Folia Psychiatrica et Neurologica Japonica, Vol. 31, No. 2, 1977
Electron Microscopic Study of Experimental Thallotoxicosis Motonori Deshimaru, M.D.* Taihei Miyakawa, M.D. * Shiro Sumiyoshi, M.D. * * Fumie Yasuoka, M.D. * * Kosuke Kawano, M.D. * * and Megumi Kuramoto, M.D. * * *Department of Neurology, Toxicology Institute, Kumamoto University School of Medicine, Kumamoto **Department of Neuropsychiatry, Kumamoto University School of Medicine, Kumamoto
INTRODUCTION In the late 19th century, thallium salts were used in treating gonorrhea, syphylis, tuberculosis and ringworm. But therapeutic use of this drug was discontinued because of its toxity to human. Recently, thallium salts have come to be widely used as pesticides and rodenticides. There are many reports of human and experimental animals afflicted with thallotoxicosis caused by suicidal injection of rodenticides.13 .1 0 0 11 14 15 18 10 22 23 The clinical symptoms of thallotoxicosis are described as polyneuropathy, gastrointestinal symptoms, alopecia, retrobulbar optic neuritis, encephalopathy (unconsciousness, convulsion, ataxia, choreoathetosis) and muscular weakness.l l4 l5 l8 21 Pathologically, changes in the peripheral nerves are the chiefly observed symptoms, and this type of changes is of the dying-back type that is found most widespread in the distal regions. The damage to the peripheral nerves is considered to be primary disorder of the axon with secondary changes of the myelin sheaths.3 l7 21 The changes of CNS are edema and vascular enlargement in the cerebral hemispheres and brainstem, and there are chromatolytic changes of neurons in the morter cortex, the nuclei of the oculomotor or I11 cranial Received for publication Jan. 6, 1977.
nerve, the substantia nigra, and the globus pallidus and anterior horn cells of the spinal cord.3 I) 11 14 In the present experiment, we examined the nervous system, muscles and visceral organs of rats poisoned with thallium salts. MATERIALS AND METHODS Adult male rats (body weight 100-120 g) were used. The experimental group consisted of five rats and the controls of five rats. To the experimental group 2 mg per rat of thallium acetate: CH, COOT1 was given orally daily. Alopecia began on the 2 1st day after administration and gradually spread over the whole body. During administration, we did not observe any kind of neurological signs. On the 180th day of administration of thallium acetate, the experimental animals were killed with controls. Fixation was carried out by perfusion through the heart with 3% glutaraldehyde in pH 7.4 phosphate buffer. Blocks from cerebrum, cerebellum, spinal cord, peripheral nerves, skeletal muscles (lower limbs), liver and kidneys were postfixed in 2% phosphate-buffered osmic acid for two hours, then dehydrated and embedded in epon. Slices for optical microscopy were prepared with a Porter-Blum T microtome. After observation of toluidine blue preparations, the ultrathin sections were studied with a Hi-
270
M. Deshimaru, et al.
Fig. 1 : Cross striations of skeletal muscle are irregular in shape and have partially disappeared. Toluidine blue stain. ( X 380)
Fig. 2: The sciatic nerve are intact. Toluidine blue stain. ( X 300)
Fig. 3 : Sarcoplasmic reticulums are degenerated and atrophic. Myofilaments of Zband are irregular in shape and partially destroyed. Glycogen granules are not observed. ( X 9 , O O O )
Study of Experimental Thallotoxicosis
Fig. 4: Mitochondrias are swollen, destroyed and have disappeared in the cristae and show vacuolation. The mitochondria1 membranes are sometimes ruptured. ( X 8,000)
Fig. 5: A great number of mitochondria were accumulated in the peripheral parts of the muscle fibers. ( X9,OOO)
Fig. 6: In the remarkably damaged regions, myofilaments are destroyed. ( x 12,000)
