J Neural Transm [P-D Sect] (1991) 3:133-142
__ Journal of Neural Transmission 9 Springer-Verlag 1991 Printed in Austria
Biogenic amines and metabolites in spinal cord of patients with Parkinson's disease and amyotrophic lateral sclerosis E. Sofic 1, P. RiedererI, W. Gseli1, M. Gavranovic2, A. Schmidtke1, and K. JelHnger3 1Department of Psychiatry, Clinical Neurochemistry, University of W/irzburg, Federal Republic of Germany 2Department of Neurosurgery, University Medical Center, Sarajevo, Jugoslavia 3Ludwig Boltzmann Institute for Clinical Neurobiology, Lainz Hospital, Vienna, Austria Accepted March 25, 1991
Summary. In four human controls, four cases of Parkinson's disease and three cases of amyotrophic lateral sclerosis analysis of dopamine, noradrenaline, serotonin and the metabolites 3,4-dihydroxyphenylacetic acid, homovanillic acid and 5-hydroxyindoleacetic acid was performed in various segments of postmortem spinal cord. In controls the concentrations of dopamine are about 1/3 to 1/4 that of noradrenaline; the significantly highest content of noradrenaline was found in the lumbar, and dopamine in thoracic, lumbar and sacral segments of the spinal cord. Intersegmental distribution of monoamines was only present in spinal cord of controls, while in the spinal cord of parkinsonian patients such a difference was not found. Otherwise, biogenic amine and metabolite concentrations in spinal cord segments of parkinsonian patients did not differ significantly from those in the control subjects. However, it cannot be excluded that these segments are sensitive to drugs including neuroleptics and combined L-DOPA treatment. In subjects with amyotrophic lateral sclerosis significantly lower concentrations of noradrenaline in the cervical and thoracic, and of dopamine and homovanillic acid in the thoracic and lumbar segments were found in comparison with controls. The concentrations of serotonin and 5-hydroxyindoleacetic acid in the thoracic segments of amyotrophic lateral sclerosis were significantly lower than that of controls. Differences in the inter-segmental distribution of noradrenaline in lumbar, lumbar-sacral, and serotonin in lumbar segments of spinal cord were found in this group. Keywords: Spinal cord, biogenic amines, Parkinson's disease, amyotrophic lateral sclerosis.
134
E. Sofic etal. Introduction
Parkinson's disease (PD) is characterized by degeneration of the melanin-containing mesencephalic neurons accompanied by variable damage to both the ascending (nigrostriatal, mesencephalocortical and less, hypothalamic) and descending (to spinal cord) dopaminergic pathways arising from the zona compacta of the substantia nigra and ventral tegmental area. Additional deficiencies of the noradrenergic and serotonergic system due to degeneration of locus ceruleus and dorsal raphe nucleus are known to occur in PD (for review see Agid et al., 1990; Jellinger et al., 1989, 1990). Recent postmortem studies of the lumbar spinal cord of PD subjects revealed considerable decrease of noradrenaline (NA), serotonin (5-HT) and its metabolite 5-hydroxyindoleacetic acid (5HIAA), while spinal cord levels of dopamine (DA) and its metabolite homovanillic acid (HVA) were within control values (Scatton et al., 1986). Although in this study the postmortem intervals in PD subjects (19 4- 3 hrs) were considerably longer than in controls (7 4- 2 hrs), and most PD patients had been treated with L-DOPA prior to 24 hours before death, these authors suggest that the descending DA pathways which project to the lumbar spinal cord are spared in PD. On the other hand, immunohistochemical studies, using an antibody against tyrosine hydroxylase (TH), the rate-limiting enzyme of the catecholamine biosynthesis, showed decreased densities of TH-immunoreactivity (IR) in all levels of the spinal cord of PD subjects, particularly of TH-IR fibres in the superficial zone of the dorsal horn and in the pericommissural gray matter (Dietl, 1985; see Jellinger, 1986). 