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Kainate resistant neurons in peripheral ganglia of the rat
Neuroscience Letters, 43 (1983) l-5 Elsevier Scientific Publishers Ireland Ltd.
KAINATE RESISTANT NEURONS IN PERIPHERAL RAT
1
GANGLIA OF THE
GERALD WOLF and GERBURG KEILHGFF
Dept. of Biology of the Institute for Anatomy and Biology, Medical Academy of Magdeburg, DDR 3090 Magdeburg, Leipziger Str. 44 (D.D. R.) (Received September 8th, 1983; Accepted October 14th, 1983)
words: kainate - neurodegeneration - glutamatergicstructures- dorsal root ganglia - superior cervical ganglia - hippocampus
Neurons of dorsal root ganglia and of superiorcervicalganglia did not display any morphological signs of degeneration when kainic acid (KA) was administeredeither systemically (20 and 40 mg/kg) or when it was directly injected (1 m). The KA doses used were sufficient to result in heavy destruction of hippocampal CA3/CA4 neurons and neurodegeneration of various brain regions after intracerebroventricular or local application, respectively. The resistance of the peripheralneurons to KA is discussed as a consequence of lacking glutamate inputs.
There is some controversy as to the mechanism of kainate (kainic acid, KA)induced neurotoxicity. Close structural similarities between KA and glutamate, as well as the excitatory and neurodegenerative potency of both amino acids* have initially led to the assumption that KA exerts its neurotoxic effect by overexcitation of neurons which are provided with postsynaptic glutamate receptors [ 161. Correspondingly, destruction of glutamate inputs markedly decreases KA neurotoxicity [2, 91. But there is also evidence that KA induces convulsions and brain damage in
connection with neurotransmitters other than glutamate [ 13, 201. To date at least two different types of glutamate receptors have been postulated [3, 6, 213, one of them prefers KA. The mode of their interaction, as well as their correlation with other receptor types, remains to be clarified. Referring to the particular vulnerability of hippocampal CA3/CA4 neurons to KA we have proposed recently (221that in addition to glutamatergic structures an interaction with opioid peptides might be responsible and, moreover, we could demonstrate in rats that the KA-induced damage of such neurons increases in parallel to the postnatal development of glutamatergic structures. In the present study we examined the effect of systemic and local administration of KA on neurons of superior cervical and dorsal root ganglia in which glutamate inputs do not obviously occur. Male Wistar rats (170-19Qg) derived from VELAZ (Prague) were used. A group of 8 animals was injected intraperitoneally with 20 or 40 mg KA/kg (KA was pur0304-3940/83/$ 03.00 @ 1983 Elsevier Scientific Publishers Ireland Ltd.
chased flom Sigma), solved in phosphate (0.01 M)-buffered saline (PBS) and adjusted to pH 7.35. In a second group of animals under hesobarbital anesthesia the left side superior cervical ganglion (8 rats) and 2 dorsal root ganglia of the lumbar region (8 rats) were exposed and, using a stereomicroscope, KA solution was directly injected into the ganglia and its immediate proximity by means of a Hamilton micros:. .rge. In each ganglion an amount of 1 pg KA diluted in 1 ~1 PBS was administered. Control animals received PBS only. After a survival time of 5 h and 7 or 14 days, respectively, the rats were decapitated, superior cervical and dorsal root ganglia and (in case of systemic KA administration) the brain were quickly removed and fixed in Bouin’s fIuid for 4 days. After embedding in paraffin and sectioning (10 !~rn) the material was stained with hematoxylin and eosin or with cresyl violet. Only when applied systemically did KA induce typical behavioral phenomena such as ‘wet dog shake’, trembling, reeling and cotis+lsions to a degree which was dependent on the dosage used. As described previously by many other authors (7, 14, 171, in brain preferentially pyramidal neurons of hippocampa! fields CA4 and CA3 already displayed signs of heavy degeneration after a surviv;;l time of 5 h, in particular when a KA dose of 40 mg/kg was used (Fig. 1). while neurons of other brain regions showed no or merely little histopathological alterationi. On the other side, neurons of the peripheral ganglia studied did not provide any evicknt signs of
3
Fig. 2. Sections through a dorsal root ganglion (a) and a superior cervical ganglion (h) of rats 7 days after local injection of 1 leg kainate. * shows areas of local damage caused by the tip of the cnnnula. c‘resyl ~~ioletGaining (a, x 60; b s, 40).