27 1
M. Deshimaru, et al.
212
tachi 1 1A electron microscope. RESULTS
Light Microscopy Cross striation of skeletal muscles was irregular in shape and had partially disappeared (Fig. 1). The sciatic nerves were not different from those of the controls (Fig. 2). Toluidine-blue stained sections did not reveal any changes in the frontal, parietal lobes, thalamus, hypothalamus, cerebellum, spinal cord, liver and kidney. Electron Microscope In the skeletal muscles, sarcoplasmic reticulum was degenerated and atrophic. Myofilaments of Z-band were irregular in shape and partially destroyed. Moreover, no glycogen granules were observed (Fig. 3). Some of the rnyofilaments were destroyed and disappeared, and remarkable changes were observed in the mitochondria. They were swollen, destroyed and disappeared in cristae, and showed vacuolation. The mitochondrial membranes were sometimes ruptured (Fig. 4). A great number of mitochondria accumulated at the peripheral parts of the muscle fibers and they also showed similar changes (Fig. 5 ) . In the prominently damaged regions, myofilaments were destroyed (Fig. 6). In CNS, changes of thalamus and hypothalamus were remarkable in comparison with the other parts of the brain. Slight hypertrophy and vacuolation of mitochondria were observed in the cytoplasm of the nerve cells in the hypothalamus, along with dilatation of the Golgi cisterns (Fig. 7).
DISCUSSION Among the described-above pathological findings, the changes of the skeletal muscles were the most pronounced among all animals of the experimental group. The changes in the muscle were observed chiefly in the mitochondria, sarcoplasmic reticulum
and myofilament. In CNS, changes were observed chiefly in the thalamus and hypothalamus. These changes were vacuolation of the mitochondria and dilatation of the Golgi cisterns in the cytoplasm of the nerve cells. According to pathological findings in autopsy cases and experimental animals that have been reported so far, the peripheral nerves showed noticeable l1 l4 but changes in muscles changes1 have not been described. Spencer et aLZg reported that in examining the effect of thallium on organotypic cord-ganglia-muscle combination cultures, no changes were observed in the skeletal muscle tissue after a week of exposure to thallous sulfate. Thallous ions, biochemically, are considerably similar to potassium ions, and their ionic radii are very similar.2oTI' ions substituted for K' ions in (Na' +K') activated ATPaseqGT1' ions has 10 times greater affinity for activating ATPase than K' ions.2 Mullins et reported that a high concentration of T1' depolarized the membrane. And, if a high concentration of T1' was allowed to act for an appreciable period of time, TI' ions led to an irreversible damage to the muscle. Miyakawa et u1.12 administered lithium carbonate to rats and reported electron microscopic findings. The changes they found were very similar to our findings in the muscles. The changes in the muscles were considered to be due to the disturbances of electrolyte by replacement of Na' ions to Li' ions. We speculate that our findings in the muscles, as a result of our experiment, were also caused by the disturbances of the electrolyte due to replacement of K' ions to T1' ions. As the changes of mitochondria in the muscles were severer than the changes of myofilament, the changes in the muscles might be one of primary disorder in the mitochondria with secondary changes in the myofilament. Hermann et almsand Spencer et al." pointed to the changes of mitochondria in experimental thallotoxicosis. Spencer et d.
Study of Experimental Thallotoxicosis
273
Fig. 7: Nerve cell in the hypothalamus. Slight hypertrophy and vacuolation of mitochondria and dilatation of Golgi cisterns are observed. ( X7,OOO)
examined the ultrastructural changes in nerve tissue that has been exposed to thallium salts in vitro and reported the enlargement, destruction of cristae, vacuolation of the axonal mitochondria and partial rupture of the outer mitochondrial membrane. It is very interesting that our findings regarding the mitochondria were similar to these of Spencer's experiment, in spite of the different techniques employed in the experiments. The mechanism by which the mitochondria is affected selectively by T1' ions is still unsolved. However, Hermann et al. suggested the possibility of thallium becoming bound to the mitochondrial membranes. Spencer et d.,using electron microprobe mass spectrometry, detected thallium in the outer mitochondrial membranes. The changes of CNS reported up to the present are edema, vascular enlargement and chromatolytic changes of neurones.
