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A 24-hour rhythm in the content of norepinephrine in the pineal and salivary glands of the rat
5,
Life 8cienoes Vol . pp . Printed in Great Britain.
665-669, 1966 .
Pergamoa Prosa Ltd .
e a~-aoa~ asx~rmc Ix T~ co~rr a~ ~aosrl~sna n~ T~ tvosL !!1® BALIYABY QI~d ~ TSe BAT Biohard J. "m~tcran acrd Jw].ins A~telrod Is
(1), rant' o! which tercriaate directly as
pi.aeal parenohycral sells (a) .
It aLo contaias relatively high
concentrations of naa~epinephrine (3,4), which are localized in the deale core vesicles o! the nerve eadiags (9, d) .
It has
recently been shows that the eycrpathetic serves to the pineal ezercise a crajor iatlueace oa its bioâhecrioal fs~matioas :
They
transcrit signals (generated by environrental lighting) which regulate pineal weight (8)
and the activity o! an enzycra
localized within this organ, hydrozyindole-O-aethyl traaslerase (HIQQ)
(8) .
The sycrpathetic nerves also
c~7 crsssages
free the
bleu which are responsible for two intrapiaeai rhythcrs of about 24 hours
d:sation, those o! changes in HIaIT activity (Z) cad o!
changes in the content o! serotonin (8) . 8t~ies described in this report will show that the level of norepinephrine in the rat pineal varies over a 9-fold range during each 24-hour period .
A rhythcr with sicrilar te~paral
charaa~teristios is described is the subcrazillary gland .
This
suggegts that the activity of part o! the sycrpathetic nervous system is related to eavironcreatal lighting cad the ticre o! day . 665
666
RHYTHM IN NE CONTENT
Vol . 5, No .
Materials and Methods Adult 8pragne-Dawley female rate, weighing 180-~00 g were kept under controlled lighti.ag conditions (lights on from 7 AY to 7p1I= darlmess lraa 7 PII to 7 Ain for a weei prior to use. =ach animal was ezposed to ßb-60 loot-candles o! cool-white llucresceat illurinatioa .
Groups o! 30 rats were used to
deterriae the pineal norepiaephrine content at any particular tire .
The spirals were ülled by aeci fracture, and the pineal
glade were removed se described before (7) and placed on filter paper impregnated with chilled isotonic saline .
Five
pinesls were pooled, weighed, and harogeaized in 5 ml of O .dN perchloric acid .
The aorepiaephriae was separated by alumna
colas chra~atography, and assayed by the rethod of von 8'uler and Lishajio (9) .
One salivary gland was also taten fro® each
aniralj four glands were pooled and assayed as above. Results Rat pineal glands were found to contain 3 .8 - 10 .8 nano~ars o! norepiaephrine per organ, ar about 3.3 - 9.8 ricrogrars per~grrar of tissue . reported previously (4) .
This is is agreereat with values
The norepinephriae content was
highest at the end o! the dart period, and tell during the light period, reaching a nadir at 7 PII (Figure 1) . The norepinephriae content o! the rat subrazillary gland was about twice as high at the end of the dart period as it was twelve hours later (Table 1) . Discussion These data provide the first direct evidence that the content of the neurotransritter norepinephrine varies with a regular 2~L-hour cycle in sympathetically-innervated structures .
Vol . 5, No . 7
RHYTHM IN NE CONTENT
66~
Iz .o ,g IQO z â z 8.0 x a w ~ 60 w o_ 0 z rn 4.0 ï a c~ 0 2.0 z
cw~ar ÎpY
YIDNIOHT
tuaK 6AY
T-i NDDN 6PY
YIDNIWiT
7AY
1~IG, I Pineal aorepinephriae content at di!lereat times o! day. Lights were oa lraa 7 AY to 7 PY . Bach point represents 8 groups o! ô pooled pineal glass . TAßLE I Norepiaephriae Content o! the ßat Subaazillary Gland at Di!lereat Tines o! Day ? -8AM
8-?PM
N
Bzperiaeat 1:
2.27 t 0,17
1.88 t 0.10*
~L8
Bzperimeat 2:
1 .8d t 0.19
0.82 t 0.10*
d8
Data are ezp~reseed as micrograms o! norepinephriae per grate o! tissue, t standard error o! the mena . Bsch ezperimeatal group contained 8 pools o! d submazillary glande each . Lights were oa Tram ? AlI to 7 PM . *P~0 .001 Di!lers irae nosing value. They are consistent with the hypothesis that the daily rhythms is pineal HI01[r activity (7) and serotonia content (8) are mediated by cyclic alterations is sympathetic nervous activity .
The close
similarity between the changes in narepiaephriae content is the submazillary and pineal glands suggests that the activity o!
668
RHYTTHIII IN NE CON~HT
Qol .
5, No .
whole omits of the sympathetic aervaas system varies with evairoamentsl lighting and the time o! day. Almost all daily rhythms is aaamalr appear to be gensrated by
biologic cloa]rs^ originating within the animal .
