THE JOURNAL OF COMPARATIVE NEUROLOGY 304:419-434 (1991)

Distribution of Catecholamine Uptake Sites in Human Brain as Determined by Quantitative C3H]Mazindol Autoradiography GEOFFREY A. DONNAN, STAN J. KACZMARCZYK, GEORGE PAXINOS, PETER J. CHILCO, RENATE M. KALNINS, DIANNE G. WOODHOUSE. AND FREDERICK A.O. MENDELSOHN Departments of Neurology (G.A.D.),Medicine (S.J.K., P.J.C., D.G.W., F.A.O.M.), and Pathology (R.M.K.),Austin Hospital, Melbourne 3084, and School of Psychology, University of New South Wales, New South Wales 2033 (G.P.),Australia

ABSTRACT Because of the importance of the catecholamine system in Parkinson's disease and its relevance to a variety of clinical movement disorders, catecholamine uptake sites were mapped in the human brain using r3H1 mazindol autoradiography. Displacement studies with known dopamine (DA) and noradrenaline (NA) uptake blockers showed that binding in the striatum was to dopamine uptake sites; binding in the locus coeruleus was to noradrenergic uptake sites. By using the selective noradrenergic uptake blocker desmethylimipramine (DMI),a comprehensive map of both DA and NA uptake sites was generated. In general, catecholamine uptake sites were better seen in terminals than in cells of origin or axonal projections. In some areas, such as the locus coeruleus, punctate binding could be seen over individual pigmented cells. A variegated pattern of binding was seen in caudate nucleus and putamen and some correspondence of patches of low binding with striosomes was observed in the caudate. The highest levels of binding to DA uptake sites was observed in the striatum, where regional differences in binding occurred. The most dense binding was seen in the ventral striatum, and a rostral-tocaudal decrement in binding levels in caudate nucleus and putamen was evident. Binding was more intense in the putamen compared to the caudate and within the caudate lower values were seen laterally. The highest levels of binding to noradrenergic uptake sites were in the locus coeruleus and dorsal raph6, although these sites may be on terminals from other projections. Whereas uptake sites were more often evident in known catecholamine pathways, [3Hl mazindol binding was seen in some areas where catecholamine neurons or terminals had not been identified previously. These maps of the catecholamine uptake system add further information concerning the nature of the distribution of catecholamines in human brain and provide an important baseline for the study of disease and ageing processes. Key words: dopamine, noradrenaline, central nervous system, neurotransmitter, uptake inhibitors

The presynaptic catecholamine uptake system is receiving increased attention in the overall assessment of the catecholamine system in movement disorders, particularly Parkinsonism. This applies to postmortem neurochemical studies (Maloteaux et al., '88; Joyce et al., '89; Hurtig et al., 'go), as well as to in vivo studies using Positron Emission Tomography (PET) (Tedroff et al., '88; Brooks et al., '89). Mazindol is a potent inhibitor of catecholamine uptake and binds presynaptically to both high and low affinity uptake sites (Javitch et al., '83, '84;Angel et al., '87). L3H]mazindol autoradiography has been used to determine the regional o 1991 WILEY-LISS, INC.

distribution of catecholamine uptake sites in rodents in normal and disease model states (Javitch et al., '85a,b; Donnan et al., '86, '87). By the addition of desmethylimipramine (DMI) to the incubation medium to block NA uptake sites during the generation of autoradiographs, a clear distinction between dopaminergic and noradrenergic uptake sites may be made while using a single radioligand. Accepted October 3,1990. Address reprint requests to Dr. G.A. Donnan, Department of Neurology, Austin Hospital, Heidelberg,Victoria, 3084, Australia.

G.A. DONNAN ET AL.

420

Abbreviations

Aq BST Cb Cd CG CLi CP DR DTg EW f GP GPe GPi Hi HY IC ic IF IOD IOPr IP Jx LC

IfP LG MB mfi mlf MnR opt PBP PMR PN Pu R RLi k g Rob RPa RRF

anterior commissure nucleus accumbens anterior commissure, posterior part amygdala aqueduct bed nucleus of the strial terminalis cerebellum caudate nucleus central gray caudal linear nucleus cerebral peduncle dorsal raphe nucleus dorsal tegmental nucleus Edinger-Westphal nucleus fornix globus pallidus globus pallidus, pars externa globus pallidus, pars interna hippocampus hypothalamus inferior colliculus internal capsule interfascicular nucleus inferior olive, dorsal nucleus inferior olive, principle nucleus interpeduncular nucleus juxtacapsular portion of BST locus coeruleus

sc

SN SPP Th Tu

VP

vs * I 10 12

(Javitch et al., '85a) We have previously mapped the distribution of catecholamine uptake sites in the mouse brain using this technique (Donnan et al., '89),but no such information exists for the human brain. The regional distribution of catecholamine uptake sites in the human brain should form a baseline for comparative studies of disease states in vitro and in vivo as well as provide greater insight into neuroanatomical and functional relationships of the catecholamine system.

MATERIALS AND METHODS Five human brains were studied, mean age 66 years (range 59-71 years). None of these suffered from known neurological or psychiatric disease. Causes of death and postmortem delay are listed in Table 1. The cerebral hemispheres were cut coronally into 2-cm sections and the remaining brain divided into midbrain, pons, and medulla prior to storage a t -80°C. Coronal sections were then cut at -2o"C, each of 20 pm, and mounted on gelatine-coated slides. Conditions for binding were established by incubating [3H] mazindol with sections from 0 nM to 85 nM. Scatchard plots were then constructed. The sections were preincubated for 5 minutes at 4°C in 50 mM Tris buffer, pH 7.4, containing 300 mM NaC1,5 mM KCL, and 0.2% bovine serum albumin. Incubation was then performed in the same buffer for 60 minutes with 4 nM [3H] mazindol

TABLE 1. Details of Brains Studied Brain bank number

Post mortem delay (hrs)

Age (years)

1 10 11 13

16.0 36.0 6.5 18.5 6.0

61 71 70 59 70

14

Sex

Cause of death

M F

Gastmintest. hemorrhage Acute renal failure Renal adenocarcinoma Chronic renal failure Left ventricular failure

F

M F

longitudinal fasciculus of the pons lateral geniculate mammillary body medial forebrain bundle medial longitudinal fasciculus median raphe nucleus optic tract parabrachial pigmented nucleus paramedian raphe nucleus paranigral nucleus putamen red nucleus rostral linear nucleus of the raphe raphe magnus nucleus raphe obscurus nucleus raphe pallidus nucleus retrorubral field superior colliculus substantia nigra subpeduncular pigmented nucleus thalamus olfactory tubercle ventral pallidum ventral striatum supracapsular strip facial nucleus dorsal motor nucleus of the vagus hypoglossal nucleus