But, in our experiment, the toluidine-blue stained sections did not reveal any changes in CNS. However, electron microscopically, changes in the mitochondria and dilatation of the Golgi cisterns were observed in the cytoplasm of the nerve cells-chiefly in the thalamus and hypothalamus. Hendleman7 reported a marked swelling of perikaryal granular endolplasmic reticulum when thallous acetate was exposed to cultures of rat dorsal root ganglia. As mentioned above, no changes in peripheral nerves were found in our experi. ~ ment. However, Hermann et ~ 1 studied the morphologic changes induced by the subcutaneous injection of varying doses of thallium acetate in rats and reported that no abnormalities were noted in the peripheral nerves. But, according to the histopathological studies of autopsy cases and in experimental animals (using mice and cats), the peripheral nerves have shown some
M. Deshimaru, et al.
274
changes. Thus, it is necessary to try an experiment under the same conditions to determine whether the difference of races or quantity of administration could be the reason. SUMMARY
To adult male rats 2 mg per rat of thallium acetate: CH, COOT1 was given orally daily for six months. Clinically, the experimental rats revealed only alopecia and showed no neurological signs. Pathological findings were noted in muscle and cerebrum. Marked changes were observed in muscles and were as follows: swelling and vacuolation of mitochondria, destruction of cristae, ruptures of mitochondrial membranes, degeneration of sarcoplasmic reticulum and destruction of myofilaments. In the cerebrum, there were vacuolation of mitochondria, dilatation of Golgi cisterns in hypothalamus and thalamus. Sciatic nerve, liver and kidney were intact. From this, we consider that T1' ions selectively affect the muscle and its mitochondria. REFERENCES Bank, R. J., Pleasure, D. E., Suzuki, K., Nigro, M. and Katz, R.: Thallium poisoning, Arch Neurol, 26: 456-464, 1972. Britten, J. S. and Blank, M.: Thallium activation of the (Na+-K+) activated ATPase of rabbit kidney, Biochem Biophys Acta 159: 160-166, 1968. Cavanagh, J. B., Fuller, N. H., Johnson, H.R.M. and Rudge, P.: The effects of thallium salts, with particular reference to the nervous system changes, Quart J Med, 43: 293-319, 1974. Chamberlain, P. H., Stavinoha, W. B., Davis, H., Kniker, W.T. and Panos, T. C.: Thallium poisoning, Pediatrics, 22: 1170-1 182, 1958. Gehring, P. and Hammond, P.: Mechanism for the increase in thallium excretion during the administration of potassium, Pharmacologist, 8: 262, 1969.
6 Grunfeld, 0. and Hinostroza, G.: Thallium poisoning, Arch Intern Med, 114: 132138, 1964. 7 Hendleman, W.: The effect of thallium on
peripheral nervous tissue in culture: A light and electron microscopic study, Anat Rec, 163: 198A, 1969. 8 Hermann, M. M. and Bench, K. G.: Light and electron microscopic studies of acute and chronic thallium intoxication in rats, Toxicol Appl Pharmacol, 10: 199-222, 1967. 9 Kakehashi, K.: Experimental studies on the pathogenesis of the Minamata disease: Studies of delayed thallium intoxication, Kumamoto Med J, 33, suppl, 3: 704-713, 1959 (in Japanese). 10 Lund, A.: Distribution of thallium in the organism and its elimination, Acta Pharmacol, 12: 251-259, 1956. 1 1 Matsumura, K.: Experimental studies on the pathogenesis of the Minamata disease: Studies of acute and chronic thallium intoxication, Kumamoto Med J, 34, suppl, 3: 655-669, 1960 (in Japanese), 12 Miyakawa, T., Sumiyoshi, S., Deshimaru, M., Hattori, E. and Miyakawa, M.: Electron microscopic studies on the effects of the administration of Lithium carbonicum in rats, Ann Rep Pharmac Res Foun, 5: 9-14, 1973 (in Japanese). 13 Mullins, L. T. and Moore, R. D.: The movement of thallium ions in muscle, J gen Physiol, 43: 759-773, 1960. 14 Munch, J.C., Guinsbeg, H. M. and Nixon, C. E.: The 1932 thallotoxicosis outbreak in California, JAMA, 100: 1315-13 19, 1933. 15. Oguchi, K., Nishida, M. and Tsubaki, T. Thallotoxicosis: Report of two cases, Clin Neurol, 14: 42-47, 1974, (in Japanese). 16 Peterson, E. R. and Murray, M. R.: Patterns of peripheral degeneration in vitro, A m NY Acad Sci, 122: 39-51, 1965. 17 Peterson, E. R. and Grain, S. M.: Regeneration and innervation in cultures of adult mammalian skeletal muscle coupled with fetal rodent spinal cord, Exp Neurol, 36: 136, 1972. 18 Prick, J.J.G., Smith, W.G.S. and Miller, L.: Thallium poisoning, Amsterdam, 1955. 19 Reed, D.C.J., Faro, S.N., Pieper, S. J. and Kurland, L. T.: Thallotoxicosis: Acute manifestations and sequelae, JAMA, 183: 516-522, 1963.