5-HT, NA and DA-containing neurons have been shown to project on the mammalian spinal cord, although the exact details of areas of termination are not yet fully understood (see Hunt, 1985). While NA-ergic and 5-HT-ergic spinal systems originate from the locus ceruleus, lateral medullary and pontine nucleus (Hunt, 1985; Moore and Card, 1984), and terminate in the dorsal horn (for NA and 5-HT), ventral horn (for 5-HT; Pelletier et al., 1981), lateral sympathetic columns and motor neurons of the ventral horn (for NA) (Hunt, 1985; Moore and Card, 1984), recent studies also demonstrated the existence of DA containing fibres in the mammalian spinal cord (Commissiong and Neff, 1979; Mouchet et al., 1986; Lindvall etal., 1983; Bj6rklund and Lindvall, 1984). The cell bodies of this system originate periventricularly in the dorsal hypothalamus, caudal thalamus and diencephalic A-11 and A-13 DA cell groups (Bj6rklund and Lindvall, 1984). The terminal innervations were found at all spinal cord levels, with highest densities of DA fibres in superficial layers of the dorsal horn, the area surrounding the central canal, the intermediolateral cell column of the thoracolumbar spinal cord and around the preganglionic parasympathetic neurons of the sacral spinal cord (Lindvall et al., 1983; Bj6rklund and Lindvall, 1984). In addition, DA-ergic cell bodies have been described in the spinal cord (Dietl et al., 1985; Mouchet et al., 1986). A similar regional distribution of DA receptors
Biogenic amines in human spinal cord
135
in the mammalian spinal cord has been observed by autoradiography (Commissiong and Neff, 1979; Palacios and Wamsley, 1984; Scatton etal., 1984). While the involvement of the cerebral monoaminergic systems in PD is now well documented, the possible alterations of the spinal cord monoaminergic systems in this and other degenerative disorders like amyotrophic lateral sclerosis (ALS) are poorly understood (Scatton et al., 1986; Lackovic et al., 1988; Ohsugi et al., 1987). Here we present data on the monoamine levels and their metabolites in different parts of the spinal cord in four cases of PD, three cases of ALS and in four control subjects. Materials and methods
Portions of cervical (C), thoracic (T), lumbar (L) and sacral (S) segments of human spinal cords were obtained at autopsy from eleven subjects. Essential clinical and (neuro)pathological data of patients with PD, ALS and controls are given in Table 1. Tissue samples were frozen at - 80 ~ prior to analysis. There were no differences for storage time at - 80 ~ among the three groups. DA, NA, 5-HT, 3,4-dihydroxyphenylacetic acid (DOPAC), HVA and 5-HIAA were analyzed by reversed-phase high-performance liquid chromatography with electrochemical detection [(5-HT, 5-HIAA, HVA, DOPAC; Sperk, 1982) and (NA and DA; Sofic, 1986)]. Statistical analysis of the data was performed using Student's t-test, including Benferroni (~) adjustment. [a* (C) = 0.00119 (7 segments, 6 parameters), a* (T) = 0.000694 (12 segments, 6 parameters), a* (L) = 0.00167 (5 segments, 6 parameters), a* (L-S) = 0.0083 (1 segment, 6 parameters), a* (S) = 0.0083 (1 segment, 6 parameters)l. Results
Data of the biochemical analysis in spinal cord are displayed in Tables 2 to 7. In controls, D A concentrations were about three to four times lower than that of N A (Tables 2 and 3). The significantly highest content of N A was found in the L segment (Table 2) and of D A in L and S segment (Table 3). The concentrations of D A in the T segments of controls were significantly lower than in C segments (Table 3). A n inter-segmental distribution of monoamines was only present in spinal cord of controls. In contrast, in the spinal cord of PD such inter-segmental differences were not found (Tables 2 to 7), despite D O P A C in the L segments (Table 4). Furthermore, spinal cord biogenic amines and their metabolite concentrations did not differ significantly from those measured in controls (Tables 2 to 7). In the L spinal cord of ALS subjects N A contents were significantly higher than in the C and T segments (Table 2). The concentrations of 5-HT in the L segments of ALS were significantly lower than in C segments (Table 6). The concentrations of N A in the C, T (Controls/ALS) and L-S (PD/ALS) segments of patients with ALS were significantly lower than compared to controls or PD (Table 2). Furthermore, significantly lower D A and HVA levels were found in the