(specific activity 15-24 Ci/mmol, New England Nuclear). Consecutive sections were incubated with (1)[3H]mazindol (for total catecholamine uptake site binding), (2) [3Hl mazindol with 1 pM unlabeled mazindol (nonspecificbinding), and (3) [3H] mazindol with 300 nM DMI to block NA uptake sites. The slides were then given two consecutive washes with 50 mM Tris buffer containing 300 mM NaCl and 5 mM KCL at O T , and then dipped in ice cold distilled water for 1 second and then dried under a stream of cold air. Slides were exposed to Amersham Hyperfilm-3H for 6 weeks. Sections were later stained with 0.1% thionin for histological examination. Calibration curves were constructed from tritium standards (Amersham, UK) and [3Hl mazindol binding was quantitated (fmollmgof tissue) using Eyecom Model 850 image processor and DEC 11/23 computer. A minimum of six readings were taken for each anatomical region. On sections incubated with DMI, DA uptake sites were quantitated and values for NA uptake sites were obtained by subtracting these from total catecholamine binding values. Drug displacement studies were carried out on sections of caudate/putamen and locus coeruleus. Sodium ion concentration experiments were carried out on caudate/putamen sections. In two brains, adjacent sections of caudate-putamen were stained for acetylcholinesterase (AChE) and tracings of the distribution of the striosomes generated by overhead projection. Similar tracings of the adjacent section of [3H] mazindo1 binding were then overlaid to compare the patterns of binding to determine if DA uptake site distribution conformed to a striosomal pattern (Graybiel and Ragsdale, '78). A detailed topographical quantitative analysis of [3H] mazindol binding in the corpus striatum was undertaken by dividing the caudate nucleus, putamen and nucleus accumbens into sections dorsoventrally and medial laterally (see Fig. 5). Representative sections were also taken from rostral, central, and caudal regions of the striatum.

CATECHOLAMINE UPTAKE SITES IN HUMAN BRAIN

A

421 Locus coeruleus

B

Putamen

c

0

ae

0

I .

'

.

'

.

-12 -11 -10

log(drug

'

- 9

.

I

I

.

.

- 8

- 7

-6

- 5

- 4

log(dru g concentration) ,M

concentration),M

Fig. 1. Displacement curves of t3Hl mazindol binding by various catecholamine uptake blockers on human brain sections. 1 = mazindol; 2 = GBR 12909;3 = benztropine;4 = mianserin, 5 = chlorimipramine; 6 = desmethylimipramine; 7 = citalopram. Specific binding was determined at 4 nM [%I mazindol in the presence of 1 p,M mazindol.

Displacement studies were performed on caudate/putamen (A)and locus coeruleus (B).Each point represents the mean vaiue of three experiments with at least six readings taken at each concentration of displacingdrug.

TABLE 2. IC 50 Values bM)'

TABLE 3. PublishedIC 50 Values (nM)'

1 . Mazindol 2. GBR 12909 3.Benztropine 4.Mianserin 5. Chlorimipramine 6. Desmethylimipramine 7. Citalopram

DA

NA

4.7 125.8 398.1

-

-

-

17.8

5.6 56.2

-

DA Mazindol GBR 12909 Benztropine DMI Cblorimipramine Mianserin Citalopram

18

51 110 2,200 4,300 40,000 41,000

NA

5HT

0.65 2,600 150 0.46 24

13,000

23 8,800

30

-

41

1.5 1,200 1.8

'IC 50 values determined from current experiments (Fig. 2) using autoradiographic methods (displacementof 4 nM [3H1 mazindol).

'All figures relate to uptake of tritiated amine into rat brain synaptosomes (Hyttel, '81), exmpt for GBR 12909 where uptake is into rat brain slices (Heikkila and Manzino, '84).

RESULTS Drug displacement studies

appropriate for published IC 50 values for NA uptake into rat brain synaptusomes or brain slices (Table 3). This would suggest that binding in this region is entirely to NA uptake sites.

[3H] mazindol was found to bind to a single class of binding sites with a Kd of 18.5 & 2.8 nM and Bmax of 1.6 c 0.2 fmol/mm*of tissue. The Kd was similar to values found in rodents (Javitch et al.,'85a; Donnan et al., '89), although Bmax is more difficult to compare, since in our current study the value has been determined autoradiographically and expressed as a function of tissue area. Binding was enhanced by sodium ion with optimal binding occurring at concentrations of 250-350 mM (Km 340 mM). Drug displacement studies performed in the striatum confirmed that uptake sites in this region are dopaminergic, since [3H]mazindol was completely displaced by the known DA uptake blockers mazindol, GBR 12909, and benztropine. No displacementwas seen with the noradrenergicuptake blockers DMI and chlorimipramine(Fig. 1A). The rank order of potency of these drugs in displacing i3H1 mazindol (Table 2) was consistent with their published IC 50 values for inhibition of uptake of DA into rat brain synaptosomes (Table 3). L3H1 mazindol also has some affinity for serotonin (5HT) uptake sites, but the 5HT blocker citalopram (Hyttel, '82) (1fl)did not displace [%I mazindol in either corpus striatum or median raph6 in our material. In the locuscoeruleus,displacementwas seen with the NA uptake blockers DMI, chlorimiprimine, and mianserin but no displacement with citalopram (Fig. lb). Agam, the rank order of potency for drug displacement was

Autoradiographic distribution Representative autoradiographs are shown in Figures 2 and 3 and quantitive values for [3Hl mazindol binding in Tables 4 and 5. The areas of most intense binding were found to dopamine uptake sites in the striatum and noradrenergic uptake sites in the locus coeruleus. Nonspecific binding was seen in many cellular areas, but was most marked in the inferior olivary nucleus, cerebellar cortex, pontine nuclei, and grey matter of the spinal cord. In general, catecholamine fibre pathways did not show any binding, with the exception of the ascending nigrostriatal tract and medial forebrain bundle. Binding was most evident in areas of termination of catecholamine projections, such as the corpus striatum. In other areas, for example the locus coeruleus, intense binding was seen over cell bodies, but whether this represents terminations of catecholamine projections or uptake sites on intrinsic neurons is less clear and is discussed below. Forebrain. [3H] mazindol binding was entirely to dopamine uptake sites in the caudate nucleus, putamen, and nucleus accumbens (Fig. 2A-P). Rostrally, binding levels were highest in the ventral striatum, and a significant decrement in values occurred from putamen to caudate

G.A. DONNAN ET AL.

422 TABLE 4. 13H1Mazindol Binding in Human Brain (fmolimg tissue) ~~~~

~

~

DA+NA' Area Cd head, body tail Pu Ach

GP

vs

VP Tu Jx BST Th HY Hi LG mfb R SN compacta reticulata EW RLi CLi PBP PN CG DR MnR RMg Rob RPa PMR LC Ch DTg

SPP

7 10 rostral caudal 12

DA'

NA3

mean

s.e.m.

mean

s.e.m.

51.5 30.1 63.2 47.6 18.5 108.3 13.9 46.7 57.4 25.5 12.1 25.6 45.8 15.0 14.9 15.0 15.9 9.6 32.3 17.6 23.7 24.5 50.5 8.1 17.5 36.2 60.8 47.3 9.3 21.0 10.2 68.6 74.1 6.6 27.5 25.0 14.9 73.1 46.7 31.1

2.9 2.5 3.1 1.9 1.4 1.9 0.4 2.6 1.8 4.3 1.2 1.4 2.5 0.6 0.5 0.7 0.3 0.4 1.3 1.6 1.7 2.2 4.9 0.2 0.5 2.5 3.0 3.6 0.2 0.9 0.3 1.8 4.1 0.3 0.8 2.0 0.5 2.3 2.1 1.8

51.5 30.1 63.2 47.6 9.9 108.3 11.9 46.7 57.4 17.5 12.1 15.1 13.1 15.0 14.9 15.0 15.9 2.0 32.3 12.2 5.3 17.3

2.9 2.5 3.1 1.9 1.3 1.9 0.2 2.6 1.8 1.4 1.2 1.2 0.7 0.6 05 0.7 0.3

7.0 12.9 18.2

0.2 0.4 0.9

-

2.2 3.3 4.7 -

-

1.3 07 0.2 1.7

-

-

0.1

-

0.1

-

-

mean

s.e.m.

