Epinephrine distribution in human brain

~reuroscience Letters, 9 (1978) 227--231 ~ Elsevier/North,Holland Scientific Publishers Ltd.

227

EPINEPHRINE DISTRIBlYrlON IN HUMAN BRAIN

I. MEFFORD, A. OKE, R. KELLER, R.N. ADAMS and G. JONSSON* D~;partment of Chemistry, University of Kansas, Lawrence, Kans. 66045 (U.S.A.) and *1~rtrn~.nt o f Histology, Karolinska lnstitutet, S-104 Ol Stockholm 60 (Sweden)

(Received April 3rd,I978) ( R e ~ d r e . o n ~ i ~ d May 5th, 1978) (Accepted May 8th, 1978)

SUMMARY

The distributionof epinephrine (E) in human brain has been analyzed by a pr,~viouslydescribed [11 ] liquidchromatographic method. E, though detectable in most areas sampled, was found to be highly localizedin those areas pr,~viouslyknown to contain highest norepinephrine (NE) concentrations [4]. The regions found to be highest in E concentration were the ventromediM, dcrsomedial, supraoptic and paraventriculaz nuclei of the hypothalamus.

Phenylethanolamine,N-methyltransferase (PNMT), the enzyme which converts N E to E, has often been used as a marker for the presence of E in mammalian and human brain [6,9,13,14,16]. The presence and localizationof E itselfhave also been demonstrated in mamm~dian brains [~.,5,8,12,17,18]. W e recently reported the detailed distribution of E in human brainstem [11]. Although immunofluorescence techniques have shown that P N M T is present in nerve terrains and neurons in specific nuclei of the rat brain, and that these neurons appear morpholo~cally similar to N E neurons [6], it has not yet been proven that E acts as a neurotransmitter in mammalian CNS. Several possible C N S functions orE have been suggested [6,10,15] and.among the~e isa role in blood pressure relation [3,6]. The existing data on human brains tend ~ support the5hypothesis since both P N M T and E are highly localized in ~ e region of the nucleus of the solitarytract [9,11 ]. As a step to further elucidate the functional and beha'~ioral~mplications in hummls, more information on the l o ~ ~ t i o n of E beyond the brainstem region is needed. This repot, together with previous data, provides an overall, reasonably det~dled picture of the dis~bution o:~Ein human brain. Five human b ~ n s were analyzed i n ~ s study, All brains were obt~dned from r o u ~ e au~psy specimens. The clinicalhistoriesindicated no abnormal psychoses or neurological diso~em, nor did they indicate any hypertensive dise~e, as is shown in Table I.

228

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4

TABLE I DATA FOR AUTOPSY BRAIN SPECIMENS .

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.

,.

,

,,

.

.

.

.

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.

.

w

Autopsy specimen number

Sex

Age

Lengthof time from death to autopsy

Cause of death ~....

1

Male

52

12.5 h

2 3 4 5

Male Male Male Male

58 70 60 63

8.75 h 10.5 h 18 h 9.75 h

T e ~ n a l liver c ~ i n o m a compliea~ byemphysenm and chronic bronc~tb L u ~ careinO~ ' Respira~ imp~nt

:

