53 1st A4 EETING , LAN CASTE,R
111
As one-half of the total body Mg2+is deposited in bones, the proposed mechanism
for the Li+-induced changes in Ca2+turnover might also be relevant for MgZ+.Fig. 1
shows that bone radioactivity was decreased after injection of isotopically labelled Mg2+
together with Li+, whereas serum radioactivity was increased; the results were thus
qualitatively similar to those found with the radioactive Caz+.In accordance with this
finding it has been reported that Li+-treated manic patients had an increased serum
Mg2+concentration (Nielsen, 1964).
The intimate relationship between glucose, phosphate, Caz+ and Mg2+metabolism
may therefore constitute the basis for the effects of Li+ on each of the four metabolites,
enzymes in glucose metabolism being the direct sites of action. However, such a concept
implies that other drugs or hormones that influence overall carbohydrate metabolism
in the same way as Li+ also may produce similar effects in phosphate, Ca2+and Mg2+
metabolism.
The most obvious choice in this respect was insulin, which increases glucose uptake
and glycogen synthesis, decreases serum phosphate concentration (Harrop & Benedict,
1924) and increases serum Ca2+concentration (Brougher, 1927). By using radioactive
phosphate, Ca2+ or Mg2+ it was possible to show that many similar results were
obtained with insulin and Li+.
Balan, G., Cernitescu, D., Trandafirescu, M. & Ababei, L. (1970)Proc. Coii'gr. Collegiwii
Internationale Neuro-Psjc~ioyharniacolo~rii~in
7th 19
Bhattacharya, G. (1959) Nature (London) 183, 324325
Bhattacharya, G. (1964) Biochinz. Biophys. Acta 93, 644
Brougher, J. C. (1927) Amer. J . Physiol. 80, 41 1-415
Harrop, G. A. & Benedict, E. M. (1924) J . Biol. Chem. 59, 683-697
Kachmar, J. F. & Boyer, P. D. (1953) J . Bid. Chem. 200, 669-682
Mellerup, E. T., Plengc, P., Ziegler, R. & Rafaelson, 0. J. (1970) Int. Plirtrinacof~s,~c/ii~it.
5,
258-264
Mellerup, E. T., Thonisen, H. G., Bjmum, N. & Rafaelsen, 0. J. (1972) Acra Psychiut. S c a d
in the press
Nielsen, J. (1964) Acta Psjchiat. Scotid. 40, 19C196
Plenge, P., Mellerup, E. T. & Rafaelsen, 0. J. (1970)J. Psjchiut. Res. 8, 29--36
Plenge, P., Mellerup, E. T. & Rafaelsen, 0. J. (1971) Int. Pharmacopsychiot. 6 , 52-59
Walaas, E., Walaas, 0. & Horn, R. S . (1970) Mech. Insulin Actiorz Synip. 93-98
Effects of Lithium Ions in a Pharmacological Concentration on
Potassium and Sodium Ions in Rat Brain-Cortex Slices
CHRISTEL S. KJELDSEN, HENRIK LUND-ANDERSEN and LEIF HERTZ
Biokeniisk Itistitiit A, Kabeiihacris Utiioeusitet, 30 Jiilinnc Marieswj,
2 100 Kaberilic~r~ti
0,Dcnnmrii
The Li+ ion is placed in the Periodic Tablc near the biologically important ions Na'
and K+. Li+ is therefore used as an important tool in investigations on Na' and K +
metabolism in nervous tissue (Hodgkin & Katz, 1949; Pappius et d.,
1958; Keynes &
Swan, 1959; Hertz & Schou, 1962; Bourke & Tower, 1966); relatively high concentrations of Li+ have been used in these studies. In addition, Li+ plays an important role in
psychopharniacology in the therapy of manic-depressive disease (Cade, 1949; Schou,
1957; Baastrup & Schou, 1967), and interest has therefore also been directed towards
~ ) brain metabolism
the effect of Li+ in pharmacological concentrations ( ~ 2 n 1 on
1966; Schou, 1969; Baer et a / . , 1970).The present study
(Davenport, 1950; Israel ct d.,
iir uiWo deals with the eff'ects of I .9iii~-LiCIon the K ',N a ' , Li ' and Cl- concentrations
in rat brain-cortex slices.
VOI.
1
I12
BIOCHEMICAL SOCIETY TRANSACTIONS
Table 1. Cotzcetitratioiis of K I, Nu+, Cl arid Li+ and coritetits of solids in rat brain-cortex
slices incirhated for 111 icnder vigorous oxygenation iti a ‘balanced’ medium or in corresponding media containing 2mM-Lif, -Rb+ or Cs’
Results are given as means&s.e.ki. for four to 17 experiments.
‘Balanced’
Li+ (1.9mM) Rb+ (2.0111~) Cs+ ( 2 . 0 m ~ )
medium
medium
medium
medium
KC(pmollg final
69.5 f0.7
61.3k 1.2
65.4 k 1.9
54.6 d: 2.4
wet wt.)