8.7 -

0.9

-

2.0

0.1

8.0 -

-

10.5 36.7

1.1 2.0

-

-

-

-

1.0

-

-

-

-

7.6 0 5.4 18.4 7.2 50.5 1.0 4.6 18.0 60.8 47.3 7.1 21.0 10.2 68.6 70.8 6.6 27.5 20.3 14.9 73.1 46.7 31.1

0.2 0 0.2 1.1 0.7 4.9 0.1 0.1 1.1 3.0 3.6 0.1 0.9 0.3 1.8 3.0 0.3 0.8 1.8 0.5 2.3 2.1 1.8

'Both dopamine and noradrenaline uptake sites represented by specific 13H1 Mazindol binding (Total minus non-specific with 1pm unlabeled Mazindolj. 'Dopamine uptake sites represented by total l3H1Mazindol binding incubated with 300 nM DM1 to block NA uptake sites. 3Noradrenalineuptake sites obtained hy subtracting DA from DA+NA. Dash (-4= undetedable binding (not different from background).

nucleus to nucleus accumbens (p < 0.001, ANOVA) (Table 5). The caudate-putamen difference was lost in the central region but re-emerged posteriorly (p < 0.05,2 tailed t-test). Within the caudate nucleus there was a lower binding laterally compared to the remainder and levels diminished significantly from rostral to caudal. Dorsalhentral comparisons for the caudate revealed little difference except for caudally, where dorsal values were lower. In the putamen there was also a significant reduction in binding from rostral to caudal, but values were otherwise fairly constant except for some reduction dorsomedially in the rostral region. In both caudate nucleus and putamen the binding was clearly heterogeneous with patches of lower activity. (Fig. 2A-H). Matching of adjacent sections of the caudate nucleus stained for acetylcholine esterase (AChE) revealed a confluence between these hypodense regions and the AChE poor striosomal patches (Graybiel and Ragsdale, '78). However, this occurred in only approximately 50% of instances in the caudate nucleus and was not apparent in the putamen. Bridges of binding connected the striatum with the olfactory tubercle, which supports the concept that the olfactory tubercle is a ventral extension of the striatum (Heimer and Wilson, '75; Alheid et al., '90) (Fig. 2D). The juxtacapsular portion of the bed nucleus of the striae terminalis showed binding to DA uptake sites (not displaced by DMI) (Fig. 2D), which was surprising in view of the known substantial projection of the locus coeruleus to this structure in rodents (Moore and Bloom, '79; Moore and Card, '84). Neocortical structures showed no binding to catecholamine uptake sites. Although the amygdala and hippocampus receive catecholaminergic inervation, in the human brain (Pearson et al., '90) we could demonstrate low levels of binding to DA uptake sites only (Fig. 2 G J). Both DA and NA uptake sites were observed in the hypothalamus and this was most evident in the midhypothalamic levels (Fig. 2M,N).All dopamine cell groups known to exist in the rat have now been demonstrated in the human (Pearson et al., '90) and the hypothalamus is known to receive terminals from the ventral noradrenergic bundle

TABLE 5. Specific [3H] Mazindol Binding to DA uptake Sites in Regions of the Striatum (fmolimg tissue) ~~

Caudate mean Rostral

d'

Putamen s.e.m.

52.5

5.2

60.9

4.8 4.5 5.2 3.2 3.4 3.9

mean

s.e.m.

C'

v3 m' C

15

Central

Total d

75.7 69.0 52.5 57.1

4.5

Caudal

1 Total d C

V

m C

1 Total

39.4

3.7 4.2 2.9 2.5

~

Accumbens

46.9 47.1 44.1

'Dorsal. 'Central. Wentrd; *Medial:as shown in Figure 5. 'Lateral. For ANOVA, 'P < 0.05, ** P 4 0.01, *** P < 0.001 andasterisks areplacedoppositelowest values. Two-tailed t-test, t P < 0.05.

4.5 3.7 1.8 3.9 3.7 3.7 24 4.0 3.9 4.6 4.9 4.5 4.6 3.8 4.9 1.8 1.2 3.7 3.3 2.7 2.2

mean

6.e.m. 1.7

49.7

4.0 4.5

46.0 47.8

2.0 2.4

CATECHOLAMINE UPTAKE SITES IN HUMAN BRAIN (Loughlin and Fallon, '85). Some low level binding to both DA and NA uptake sites was seen in the thalamus (Fig. 2H-P), but the discrete nuclear delineation seen in the mouse (paraventricular and anteroventral nuclei) was not present. Modest binding to DA uptake sites was seen in the lateral geniculatenucleus. Lamination was seen that corresponded roughly to the lateral granular layers with increased binding in cellular regions (Fig. 2L). Midbrain. Moderate [3Hl mazindol binding was seen in substantia nigra pars compacta and a continuum was formed with the ventral tegmental area medially (Fig. 2Q-S). Binding was mainly to DA uptake sites, although low levels of NA uptake sites were present, consistent with the knowledge that low levels of NA are present in substantia nigra (Farley and Hornykiewicz, '77). Low levels of binding were also seen in the substantia nigra pars reticulata. In addition to the diffuse binding seen in the substantia nigra, there was punctate binding that we were able to correlate with pigmented cells on a number of occasions. The binding in the substantia nigra extended dorsomedially with little diminition to engulf the parabrachial pigmented nucleus (Fig. 2Q). The paranigral nucleus, which in the human is located at the medial tip of the substantia nigra, showed as intense binding as the substantia nigra pars compacta (Fig. 2Q). The catecholamine projection of the medial forebrain bundle was identified at its caudal levels near its interface with the internal capsule (Fig. 2N). The binding was to DA uptake sites, suggesting that the nigrostriatal projection has been identified. A supracapsular strip was identified that showed [3H] mazindol binding to DA uptake sites (* in Fig. 2N) and may represent a branch of the main ascending catecholamineprojection. The caudal linear and interfascicular nuclei displayed modest binding to DAuptake sites (Fig. 2s). The binding to the interfascicular nucleus was clearly higher than that of the subjacent interpeduncular nucleus (Fig. 2s).The retrorubal fields (A8) showed low binding (Fig. 2s). The rostral linear nucleus, the remaining dopaminergic cell group of the mesencephalon and consisting of scattered neurons dorsal and medial to the red nucleus and at times within the superior cerebellar peduncle, also showed modest levels of binding to DA uptake sites (Fig. 2Q). These modest levels of binding may be expected because of the paucity and diffuseness of cells, although occasional punctate binding was seen over individual cells. The EdingerWestphal nucleus showed moderate binding, mainly to NA uptake sites, although this could not be separated reliably from dorsal aspects of the occulomotor nucleus and further clarification may be required (Fig. 2Q). The red nucleus, especially at its rostral pole, displayed low levels of binding to both DA and NA uptake sites (Fig. ZQ). The central grey showed binding mainly to NA and DA uptake sites, which was somewhat surprising given the presence of only a few tyrosine hydroxylase immuno reactive cells in the central grey of the human (Pearson et al., '90) and the lack of binding found in similar areas of our mouse study (Donnan et al., '89). Pons and medulla. The locus coeruleus displayed the highest [3H] mazindol binding of all structures identified and binding was entirely to NA uptake sites (Figs. lB, 3U,U'). The binding was both diffuse and punctate and the puncta often registered over the pigmented cells. The correspondence between puncta and cell bodies is displayed in Figure 4. A pigmented nucleus associated with the

