Brain Research, 235 (1982) 233-243 233

Elsevier Biomedical Press

EVIDENCE THAT SEROTONERGIC NEURONS IN THE DORSAL RAPHE

N U C L E U S E X E R T A S T I M U L A T O R Y E F F E C T O N T H E S E C R E T I O N OF
R E N I N B U T N O T OF C O R T I C O S T E R O N E

LOUIS D. VAN DE KAR*, CHARLES W. WILKINSON, YOANNA SKROBIK, MARK S.

BROWNFIELD and WILLIAM F. GANONG
Department of Physiology, University of California, San Francisco, CA 94143 (U.S.A.)
(Accepted August 5th, 1981)

Key words: serotonin - - renin secretion - - corticosterone secretion - - dorsal raphe nucleus - -
median raphe nucleus

SUMMARY

Data from previous experiments in rats indicate that release of serotonin in the
central nervous system increases renin and corticosterone secretion a~. To determine
which serotonergic neurons are involved, lesions of the dorsal or median raphe nuclei
were produced by local injections of 5,7-dihydroxytryptamine (5,7-DHT) in rats, and 2
weeks later, the renin responses to parachloroamphetamine (PCA) were determined.
Plasma corticosterone was also measured. PCA produced significant increases in
plasma renin activity and plasma corticosterone in sham-lesioned animals and animals
with median raphe lesions. The plasma corticosterone response to PCA was also
normal in rats with dorsal raphe lesions but the renin response was significantly
reduced. The data support the hypothesis that serotonergic neurons in the dorsal
raphe nucleus are part of a neural pathway mediating increased renin secretion, and
that the stimulatory effect of serotonin on corticosterone secretion is mediated by a
different pathway.

INTRODUCTION

The involvement of serotonin in the regulation of renin secretion has been

studied in various species. Epstein and Hamilton 10 reported that in human subjects,
the serotonin antagonist cyproheptadine inhibited the increase in plasma renin activity
produced by furosemide, and Modlinger et al. 20 reported that the serotonin precursor
tryptophan caused an increase in plasma renin activity. Z i m m e r m a n n and Ganong 34
reported that in anesthetized dogs, tryptophan and 5-hydroxytryptophan caused an

* To whom correspondence and reprint requests should be addressed at: Department of Pharma-
cology, Loyola University, Stritch School of Medicine, 2160 S. First Avenue, Maywood, IL 60153,
U.S.A.

0006-8993/82/0000--0000/$02.75 © Elsevier Biomedical Press

234

increase in plasma renin activity that was central in origin and blocked by the
serotonin antagonist metergoline.
Recently, we administered the serotonin agonist quipazine and the serotonin
releasing agent p-chloroamphetamine (PCA) to conscious rats and observed dose-
dependent increases in plasma renin activityaL The effect of PCA on renin was
abolished by pretreatment with the serotonin synthesis inhibitor p-chlorophenyl-
alanine, suggesting that serotonin stimulates renin secretion. The effect appeared to be
central because intraventricular injection of 5,7-dihydroxytryptamine (5,7-DHT, a
neurotoxin which destroys serotonergic neurons 3) lowered plasma renin activity.
The serotonergic system in the brain can be roughly divided into ascending and
descending pathways originating in the brain stem 9. The ascending serotonergic
pathways originate mainly in the dorsal and median raphe nuclei in the midbrain 2 and
send nonoverlapping projections to various foreblain areas 12,13,17,a°,31.
The present studies were designed to localize the serotonergic neurons that
mediate the stimulatory influence on renin secretion. For this purpose, discrete
chemical lesions were produced in the dorsal or median raphe nuclei by stereotaxic
intracerebral microinjections of 5,7-DHT after pletreatment with the norepinephrine
uptake inhibitor desmethylimipramine 3. The serotonin releaser PCA is was then
administered to the rats to see whether the lesions would inhibit the drug-induced
increase in renin secretion.
In a previous study 32 we observed that PCA increased plasma corticosterone
concentration as well as plasma renin activity, and that pretreatment with the
serotonin synthesis inhibiting drug p-chlorophenylalanine prevented this response as
well as the renin response. Fuller and Snoddy 11 reported similar results with respect to
corticosterone. We therefore measured plasma corticosterone as well as plasma renin
activity in the present study to determine whether the corticosterone response was
mediated by the same or different pathways.

