The aim of this study was to determine the subcellular distribution of bis(N-ethoxy N-ethyl)dithiocarbamato nitrido technetium(V) (99mTcN-NOEt) in rat heart by differential centrifugation techniques. Extraction of the activity from homogenized rat heart tissue was also performed to assess whether myocardial retention might induce changes in the chemical identity of the complex.
Methods: Anesthetized rats were intravenously injected with 99mTcN-NOEt, the heart tissue was extracted and homogenized and tissue fractions were obtained by differential centrifugation. The efficiency of organelle separation was determined by assay of each centrifugal fraction using enzyme markers. Lactate dehydrogenase (LDH), acid phosphatase (ACP), alkaline phosphatase (ALP) and 5'-nucleotidase (5'ND) activities were assayed using standard spectrophotometric methods. Succinic dehydrogenase (SDH) activity was determined using a p-iodo-nitrotetrazolium-linked assay. Severe cell membrane and organelle disruption were induced by prolonging the homogenization time and their effect on the subcellular distribution of 99mTcN-NOEt was studied. The activity from homogenized heart tissue was extracted using the Folch technique and analyzed by TLC and HPLC.
Results: Most of the 99mTcN-NOEt activity was found to be associated with the hydrophobic components of the cell. No evidence of specific association of activity with the cytosolic and mitochondrial components was observed. Organelle and membrane cleavage did not cause release of activity into the cytosol. Approximately 90% of 99mTcN-NOEt activity was extracted from ventricular tissue and the chemical nature of 99mTcN-NOEt was not altered by uptake by myocardium.
Conclusion: Cell membranes are the most apparent site of localization of 99mTcN-NOEt in heart tissue.