423

ventrolateral aspects of the superior cerebellar peduncle has recently been identified in the human and named the subpeduncular pigmented nucleus (Fix, '80; Ohm et al., '87; Paxinos et al., '90). This nucleus was associated with low levels of binding, mainly to NA uptake sites (Fig. 3U,U'). The subcoeruleus nucleus (A7 and A5) constitutes the other noradrenergic group in the pons and connects the locus coeruleus with the dorsal aspect of the superior olive. Pigmented cells were sparsely distributed in this large area, but our techniques did not reveal increased binding. Similarly, the noradrenergic cell group found in the caudal part of the intermediate reticular zone (A1 or caudoventrolatera1 medulla), did not reveal enhanced binding. High levels of binding to NA uptake sites was seen in the rostral pole of the dorsal motor nucleus of the vagus with lower levels of binding observed elsewhere in the nucleus (Figs. 3W,3W', 3X,3X'). The adrenergic C 1 cell group is found in the rostral part of the intermediate reticular zone of the ventrolateral medulla (Paxinos et al., '90; Pearson et al., 'go), but we found no clear evidence of increased binding here, though the diffuse organization of the nucleus may account for this. The greatest concentration of uptake sites in the raphe system was associated with the dorsal raphe nucleus and this binding was to noradrenergic uptake sites (Fig. 2R,S). The binding associated with the dorsal raph6 extended to the caudal pole of the raphe where it was then confined to the midline by the development of the dorsal tegmental nucleus laterally (Fig. 2s). The median raphe also showed binding to NA uptake sites (Figs. 2T, 3V,V'). Rostrally, this binding was associated with the magnocellular median raphe proper as a midline strip as well as with the more diffuse paramedian raphe nucleus as recently identified in the rat and human (Paxinos and Watson, '86; Tork and Hornung, '90). In the midline strip the binding corresponded roughly with the distribution of large cells. Posterially, binding was to two slender paramedian strips that have been thought previously to belong raphe pontis, but has recently been shown to be an extension of the median raph6 (Tork and Hornung, '90). The raphe obscurus nucleus was associated with moderate binding (Fig. 3W,W'). Unlike the mouse and rat, the human facial nucleus displayed moderate binding to NA uptake sites and similar binding levels were found in the hypoglossal nucleus (Fig. 3V,V',X,X').

DISCUSSION A number of radioligands have now been used to label catecholamine uptake sites. These include tritiated cocaine (Kennedy and Hanbauer, '831, nomifensine (Scatton et a]., '85), mazindol (Javitch et al., '83, ,841, threo (+)-methyl phenidate (Janowski et al., '85; Schweri et al., '851, desmethylimipramine (DMI) (Langer et al., '81, Lee et al., %l), GBR 12935 (Berger et al., '85; Janowski et al., '861, and GBR 12783 (Bonnet et al., '86). GBR12935 is the most specific for dopamine uptake sites, but it may also bind to piperazine sites (Andersen, '87; Andersen et al., '87) and has at least some affinity for noradrenergic uptake sites (van der Zee et al., '80). Mazindol has a comparable affinity to GBR 12935 for dopamine uptake sites together with a high affinity for noradrenergic uptake sites; by the addition of DMI to block noradrenergic uptake sites, [3Hl mazindol can be used to autoradiographically identify dopaminergic uptake sites alone; by subtracting these values from the

424

total catecholamine binding, noradrenergic uptake sites may be determined while using a single radioligand (Javitch et al., '85a). This technique has already proven to be useful in determining the effect of neurotoxins in the presynaptic catecholamine system in animal models (Javitch et al., '85b; Donnan, '86, '87) and human disease processes (Joyce et al., '89) and a detailed map of the catecholamine uptake system in mouse has been generated (Donnan et al., '89). There is good evidence that [3H] mazindol binds to presynaptic dopaminergic and noradrenergic uptake sites. In particular, binding has been shown to be saturable, sodium dependent (Javitch et al., '83, '841, and compounds that displace [3H] mazindol from rat brain striatal membranes and mouse brain striatal sections are highly correlated with their ability to inhibit both dopamine and noradrenaline uptake into synaptosomal preparations (Javitch et al., '83; Donnan et al., '891.. Our displacement studles in both the mouse and human brain confirm that uptake sites are dopaminergic in the caudate nucleus and putamen and noradrenergic in the locus ceoruleus. Further, the distribution in the rat, mouse, and human brain correlates well with known areas of catecholamine distribution (Javitch et al., '85; Donnan et al., '89, this study). Lesioning experiments of the nigro striatal tract; surgically in rats (Javitch et al., '85af and with MPTP in mice (Donnan et al., '86, '87) confirm that the uptake sites are located presynaptically. In the current study we present the first detailed maps of catecholamine uptake sites in the human brain. By obtaining sections at closely spaced intervals from all brains, we were able to simplify the determination of anatomical relationships, particularly with ascending pathways such as the nigrostriatal tract. Whereas the distribution of uptake sites is presented visually for most structures, some allowance should be made for the "quenching effect" known to occur with tritium and may result in up to 30% error over white matter structures (Rainbow et al., '84, Kuhar and Unnerstall '85). However, for practical reasons, uncorrected images are commonly used, particularly since error rates are low over grey matter structures where the majority of [3H] mazindol binding is found. Whereas the general distribution of catecholamine uptake sites followed the known immunohistochemical distribution of catecholamines in humans (Pearson et al., '90) and rodents (Moore and Bloom, '78; Moore and Bloom, '79; Bjorklund and Lindvall, '84; Moore and Card, '84),some differences did exist. The main reasons for this may be as follows: 1. Uptake sites are located predominantly on terminals, whereas immunohistochemical localization of tyrosine hydroxylase is predominantly in cell bodies with little visualization of terminals. 2. Some essential differences between rodent and human neurochemical anatomy. 3. Lower sensitivity of t3Hl mazindol autoradiography compared to immunohistochemical techniques generally.

The pattern of intensity of OH1 mazindol binding was also similar to our findings in the mouse brain in that, for example, within the dopaminergic system, binding to uptake sites was more intense in striatal terminals rather than the substantia nigra cells of origin, or in the nigrostriatal projection pathway. This raises the interesting question as to the precise cellular location of the mapped catecholamine uptake sites found in a particular nucleus.