4
necrosis when KA was administered either systemically, or when it was directly injected (Fig. 2). Sometimes darkly staining neurons were observed; however, such nerve cells occurred likewise in control animals. The KA dose used locally largely exceeded the quantity which results in degeneration of hippocampal CA3KA4 neurons, when we had administered the agent intracerebroventricularly [22]. Also, the same KA dose has been proven to be adequate for nerve cell destruction in various brain regions following a single local injection (cf. review in ref. 11). Concerning the dorsal root ganglia, the negative results appear to parallel findings on unipolar neurons of the mesencephalic trigeminal nucleus [4] which likewise failed to be affected by local KA application, possibly as a consequence of lacking synaptic structures which are considered to be responsible for the neurotoxic action of KA. Dorsal root ganglia contain no identifiable synapses at all [ 121. By contrast with dorsal root ganglia, neurons of cervical ganglia, which also failed to show morphological signs of KA-induced neurotoxic actions, are well provided with synaptic inputs, but there are no criteria for transmission processes utilizing acidic amino acids [ 15, 191. For this reason, both dorsal root ganglia and cervical ganglia may be considered to be suitable as models for nerve tissue which is devoid of synapses in general and, respectively, of glutamatergic (and GABAergic ?) structures in particular. Our present results suggest a connection between KA-induced vulnerability of nerve cells and the presence of appropriate synaptic receptors. In the first place we have to consider glutamate receptors as outlined above. Transmitters other than glutamate may also be taken into account; however, many of them, such as acetylcholine, catecholamines [ 1,8] and peptides such as substance P and enkephalins [ 10, 181, occur in cervical ganglia as well as GABA in dorsal root ganglia [ 121. As to a role of GABA in KA neurotoxicity there are reasonable doubts anyway. Very recently Contestabile et al. [5] have reported that the pharmacologically manip. ted increase in GABA content of the goldfish optic tectum failed to protect tectal neurons from the neuroexcitatory and neurotoxic actions of KA. The expert technical assistance by Mrs. U. Schulz is gratefully acknowledged.
Archakova, L.I., Bulygin, I.A. and Netukova, N.I., The ultrastructural organization of sympathetic ganglia of the cat, J. auton. Nerv. Syst., 6 (1982) 83-93. Biziere, K. and Coyle, J.T., Influence of cortico-striatal afferents on striatal kainic acid neurotoxicity, Neurosci. Lett., 8 (1978) 303-310. Collins, G.G.S., Anson, J. and Sutees, L., Presynaptic kainate and N-methyl-o-aspartate receptors regulate excitatory amino acid release in the olfactory cortex, Brain Res., 265 (1983) 157-159. Colonnier, M., Steriade, M. and Landry, P., Selective resistance of sensory cells of the mesencephalic trigeminal nucleus to kainic acid-induced lesions, Brain Re;., 172 (1979) 552-556. Contestabile, A., Migani, P., Poli, A., Villani, L. and Barnabei, O., Pharmacological manipulation of GABA system does not protect the goldfish optic tectum from the neuroexcitatory and neurotoxic action of kainic acid, Brain Res., 262 (1983) 339-343.
5
6 Davies, J., Evans, R.H., Jones, A.W. and Smith, D.A., Differential activation and blockade of excitatory amino acid receptors in the mammalian and amphibian central .nervous systems, Camp. Biochem. Physiol., 72 (1982) 211-224. 7 Heggli, D.E., Aamodt, A. and Malthe-Ssrenssen, D,, Kainic acid neurotoxicity; effect of systemic injection on neurotransmitter markers in different brain regions, Brain Res., 230 (1981) 253-262. 8 Heym, C., Kiinig, R., Schroder, H. and Gerold, N., lmmunofluorescent and biochemical studies on dopamine-&hydroxylase and catecholamines in SIP cells of the superior cervical ganglia, Acta histochem., Suppl. 24 (1981) 123-129. 9 Kohler, C., Schwartz, R. and Fuxe, J., Perforant path transsections protect hippocampal granule cells from kainate lesion, Neurosci. Lett., 10 (1978) 241-246. 10 Konishi, S., Tsunoo, A. and Yanaihara, N,, Peptidergic excitatory and inhibitory synapses in mammalian sympathetic ganglia: roles of substance P and enkephalin, Biomed. Res., 1 (1980) 528-536. 11 McGeer, P.L. and McGeer, E.G., Kainate as a selective lcsioning agent. In P.J. Roberts, J. StormMathisen and G.A.R. Johnston (Eds.), Glutamate: Transmitter in the Central Nervous System, 9. Wiley and Sons, Chichester, 1981, pp. 55-75. 12 Minchin,. M.C.W. and Beart, P.M., Compartmentation of amino acid metabolism in the rat dorsal root ganglion; a metabolic and autoradiographic study, Brain Res., 83 (1975) 437-449. 