MATERIALS AND METHODS

Male Sprague-Dawley rats weighing 200-250 g were obtained from Simonsen
Laboratories (Gilroy, CA) and housed, 2 per cage, in a temperature controlled room
at 20-22 °C with a 12:12 light-dark cycle (lights on at 07.00 h). Purina Rat Chow
(Deans, Belmont, CA) and water were available ad libitum. 5,7-DHT and PCA were
purchased from Regis (Morton Grove, IL). Desmethylimipramine was a generous
gift from Merrel National Laboratories (Cincinnati, OH).
Discrete intracerebral injection of 5,7-DHT or vehicle into the median or dorsal
raphe nuclei of desmethylimipramine pretreated rats was performed stereotaxically as
described previously 31. The needle angle was 40 ° caudal to the vertical plane and the
dose of 5,7-DHT was 5 #g, injected (at a rate of 0.1 #l/min) over a period of 10 min in
1/zl of a solution containing 0.1 ~ ascorbic acid in 0.9 ~ NaCl. The needle was left in
situ for an additional 10 min to prevent dorsal diffusion. This mode of 5,7-DHT
injection was used because in a previous study 31, it was shown not to affect the
norepinephrine and dopamine content of the hypothalamus yet it caused a relatively
selective lesion of the designated nucleus.
TABLE I
Effect of 5,7-dihydroxytryptamine (5,7-DHT) produced lesions of the dorsal and median raphe nuclei on the serotonin content of the hippocampus and caudate
nucleus
5,7-DHT or vehicle injected 2 weeks before sacrifice. PCA, 10 mg/kg, or saline injected i.p. 2 h before sacrifice.

Injection site Injection Serotonin concentration (ng/g wet weight)

Hippocampus Caudate nucleus
Saline PCA Saline PCA

Dorsal raphe Vehicle 296.6 4- 23.3 134.7 ± 9.9** 675.9 ± 27.3 345.7 q- 18.5"*
Dorsal raphe 5,7-DHT 242.0 4- 38.8 100.7 4- 8.0** 255.3 ± 39.2* 151.0 4- 11.2",**
Median raphe Vehicle 377.9 4- 32.5 200.2 4- 28.7** 380.0 4- 39.2 223.0 ± 30.0**
Median raphe 5,7-DHT 74.5 ± 10.9" 34.8 ± 9.7* 249.8 4- 26.1 * 139.3 4- 7.3*,**
Dorsal and median raphe Vehicle 328.2 4- 20.8 158.3 4- 19.5"* 558.0 q- 42.7 308.9 -4- 20.1"*
Dorsal and median raphe 5,7-DHT 62.6 4- 11.7" 56.7 4- 8.2* 135.4 ± 36.1" 94.8 4- 13.2"

* P < 0.01 vs vehicle (two-way ANOVA and Duncan's multiple-range test).