G.A. DONNAN ET AL. The possibilities include: (1) location on cell body membranes, (2) location on dendrites from adjacent neurons, (31 location on terminals of axonal projections making synaptic connection with neurons of the nucleus under consideration. In the striatum it seems likely that the DA uptake sites are located on the neuronal terminals of the nigrostriatal projection in view of the well-documented reuptake system at this site and the marked reduction in [3Hl mazindol binding when the nigrostriatal system is lesioned surgically or chemically in rodents (Javitch et al., '85b; Donnan et al., '86, '87), or pathologically in humans (Hurtig et al., '89; Joyce et al., '89; Donnan et al., '91). However, in the case of binding seen on cell bodies, such as within the nigrostriatal tract and locus coeruleus, a similar arrangement is far less certain. Punctate [3H] mazindol binding was seen to be associated with individual pigmented cell bodies in both of these regions. This may have been associated with either intrinsic uptake sites, presynaptic sites on dendrites of nearby catecholaminergic cells, or terminals from distant axonal projections. Both axons and dendrites of catecholaminergic cells have been shown to have active reuptake and release systems in rodents (Geffen et al., '76, Cheramy et al., %l), although little release of DA from human synaptosomal preparations of substantia nigra may occur (Hardy et al., '87). The subregional differences seen in [3H] mazindol bindings to DA uptake sites within the striatum were quite marked and somewhat different to changes reported in DA levels in other studies (Hortnagl et al., '83; Kish et al.,'88) Within the striatum we found that the highest levels of binding were seen in the ventral striatum and, rostrally, binding in the putamen was higher than in the caudate nucleus and nucleus accumbens. However, centrally and caudally there was little difference in binding levels between caudate nucleus and putamen, but a significant rostral to caudal decrement was present. This pattern is unlike what we saw in the mouse where higher levels were seen in the caudal caudate-putamen. Although it is difficult to compare our findings with other markers of the human striatal system because [3Hl mazindol autoradiography allows more precise regional quantitation, an analysis of the regional dopamine values in the control patients of Kish et al. ('88) failed to reveal a similar pattern of gradation within the striatum. In fact, dopamine levels in the putamen were lower than in the caudate nucleus and little dorsalhentral or rostrakaudal differences in levels were observed. However, in an earlier study the same group found a rostrallcaudal decrement in dopamine levels within the caudate nucleus (Hortnagl et al., '83). Tyrosine hydroxylase immunoreactivity also tends to be lower in the putamen, although more intense staining is found in the nucleus accumbens (Pearson et al., '90). Documentation of the pattern of the distribution of DA uptake sites in the normal striatum is important to allow more precise interpretation of PET studies using "C nomifensine, and changes seen in Parkinson's disease where DA uptake site loss appears to preferentially effect the dorsolateral regions of the striatum (Donnan et al., '91). In studies of pathological states, the importance of carefully matching sample and

Fig. 2. Autoradiographs of total r3H1 mazindol binding in coronal sections of human brain showing representative sections from caudate nucleus and putamen to hypoglossal nucleus. These autoradiographs represent DA plus NA uptake sites. (magnification x 3.5)

CATECHOLAMINE UPTAKE SITES IN HUMAN BRAIN

Figure 2 A-D

425

G.A. DONNAN ET AL.

Figure 2 E-H

CATECHOLAMINE UPTAKE SITES IN HUMAN BRAIN

Figure 2 I-L

427

G.A. DONNAN ET AL.

428

Figure 2 M-P

CATECHOLAMINE UPTAKE SITES IN HUMAN BRAIN

Figure 2 Q-T

429

G.A. DONNAN ET AL.

430

Figure 3

CATECHOLAMINE UPTAKE SITES IN HUMAN BRAIN

431

Fig. 4. Punctate [3Hl mazindol binding (lower panel) seen over individual pigmented neurons (upper panel, thionin stain) of the locus coeruleus. Corresponding cells and puncta are identified by arrows. The main body of the locus coeruleus is seen inferiorly in both panels (magnification x 10).

Fig. 3. Autoradiographs of adjacent sections showing total L3H1 mazindol binding representing both DA and NA uptake sites (left-hand panels) and in the presence of 300 nM DMI to adjacent sections to block NA uptake sites (right-hand panels U'-X'). NA uptake sites represent the DMI displaceable component of mazindol binding (left-band panels vs right-hand panels). (magnification X 5)

control areas anatomically within the striatum becomes obvious in view of the regional differences we have demonstrated. A patchy distribution of [3H] mazindol binding in caudate nucleus and putamen was observed and a limited correspondence between areas of low binding and striosomes was seen in caudate nucleus. These findings are in accord with those of Graybiel and Moratalla ('89) who observed an even greater confluence in the cat and monkey caudate nucleus. Lowenstein et al. ('901,in a short commu-

G.A. DONNAN ET AL.

432 Medial-Lateral Divisions

Dor sa I -Vent ra I Divisions

Caudal

Fig. 5. Topographical divisions of the striatum for quantitative analysis of 13Hl mazindol binding presented in Table 6 .

nication, also found that the pattern of [3H] mazindol sites conformed to a patchlmatrix organization in human caudate and that the denser binding was i a the matrix component. Given the potential importance of the variability in arrangement in the striatum of dopamine uptake sites as an explanation for selective potencies of drug and toxins, a more detailed study is indicated. Not all structures within the catecholamine system were identified. Termination sites of known projections in experimental animals to the cortex and thalamus were not seen (Mooreand Bloom, '79), unlike in the mouse, where binding to the paraventricular, anteroventral thalamic nuclei, and cerebral cortex was clearly seen using the same techniques. In a study restricted to the telencephalon and cerebrum, where [3H] DMI was used as the radioligand to identify noradrenergic uptake sites, binding to these structures was demonstrated (Gross-Isseroffet al., '88). This is surprising given that DMI and mazindol have an almost identical affinity for NA uptake sites based on inhibition of uptake into rat brain synaptosomes (Table 3). However, the two studies are difficult to compare, since in the DMI study