13 Nadler, J.V. and Cuthbewison, G. J., Kainic acid neurotoxicity toward hippocampus: dependence on specific excitatory pathways, SOL’.Neurosci. Abstr., 5 (1979) p. 280. 14 Nadler, J .V., Perry, B.W. and Cotman, C.W., lntraventricular kainic acid preferentially destroys hippocampal pyramidal cells, Nature (Lond.), 271 (1978) 676-677. 15 Nagata, Y., Yokoi, Y. and Tsukada, Y., Studies on free amino acid metabolism in excised cervical sympathetic ganglia from the rat, J. Neurochem., 13 (1966) 1421-1431. 16 Glney, J.W., Excitotoxic amino acids: research applications and safety implications. In L.J. Filer, Jr. et al. (Eds.), Glutamic Acid: Advances in Biochemistry and Physiology, Raven Press, New York, 1979, pp. 287-319. 17 Olney, J.W., Fuller, T. and de Gubareff, T., Acute dendrotoxic changes in the hippocampus of kainate treated rats, Brain #es., 176 (1979) 91-100. 18 Otsuka, M., Konishi, S., Yanagisawa, M., Tsunoo, A. and Akagi, H., Role of substance P as a sensory transmitter in spinal cord and sympathetic ganglia. In R. Porter and M. Connor (Eds.), Substance P in the Nervous System, Ciba Foundation Symposia, Vol. 91, Pitman, London, 1982, pp. 13-24. 19 Rothe, F., Schmidt, W. and Wolf, G., Postnatal changes in the activity of glutamate dehydrogenase and aspartate aminotransferase in the rat nervous system with special reference to the glutamate transmitter metabolism, Develop. Brain Res., in press. 20 Schwartz, R. and Kohler, C., Evidence against an exclusive role of glutamate in kainic acid neurotoxicity, Neurosci. Lett., 19 (1980) 243-249. 2 1 Watkins, J .C., Pharmacology of excitatory amino acid receptors. In P. 3. Roberts, J. Storm-Mathisen and G.A.R. Johnston (Eds.), Glutamate: Transmitter in the Central Nervous System, J. Wiley and Sons, Chichester, 1981, pp. l-24. 22 Wolf, G. and Keilhoff, G., Kainate and glutamate neurotoxicity in dependence on the postnatal development with special reference to hippocampal neurons, Develop. Brain Res., submitted.
KAINATE RESISTANT NEURONS IN PERIPHERAL RAT
1
GANGLIA OF THE
GERALD WOLF and GERBURG KEILHGFF
Dept. of Biology of the Institute for Anatomy and Biology, Medical Academy of Magdeburg, DDR 3090 Magdeburg, Leipziger Str. 44 (D.D. R.) (Received September 8th, 1983; Accepted October 14th, 1983)
words: kainate - neurodegeneration - glutamatergicstructures- dorsal root ganglia - superior cervical ganglia - hippocampus
Neurons of dorsal root ganglia and of superiorcervicalganglia did not display any morphological signs of degeneration when kainic acid (KA) was administeredeither systemically (20 and 40 mg/kg) or when it was directly injected (1 m). The KA doses used were sufficient to result in heavy destruction of hippocampal CA3/CA4 neurons and neurodegeneration of various brain regions after intracerebroventricular or local application, respectively. The resistance of the peripheralneurons to KA is discussed as a consequence of lacking glutamate inputs.
There is some controversy as to the mechanism of kainate (kainic acid, KA)induced neurotoxicity. Close structural similarities between KA and glutamate, as well as the excitatory and neurodegenerative potency of both amino acids* have initially led to the assumption that KA exerts its neurotoxic effect by overexcitation of neurons which are provided with postsynaptic glutamate receptors [ 161. Correspondingly, destruction of glutamate inputs markedly decreases KA neurotoxicity [2, 91. But there is also evidence that KA induces convulsions and brain damage in
connection with neurotransmitters other than glutamate [ 13, 201. To date at least two different types of glutamate receptors have been postulated [3, 6, 213, one of them prefers KA. The mode of their interaction, as well as their correlation with other receptor types, remains to be clarified. Referring to the particular vulnerability of hippocampal CA3/CA4 neurons to KA we have proposed recently (221that in addition to glutamatergic structures an interaction with opioid peptides might be responsible and, moreover, we could demonstrate in rats that the KA-induced damage of such neurons increases in parallel to the postnatal development of glutamatergic structures. In the present study we examined the effect of systemic and local administration of KA on neurons of superior cervical and dorsal root ganglia in which glutamate inputs do not obviously occur. Male Wistar rats (170-19Qg) derived from VELAZ (Prague) were used. A group of 8 animals was injected intraperitoneally with 20 or 40 mg KA/kg (KA was pur0304-3940/83/$ 03.00 @ 1983 Elsevier Scientific Publishers Ireland Ltd.