E. Sofic etal.
136
Table 1. Clinical notes Code
Sex
Age (years)
Clinical diagnosis
Neuropathology
466/84
f
86
lung carcinoma
diffuse, mild brain atrophy, no other lesions
470/84
f
81
chronic alcoholism, liver cirrhosis
moderate brain atrophy, no other lesions
__ Journal of Neural Transmission 9 Springer-Verlag 1991 Printed in Austria
Biogenic amines and metabolites in spinal cord of patients with Parkinson's disease and amyotrophic lateral sclerosis E. Sofic 1, P. RiedererI, W. Gseli1, M. Gavranovic2, A. Schmidtke1, and K. JelHnger3 1Department of Psychiatry, Clinical Neurochemistry, University of W/irzburg, Federal Republic of Germany 2Department of Neurosurgery, University Medical Center, Sarajevo, Jugoslavia 3Ludwig Boltzmann Institute for Clinical Neurobiology, Lainz Hospital, Vienna, Austria Accepted March 25, 1991
Summary. In four human controls, four cases of Parkinson's disease and three cases of amyotrophic lateral sclerosis analysis of dopamine, noradrenaline, serotonin and the metabolites 3,4-dihydroxyphenylacetic acid, homovanillic acid and 5-hydroxyindoleacetic acid was performed in various segments of postmortem spinal cord. In controls the concentrations of dopamine are about 1/3 to 1/4 that of noradrenaline; the significantly highest content of noradrenaline was found in the lumbar, and dopamine in thoracic, lumbar and sacral segments of the spinal cord. Intersegmental distribution of monoamines was only present in spinal cord of controls, while in the spinal cord of parkinsonian patients such a difference was not found. Otherwise, biogenic amine and metabolite concentrations in spinal cord segments of parkinsonian patients did not differ significantly from those in the control subjects. However, it cannot be excluded that these segments are sensitive to drugs including neuroleptics and combined L-DOPA treatment. In subjects with amyotrophic lateral sclerosis significantly lower concentrations of noradrenaline in the cervical and thoracic, and of dopamine and homovanillic acid in the thoracic and lumbar segments were found in comparison with controls. The concentrations of serotonin and 5-hydroxyindoleacetic acid in the thoracic segments of amyotrophic lateral sclerosis were significantly lower than that of controls. Differences in the inter-segmental distribution of noradrenaline in lumbar, lumbar-sacral, and serotonin in lumbar segments of spinal cord were found in this group. Keywords: Spinal cord, biogenic amines, Parkinson's disease, amyotrophic lateral sclerosis.
134
E. Sofic etal. Introduction
Parkinson's disease (PD) is characterized by degeneration of the melanin-containing mesencephalic neurons accompanied by variable damage to both the ascending (nigrostriatal, mesencephalocortical and less, hypothalamic) and descending (to spinal cord) dopaminergic pathways arising from the zona compacta of the substantia nigra and ventral tegmental area. Additional deficiencies of the noradrenergic and serotonergic system due to degeneration of locus ceruleus and dorsal raphe nucleus are known to occur in PD (for review see Agid et al., 1990; Jellinger et al., 1989, 1990). Recent postmortem studies of the lumbar spinal cord of PD subjects revealed considerable decrease of noradrenaline (NA), serotonin (5-HT) and its metabolite 5-hydroxyindoleacetic acid (5HIAA), while spinal cord levels of dopamine (DA) and its metabolite homovanillic acid (HVA) were within control values (Scatton et al., 1986). Although in this study the postmortem intervals in PD subjects (19 4- 3 hrs) were considerably longer than in controls (7 4- 2 hrs), and most PD patients had been treated with L-DOPA prior to 24 hours before death, these authors suggest that the descending DA pathways which project to the lumbar spinal cord are spared in PD. On the other hand, immunohistochemical studies, using an antibody against tyrosine hydroxylase (TH), the rate-limiting enzyme of the catecholamine biosynthesis, showed decreased densities of TH-immunoreactivity (IR) in all levels of the spinal cord of PD subjects, particularly of TH-IR fibres in the superficial zone of the dorsal horn and in the pericommissural gray matter (Dietl, 1985; see Jellinger, 1986). 