Study of Experimental Thallotoxicosis 20 Ruszynyack, I., Gyorgy, L. and Ormari, S.: On some potassium-like qualities of the thallium ion, Experimentia, 24: 809910, 1968. 21 Smith, D. H. and Doherty, R. A.: Thallotoxicosis: Reports of three cases in Massachusetts, Pediatrics, 34: 480-490, 1964. 22 Spencer, P. S., Peterson, E. R., Madrial, R. A. and Raine, C. S.: Effects of thallium salts on neuronal mitochondria in organo-
275
typic cord-ganglia-muscle combination cultures, J Cell Biol, 58: 79-95, 1973. 23 Thyresson, N.: Experimental investigation on thallium poisoning: Influence of thallium on tissue metabolism, Acta Dermato Vener, 30: 417-441, 1950. 24 Whetsell, W. 0. and Bunge, R. P.: Reversible alterations in the Golgi complex of cultured neurons treated with an inhibition of active Na and K transport, J Cell Biol, 42: 490-500, 1969.
Electron Microscopic Study of Experimental Thallotoxicosis Motonori Deshimaru, M.D.* Taihei Miyakawa, M.D. * Shiro Sumiyoshi, M.D. * * Fumie Yasuoka, M.D. * * Kosuke Kawano, M.D. * * and Megumi Kuramoto, M.D. * * *Department of Neurology, Toxicology Institute, Kumamoto University School of Medicine, Kumamoto **Department of Neuropsychiatry, Kumamoto University School of Medicine, Kumamoto
INTRODUCTION In the late 19th century, thallium salts were used in treating gonorrhea, syphylis, tuberculosis and ringworm. But therapeutic use of this drug was discontinued because of its toxity to human. Recently, thallium salts have come to be widely used as pesticides and rodenticides. There are many reports of human and experimental animals afflicted with thallotoxicosis caused by suicidal injection of rodenticides.13 .1 0 0 11 14 15 18 10 22 23 The clinical symptoms of thallotoxicosis are described as polyneuropathy, gastrointestinal symptoms, alopecia, retrobulbar optic neuritis, encephalopathy (unconsciousness, convulsion, ataxia, choreoathetosis) and muscular weakness.l l4 l5 l8 21 Pathologically, changes in the peripheral nerves are the chiefly observed symptoms, and this type of changes is of the dying-back type that is found most widespread in the distal regions. The damage to the peripheral nerves is considered to be primary disorder of the axon with secondary changes of the myelin sheaths.3 l7 21 The changes of CNS are edema and vascular enlargement in the cerebral hemispheres and brainstem, and there are chromatolytic changes of neurons in the morter cortex, the nuclei of the oculomotor or I11 cranial Received for publication Jan. 6, 1977.
