Histochemical localization of primary and secondary alcohol dehydrogenases in whole-body, freeze-dried sections of mice

1984 ◽  
Vol 16 (9) ◽  
pp. 931-940 ◽  
Author(s):  
Toru Egashira ◽  
William J. Waddell
2008 ◽  
Vol 43 (4) ◽  
pp. 265-274 ◽  
Author(s):  
Claude Rouleau ◽  
Jagmohan Kohli

Abstract Nonpersistent contaminants represent thousands of chemicals used as pesticides, pharmaceuticals, personal care products, additives, etc. Because of this diversity, the assessment of the environmental risks they may pose for the environment represents a formidable task. Identification of target organs is key information needed to orient further research on newlyinvestigated organic xenobiotics. We used whole-body autoradiography to visualize the distribution of 14C-labelled atrazine, methoxychlor, glyphosate, and bisphenol-A in goldfish (Carassius auratus) and identify target organs. Fish were exposed for 2 days (glyphosate and bisphenol-A) and 7 days (atrazine and methoxychlor) to the radiolabelled compounds at a concentration of 15 nM. They were then frozen, embedded in carboxymethylcellulose gel, 20-μm-thick cryosections were collected, freeze-dried, and exposed to phosphor screens to visualize the tissue distribution of radioactivity. Goldfish did not accumulate glyphosate. The three other compounds were accumulated, mostly in the gall bladder. Nevertheless, unforeseen accumulation sites were observed; atrazine accumulated in the uveal tract of the eye, high levels of radioactivity were found in the cerebrospinal fluid of goldfish exposed to methoxychlor, and an important accumulation of bisphenol-A was seen in urine, oral mucosa, esophagus, and intestinal lumen. The potential toxicological consequences of the accumulation of these chemicals at very specific locations within the fish body are discussed and further research suggested.


1987 ◽  
Vol 1 (2) ◽  
pp. 236-244 ◽  
Author(s):  
T. Kawamoto ◽  
M. Shimizu

The distribution of calcium and phosphate in the cells of the enamel organ of the rat lower incisors was investigated by autoradiography and energy-dispersive x-ray spectrometry (EDS). Radioactive calcium or phosphate was injected i.p. into seven-day-old rats of the Wistar strain. The animals were frozen 0.5, 1, and 10 min after injection, and embedded in 5% carboxymethyl cellulose. Sagittal sections of 10 μm thickness were made in which the lower incisor was included as a part of the whole-body section. For autoradiography, the sections were freeze-dried and placed in contact with dry thin films prepared from autoradiographic emulsion. For EDS, sections were mounted on carbon stubs, freeze-dried, coated with carbon, and examined by EDS in a SEM. 45Ca and 32P autoradiograms showed that the radioactivity was located over the papillary layer cells adjacent to the secretory stage ameloblasts and was much higher here than in the ameloblastic layer. On the other hand, there was no significant difference between the amount of radioactivity of these two cell layers in the maturation stage, although higher radioactivity was detectable in the maturation stage enamel than in the secretory stage enamel. Pronounced Ka x-ray peaks were obtained for P, S, Cl, and K originating from the cells of the papillary and ameloblastic layers in the secretory stage, but only very low peaks were obtained for Ca. On the other hand, in addition to these elements, remarkably high Ca and Fe peaks could be detected in the ameloblastic layer of the maturation stage.


1984 ◽  
Vol 32 (1) ◽  
pp. 1-6 ◽  
Author(s):  
S Sjögren

Freeze-dried sagittal, whole-body sections of 10-day-old rats were incubated for lactic dehydrogenase (LDH) using different media in the presence of the inhibitors urea and fluoropyruvate. Phenazine methosulfate (PMS) and menadione, which are regularly used in current histochemical media and are believed to promote the demonstration of LDH activity, were also added and shown to be insufficient for the demonstration of total LDH activity, and PMS even seemed to have an inhibitory effect on LDH activity in oral epithelium. However, cumulated data from the different incubations show that the oral epithelium of developing rats may contain two different types of LDH, one in the basal cells with possibly aerobic characteristics, and another in the spinosum/granulosum cells with anaerobic characteristics.


1978 ◽  
Vol 26 (11) ◽  
pp. 967-972 ◽  
Author(s):  
L Heywood ◽  
A Blackshaw

The activity of lactate dehydrogenase (LDH) in freeze-dried sections of rat testes was determined by using a fluorometric assay method and found to be 4.47 +/- 0.23 moles/Kg dry weight/hr (MKDH +/- S.E.M.) in whole sections, 3.31 +/- 0.16 in tubules and 12.0 +/- 1.9 in interstitial tissue. The activities and regional variation are similar to those measured in nervous tissue and are well correlated with the histochemical localization of LDH activity when phenazine methosulphate (PMS) is not used as an electron carrier. LDH and lipoamide dehydrogenase activity have the same histochemical distribution and there is no nonspecific staining with either method. The use of PMS results in reduced dependence on substrate and coenzyme and does not indicate higher interstitial activity but may provide an indication of developing lactate metabolism in maturing sperm. It is recomended that methods with and without PMS be used in studies of LDH activity in the testis.


2004 ◽  
Vol 20 (1-5) ◽  
pp. 9-19 ◽  
Author(s):  
Sam Jacob ◽  
Ahmed E Ahmed

Previous studies from this and other laboratories have indicated the role of species difference in acrylonitrile (VCN) toxicity and its metabolism to cyanide. Our recent studies also indicated a more pronounced elimination of VCN following oral as compared to i.v. administration. To further characterize the mechanism of these differences on the distribution of VCN, quantitative whole-body autoradiographic distribution and elimination studies were conducted at various time points (0.08, 8, 24, 48 h) following the administration of an equivalent i.v. dose of 2-[14C]-VCN to male Fischer rats and male CD-1 mice. Whole-body autoradiographs obtained from freeze-dried and acid-extracted sections of rats and mice demonstrated a rapid uptake of 14C in liver, lungs, spleen and bone marrow at early time intervals. Quantitatively, the uptake, retention and covalent interaction of 14C were higher in organs of rats as compared to mice, over 48 h. Mice eliminated 74% of the total administered dose of 2-[14C]-VCN (expired air 4%, urine 16% and feces 54%), while rats eliminated only 26% of the dose (expired air 2%, urine 4% and feces 20%). Species differences in VCN toxicity seem to be correlated with its rate of elimination. The distribution and elimination data demonstrated that mice divest VCN more rapidly than rats. The study also demonstrated that administration of VCN in rats resulted in covalent interactions and retention of 2-[14C]-VCN/metabolites in the tissues thus exerting more chronic toxicity to rats than to mice.


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