Myocardial infarctiOn Pulmor~7 fsUure, massive pulmonary embolism ,

,,

i

_ _

|,,,

i

i,

The elapsed time between death and autopsy varied from 10--20 h. U p o n removal the brains were immediately frozen at -70°C until sectioning~and punching could be performed. At the time of sectioning, brains ~ r e sliced coronally in approx 2 mm dices while still frozen, ar~;J the slices were placed on glass plates over dry ice. The coronal slices were then photographed and punching was performed in much the same manner as previously described [ 11]. 2 0 - 4 0 mg tissue punches were taken for most regions, using a hollow cylindrical punch approx 3 - 5 mm in diameter. For more discrete punching (i.e., supraoptic nuclei), a sharpened 17 gauge needle was used. Immediately after punching, the coronal slice was re-photographed and exact neuroanatomical locations were confirmed for each punch using a standard human brain atlas [ 2 ]. Tissue punches were immediately returned to the -70°C environment until analyzed. Analysis of catecholamines was performed by the liquid chromatographic method previously described [7,11]. Frozen tissue samples were weighed into a 1.5 ml conical centrifuge tube and homogenized in 250 ~1 of 0.1 M HCIO4 plus internal standard (~ 1 X 10 °9 mole ~-methyI dopamine, MSD). To this ~s added 10 mg A1203 and the pH raised to 8.6 ~ t h 0.5 M Tris buffer. Catecholamim,~ are desorbed into 50 #1 of 0.1 M HCIO4 and aportion of this is injected int~ the chromatographic system. The catecholamines are then separated on the ion exchange column and electrochemicldly oxidized as they pass the carbon paste detector electrode. The oxidation current thus obtained is proportional to the amount of amine injected. The observed trend in E distribution is shown by Fig. 1. We wish to emphasize that the distribution patterns in all 5,brains were remarkably similar, although considerable variation in absolute content was f o ~ d b e t ~ e n brains. Table H is a summary of the data from the brains a ~ y z e d , The individual values for a given region or subnucleus are the mean values in ng/g t ~ u e (wet wt.). (The number of punches contributing to a pa~ictflar m e ~ ~ u e is given in parenth~ses.) The punching did not necessarily sample ~ e n t r e structure

229 A

B

H

C

I

D

J:

"

f'

[

K

L

G

"M

Fig. 1. Relative distribution o f E in h u m a n brain. Slices A--M represent serial coronal slices from human brain, from the level! of the anterior caudate (slice A) •through the level of the posterior pulvinar in the thalamus and occulomotor region of the brainstem (slice M). Density shading indicates relative values of E concentration. Absolute E values can be found for each region in Table I.

and, in some instances, punches were taken in a random fashion throughout a structure. Nevertheless, the mean values can be considered representative of each region named. As Table K shows, there is a wide variation in the regional E content among the 5 brains. There appears to be no obvious correlation of E concentration with age or other data from the clinical histories, nor does there appear to be any regular decrease in E content with the time interval between death and autopsy (Table I). The E concentrations in all regions of brain 5 (and to a lesser extent in brain 4) are seriously out of line with those in brains 11--3. We were, however, unable to con'elate rids with any of the obvious clinical data. As is demonstrated by Fig. 1, E was found to be concentrated in the hypothalamus. As one proceeds latelmlly from the hypothalamus, the concentration of E decreases rapidly. High levels are also found around the third ventricle and in the septal region. E levels are relatively l~xge in the more medial portions of the thalamus. As one proceeds to the level of the red nucleus ~ld occulomotor complex, the E concentrations decrease and approach the levels previously reported for brainstem [ 11 ]. Very low concentrations of E are found in the hippocampus. Similarly, levels are very low in the neocortex - e v e n below detectable limits (~ 0.2 ng/g for 50 mg of tissue) for several cortical regions. (Cortex was sampled somewhat randomly and not in terms of specific areas.) Generally speaking, braiin areas rich in d o ~ d n e (caudate, putamen) were found to have very low or non-detectable a~,ount of E. T h ~ e data, t~)gether wit,h those reported previously [ 11 ], Show E

. -

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,

ND ND 1.0 0.3 ± 15.1 ± 40.2 -~ 14.4 z 2.4± 1.4 ~ 6.9 *11.9± ND ~3.0± .;.2 * 13.5 + 0.~ *-

.