nerve, the substantia nigra, and the globus pallidus and anterior horn cells of the spinal cord.3 I) 11 14 In the present experiment, we examined the nervous system, muscles and visceral organs of rats poisoned with thallium salts. MATERIALS AND METHODS Adult male rats (body weight 100-120 g) were used. The experimental group consisted of five rats and the controls of five rats. To the experimental group 2 mg per rat of thallium acetate: CH, COOT1 was given orally daily. Alopecia began on the 2 1st day after administration and gradually spread over the whole body. During administration, we did not observe any kind of neurological signs. On the 180th day of administration of thallium acetate, the experimental animals were killed with controls. Fixation was carried out by perfusion through the heart with 3% glutaraldehyde in pH 7.4 phosphate buffer. Blocks from cerebrum, cerebellum, spinal cord, peripheral nerves, skeletal muscles (lower limbs), liver and kidneys were postfixed in 2% phosphate-buffered osmic acid for two hours, then dehydrated and embedded in epon. Slices for optical microscopy were prepared with a Porter-Blum T microtome. After observation of toluidine blue preparations, the ultrathin sections were studied with a Hi-
270
M. Deshimaru, et al.
Fig. 1 : Cross striations of skeletal muscle are irregular in shape and have partially disappeared. Toluidine blue stain. ( X 380)
Fig. 2: The sciatic nerve are intact. Toluidine blue stain. ( X 300)
Fig. 3 : Sarcoplasmic reticulums are degenerated and atrophic. Myofilaments of Zband are irregular in shape and partially destroyed. Glycogen granules are not observed. ( X 9 , O O O )
Study of Experimental Thallotoxicosis
Fig. 4: Mitochondrias are swollen, destroyed and have disappeared in the cristae and show vacuolation. The mitochondria1 membranes are sometimes ruptured. ( X 8,000)
Fig. 5: A great number of mitochondria were accumulated in the peripheral parts of the muscle fibers. ( X9,OOO)
Fig. 6: In the remarkably damaged regions, myofilaments are destroyed. ( x 12,000)
27 1
M. Deshimaru, et al.
212
tachi 1 1A electron microscope. RESULTS
Light Microscopy Cross striation of skeletal muscles was irregular in shape and had partially disappeared (Fig. 1). The sciatic nerves were not different from those of the controls (Fig. 2). Toluidine-blue stained sections did not reveal any changes in the frontal, parietal lobes, thalamus, hypothalamus, cerebellum, spinal cord, liver and kidney. Electron Microscope In the skeletal muscles, sarcoplasmic reticulum was degenerated and atrophic. Myofilaments of Z-band were irregular in shape and partially destroyed. Moreover, no glycogen granules were observed (Fig. 3). Some of the rnyofilaments were destroyed and disappeared, and remarkable changes were observed in the mitochondria. They were swollen, destroyed and disappeared in cristae, and showed vacuolation. The mitochondrial membranes were sometimes ruptured (Fig. 4). A great number of mitochondria accumulated at the peripheral parts of the muscle fibers and they also showed similar changes (Fig. 5 ) . In the prominently damaged regions, myofilaments were destroyed (Fig. 6). In CNS, changes of thalamus and hypothalamus were remarkable in comparison with the other parts of the brain. Slight hypertrophy and vacuolation of mitochondria were observed in the cytoplasm of the nerve cells in the hypothalamus, along with dilatation of the Golgi cisterns (Fig. 7).