brainstem structures were not examined and this region is where the majority of NA uptake sites were found as identified by [3H] mazindol. Several other important catecholamine groups were not associated with binding to uptake sites. Specifically,the A1 (noradrenergic) and C1 (adrenergic) regions were not identified. Presumably, these groups receive few catecholaminergic afferents from elsewhere and the cell bodies are too sparse to register the presence of uptake sites. Some specific structures where [3Hl mazindol binding was seen deserve further comment, particularly with reference to the known distribution of the catecholamine system in the human brain and our findings of the distribution of catecholamine uptake sites in the mouse brain. The identification of the nigrostriatal projection within the medial forebrain bundle was based on the finding of a discrete bundle of dopamine uptake sites lying between the zona incerta and subthalamus, which were able to be traced back to the substantia nigra. In the human as well as the mouse, binding was stronger at caudal levels where the projections are most compact (Bjorklund and Lindvall, '841,particularly at the levels of the subthalamus. At these levels in the rat, Javitch suggested that there was binding in the zona incerta and subthalamus (Javitch et al., '85a). Neither of these structures showed any binding in our material (human and mouse). The supracapsular strip we have identified (labelled*in Fig. 2N) has not been described previously and may represent a branch of the main ascending catecholamine projection. The occurrence of uptake sites in the red nucleus is puzzling in that no tyrosine hydroxylase immunoreactive terminals or cell bodies are present here in the rat (Hokfelt et al., '84) or the human (Pearson et al., '90) and only the occasional ectopic rostral linear nucleus cell body may be seen within the red nucleus itself. Furthermore, no binding in this region was found in our mouse material using the same techniques. Similarly, the central grey, which showed moderate binding of mixed NA and DA uptake sites in the human, shows only a few tyrosine hydroxylaseimmunoreactive positive cells (Pearson et al., '90) and no binding to catecholamine uptake sites was shown in the mouse. Noradrenergic uptake sites were found in the dorsal motor nucleus of the vagus in both human and mouse preparations. Modest levels of NA have been found on chemical assay in human dorsal motor nucleus of vagus (Farley and Hornykiewicz, '77; Mefford et al., '78), but immunohistochemical techniques have not revealed an extensive network of catecholamine cells or terminals (Pearson et al., '79; Pearson et al., '83; Kitahama et al., '85, '88; Kemper et al., '87; Arango et al., '88; Pearson et al., '90). As described by Pearson et al. ('901, a column of dopamine beta hydroxylase-immunoreactive and tyrosine hydrosylase immuno reactive (TH-IR) neurons accompanies the dorsal motor nucleus of the vagus for all but its most rostral extent (their figures 2,3,4).A few of the neurons lie within the nucleus itself, but most catecholamine cells lie medially, ventrally, and laterally. Catecholamine cell bodies are present in the adjacent solitary nucleus cell complex with the greatest number in the nucleus gelatinosus. The catecholamine elements in the vagal-solitary complex correspond to the A2/C2 cell groups in the rat that are immunoreactive for tyrosine hydrosylase, as depicted in Hokfelt et al. ('84). We are unable to explain why binding sites are found only in the dorsal motor nucleus of the vagus and not other areas

CATECHOLAMINE UPTAKE SITES IN HUMAN BRAIN of the vagal-solitary complex that also feature catecholamine cells. Possibly, the vagus nucleus receives terminals from the adjacent cell bodies described. Our finding of very high levels of binding to NA uptake sites in the dorsal raphe of human brain was similar to our finding in mouse brain but at variance with studies in human brain using immunohistochemicaltechniques. Scattered TH-IR cells have been demonstrated in the raphe pallidus, obscuris, and magnus (Halliday et al., '88) and projections of TH-IR neurons have been traced to raphe obscuris and pallidus (Arango et al., '881, but Pearson et al. ('83, '90) found no TH-IR neurons in dorsal raphe. However, NA has been detected in dorsal raphe by chemical assay (Farley and Hornykiewiez, '77; Kopp et al., '79) and a rich network of terminals to dorsal raphe (presumably from locus coeruleus, see Tork '85) has been documented in the rat brain (Hokfelt et al., '84, plates 93-96). The most likely explanation for the discrepancy between our findings of extremely high levels of NA uptake sites in dorsal raphe and the paucity of terminals documented immunohistochemically is the low sensitivity of the latter technique in the human postmortem brain. In view of the low number of TH-IR cells observed in the dorsal raphe, the most likely origin of the dense binding to NA uptake sites in this region is terminals of projections from the locus coeruleus. The presence of NA uptake sites in the facial and hypoglossal nuclei was surprising in view of the fact that no binding was seen in these regions in the mouse brain and no catecholamine cells or terminals have been identified in either nuclei in the rat (Hokfelt et al., '84) or human brain immunohistochemically (Pearson et al., '79, '90; Kitahama, '85, '88;Kemper et al., '87; Arango et al., '88; Halliday, '88). However, assayed levels of NA have been detected in the human brain seventh and twelvth nuclei (Farley and Hornykiewicz, '77; Mefford et al., '78) and the lack of immunohistochemically detected catecholamine terminals should again be tempered by the observation that such terminals are not readily detected by these techniques in the human postmortem brain. In studies of the rat brain, Levitt and Moore ('79) have proposed that noradrenergic projections to the facial and hypoglossal nuclei exist that arise from the Kollieker-Fuse nucleus located laterally and superiorly in ventrolateral aspects of the parabrachial nuclei. Perhaps similar projections are responsible for the NA uptake sites we have observed in human facial and hypoglossal nuclei. The recently described subpeduncular nucleus (Fix, '80) has pigmented cells present (Halliday et al., '88)' although no major efferent or afferent catecholamine connections have yet been identified. Our finding of moderate binding to predominantly NA uptake sites in this nucleus confirms its catecholaminergic association, although the cellular location of the uptake sites is unclear.

ACKNOWLEDGMENTS This research was supported by the Australian National Health and Medical Research Council, the Australian Brain Foundation, and the Parkinson's Disease Association of Victoria. We are grateful for helpful discussions with Dr. P.M. Beart.

LITERATURE CITED Alheid, G.F., L. Heimer, and R.C. Swifzer (1990) The basal ganglia. In G. Paxinos (ed):The Human Nervous System. San Diego: Academic Press, pp. 483-582.