** P < 0.01 vs saline (two-way ANOVA and Duncan's multiple-range test).
236

Two weeks later the rats were given PCA (10 mg/kg i.p.) and sacrificed 2 h later
by decapitation. Both rats in each cage were sacrificed within 15 s after opening of the
cage. The brains were quickly removed and the caudate nuclei and hippocampi were
dissected and stored at - - 8 0 °C for serotonin determination. The midbrain was
dissected, stored in formalin, sectioned, and stained with cresy[ violet for histological
verification of the lesion placement. Blood from the decapitated rats was collected into
centrifuge tubes containing 0.5 ml of 0.3 M E D T A and plasma was stored a t - - 1 0 °C
until assayed for plasma renin activity (PRA) as previously described24, 29, and
corticosterone using procedures and corticosterone antisera from Radioassay Systems
Laboratories (Carson, CA), on unextracted plasma in which binding proteins had
been heat-denatured 1. Serotonin was determined according to Saavedra et al. 25 with
the exception that N-acetyl transferase was isolated from dorsophila according to
McCaman et al. 19. Two criteria were used to select lesion animals for analysis of their
PRA and corticosterone concentrations: the needle track had to descend and
terminate within 0.5 m m of the midline in the designated nucleus; and the dorsal and
median raphe lesions had to reduce serotonin content to values that were more than 2
standard deviations below the control mean in the hippocampus in the case of median
raphe lesions and in the caudate nucleus in the case of dorsal raphe lesions. Animals
with large decreases in the serotonin content of both the hippocampus and caudate
nucleus were considered separately as animals with combined lesions of the dorsal and
median raphe nuclei 1zA~,17,3°,31. PCA caused a significant reduction in the serotonin
concentration in both the caudate nucleus and the hippocampus (Table I), and
therefore the saline and PCA vehicle control groups were considered separately.
Statistical analysis was performed by two-way analysis of variance and Duncan's
new multiple range test by a computer statistical package according to Nie et al. z2.

RESULTS

Histological analysis revealed that in 13 rats meeting our criteria (see above),
5,7-DHT injections produced lesions in the dorsal raphe nucleus about 400/~m in
diameter, with glial scarring surrounding the lesion (Fig. 1). Injections of the same
volume of vehicle failed to produce visible lesions. In one rat injected with 5,7-DHT,
the needle track ended in the aqueduct, and in 9 rats, the needle descended to the
periaqueductal gray and ended in or ventral to the superior cerebellar peduncle or in
the dorsal part of the median raphe nucleus. These rats were considered unsuccessful
dorsal raphe lesions. In 13 rats, injections aimed at the median raphe produced lesions
restricted to this nucleus and spared the dorsal raphe nucleus (Fig. 2). In 10 rats,
injections produced lesions that included the caudal part of the dorsal raphe nucleus at
the level of the trochlear nucleus and in some cases the pontine B6 area as well as the
median raphe (Fig. 3). These rats were combined for statistical analysis with the rats in
which injections aimed at the dorsal raphe nucleus caused damage to the median raphe
nucleus and were considered together as a dorsal and median raphe lesion group.
PCA caused a significant decrease in the serotonin content of the caudate
nucleus and hippocampus (Table I). Therefore, for the analysis of decreases in
 237

Fig. 1. Dorsal raphe lesions produced by intracerebral 5,7-DHT injections in 3 rats. A: rostral, B:
medial, C: caudal dorsal raphe lesions.

serotonin due to the 5,7-DHT injections, two control groups were considered
separately. The vehicle/saline group was the control group for the 5,7-DHT/saline
group and the vehicle/PCA group was the control group for the 5,7-DHT/PCA group.
All the rats in which histological analysis revealed a lesion in the dorsal raphe nucleus
due to injection of 5,7-DHT had a decreased serotonin content of the caudate nucleus
238

by more than 2 × the standard deviation below the control mean. There was 62
depletion in the saline-treated rats and 56 ~ depletion in the PCA-treated rats. The
serotonin content of the hippocampus was reduced 18 ~ in the saline-treated rats and
25 ~ in the PCA-treated rats.

Fig. 2. Median raphe lesions produced by intracerebral 5,7-DHT injections.

 239

Fig. 3. Combined median and dorsal raphe lesions produced by intracerebral 5,7-DHT injections.