chased flom Sigma), solved in phosphate (0.01 M)-buffered saline (PBS) and adjusted to pH 7.35. In a second group of animals under hesobarbital anesthesia the left side superior cervical ganglion (8 rats) and 2 dorsal root ganglia of the lumbar region (8 rats) were exposed and, using a stereomicroscope, KA solution was directly injected into the ganglia and its immediate proximity by means of a Hamilton micros:. .rge. In each ganglion an amount of 1 pg KA diluted in 1 ~1 PBS was administered. Control animals received PBS only. After a survival time of 5 h and 7 or 14 days, respectively, the rats were decapitated, superior cervical and dorsal root ganglia and (in case of systemic KA administration) the brain were quickly removed and fixed in Bouin’s fIuid for 4 days. After embedding in paraffin and sectioning (10 !~rn) the material was stained with hematoxylin and eosin or with cresyl violet. Only when applied systemically did KA induce typical behavioral phenomena such as ‘wet dog shake’, trembling, reeling and cotis+lsions to a degree which was dependent on the dosage used. As described previously by many other authors (7, 14, 171, in brain preferentially pyramidal neurons of hippocampa! fields CA4 and CA3 already displayed signs of heavy degeneration after a surviv;;l time of 5 h, in particular when a KA dose of 40 mg/kg was used (Fig. 1). while neurons of other brain regions showed no or merely little histopathological alterationi. On the other side, neurons of the peripheral ganglia studied did not provide any evicknt signs of
3
Fig. 2. Sections through a dorsal root ganglion (a) and a superior cervical ganglion (h) of rats 7 days after local injection of 1 leg kainate. * shows areas of local damage caused by the tip of the cnnnula. c‘resyl ~~ioletGaining (a, x 60; b s, 40).
4
necrosis when KA was administered either systemically, or when it was directly injected (Fig. 2). Sometimes darkly staining neurons were observed; however, such nerve cells occurred likewise in control animals. The KA dose used locally largely exceeded the quantity which results in degeneration of hippocampal CA3KA4 neurons, when we had administered the agent intracerebroventricularly [22]. Also, the same KA dose has been proven to be adequate for nerve cell destruction in various brain regions following a single local injection (cf. review in ref. 11). Concerning the dorsal root ganglia, the negative results appear to parallel findings on unipolar neurons of the mesencephalic trigeminal nucleus [4] which likewise failed to be affected by local KA application, possibly as a consequence of lacking synaptic structures which are considered to be responsible for the neurotoxic action of KA. Dorsal root ganglia contain no identifiable synapses at all [ 121. By contrast with dorsal root ganglia, neurons of cervical ganglia, which also failed to show morphological signs of KA-induced neurotoxic actions, are well provided with synaptic inputs, but there are no criteria for transmission processes utilizing acidic amino acids [ 15, 191. For this reason, both dorsal root ganglia and cervical ganglia may be considered to be suitable as models for nerve tissue which is devoid of synapses in general and, respectively, of glutamatergic (and GABAergic ?) structures in particular. Our present results suggest a connection between KA-induced vulnerability of nerve cells and the presence of appropriate synaptic receptors. In the first place we have to consider glutamate receptors as outlined above. Transmitters other than glutamate may also be taken into account; however, many of them, such as acetylcholine, catecholamines [ 1,8] and peptides such as substance P and enkephalins [ 10, 181, occur in cervical ganglia as well as GABA in dorsal root ganglia [ 121. As to a role of GABA in KA neurotoxicity there are reasonable doubts anyway. Very recently Contestabile et al. [5] have reported that the pharmacologically manip. ted increase in GABA content of the goldfish optic tectum failed to protect tectal neurons from the neuroexcitatory and neurotoxic actions of KA. The expert technical assistance by Mrs. U. Schulz is gratefully acknowledged.
Archakova, L.I., Bulygin, I.A. and Netukova, N.I., The ultrastructural organization of sympathetic ganglia of the cat, J. auton. Nerv. Syst., 6 (1982) 83-93. Biziere, K. and Coyle, J.T., Influence of cortico-striatal afferents on striatal kainic acid neurotoxicity, Neurosci. Lett., 8 (1978) 303-310. Collins, G.G.S., Anson, J. and Sutees, L., Presynaptic kainate and N-methyl-o-aspartate receptors regulate excitatory amino acid release in the olfactory cortex, Brain Res., 265 (1983) 157-159. Colonnier, M., Steriade, M. and Landry, P., Selective resistance of sensory cells of the mesencephalic trigeminal nucleus to kainic acid-induced lesions, Brain Re;., 172 (1979) 552-556. Contestabile, A., Migani, P., Poli, A., Villani, L. and Barnabei, O., Pharmacological manipulation of GABA system does not protect the goldfish optic tectum from the neuroexcitatory and neurotoxic action of kainic acid, Brain Res., 262 (1983) 339-343.