433 Andersen, P.H. (1987) Biochemical and pharmacological characterization of [3Hl GBR 12935 binding in vitro to rat striatal membranes: Labelling of the dopamine uptake complex. J. Neurochem. 48:1887-1896. Andersen P.H., J.A. Jansen, and E.R. Nielsen (1987) [3Hl GBR 12935 binding in vivo in mouse brain: Labelling of a piperazine acceptor site. European J. Pharmacolf44: 1-6. Angel, I., M.D. Luu, and S.M. Paul (1987) Characterization of Mazindol binding in rat brain: Sodium-sensitive binding correlates with the anorectic potencies of phenyl-ethylamines. J. Neurochem. 48:495-497. Arango, V., D.A. Ruggiero, J.L. Callaway, M. Anwar, J.J. Mann, and D.J. Reis (1988) Catecholaminergic neurons in the ventrolateral medulla and nucleus of the solitary tract in the human. J. Comp. Neurol. 273924240. Baker, K.G., I. Tork, J.-P. Hornung, and P. Halasz (1989) The human locus coeruleus complex: An immunohistochemical and three dimensional reconstruction study. Exp. Brain Res. (in press). Berger, P., F. Janowski, F. Vocci, P. Skolnick, M.M. Schwari, and S.M. Paul (1985) [3H1 GBR 12935: A specific high affinity ligand for labelling the dopamine transport complex. Eur. J. Pharmacol. 107:289-290. Bjorklund, A., and 0. Lindvall (1984) Dopamine-containing systems in the C.N.S. In A. Bjorklund and T. Hokfelt (eds): Handbook of Chemical Neuroanatomy. Elsevier, pp. 55-111. Bonnet, J.J., A. Protaix, A. Chagraovi, and 3. Costentin (1986) High af€inity [3Hl GBR 12783 binding to a specific site associated with the neuronal uptake complex in the central nervous system. Eur. J. Pharmacol. 126.921-222. Brooks, D.J., E.P. Salmon, R. Bannister, C. Mathias, and R.S.J. Frackowiak (1989) The integrity of the dopaminergic system in multiple system atrophy and pure autonomic failure studied with PET. In A.R. Crossman, and M.A. Sambrook (eds): Neural Mechanisms in Disorders of Movement. John Libbey, pp. 419-425. Cheramy A,, V. Leriel, and J. Glowinski (1981) Dendritic release ofdopamine in the substantia nigra. Nature 289:537-542. Donnan, G.A., S.J. Kaczmarczyk, P.J. Rowe, R. Figdor, and F.A.O. Mendelsohn (1986) The effects of l-methyl-4-phenyl-l,2,3,6-tetrahydropyridine on presynaptic dopamine uptake sites in the mouse striatum. J. Neurol. Sci. 74:lll-119. Donnan, G.A., S.J. Kaczmarczyk, J.S. McKenzie, R.M. Kalnins, and F.A.O. Mendelsohn (1987) The regional and temporal effects of l-methyl-4phenyl-1,2,3,6-tetrahydropyridineon dopamine uptake sites in mouse brain. J. Neurol. Sci. 81261-271. Donnan, G.A., S.J. Kaczmarczyk, J.S. McKenzie, R.M. Kalnins, P.J. Chilco, and F.A.O. Mendelsohn (1989) Catecholamine uptake sites in mouse brain: Distribution determined by quantitative [3H] mazindol autoradiography. Brain Res. 504:64-71. Donnan, G.A. S.J.Kaczmarczyk, G. Paxinos, R.M. Kalnins, P.J.Chilco, D.G. Woodhouse,and F.A.O. Mendelsohn (1991) The topography of catecholamine uptake site loss in Parkinsons disease (in prep). Farley, I.J. and 0. Hornyklewicz (1977) Noradrenalin distribution in subcortical areas of the human brain. Brain Res I26:53-62. Fix, J.D. (1980) A melanin containing nucleus associated with the superior cerebellar nucleus in man. J. Hirnforsch. 21:429-436. Geffen, L.B., T.M. Jessell, A.C. Cuello, and L.L. Iversen (1976) Release of dopamine from dendrites in rat substantia nigra. Nature 260:258-260. Graybiel, A.M., and R. Moratalla (1989) Dopamine uptake sites in the striatum are distributed differentiallyin striosome and matrix compartments. Proc. Natl. Acad. Sci. 86:9020-9024. Graybiel, A.M., and C.W. Ragsdale (1978) Histochemically distinct compartments in the striatum of human, monkey and cat demonstrated by acetylthiocholinesterasestaining. Proc. Natl. A d . Sci. 75:5723-5726. Gross-Isseroff, R., M. Israeli, and A. Biegon (1988). Autoradiographic analysis of r3H] desmethylimipramine binding in the human brain postmortem. Brain Res. 456120-126. Halliday, G.M.,Y.W. Li, T.H. Joh, R.G.H. Cotton, P.R.C. Howe, L.B. Geffen, and W.W. Blessing (1988) Distribution of monoamine-synthesizing neurons in the human medulla oblongata. J. Comp. Neurol. 273:301317. Hardy, J.A., P. Webster, I. Blackstrom, J. Gottfries, L. Oreland, A. Streustrom, B. Winblad. (1987). The regional distribution of dopamine and serotonin uptake and transmitter concentrations in man. Neurochem. Int. 10:445-449. Heikkila, R.E., and L.Manzino (1964) Ekhavioral properties of GBR 12909, GBR 13069 and GBR 13098: specific inhibitions of dopamine uptake. Eur. J. Pharmacol103r241-248.

434

G.A. DONNAN ET AL.