These
mechaair~ms operate with a periodicity apprasimatiag, but not egtrshling; s4 boars ; hence they have been termed °circadinaM(10) . Such rhythms are o~diaarily entrained by the ezternal day-nicht cycle o! eaviraamental lighting however, they persist when animals are blinded or are iept is continuous darhaesa . ezcsptioa is the HI01lP cycle is the rat pineal :
Aa
This appears
to depend entirely upon the animal's ability to perceive rhythmic changes in en.iroameatal lighting (7,11) .
It has not
yet been determined whether the rhythms in pineal a~ salivary gland narepiaephrine content described above are endogenous or ezogenoas.
Studies are in progress to establish their mechanism
o! control. The iacreaae in pineal aorepiaephriae content during dariaess could be due primarily to as increase is the synthesis of the amine, a decrease in its iatranevrsl destruction (by monwmiae azidase), or a deosease in its rate o! release.
It is
also possible that darhaess enchaaces sympathetic nervous tone is rats, psoduciag both a slight increase in the rate o! release o! narepinephriae, and a greater elevation of its rate of biosynthesis .
=videace has already been presented that
preganglioaic usual impulses can inllueace the rate at which sympathetic nerve endings lama narepinephrins (1S) . 8urary The nasepinephrine content of the pineal sad submazillsry glands varies with a
-hour rhythm ; it is greatest at the end
of the dart period, cad !ails during the'day.
Yol . 5, No . ~
RHYTf~ IN NE CONTENT
669
References 1. Z.
J, A, HAPPERS, Z, Zellforsch. ~ 183 (1981), D, E, wOd~E, L,_T, POITEB, a . C, RICHARDSON and J, A~LROD, 8cieace 138, 4ß0 (1982),
3,
N. J, GIARYAN aad Y, DAY, Bioohem, Pharaacol . ~ 236 (196ß),
4.
A, PELLEGRINO DE ZltALDI anâ L, lI, ZZEH~, Life Sciences ia press (198b) .
6,
A . PELLEGRINO DE IItALDI, L, Y, ZIBHER aad E, DIE ROBERTIS Prog. Hraia Res . 10, sß9 (198x) .
8,
R . J, ~tTYAN, J. AIBLROD and J. E, FISCH88, Scieace 1d3, (1984),
7.
J, AIBLROD, R. J, wIIftTYAN and 8, H, SNYDER, J. Biol. Chea. 949 (198b) .
8.
8, H, SNYDER, J. A~LROD aad Y, ZWEIG, Prac, Natl, Acad,Sci , b~,
9,
aol
(19ßb) .
II, S . Y~ EULBTt snd F, LISHAJSO, Acta Physiol . Scand , 61 348 (1981) .
10,
F, HA7~RG, J, Leaoet T3, 20 (1963),
11 .
R . J, wIIRfYAN and J. A~LROD
Scientific Americaa 213, 60
(198b) . 7.9,
J, IIDSACCHIO and V, IIEISE, P'hattiacologist ~ I68 (1886),
Life 8cienoes Vol . pp . Printed in Great Britain.
665-669, 1966 .
Pergamoa Prosa Ltd .
e a~-aoa~ asx~rmc Ix T~ co~rr a~ ~aosrl~sna n~ T~ tvosL !!1® BALIYABY QI~d ~ TSe BAT Biohard J. "m~tcran acrd Jw].ins A~telrod Is
(1), rant' o! which tercriaate directly as
pi.aeal parenohycral sells (a) .
It aLo contaias relatively high
concentrations of naa~epinephrine (3,4), which are localized in the deale core vesicles o! the nerve eadiags (9, d) .
It has
recently been shows that the eycrpathetic serves to the pineal ezercise a crajor iatlueace oa its bioâhecrioal fs~matioas :
They
transcrit signals (generated by environrental lighting) which regulate pineal weight (8)
and the activity o! an enzycra
localized within this organ, hydrozyindole-O-aethyl traaslerase (HIQQ)
(8) .
The sycrpathetic nerves also
c~7 crsssages
free the
bleu which are responsible for two intrapiaeai rhythcrs of about 24 hours
d:sation, those o! changes in HIaIT activity (Z) cad o!
changes in the content o! serotonin (8) . 8t~ies described in this report will show that the level of norepinephrine in the rat pineal varies over a 9-fold range during each 24-hour period .
A rhythcr with sicrilar te~paral
charaa~teristios is described is the subcrazillary gland .
This
suggegts that the activity of part o! the sycrpathetic nervous system is related to eavironcreatal lighting cad the ticre o! day . 665
666
RHYTHM IN NE CONTENT
Vol . 5, No .
Materials and Methods Adult 8pragne-Dawley female rate, weighing 180-~00 g were kept under controlled lighti.ag conditions (lights on from 7 AY to 7p1I= darlmess lraa 7 PII to 7 Ain for a weei prior to use. =ach animal was ezposed to ßb-60 loot-candles o! cool-white llucresceat illurinatioa .