5-HT, NA and DA-containing neurons have been shown to project on the mammalian spinal cord, although the exact details of areas of termination are not yet fully understood (see Hunt, 1985). While NA-ergic and 5-HT-ergic spinal systems originate from the locus ceruleus, lateral medullary and pontine nucleus (Hunt, 1985; Moore and Card, 1984), and terminate in the dorsal horn (for NA and 5-HT), ventral horn (for 5-HT; Pelletier et al., 1981), lateral sympathetic columns and motor neurons of the ventral horn (for NA) (Hunt, 1985; Moore and Card, 1984), recent studies also demonstrated the existence of DA containing fibres in the mammalian spinal cord (Commissiong and Neff, 1979; Mouchet et al., 1986; Lindvall etal., 1983; Bj6rklund and Lindvall, 1984). The cell bodies of this system originate periventricularly in the dorsal hypothalamus, caudal thalamus and diencephalic A-11 and A-13 DA cell groups (Bj6rklund and Lindvall, 1984). The terminal innervations were found at all spinal cord levels, with highest densities of DA fibres in superficial layers of the dorsal horn, the area surrounding the central canal, the intermediolateral cell column of the thoracolumbar spinal cord and around the preganglionic parasympathetic neurons of the sacral spinal cord (Lindvall et al., 1983; Bj6rklund and Lindvall, 1984). In addition, DA-ergic cell bodies have been described in the spinal cord (Dietl et al., 1985; Mouchet et al., 1986). A similar regional distribution of DA receptors
Biogenic amines in human spinal cord
135
in the mammalian spinal cord has been observed by autoradiography (Commissiong and Neff, 1979; Palacios and Wamsley, 1984; Scatton etal., 1984). While the involvement of the cerebral monoaminergic systems in PD is now well documented, the possible alterations of the spinal cord monoaminergic systems in this and other degenerative disorders like amyotrophic lateral sclerosis (ALS) are poorly understood (Scatton et al., 1986; Lackovic et al., 1988; Ohsugi et al., 1987). Here we present data on the monoamine levels and their metabolites in different parts of the spinal cord in four cases of PD, three cases of ALS and in four control subjects. Materials and methods
Portions of cervical (C), thoracic (T), lumbar (L) and sacral (S) segments of human spinal cords were obtained at autopsy from eleven subjects. Essential clinical and (neuro)pathological data of patients with PD, ALS and controls are given in Table 1. Tissue samples were frozen at - 80 ~ prior to analysis. There were no differences for storage time at - 80 ~ among the three groups. DA, NA, 5-HT, 3,4-dihydroxyphenylacetic acid (DOPAC), HVA and 5-HIAA were analyzed by reversed-phase high-performance liquid chromatography with electrochemical detection [(5-HT, 5-HIAA, HVA, DOPAC; Sperk, 1982) and (NA and DA; Sofic, 1986)]. Statistical analysis of the data was performed using Student's t-test, including Benferroni (~) adjustment. [a* (C) = 0.00119 (7 segments, 6 parameters), a* (T) = 0.000694 (12 segments, 6 parameters), a* (L) = 0.00167 (5 segments, 6 parameters), a* (L-S) = 0.0083 (1 segment, 6 parameters), a* (S) = 0.0083 (1 segment, 6 parameters)l. Results
Data of the biochemical analysis in spinal cord are displayed in Tables 2 to 7. In controls, D A concentrations were about three to four times lower than that of N A (Tables 2 and 3). The significantly highest content of N A was found in the L segment (Table 2) and of D A in L and S segment (Table 3). The concentrations of D A in the T segments of controls were significantly lower than in C segments (Table 3). A n inter-segmental distribution of monoamines was only present in spinal cord of controls. In contrast, in the spinal cord of PD such inter-segmental differences were not found (Tables 2 to 7), despite D O P A C in the L segments (Table 4). Furthermore, spinal cord biogenic amines and their metabolite concentrations did not differ significantly from those measured in controls (Tables 2 to 7). In the L spinal cord of ALS subjects N A contents were significantly higher than in the C and T segments (Table 2). The concentrations of 5-HT in the L segments of ALS were significantly lower than in C segments (Table 6). The concentrations of N A in the C, T (Controls/ALS) and L-S (PD/ALS) segments of patients with ALS were significantly lower than compared to controls or PD (Table 2). Furthermore, significantly lower D A and HVA levels were found in the
E. Sofic etal.
136
Table 1. Clinical notes Code
Sex
Age (years)
Clinical diagnosis
Neuropathology
466/84
f
86
lung carcinoma
diffuse, mild brain atrophy, no other lesions
470/84
f
81
chronic alcoholism, liver cirrhosis
moderate brain atrophy, no other lesions