Na+ (pmol/g final
87.7 k 1.7
8 4 S k 1.1
83.4+ 5.0
86.1 k 2.9
wet wt.)
CI- (pmol/g final
62.7 k 2.2
58.2k 1.7
56.6zt 3.3
61.4 :t 2.1
wet wt.)
Li+ (pmol/g final
3.6 k 0.I
_wet wt.)
Solids (% of final
15.7k0.13
15.8 kO.1
16.0 k 0. I
16.0 k 0.2
wet wt.)
Adult Wistar rats were decapitated without previous anaesthesia. One slice of
0.5 mm thickness was cut from the lateral surface of each hemisphere as described
by Franck et al. (1968). Within 2min after decapitation of the animal each slice was
placed in a test tube containing 4ml of medium equilibrated with 0 2 + C 0 2 (95:5) at
37°C; the aeration was made sufficiently vigorous for the brain slices to be floating
freely in the medium (Lund-Andersen & Hertz, 1970; Arnfred et al., 1970). After
60min incubation the slices were drained on a Petri dish to remove adherent fluid,
weighed, dried overnight at 115°C and reweighed. Ion concentrations were determined
as described by Schousboe & Hertz (1971).
The ‘balanced’ medium contained (final concentrations) 115 m ~ - N a C l ,2 0 m ~ NaHC03, 5 m ~ - K C l 1.5m~-CaC1,,
,
1 .OmM-MgSO, and 6m~-glucose.The K’~-,Li+-,
Rb+- and Cs+-enriched media were obtained by the addition of KCI, LiCI, RbCl or
CsCl to the ‘balanced’ medium.
Table 1 shows that the presence of l.9ni~-LiCIit1 the ‘balanced’ medium causes a
significant decrease of 8.2pmol/g final wet wt. in the K + concentration, and seems to
cause a slight decline of the Na+ concentration. The Li+ concentration in the tissue
(3.6pmol/g final wet wt.) is much lower than the decrease in the K+ concentration, and
the C1- concentration is slightly decreased. The content of solids (and thus the swelling)
is unaffected. No corresponding phenomena were observed after incubation in Warburg
manometers (results not presented).
The Li+-induced decrease in the K+ concentration is counteracted by increasing the
K+ concentration of the medium. This is indicated in the lower part of Fig. 1, which
shows that the difference in K+ concentration between slices incubated in the Li ‘-free
medium and in the Li+-containing medium is gradually decreased to near zero as the
K+ concentration in the medium is increased to 8 5 m M ; concomitantly the Li’ conce1:tration in the tissues decreases from 3.6 k 0.07 to 1.9 -I- 0.1 1 ptnol/g final wet wt. and the
decrease in the C1- concentration is abolished (results net presented). The tendency
towards a decrease of the Na+ concentration is similarly counteracted by very high concentrations of K+ (upper part of Fig. I ) , but the Li+-induced decrease in the Na+ concentration is especially pronounced (and statistically significant) at a K+ concentration
of about 1 5 m in
~ the medium. The ‘segmentation’ of the curve showing the Na+ concentration in the tissue as a function of the K+ concentration in the medium (LundAndersen & Hertz, 1970) is thus, if anything, accentuated in the presence of I?.
The possibility of competition between Li+ and Na+ was studied in experiments i n
which the NaC concentration of the medium was varied between 78 and 2 5 0 n i ~(results
not presented): in the absence of Li’~even a deviation of the Na’ concentration from
1973
I13
531st MEETING, LANCASTER
0
10
20
30
40
50
60
70
80
90
External concn. of K+ (mM)
Fig. I . Coticoitrations of Nu+ (a and A) and K+ ( o arid 0 ) in rat brain-cortex slices as a
jiitictiori of the external K+ concentratiotz after 1 h of incubation iii un otherwise ‘balairced’
oxygetiated merliiiin (,I atid 0 ) or it1 a ‘balancetl’oxj~etiatc~d
medium to which 1.9tnM-LiCI
l i d beeti nrlded (A and e)
Results are given as means of five to 17 experiments, with S.E.M. values indicated by
vertica I ha r9
the ‘balanced’ 1 1 5 m ~to 100 or 1 3 0 m ~was found to cause a slight fall in the K+
concentration, and the addition of Li+ caused no or only very little further decrease.
The observed decrease in the Kf concentration induced by Li+ is not specific for this
ion, since a similar effect is observed in the presence of Rb+ or Cs+ (Table 1). The
effect exerted by Rb+ seems to be less pronounced than that evoked by Li+, but Cs’
causes an even larger fall in the K+ concentration. The concentrations of Na’ and of
CI- and the swelling are largely unaffected, however.
The present report has demonstrated that a pharmacological concentration of Lit
causes decreases of the concentrations of K+, Na+ and C1- in brain-cortex sliccs. The
concentration of Li+ in the tissue is so low that the decrease of the K+ and Na+ concentrations cannot be quantitatively explained by replacement with Li’. That the
slice/niedium Li’ concentration ratio, on the other hand, exceeds 1 .O is in accordance
with findings obtained in oioo by Schou (1958).
