterize these structures as lysosomes. Smaller (0.1 µ) granules also present in the perikarya are generally unreactive towards enzyme activity and resemble in form the neurosecretory granules in the neurohypophysis.

1962 ◽  
Vol 12 (2) ◽  
pp. 385-410 ◽  
Author(s):  
Sanford L. Palay ◽  
S. M. McGee-Russell ◽  
Spencer Gordon ◽  
Mary A. Grillo

This paper describes in detail a method for obtaining nearly uniform fixation of the nervous system by vascular perfusion with solutions of osmium tetroxide. Criteria are given for evaluating the degree of success achieved in the preservation of all the cellular components of the nervous system. The method permits analysis of the structural relations between cells at the electron microscopic level to an extent that has not been possible heretofore.


Author(s):  
J. R. Hillman ◽  
W. G. Seliger ◽  
P. E. Burk

In a previous histochemical study, high levels of acid phosphatase were found in the developing adrenal cortex of the golden hamster. The present study describes the cytochemical localization of acid phosphatase and thiamine pyrophosphatase in the adult animal. These localizations are then related to protein uptake by adrenal cortical cells using the exogenous horseradish peroxidase as a tracer protein. Animals were sacrificed at times ranging from 15 minutes to 24 hours after injection of peroxidase. The techniques described by Novikoff et al. were utilized for localization of acid phosphatase and thiamine pyrophosphatase.In acid phosphatase preparations (Fig. 1), dense bodies containing reaction product are found in the cytoplasm of most cortical cells and are often associated with small coated vesicles near the Golgi area. Acid phosphatase activity is present in cells of the zona glomerulosa as well as those of the other two cortical zones.


2021 ◽  
Vol 25 (3 (99)) ◽  
pp. 25-32
Author(s):  
R. Bulyk ◽  
T. Bulyk ◽  
O. Smetanuik

The aim: to study the effect of melatonin on the ultrastructural state of the supraoptic nuclei of the hypothalamus of rats under immobilization stress.Materials and methods. The experiments were performed on non-linear male white rats weighing 200-220 g. The animals were divided into 3 study series, in each of which the biomaterial was collected at 2 p.m. and at 2 a.m. using electron microscopic method. Long immobilization stress was simulated by keeping rats in special plastic penal cages for 6 hours daily for 7 consecutive days. Melatonin (Sigma, USA, 99.5% purification degree) at a dose of 0.5 mg/kg, in 1.0 ml of solvent (0.9% ethanol solution on physiologic saline) was injected daily, intraperitoneally.Results. When the animals were kept under the standard light regime, the ultrastructural organization of the hypothalamic nuclei at 2 p.m. indicated their low functional activity in comparison with the studies carried out at 2 a.m. Prolonged exposure of rats to immobilization stress was reflected in a significant rearrangement of the ultrastructural organization of supraoptic nuclei of the hypothalamus. The established changes can be considered as a manifestation of neurosecretory activity suppression, a decrease in neurosecretase production by hypothalamic neurons. Melatonin injections against the background of immobilization stress resulted in relative normalization of ultrastructural state of neurons of supraoptic nuclei of the hypothalamus of animals. In particular, studies at 2 a.m. revealed light neurosecretory cells containing a large nucleus, it was pyknotically altered. Karyolema invaginations, euchromatin dominance in the nucleus were observed. Heterogeneous changes were observed on the part of mitochondria. Enlarged tubules of granular endoplasmic reticulum were seen. At the same time, a small number of ribosomes and few hormonal granules were noticeable in neuroplasm. The mentioned picture of neurosecretory cells reflects a relative improvement in their electron microscopic state, which is evidenced by the appearance of neurosecretory granules. However, the ultrastructure of other organelles of the studied neurons indicates a depleted state caused by prolonged immobilization.Conclusions. 1. In animals under standard photoperiod conditions, the structural organization of supraoptic neurons of the hypothalamic nuclei during the nighttime of the experiment reflects the intensity of intracellular synthesizing processes (at 2 a.m.). A decrease in the activity of the structures under study is noted during the daytime. 2. Under immobilization stress, the ultrastructural organization of the above neurons indicates a pronounced disturbance of reactive nature with the signs of decreased functional ability of the structures and the phenomena of edema and destruction during the period of observation. 3. Melatonin injections against the background of immobilization stress led to a relative improvement in the ultrastructural state of the animals’ hypothalamic nuclei neurons, which is evidenced by the appearance of neurosecretory granules. However, the ultrastructure of other organelles of the studied neurons indicated a depleted state caused by prolonged immobilization.