(!4) ~3) (I) 0.3 (3) 9.7 (4) 8.0 (3) 0.8 (3) 0.7 (2) 0.2 (4) 2.0 (6) 0.7 (2) (13) 1.8(15) 0.7 (2) 5.9 (2) 0.1 (29)

± 4.2 (2) ± 6.0 (2) ± 25,6 (~) ± 30.8 ( 2 ) -+ 0.2 (2) 5 6 . 6 ± 22.4 (2) "8.6 ± 1.9 (2)

88.3 20.5 69.~ 118,5 8.0

139.5 ~- 7.6 (2) a 115.5 ± 14.6 (2)

± 1.7 _+ 38.2 +- 19.9 ± 3.1 ± 11.5 ± 1.8

(2) (4) (4) (6) (2) (2)

3.0 (1) 9.6 ± 6.3 (2) 15.3 ~- 2.7 (2) 0.3 ± 0 . 0 ( 2 ) 5.4 (1) 3.9 ± 1.4 (2) 12.4 (1)

10.7 -~ .0.A (2) 113.0 -* 31.8 (5) 41.5 ± 30.2 (2)

1.5 (1) 25.4 ± 13.5 (3) 4.0 ± 1.6 (5) 10.7 ± 5.4 (4)

56.6 247.0 108.0 i0.2 36.5 6.5

332.0 ± 3.5 (2) 239.0 *- 11.5 (4) 284.0 ± 13.6 (3)

Brain 2

(1)

0.? 0.4

1.1 ± 0.4 (2)

1.3± 0 . 6 ( 2 ) 17.3 (1) 11.9 ± 2.7 ( 5 )

7.0 +- 0 . 0 ( 2 ) 8.6 +- 0.6 (2)

50.~

(1)

~1) (1) (1)

0.2

0.6

12.1 ± 3.7 (4) 7.2 ± 1.1 (4) 3.8 ± 0.7 (4)

141.3 ± 27.~ (2) 123.6 ± 8.1 (2) 141.1 ± 5.7 (4) 113.7 18.7 ± 4.1 (2) 180.8 ~ 27.1 (2) 60.9 ± 15.2 (4) 12.4 ± (1) 26.5-+ 4 . 8 ( 2 )

Brain 3

(2) (5) (2) (2)

40.2 ± 12.6 6.9± 2.2 14.4 ± 4.3 2.0 ± 0.3

1.3 -* 4.5 1.2 2.6 ± 1.2± 3.5£

0.5 (2) 0.1 (4) 0.7(2)

(1)

1.3 (2) (1)

$7.4 ÷ 2.2 (3)

5.3 ± 1.3 (8) 1.6± 0.4 (8) 4.1 ± 1.6 ( 4 )

(4) (4) (4) (2) (4)

± ± ± ± ±

3.6 2.0 5.3 0.2 2.7

44.8 34.2 31.6 43.4 10.2

Brain 4

(4) (2)

(2)

(2)

(4)

(3)

,~D

ND

(21)

0.7 - 0.4 (41)

1,5 -* 1.1 (4) 7.9 -+ 1.5 (2) 6.4 (1)

(12) (11) (1)

ND ND ND

1.5-+ 1.5 (2)

8.6 ± 0.4 6.2 -+ 1.1 13.9 ± 2.3 ND

15.6 ± 0.8

19.9 ± 1.6 (4)

Brain 5

a ~ h value is the mean in ng/g ± SEM for the region named; the number in parentheses indicates the number of individual punches of r e , o n analysed. ND = not detectable.

_

~uedu~Gray H~,p~ampus Amygdala Subth~us Occulomotor N. N¢ocortex

SuperiorCoIEculus

Hypothaiamus Ventromedial N. Dorsomedial N. Paraventricular N, N. Tuber Cinereum Lateral Posterior N. Supraoptic N. Pmoptie area Olfactory area Posterior N MammCiary bodies Thalamic Nuclei Anterior N. Dorsomediai N. Ventrolaterai N. Pulvinar Caudate Putamen Globus Pailidus I Globus Pallidus H N. Accumbens Sept.! Nuclei SeptumPellucidum Re d N ~ e i Substantia Nigra