In the rats with histologically verified median raphe lesions, the reverse occurred,
i.e. hippocampal serotonin was decreased 80 ~ and 82 ~ and caudate serotonin was
decreased 34 ~o and 37 ~ . Rats with histologically verified dorsal and median raphe
lesions had decreases in hippocampal serotonin of 8 0 ~ and 6 4 ~ , and in caudate
serotonin of 75 ~ and 69 ~ .
240

TABLE II
Effect of local intracerebral injections of 5,7-dihydroxytryptamine (5,7-DHT) on the increase in plasma
renin activity produced by parachloroamphetamine (PCA )
5,7-DHT or vehicle injected 2 weeks before sacrifice. PCA, 10 mg/kg or saline injected i.p. 2 h before
sacrifice, n is indicated in parentheses.

Injection site Injection Plasma renin activity (ng Al/ml/3 h)

Saline PCA

Dorsal raphe Vehicle 9.2 4. 1.5 (12) 32.1 ± 4.4* (14)

Dorsal raphe 5,7-DHT 6.9 5:0.8 (5) 14.7 4, 2.3 ( 8 )
Median raphe Vehicle 9.0 ± 1.1 ( 8 ) 32.1 4. 7.0* ( 8 )
Median raphe 5,7-DHT 6.1 ± 0.8 ( 6 ) 33.9 4- 11.0" ( 7 )
Dorsal and median raphe Vehicle 9.0 ± 0.8 (20) 33.3 ± 3.6* (22)
Dorsal and median raphe 5,7-DHT 7.8 4. 1.3 ( 7 ) 12.8 4, 2.0 (12)

* P < 0.01 vs saline (two-way ANOVA and Duncan's new multiple-range test).

A d m i n i s t r a t i o n o f P C A to the vehicle-injected control rats caused a significant

increase in plasma r e n i n activity a n d corticosterone c o n c e n t r a t i o n (Tables II a n d III).
The effect of P C A o n plasma r e n i n activity was significantly inhibited by lesions in the
dorsal raphe b u t n o t by lesions in the m e d i a n raphe. Rats with lesions in b o t h the
dorsal a n d the m e d i a n raphe also showed a significant i n h i b i t i o n of the effect of P C A on
renin secretion. However, n o n e of the lesions inhibited the effect of P C A on
corticosterone secretion. Histological e x a m i n a t i o n showed that all the rats in which
the stimulatory effect of P C A on plasma r e n i n activity was inhibited had in c o m m o n a
lesion of the caudal part of the dorsal raphe nucleus at a n d b e h i n d the level of the
trochlear nucleus.

TABLE II1
Effect of local intracerebral injections of 5,7-dihydroxytryptamine (5,7-DHT) on the increase in plasma
corticosterone concentrationproduced by parachloroamphetamine (PC~I)
5,7-DHT or vehicle injected 2 weeks before sacrifice. PCA, 10 mg/kg or saline injected i.p. 2 h before
sacrifice, n is indicated in parentheses.

lniection site Injection Plasma corticosterone (#g/dl)

Saline PCA

Dorsal raphe Vehicle 7.4 i 2.2 (12) 26.2 i 2.2* (14)

Dorsal raphe 5,7-DHT 7.8 i 2.1 ( 5 ) 27.9 ± 5.5* ( 8 )
Median raphe Vehicle 4.0 4. 1.2 ( 8 ) 21.4 ± 3.9* ( 8 )
Median raphe 5,7-DFIT 2.6 i 1.0 ( 6 ) 24.1 i 2.6* ( 7 )
Dorsal and median raphe Vehicle 4.5 ± 1.0 (20) 24.3 :~ 2.0* (22)
Dorsal and median raphe 5,7-DHT 3.1 ± 1.1 (7) 35.9 i 3.5* (12)