DISCUSSION Among the described-above pathological findings, the changes of the skeletal muscles were the most pronounced among all animals of the experimental group. The changes in the muscle were observed chiefly in the mitochondria, sarcoplasmic reticulum
and myofilament. In CNS, changes were observed chiefly in the thalamus and hypothalamus. These changes were vacuolation of the mitochondria and dilatation of the Golgi cisterns in the cytoplasm of the nerve cells. According to pathological findings in autopsy cases and experimental animals that have been reported so far, the peripheral nerves showed noticeable l1 l4 but changes in muscles changes1 have not been described. Spencer et aLZg reported that in examining the effect of thallium on organotypic cord-ganglia-muscle combination cultures, no changes were observed in the skeletal muscle tissue after a week of exposure to thallous sulfate. Thallous ions, biochemically, are considerably similar to potassium ions, and their ionic radii are very similar.2oTI' ions substituted for K' ions in (Na' +K') activated ATPaseqGT1' ions has 10 times greater affinity for activating ATPase than K' ions.2 Mullins et reported that a high concentration of T1' depolarized the membrane. And, if a high concentration of T1' was allowed to act for an appreciable period of time, TI' ions led to an irreversible damage to the muscle. Miyakawa et u1.12 administered lithium carbonate to rats and reported electron microscopic findings. The changes they found were very similar to our findings in the muscles. The changes in the muscles were considered to be due to the disturbances of electrolyte by replacement of Na' ions to Li' ions. We speculate that our findings in the muscles, as a result of our experiment, were also caused by the disturbances of the electrolyte due to replacement of K' ions to T1' ions. As the changes of mitochondria in the muscles were severer than the changes of myofilament, the changes in the muscles might be one of primary disorder in the mitochondria with secondary changes in the myofilament. Hermann et almsand Spencer et al." pointed to the changes of mitochondria in experimental thallotoxicosis. Spencer et d.
Study of Experimental Thallotoxicosis
273
Fig. 7: Nerve cell in the hypothalamus. Slight hypertrophy and vacuolation of mitochondria and dilatation of Golgi cisterns are observed. ( X7,OOO)
examined the ultrastructural changes in nerve tissue that has been exposed to thallium salts in vitro and reported the enlargement, destruction of cristae, vacuolation of the axonal mitochondria and partial rupture of the outer mitochondrial membrane. It is very interesting that our findings regarding the mitochondria were similar to these of Spencer's experiment, in spite of the different techniques employed in the experiments. The mechanism by which the mitochondria is affected selectively by T1' ions is still unsolved. However, Hermann et al. suggested the possibility of thallium becoming bound to the mitochondrial membranes. Spencer et d.,using electron microprobe mass spectrometry, detected thallium in the outer mitochondrial membranes. The changes of CNS reported up to the present are edema, vascular enlargement and chromatolytic changes of neurones.
But, in our experiment, the toluidine-blue stained sections did not reveal any changes in CNS. However, electron microscopically, changes in the mitochondria and dilatation of the Golgi cisterns were observed in the cytoplasm of the nerve cells-chiefly in the thalamus and hypothalamus. Hendleman7 reported a marked swelling of perikaryal granular endolplasmic reticulum when thallous acetate was exposed to cultures of rat dorsal root ganglia. As mentioned above, no changes in peripheral nerves were found in our experi. ~ ment. However, Hermann et ~ 1 studied the morphologic changes induced by the subcutaneous injection of varying doses of thallium acetate in rats and reported that no abnormalities were noted in the peripheral nerves. But, according to the histopathological studies of autopsy cases and in experimental animals (using mice and cats), the peripheral nerves have shown some
M. Deshimaru, et al.
274
changes. Thus, it is necessary to try an experiment under the same conditions to determine whether the difference of races or quantity of administration could be the reason. SUMMARY
To adult male rats 2 mg per rat of thallium acetate: CH, COOT1 was given orally daily for six months. Clinically, the experimental rats revealed only alopecia and showed no neurological signs. Pathological findings were noted in muscle and cerebrum. Marked changes were observed in muscles and were as follows: swelling and vacuolation of mitochondria, destruction of cristae, ruptures of mitochondrial membranes, degeneration of sarcoplasmic reticulum and destruction of myofilaments. In the cerebrum, there were vacuolation of mitochondria, dilatation of Golgi cisterns in hypothalamus and thalamus. Sciatic nerve, liver and kidney were intact. From this, we consider that T1' ions selectively affect the muscle and its mitochondria. REFERENCES Bank, R. J., Pleasure, D. E., Suzuki, K., Nigro, M. and Katz, R.: Thallium poisoning, Arch Neurol, 26: 456-464, 1972. Britten, J. S. and Blank, M.: Thallium activation of the (Na+-K+) activated ATPase of rabbit kidney, Biochem Biophys Acta 159: 160-166, 1968. Cavanagh, J. B., Fuller, N. H., Johnson, H.R.M. and Rudge, P.: The effects of thallium salts, with particular reference to the nervous system changes, Quart J Med, 43: 293-319, 1974. Chamberlain, P. H., Stavinoha, W. B., Davis, H., Kniker, W.T. and Panos, T. C.: Thallium poisoning, Pediatrics, 22: 1170-1 182, 1958. Gehring, P. and Hammond, P.: Mechanism for the increase in thallium excretion during the administration of potassium, Pharmacologist, 8: 262, 1969.