Brain 1

REGIONAL CONCENTRATIONS OF EPINEPHRINE IN 5 HUMAN BRAINS (ng/g)

TABLE I! O

tO

231

to be highly localized in the human CNS. This high degree of localization in the 'central core' is suggestive of many possible CNS functions which, however, remain to be defined. REFERENCES 1 Bertler, ~., Carlsson, A. and Rosengren, E., A method for the fluorimetric determination of adrenaline and nol adrenaline in tissues, Acta physiol, stand., 44 (1958) 273-292. 2 De Armond, S.J., Fusco, M.M. and Dewey, M.M., Structure of the Human Brain, A Photographic Atlas, New York: Oxford University Press, 1974. 3 De Jong, W., Zandberg, ~'., Versteeg, D.H.G. and Palkovits, M., Brainstem structu~'es and catecholamines in the control of arterial blood pressure in the rat, Clin. Sci. Mol~. Med., 51 (1976) 381s---3~.~s. 4 Farley, I.J. and Hornykie~.dcz, O., Noradrenaline distribution in subcortical areas of the human brain, Brain Res., ~26 (1977) 53--62. 5 Gunne, L.M., Relative adrenaline content in brain tissue, Acta physiol, scand., 56 (1962) 324--333. 6 HOkfell~,T., Fuxe, K., Goldstein, M. and Johannson, O., Immunohistochemical evidence for the existence of adrenaline nel~rons in the rat brain, Brain Res., 661 (1974) 235--.751. 7 Keller, R., Oke, A., Mefford, I. and Adams, R.N., Liquid chromatographic analysis of catechoiamines --routine assay for regional brain mapping, Life Sci., 19 (1976) 9.95-1004. 8 Koslow, S.H. and Schlumpf, M., Quantitation of adrenaline in rat brain nuclei and areas by mare frsgmentometry, Nature (Lond.), 251 (1975) 530--531. 9 Lew, J.Y., Matsumoto, Y., Pearson, J., Goldstein, M., HOkfelt, T. and Fuxe, K°, Localization and characterization of phenylethanolamine-N-methyltransferase in the brain of various mammalian species, Brain Res., 119 (1977) 199--210. 10 Marley, E. and Stephenson, J.D., Central actions of catecholamines. In H. Blaschk~ and ~ Muscholl (Eds.), Catecholamines, Springer Vedag, Redin, 1972, pp. 463--537. 11 Mefford, I., Oke, Ao, Adams, R.N. and Jonsson, G., Epinephrine local~r.ation in human bndnstem, Neurosci. Lett., 5 (1977) 141--145. 12 Montagu, K.A., ~.drenaline and noradronaline concentrations in rat tis~mes, Bie~hem. J., 63 (1956) 559--56-. 13 Pahor~ky, L.A., Zigmond, M., Karten, H. and Wurtman, R.J., Enzymatic conversion of norepinephrine to epinephrine by ~he brain, J. Pharmacol. exp. Ther,, 165 (1969) 190--195. 14 Rei,d, J.L., Zivin, J.A., Foppen, F., H and Kopin, I.J., Catecholamine neurotransmitters and synthetic enzymes in the spinal cord of rat, Life Sci., 16 (1975) 975--984. 15 Ro~hballer, A.B., The effects of catecholamines on the central nervous system, Pharmacol. Rov., 11 (1959) 494--547. 16 Saavedra, J.M., Grobecker, H. and Axelrod, J., Adrenaline-forming enzyme in brainstem: Elevation in genetic and experimental hypertension, Science, 191 (1976) 483-48[~. 17 Vml der Gugten, J., Palkovits, M., Wijnen, H.L.J.M. and Versteeg, D.H.G., P~:gional distribution of adrenaline in rat brain, Brain Res., 107 (1976) 171--175. 18 Vogt, M., The concentration of sympathin in different parts of the nervous system under normal conditions and after the administration of drugs, J. Physiol. (Lond,), 123 (1954) 451--481.