* P < 0.01 vs saline (two-way ANOVA and Duncan's new multiple-range test).
 241

DISCUSSION

The injections of 5,7-DHT in the present experiments produced visible lesions

surrounded by gliosis, so they destroyed more than just the serotonergic neurons at the
site. Moreover, they did not necessarily destroy all the serotonergic neurons in a given
nucleus. This may be the reason that intraventricular 5,7-DHT caused a statistically
significant decline in resting plasma renin activity32, whereas dorsal raphe lesions did
not, even though they reduced the renin response to PCA.
Although the overall effects of PCA are complex and involve inhibition of
tryptophan hydroxylase, serotonin uptake, monoamine oxidase, and norepinephrine
uptake26,27 and possibly other effects, it is generally accepted that the initial effect of
the drug is release of serotonin18. The effect of PCA on renin is plevented by p-
chlorophenylalanine3z, and other evidence indicates that central release of serotonin
increases renin secretionaZ,3a. Therefore, the results of the present study provide
evidence that a serotonergic pathway originating in the dorsal raphe nucleus exerts a
stimulatory effect on renin secretion. The median raphe nucleus does not seem to play
a role in the regulation of renin secretion.
In contrast, lesions in the dorsal and median raphe nuclei failed to affect the
increase in plasma corticosterone produced by PCA. Since this effect is abolished byp-
chlorophenylalanine11,32, it appears to be due to serotonin. There is evidence that
serotonin can stimulate corticosterone secretion by a direct effect on the adrenal
cortex zl. However, we have found that the corticosterone stimulating effect of PCA is
abolished by hypophysectomy33, and by lesions of the mediobasal hypothalamus (Van
De Kar, Karteszi and Ganong, unpublished observations). Therefore, it seems
reasonable to conclude that it is mediated by ACTH. The serotonergic pathways
involved are unknown, but there is evidence that the pontine and medullary raphe
nuclei project to the hypothalamus in the cat 6, and since these nuclei were spared in the
present study, the possibility exists that these caudal raphe nuclei mediate the effect of
PCA on corticosterone secretion. Another possible mediator in a serotonin-accumu-
lating cell group in the hypothalamus4,7,s, 14.
The location of the serotonin receptors that stimulate renin secretion is not
known. The dorsal and median raphe nuclei both project to the hypothalamus and
other forebrain areas2,5,6,30,~1. In unpublished studies we found that mediobasal
hypothalamus lesions completely abolished the increase in PRA normally produced by
PCA, suggesting that the serotonergic pathway that originates in the dorsal raphe
stimulates renin secretion via this hypothalamic area (Van De Kar, Karteszi and
Ganong, unpublished observations). Quattrone et al. 23 presented evidence that the
median raphe nucleus mediates a stimulatory effect on prolactin secretion via a
serotonergic pathway to the hypothalamus, so the median raphe nucleus may also
exert selective endocrine effects.
The involvement of brain serotonergic neurons in the regulation of blood
pressure has been studied by a variety of experimental techniques15. Smits et al. 2s
suggested that serotonergic neurons in the median but not the dorsal raphe nucleus
mediate a pressor response, possibly via the anterior hypothalamus-preoptic area,
242

which is selectively i n n e r v a t e d by the m e d i a n raphe nucleus30, 31. The pressor effect of

stimulation o f the median raphe is p r o m p t 16,za. In contrast, the effect of P C A o n
plasma renin activity develops slowly z2 a n d lasts for several hours. Thus, there may be
two mechanisms by which serotonergic stimulation can increase blood pressure, a
direct pathway involving the median raphe nucleus a n d an indirect pathway by way of
renin involving the dorsal raphe nucleus.

ACKNOWLEDGEMENTS

The authors t h a n k Helen Hughes, Shirley Chi, Veronika Licko a n d A n n e t t e

Lowe for their excellent technical assistance.
This work was supported by U S P H S G r a n t AM06704. L.D.v.d.K. a n d C.W.W.
were N I H Postdoctoral Fellows. M.S.B. was a G i a n n i n i Fellow.