6 Grunfeld, 0. and Hinostroza, G.: Thallium poisoning, Arch Intern Med, 114: 132138, 1964. 7 Hendleman, W.: The effect of thallium on
peripheral nervous tissue in culture: A light and electron microscopic study, Anat Rec, 163: 198A, 1969. 8 Hermann, M. M. and Bench, K. G.: Light and electron microscopic studies of acute and chronic thallium intoxication in rats, Toxicol Appl Pharmacol, 10: 199-222, 1967. 9 Kakehashi, K.: Experimental studies on the pathogenesis of the Minamata disease: Studies of delayed thallium intoxication, Kumamoto Med J, 33, suppl, 3: 704-713, 1959 (in Japanese). 10 Lund, A.: Distribution of thallium in the organism and its elimination, Acta Pharmacol, 12: 251-259, 1956. 1 1 Matsumura, K.: Experimental studies on the pathogenesis of the Minamata disease: Studies of acute and chronic thallium intoxication, Kumamoto Med J, 34, suppl, 3: 655-669, 1960 (in Japanese), 12 Miyakawa, T., Sumiyoshi, S., Deshimaru, M., Hattori, E. and Miyakawa, M.: Electron microscopic studies on the effects of the administration of Lithium carbonicum in rats, Ann Rep Pharmac Res Foun, 5: 9-14, 1973 (in Japanese). 13 Mullins, L. T. and Moore, R. D.: The movement of thallium ions in muscle, J gen Physiol, 43: 759-773, 1960. 14 Munch, J.C., Guinsbeg, H. M. and Nixon, C. E.: The 1932 thallotoxicosis outbreak in California, JAMA, 100: 1315-13 19, 1933. 15. Oguchi, K., Nishida, M. and Tsubaki, T. Thallotoxicosis: Report of two cases, Clin Neurol, 14: 42-47, 1974, (in Japanese). 16 Peterson, E. R. and Murray, M. R.: Patterns of peripheral degeneration in vitro, A m NY Acad Sci, 122: 39-51, 1965. 17 Peterson, E. R. and Grain, S. M.: Regeneration and innervation in cultures of adult mammalian skeletal muscle coupled with fetal rodent spinal cord, Exp Neurol, 36: 136, 1972. 18 Prick, J.J.G., Smith, W.G.S. and Miller, L.: Thallium poisoning, Amsterdam, 1955. 19 Reed, D.C.J., Faro, S.N., Pieper, S. J. and Kurland, L. T.: Thallotoxicosis: Acute manifestations and sequelae, JAMA, 183: 516-522, 1963.
Study of Experimental Thallotoxicosis 20 Ruszynyack, I., Gyorgy, L. and Ormari, S.: On some potassium-like qualities of the thallium ion, Experimentia, 24: 809910, 1968. 21 Smith, D. H. and Doherty, R. A.: Thallotoxicosis: Reports of three cases in Massachusetts, Pediatrics, 34: 480-490, 1964. 22 Spencer, P. S., Peterson, E. R., Madrial, R. A. and Raine, C. S.: Effects of thallium salts on neuronal mitochondria in organo-
275
typic cord-ganglia-muscle combination cultures, J Cell Biol, 58: 79-95, 1973. 23 Thyresson, N.: Experimental investigation on thallium poisoning: Influence of thallium on tissue metabolism, Acta Dermato Vener, 30: 417-441, 1950. 24 Whetsell, W. 0. and Bunge, R. P.: Reversible alterations in the Golgi complex of cultured neurons treated with an inhibition of active Na and K transport, J Cell Biol, 42: 490-500, 1969.