Heimer, L., R.D. Wilson (1975) The subcortical projections of the allocortex: Lee, C.M., and S.H. Snyder (1981) Norepinephrine neuronal uptake binding sites in rat brain membrane labelled with L3H]desipramine. Proc. Natl. similarities in the neural associations of the hippocampus piriform Acad. Sci. USA 785250-5254. cortex and the neocortex. In: M. Santini (ed): Golgi Centennial S p p o sium Proceedings: New York: Raven Press, pp. 177-193. Levitt, P., and R.Y. Moore (1979) Origin and organization of brain stem catecholamine innervation in the rat. J. Comp. Neurol. 186.505-528. Hokfelt, T., R. Martensson, A Bjorklund, S. Kleinau, and M. Goldstein (1984) Distributional maps of tyrosine-hydrosylase immunoreactive Lowenstein, P.R., J.N. Joyce, J.T. Coyle, J.F. Marshall (1990) Striosomal neurons in the rat brain. In A. Bjorklund and T. Hokfelt (eds):Handbook organization of cholinergic and dopaminergic uptake sites and cholinof Chemical Neuroanatomy. Elsevier, pp. 277-279. ergic M, receptors in the adult human striatum: A quantitative receptor autoradiographic study. Brain Res. 510:122-126. Hortnagl, H., E. Schlogl, G. Sperk, and 0. Horaykiewicz (1983) The topographical distribution of the monoaminergic innervation of the basal Maloteaux, J.M., M.-A. Vanisterg, C. Laterre, F. Javoy-Agid, Y. &d, and ganglia of the human brain. Prog. Brain Res. 58:269-274. P.M. Laduron (1988) [3H1 GBR 12935 binding to dopamine uptake sites: Subcellular location and reduction in Parkinsons disease and progressive Hurtig, H., J. Joyce, J.R. Sladek, and J.Q. Trojanowski (1989) Postmortem supranuclear palsy. Eur. J. Pharmacol. 156t331-340. analysis of adrenal medulla autograft in a patient with Parkinson’s disease. Ann. Neurol. 25t607-614. Mefford, I., A. Oke, R. Keller, R.N. Adams, and G. Jonsson (1978). Hyttel, J. (1982) Citalopram. Basic and clinical studies. Prog. NeuropsychoEpinephrine distribution in human brain. Neurosci. Lett. 9:227-231. pharmacol. Biol. Psychiat. 6:275-295. Moore, R.Y., and F.E. Bloom (1978) Central catecholamine neuron systems: Janowsky, A.J., F. Berger, F. Vocci, R. Labarca, P. Skolmick, and S.M. Paul Anatomy and physiology of the dopamine systems. Ann. Rev. Neurosci. (1986).Characterization of sodium dependent [3H1 GBR 12935 binding 1:129-169. in brain: A radioligand for selective labelling of the dopamine transport Moore, R.Y., and F.E. Bloom (1979) Central catecholamine neuron systems. complex. J. Neurochem. 46:1272-1276. Anatomy and physiology of the norepinephrine and epinephrine systems. Janowsky, A.J., M.M. Schweri, F. Berger, R. Long, P. Skolmick, and S.M. Ann. Rev. Neurosci. 2:113-168. Paul (1985) The effects of surgical and chemical lesions on striatalL3Hl Moore, R.Y., and J.P. Card (1984) Noradrenaline containing neuron systhreo-(+)-methylpheridate binding: Correlation with [3Hl dopamine tems. In A. Bjorklund and T. Hokfelt (eds): Handbook of Chemical uptake. Europ. J. Pharmacol. 108:187-191. Neuroanatomy. Elsevier, pp. 123-153. Javitch, J.A., R.O. Blaustein, and S.H. Snyder (1983) PHI Mazindol binding Ohm, J., and J.W. Braak (1987) The pigmented subpeduneular nucleus associated with neuronal dopamine uptake sites in corpus striatum (nucleus subpeduncularis pigrnentosus) in the human brain. Normal membranes. Eur. J. Pharmacol. 90:461--162. morphology and pathological changes in Alzheimer’s disease. Neurosci. Javitch, J.A., R.O. Blaustein, and S.H. Snyder (1984) L3H]Mazindol binding 22:S705. associated with neuronal dopamine and norepinephrine uptake sites. Paxinos, G., and C. Watson (1986) The Rat Brain in Stereotaxic Coordinates, Molec. Pharmacol. 26:3544. 2nd ed. Sydney: Academic Press. Javitch, J.A., S.M. Strittmatter, and S.H. Snyder (1985a) Differential Paxinos, G.,I. Tork, G. Halliday, and W.R. Mehler (1990) Human homolgues visualization of dopamine and norepinephrine uptake sites in rat brain to brainstem nuclei identified in other animals as revealed by AChE using L3Hl mazindol autoradiography. J. Neurosci. 5r1513-1521. activity. In: G. Paxinos (ed): The Human Nervous System. San Diego: Javitch, J.A., R.J. D’Amato, S.M. Strittmatter, and S.H. Snyder (1985b) Academic Press, pp. 149-202. Parkinsonism-inducing neurotoxin, N-methyl-4-phenyl-l,2,3,6-tetrahyPearson, J., M. Goldstin, and L. Brandeis (1979) Tyrosine hydroxylase dropyridine. Uptake of the metabolite N-methyl-4-phenylpyridine by immunohistochemistry in human brain. Brain Res. 165:333-337. dopamine neurons explains selective toxicity. Proc. Natl. Acad. Sci. USA Pearson, J., M. Goldstein, K. Markey, and L. Brandeis (1983) Human brain 822173-2177. stem catecholamine neuronal anatomy as indicated by immunohistochemJoyce, J.N., N. Lexow, B. Neal, H. Hurtig, J.Q. Trojanowski, and A. Winokur istry with antibodies to tyrosine hydroxylase. Neurosci. 8:3-32. (1989) Receptor autoradiographic studies in neurodegenerative disorders of the basal ganglia. In A.R. Crossman and M.A. Sambrook (eds): Pearson, J.M., G. Halliday, N. Sakamoto, and J.P. Michell (1990) Catecholaminergic neurons. In G. Paxinos (ed): The Human Nervous System. Neural Mechanisms in Disorders of Movement. John Libbey, pp. 327336. San Diego: Academic Press, pp. 1023-1094. Pickel, V.M., L.A. Specht, K.K. Sumal, T.H. Joh, D.J. Rise, and A. Hervonen Kemper, C.M., D.T. O’Connor, andK.N. Westlund (1987) Immunocytochemical localization of dopamine-hydroxyIasein neurons of the human brain (1980) Immunocytochemical localisation of tyrosine hydroxylase in the stem. Neuroscience 23:981-989. human foetal nervous system. J. Comp. Neurol. 194:465-474. Kennedy, L.T., and I. Hanbauer (1983) Sodium-sensitive cocaine binding to Poirer, L.J., M. Giguere, and R. Marchard (1983) Comparative morphology rat striatal membrane: Possible relationship to dopamine uptake sites. J. of the substantia nigra and the ventral tegmental area in the money, cat Neurochem. 41:3544. and rat. Brain Res. Bull. 11:371-397. Kish, S.J., K. Skannak, and 0. Hornykiewicz (1988) Uneven pattern of Rainbow, T.C., A. Biegou, and D.J. Berk (1984) Quantitative autoradiogradopamine loss in the striatum of patients with idiopathic Parkinson’s phy with tritium labelled ligands: comparison of biochemical and densiDisease. N. Engl. J. Med. 318:876-880. tometric measurement. J. Neurosci. Methods 11.231-242. Kitahama, K., L. Denoroyl, M. Goldstein, M. Jouvet, and J. Pearson (1988) Scatton, B., A. Dubois, M.L. Dubocovich, N.R. Zahniser, and D. Fage (1985) Immunohistochemistry of tyrosine hydroxylase and phenylethanolQuantitive autoradiography of [3Hl nomifensine binding sites in rat amine N-methyltransferase in the human brainstem: Description of brain. Life Sci. 36:815-822. adrenergic perikarya and characterization of longitudinal catecholaminSchweri, M.M., P. Skolmick, M.F. M e r b y , K.C. Rice, A.J. Janowski, and ergic pathways. Neuroscience 25t97-111. S.M. Paul (1985) [3Hl Threo-(+)-methylphenydate binding to 3,4 dihyKitahama, K., J. Pearson, L. Denoroy, N. Kopp, J. Ulrich, T. Maeda, and M. droxyphenlethylamine uptake sites in corpus striatum: Correlation with Jouvet (1985) Adrenergic neurons in the human brain as demonstrated the stimulant properties of vitalinic acid esters. J. Neurochem. 45:1062by immunocytochemistry with antibodies to phenylethanolamine N1070. methyl transferase (PNMT): Discovery of a new group in the nucleus Tedroff, J., S.M. Aquilonius, P. Hartrig, H. Lundquist, A.G. Gee, J. Uhlin, tractus solitarius. Neurosci. Lett. 53:303-308. and B. Langstrom (1988) Monoamine re-uptake sites in the human brain Kopp, N., L. Denoroy, B. Renaud, J.F. Pujol, A. Tabib, and M. Tommasi evaluated in vivo by means of ”C nomifensine and positron emission (1979) Distribution of adrenaline-synthesizing enzyme activity in the tomography: The effects of age and Parkinson’s disease. Acta Neurol. human brain. J. Neurol. Sci. 41:397-409. Scand. 77:192-201. Kuhar, M.J., and J.R. Unnerstall(1985) Quantitative autoradiography with Tork, I., and J.P. Hornung (1990) Raphe nuclei and serotonergic systems. In tritium labelled ligands: comparison of biochemical and densitometric G. Paxinos (ed):The Human Nervous System. San Diego: Academic measurement. J. Neurosci. Methods 11:231-242. Press, pp. 221-260. Langer S.Z., E. Zasrifian, M. Briley, R. Raisman, and D. Sechter (1981) [3H] Tork, I. (1985) Raphe nuclei and serotonin containing systems. In G. Desipramine binding is associated with neuronal noradrenaline uptake Paxinos (ed): The Rat Nervous System. Volume 2. Academic Press, in the peripheral and central nervous system. European J. Pharmacol Australia, pp. 43-78. 72t423-430. Van der Zee P., H.S. Koger, J. Goofjes, and W. Hespe (1980) Aryl 1, Loughlin, S.E., and J.H. Fallon (1985) Locus coeruleus. In: G. Paxinos (ed): 4-dialk(en)ylpiperazines as selective and very potent inhibitors of dopaThe Rat Nervous System, Vol. 2. San Diego: Academic Press, pp. 79-89. mine uptake. Eur. J. Med. Chem. 15363-370.

Distribution of catecholamine uptake sites in human brain as determined by quantitative [3H] mazindol autoradiography.

Because of the importance of the catecholamine system in Parkinson's disease and its relevance to a variety of clinical movement disorders, catecholam...
8MB Sizes 0 Downloads 0 Views