REFERENCES
I Abraham, G. E., Manlimos, F. S. and Garza, R. In G. E. Abraham (Ed.), Handbook of Radio-
immunoassay, Marcel Dekker, New York, 1977, pp. 591-656.
2 Azmitia, E. C. and Segal, M., An autoradiographic analysis of the differential ascending projec-
tions of the dorsal and median raphe nuclei in the rat, J. comp. Neurol., 179 (1978) 641-668.
3 Bj6rklund, A., Baumgarten, H. G. and Rensch, A., 5,7-Dihydroxytryptamine: improvement of its
selectivity for serotonin neurons in the CNS by pretreatment with desipramine, J. Neurochem., 24
(1975) 833-835.
4 Beaudet, A. and Descarries, L., Radioautographic characterization of a serotonin-accumulating
nerve cell group in adult rat hypothalamus, Brain Research, 160 (1979) 231-243.
5 Bobillier, P., Petitjean, F., Salvert, D., Leger, M. and Seguin, S., Differential projections of the
nucleus raphe dorsalis and nucleus raphe centralis as revealed by autoradiography, Brain
Research, 85 (1975) 205-210.
6 Bobillier, P., Seguin, S., Petitjean, F., Salvert, D., Touret, M. and Jouvet, M., The raphe nuclei
of the cat brain stem: a topographical atlas of their efferent projection as revealed by autoradio-
graphy, Brain Research, 113 (1976) 449-486.
7 Brownstein, M. J., Palkovits, M., Tappaz, M., Saavedra, J. M. and Kizer, J. S., Effect of surgical
isolation of the hypothalamus on its neurotransmitter content, Brain Research, 117 (1976)
287-295.
8 Chan-Palay, V., Indoleamine neurons and their processes in the normal rat brain and in chronic
diet-induced thiamine deficiency demonstrated by uptake of ZH-serotonin, J. comp. Neurol., 176
(1977) 467-494.
9 DahlstrSm, A. and Fuxe, K., Evidence for the existence of monoamine-containingneurons in the
central nervous system. I. Demonstration of monoamines in the cell bodies of brain stem neurons,
Aeta physioL scand., Suppl., 232 (1969) 1-55.
10 Epstein, S. and Hamilton, S., Cyproheptadine inhibition of stimulated plasma renin activity, J.
clin. Endocr. Metab., 45 (1977) 1235-1237.
11 Fuller, R. W. and Snoddy, H. D., Effect of serotonin-releasing drugs on serum corticosterone
concentration in rats, Neuroendocrinology, 31 (1980) 96-100.
12 Geyer, M. A., Puerto, A., Dawsey, W. J., Knapp, S., Bullard, W. P. and Mandell, A. J., Histologic
and enzymatic studies of the mesolimbic and mesostriatal serotonergic pathways, Brain Research,
106 (1976) 241-256.
13 Jacobs, B. L., Wise, W. D. and Taylor, K. M., Differential behavioral and neurochemical effects
following lesions of the dorsal or median raphe nuclei in rats, Brain Research, 79 (1974) 353-361.
14 Kent, D. L. and Sladek, J. R., Jr., Histochemical, pharmacological and microspectrofluorometric
analysis of new sites of serotonin localization in the rat hypothalamus, J. comp. NeuroL, 180 (1978)
221-236.
15 Kuhn, D. M., Wolf, W. A. and Lovenberg, W., Review of the role of the central serotonergic
neuronal system in blood pressure regulation, Hypertension, 2 (1980) 243-255.
 243