5
6 Davies, J., Evans, R.H., Jones, A.W. and Smith, D.A., Differential activation and blockade of excitatory amino acid receptors in the mammalian and amphibian central .nervous systems, Camp. Biochem. Physiol., 72 (1982) 211-224. 7 Heggli, D.E., Aamodt, A. and Malthe-Ssrenssen, D,, Kainic acid neurotoxicity; effect of systemic injection on neurotransmitter markers in different brain regions, Brain Res., 230 (1981) 253-262. 8 Heym, C., Kiinig, R., Schroder, H. and Gerold, N., lmmunofluorescent and biochemical studies on dopamine-&hydroxylase and catecholamines in SIP cells of the superior cervical ganglia, Acta histochem., Suppl. 24 (1981) 123-129. 9 Kohler, C., Schwartz, R. and Fuxe, J., Perforant path transsections protect hippocampal granule cells from kainate lesion, Neurosci. Lett., 10 (1978) 241-246. 10 Konishi, S., Tsunoo, A. and Yanaihara, N,, Peptidergic excitatory and inhibitory synapses in mammalian sympathetic ganglia: roles of substance P and enkephalin, Biomed. Res., 1 (1980) 528-536. 11 McGeer, P.L. and McGeer, E.G., Kainate as a selective lcsioning agent. In P.J. Roberts, J. StormMathisen and G.A.R. Johnston (Eds.), Glutamate: Transmitter in the Central Nervous System, 9. Wiley and Sons, Chichester, 1981, pp. 55-75. 12 Minchin,. M.C.W. and Beart, P.M., Compartmentation of amino acid metabolism in the rat dorsal root ganglion; a metabolic and autoradiographic study, Brain Res., 83 (1975) 437-449. 13 Nadler, J.V. and Cuthbewison, G. J., Kainic acid neurotoxicity toward hippocampus: dependence on specific excitatory pathways, SOL’.Neurosci. Abstr., 5 (1979) p. 280. 14 Nadler, J .V., Perry, B.W. and Cotman, C.W., lntraventricular kainic acid preferentially destroys hippocampal pyramidal cells, Nature (Lond.), 271 (1978) 676-677. 15 Nagata, Y., Yokoi, Y. and Tsukada, Y., Studies on free amino acid metabolism in excised cervical sympathetic ganglia from the rat, J. Neurochem., 13 (1966) 1421-1431. 16 Glney, J.W., Excitotoxic amino acids: research applications and safety implications. In L.J. Filer, Jr. et al. (Eds.), Glutamic Acid: Advances in Biochemistry and Physiology, Raven Press, New York, 1979, pp. 287-319. 17 Olney, J.W., Fuller, T. and de Gubareff, T., Acute dendrotoxic changes in the hippocampus of kainate treated rats, Brain #es., 176 (1979) 91-100. 18 Otsuka, M., Konishi, S., Yanagisawa, M., Tsunoo, A. and Akagi, H., Role of substance P as a sensory transmitter in spinal cord and sympathetic ganglia. In R. Porter and M. Connor (Eds.), Substance P in the Nervous System, Ciba Foundation Symposia, Vol. 91, Pitman, London, 1982, pp. 13-24. 19 Rothe, F., Schmidt, W. and Wolf, G., Postnatal changes in the activity of glutamate dehydrogenase and aspartate aminotransferase in the rat nervous system with special reference to the glutamate transmitter metabolism, Develop. Brain Res., in press. 20 Schwartz, R. and Kohler, C., Evidence against an exclusive role of glutamate in kainic acid neurotoxicity, Neurosci. Lett., 19 (1980) 243-249. 2 1 Watkins, J .C., Pharmacology of excitatory amino acid receptors. In P. 3. Roberts, J. Storm-Mathisen and G.A.R. Johnston (Eds.), Glutamate: Transmitter in the Central Nervous System, J. Wiley and Sons, Chichester, 1981, pp. l-24. 22 Wolf, G. and Keilhoff, G., Kainate and glutamate neurotoxicity in dependence on the postnatal development with special reference to hippocampal neurons, Develop. Brain Res., submitted.