The decrease in the Na+ concentration is in agrecment with the observations made
in ciuo by Davenport (1950) and Baer et al. (1970), and might be due to activation of an
Na+-t K+-stimulated adenosine triphosphatase at the K+-sensitive site (Skou, 1960;
Whittam & Ager, 19641, as also indicated by the observed competition between Lit
and K+ and the lack of competition between Li+ and Na+. The decrease in the K+
Vol. 1
114
HIOCHEMrCAL SOCIETY TKANSACTlONS
concentration confirms results obtained by Israel et al. (1966), who suggested that the
decline might be due to an Li+-induced inhibition of the Na++K+-stimulated adenosine
triphosphatase. Such an inhibition is not, however, in accordance with the observed
effect on Na'. In addition to the activation of the Na+ tK+-stimulated adenosine triphosphatase Li' might therefore conceivably exert a specific inhibition of the linked
uptake of K' and C1-, which previously has been suggested in the brain cortex
(Bourkc, 1969; Lund-Andersen & Hertz, 1970; Hertz, 1973). The observed decline in
the CI- concentration supports this point of view. Cs+ and Rb' might possibly exert
a similar effect.
Arnfred,T.,Hertz, L.,Lolle, L. &Lund-Andersen,H.(1970)Esp. Brai/iRes.(Bcrliii) 11,373-375
Baastrup, P. C. & Schou, M. (1967) Arch. Gen. Psjvhiat. 16, 162-172
Baer, L.,Kassir, S. & Fieve, R. (1970) Psyc//op/zurniaco/ogiu17,216-224
Bourke, R. S. (1969) Exp. Brain Res. (Berlin) 8,232-248
Bourke, R. S. & Tower, D. B. (1966) J. Neurochem. 13, 1099-1117
Cade, J. F. J. (1949) Med. J . Aust. 36, 349-352
Davenport, V. D.(1950) Amer. J . Physiol. 161, 633-641
Franck, G.,Cornette, M. & Schoffeniels, E. (1968) J. Newochir. 15, 843-857
Hertz, L. (1973) Biochem. Soc. Tram. 1, 115-118
Hertz, L. & Schou, M. (1962) Biochem. J . 85,93-104
Hodgkin, A. L. & Katz, B. (1949) J . Physiol. (London) 108, 37-71
Israel, Y.,Kalant, H. & LeBlanc, A. E. (1966) Biochem. J. 100, 27-33
Keynes, R.D. & Swan, R. C.(1959) J . Physiol. 147, 626-638
Lund-Andersen, H. & Hertz, L. (1970) ESP. Brain Res. (Berlin) 11, 199-212
Pappius, 11. M.,Rosenfeld, M., Johnson, D. M. & Elliott, K. A. C . (1958) Ctrri. J . Biuchem.
Physiol. 36,2 17-226
Schou, M. (1957) Pharmacol. Rev. 9,17-58
Schou, M.(1958) Acra Pharmacol. Tosirol. 15, 115-124
Schou, M. (1969) P.~J'chophflrti?flcol.
Bull. 5, 33-62
Schousboe, A . & Hertz, L.(1971) Inf. J . Neurosci. I, 235-242
Skou, J. C.(1960) Biochim. Biophys. Acta 42,6-23
Whittam, R. & Ager, M. E. (1964) Biochem. J . 93,337-348
Erythrocyte Sodium and Potassium Ions in Mania and Depression
A. I. M. G L E N and LESLEY BELLINGEK
Metliccil Resecircli Coiirici[ Brait! Metabolism Uriif, Utiircrsity Depyrrrhiwrt of
Phnrmacoloxy, 1 George Sqirare, Edinbuvgh EN8 9JZ, U.K.
It has been suggested that there is an increase in intracellular Na' during episodes of
manic-depressive illness (Coppen & Shaw, 1963; Coppen et a/., 1966). Whole-body
studies utilizing isotope-exchange techniques have, however, not been cntirely satisfactory, and the results have often been conflicting. In view of this it was thought that
studies of cation movements across specific membranes would prove useful. Changes
in Na+ activity in the parotid saliva of manic-depressive patients, detected by using
ion-specific electrodes for measurement, have previotisly been reported (Glen et ul.,
1968, 1969). These investigations suggested that there was evidence of impaired N a +
transport. Naylor et al. (l970), using the erythrocyte as a model, observed an increase
in erythrocyte Naf concentration in certain depressed patients. Maizels (1968) and
Bradbury & Glen (1972) have shown that Li+ can stimulate Na+ efflux from the
erythrocyte. The model lends itself t o further investigation.
The present experiments comprise an investigation of erythrocyte Nn+ and K+ concentrations i n manic--depressivepatients and in age-matched controls. The patients were
under investigation in a metabolic ivard and receiving ;i standardized diet. Sequential
observations were made when patients were without medication during the ill phase and
1973