16 Kuhn, D. M., Wolf, W. A. and Lovenberg, W,, Pressor effects of electrical stimulation of the
dorsal and median raphe nuclei in anesthetized rats, J. Pharmacol. exp. Ther., 214 (1980) 403-409.
17 Lorens, S. A. and Guldberg, H. C., Regional 5-hydroxytryptamine following selective midbrain
raphe lesions in the rat, Brain Research, 78 (1979) 45-56.
18 Marsden, C. A., Conti, J., Strope, E., Curzon, G. and Adams, R. N., Monitoring 5-hydroxy-
tryptamine release in the brain of the freely moving unanesthetized rat using in vivo voltammetry,
Brain Research, 171 (1979) 85-99.
19 McCaman, M. W., McCaman, R. E. and Stetzler, J., A rapid radioenzymatic assay for dopamine
and N-acetyl dopamine, Analyt. Biochem., 96 (1979) 175-180.
20 Modlinger, R. S., Schonmuller, J. M. and Arora, S. P., Stimulation of aldosterone, renin and
cortisol by tryptophan, J. clin. Endocr., Metab., 48 (1979) 599-603.
21 M/iller, J., Regulation of Aldosterone Biosynthesis, Springer-Verlag, 1971.
22 Nie, N. H., Hull, C. H., Jenkins, J. G., Steinbrenner, K. and Bent, D. H., StatisticalPackagefor
the Social Sciences, Edn. 2, McGraw-Hill, New York, 1975.
23 Quattrone, A., Schettini, G., Di Renzo, G., Tedeschi, G. and Preziosi, P., Effect of midbrain raphe
lesions or 5,7-dihydroxytryptamine treatment on the prolactin-releasing action of quipazine and
D-fenfluramine in rats, Brain Research, 174 (1979) 71-79.
24 Reid, I. A., Stockigt, J. R., Goldfien, A. and Ganong, W. F., Stimulation of renin secretion in
dogs by theophyline, Europ. J. Pharmacol., 17 (1972) 325-332.
25 Saavedra, J. M., Brownstein, M. and Axelrod, J., A specific and sensitive enzymatic isotopic
microassay for serotonin in tissues, J. Pharmacol. exp. Ther., 186 (1973) 508-515.
26 Sanders-Bush, E., Bushing, J. A. and Sulser, F., Long-term effects ofp-chloroamphetamine and
related drugs on central serotonergic mechanisms, J. PharmacoL exp. Ther., 192 (1975) 33-41.
27 Sanders-Bush, E. and Steranka, L. R., Immediate and long-term effects of p-chloroamphetamine
on brain amines, Ann. N. Y. Acad. Sci., 305 (1978) 208-221.
28 Smits, J. F. M., Van Essen, H. and Struyker-Boudier, A. J., Serotonin mediated cardiovascular
responses to electrical stimulation of the raphe nuclei in the rat, Life Sci., 23 (1978) 173-178.
29 Stockigt, J. R., Collins, R. D. and Biglieri, E. G., Determination of plasma renin concentration by
angiotensin I radioimmunoassay, Circulat. Res., 28/29, Suppl. 1I, (1971) 175-189.
30 Van de Kar, L. D. and Lorens, S. A., Differential serotonergic innervation of individual hypo-
thalamic nuclei and other forebrain regions by the dorsal and median midbrain raphe nuclei, Brain
Research, 162 (1979) 45-54.
31 Van de Kar, L. D., Lorens, S. A., Vodraska, A., Allers, G., Green, M., Van Orden, D. E. and Van
Orden, L. S., III, Effect of selective midbrain and diencephalic 5,7-dihydroxytryptamine lesions on
serotonin content in individual preopticohypothalamic nuclei and on serum luteinizing hormone
level, Neuroendocrinology, 31 (1980) 309-315.
32 Van de Kar, L. D., Wilkinson, C. W. and Ganong, W. F., Pharmacological evidence for a role of
brain serotonin in the maintenance of plasma renin activity in unanesthetized rats. J. Pharmacol.
exp. Ther., (1981) in press.
33 Van de Kar, L. D., Wilkinson, C. W., Brownfield, M. S., Skrobik, Y. and Ganong, W. F., Seroto-
nergic neurons stimulate renin and corticosterone secretion via the pituitary gland, Neurosci.
Abstr., 7 (1981) in press.
34 Zimmermann, H. and Ganong, W. F., Pharmacological evidence that stimulation of central
serotonergic pathways increases renin secretion, Neuroendocrinology, 30 (1980) 101-107.