1971 ◽  
Vol 19 (12) ◽  
pp. 801-808 ◽  
Author(s):  
TATSUJI MATSUZAWA ◽  
H. CLARKE ANDERSON

The presence and distribution of alkaline phosphatase, adenosine triphosphatase (ATPase) and acid phosphatase activities in the epiphyseal plates of young mice were studied by electron microscopic cytochemical methods. Alkaline phosphatase and ATPase activities were associated with the plasma membranes of chondrocytes and with the investing membranes of matrix vesicles. These vesicles contain the earliest recognized deposits of hydroxyapatite and may promote calcification through an active process. Alkaline phosphatase and ATPase activities were greatest in the hypertrophic zone of the epiphysis, which is an area of beginning calcification. Acid phosphatase activity was demonstrable in the cytoplasm of chondrocytes in association with dense bodies which were larger than matrix vesicles and were devoid of alkaline phosphatase and ATPase activities. These cytoplasmic lysosome-like bodies were slightly more numerous in the hypertrophic zone but disappeared in the underlying zone of chondrocyte degeneration and matrix calcification. Our observations do not support a previous suggestion by others that matrix vesicles represent lysosomes. The presence of ATPase and alkaline phosphatase is compatible with an enzymatic calcium- and/or phosphate-concentrating mechanism in the matrix vesicles.


1971 ◽  
Vol 19 (12) ◽  
pp. 775-797 ◽  
Author(s):  
ANDRÉE TIXIER-VIDAL ◽  
RENÉE PICART

Structures demonstrating the presence of glycoproteins, acid phosphatase activity and OsO4 impregnation were localized by means of the electron microscope in duck and in quail pituitary cells. Two methods for the electron microscopic demonstration of glycoproteins were used: a chromic acid-phosphotungstic acid mixture on glycol-methacrylate-embedded tissues, and the periodic acid-thiocarbohydrazide-silver proteinate technique. Both methods showed glycoproteins in the following sites: ( a) the secretory granules in three types of cells (A, B, C) which are part of the seven different cells of the avian pituitary; ( b) the several kinds of dense bodies which are richer in reaction product than the secretory granules. A correlation with previous studies on similar species of birds is helpful in identifying each of the three positive types of cells as thyrotropic cell (A), prolactin cell (B) and gonadotropic cell (C). The presence of glycoproteins within the Golgi saccules (on condensing granules) was found with the periodic acid-thiocarbohydrazide-silver proteinate method in these gonadotropic cells only. In gonadotropic and thyrotropic cells, acid phosphatase activity is weak in the inner Golgi saccules and strong in the "Golgi Endoplasmic Reticulum Lysosomes" system, in the lysosomes, in the dense bodies and in the vacuolated dense bodies. The structures which are richest in glycoproteins are also those which have the most acid phosphatase activity. On the contrary, OsO4-stained structures in duck gonadotropic cells (nuclear pericisterna, rough endoplasmic reticulum, cisternae and outer Golgi saccules) have no glycoproteins or acid phosphatase activity.


Author(s):  
Joe Wood

The electron microscopic localization of biogenic amine neurotransmitters can provide for a definitive approach to the study of these chemical agents in relation to the neuroanatomical substrate of the nervous system. Therefore, if one can specifically identify a neurotransmitter within certain ultrastructural nervous system components then a better understanding is developed of the chemico-morphologic functional relationships. Thus one can develop further the concepts of the chemical interactions within the nervous system.The localization technique must be chemically specific for a particular compound yet must be so conducted that it produces relatively little artifact or damage within the tissue to disturb the morphological integrity. The cytochemical localization of the agent should possess contrast either in density or chemical configuration so that it can be specifically identified even after the tissue is treated with a relatively nonspecific agent such as osmium tetroxide or is stained after sectioning with conventional stains such as uranyl acetate and/or lead citrate.


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