Groups o! 30 rats were used to
deterriae the pineal norepiaephrine content at any particular tire .
The spirals were ülled by aeci fracture, and the pineal
glade were removed se described before (7) and placed on filter paper impregnated with chilled isotonic saline .
Five
pinesls were pooled, weighed, and harogeaized in 5 ml of O .dN perchloric acid .
The aorepiaephriae was separated by alumna
colas chra~atography, and assayed by the rethod of von 8'uler and Lishajio (9) .
One salivary gland was also taten fro® each
aniralj four glands were pooled and assayed as above. Results Rat pineal glands were found to contain 3 .8 - 10 .8 nano~ars o! norepiaephrine per organ, ar about 3.3 - 9.8 ricrogrars per~grrar of tissue . reported previously (4) .
This is is agreereat with values
The norepinephriae content was
highest at the end o! the dart period, and tell during the light period, reaching a nadir at 7 PII (Figure 1) . The norepinephriae content o! the rat subrazillary gland was about twice as high at the end of the dart period as it was twelve hours later (Table 1) . Discussion These data provide the first direct evidence that the content of the neurotransritter norepinephrine varies with a regular 2~L-hour cycle in sympathetically-innervated structures .
Vol . 5, No . 7
RHYTHM IN NE CONTENT
66~
Iz .o ,g IQO z â z 8.0 x a w ~ 60 w o_ 0 z rn 4.0 ï a c~ 0 2.0 z
cw~ar ÎpY
YIDNIOHT
tuaK 6AY
T-i NDDN 6PY
YIDNIWiT
7AY
1~IG, I Pineal aorepinephriae content at di!lereat times o! day. Lights were oa lraa 7 AY to 7 PY . Bach point represents 8 groups o! ô pooled pineal glass . TAßLE I Norepiaephriae Content o! the ßat Subaazillary Gland at Di!lereat Tines o! Day ? -8AM
8-?PM
N
Bzperiaeat 1:
2.27 t 0,17
1.88 t 0.10*
~L8
Bzperimeat 2:
1 .8d t 0.19
0.82 t 0.10*
d8
Data are ezp~reseed as micrograms o! norepinephriae per grate o! tissue, t standard error o! the mena . Bsch ezperimeatal group contained 8 pools o! d submazillary glande each . Lights were oa Tram ? AlI to 7 PM . *P~0 .001 Di!lers irae nosing value. They are consistent with the hypothesis that the daily rhythms is pineal HI01[r activity (7) and serotonia content (8) are mediated by cyclic alterations is sympathetic nervous activity .
The close
similarity between the changes in narepiaephriae content is the submazillary and pineal glands suggests that the activity o!
668
RHYTTHIII IN NE CON~HT
Qol .
5, No .
whole omits of the sympathetic aervaas system varies with evairoamentsl lighting and the time o! day. Almost all daily rhythms is aaamalr appear to be gensrated by
biologic cloa]rs^ originating within the animal .
These
mechaair~ms operate with a periodicity apprasimatiag, but not egtrshling; s4 boars ; hence they have been termed °circadinaM(10) . Such rhythms are o~diaarily entrained by the ezternal day-nicht cycle o! eaviraamental lighting however, they persist when animals are blinded or are iept is continuous darhaesa . ezcsptioa is the HI01lP cycle is the rat pineal :
Aa
This appears
to depend entirely upon the animal's ability to perceive rhythmic changes in en.iroameatal lighting (7,11) .
It has not
yet been determined whether the rhythms in pineal a~ salivary gland narepiaephrine content described above are endogenous or ezogenoas.
Studies are in progress to establish their mechanism
o! control. The iacreaae in pineal aorepiaephriae content during dariaess could be due primarily to as increase is the synthesis of the amine, a decrease in its iatranevrsl destruction (by monwmiae azidase), or a deosease in its rate o! release.
It is
also possible that darhaess enchaaces sympathetic nervous tone is rats, psoduciag both a slight increase in the rate o! release o! narepinephriae, and a greater elevation of its rate of biosynthesis .
=videace has already been presented that
preganglioaic usual impulses can inllueace the rate at which sympathetic nerve endings lama narepinephrins (1S) . 8urary The nasepinephrine content of the pineal sad submazillsry glands varies with a
-hour rhythm ; it is greatest at the end
of the dart period, cad !ails during the'day.
Yol . 5, No . ~
RHYTf~ IN NE CONTENT
669
References 1. Z.
J, A, HAPPERS, Z, Zellforsch. ~ 183 (1981), D, E, wOd~E, L,_T, POITEB, a . C, RICHARDSON and J, A~LROD, 8cieace 138, 4ß0 (1982),
3,
N. J, GIARYAN aad Y, DAY, Bioohem, Pharaacol . ~ 236 (196ß),
4.
A, PELLEGRINO DE ZltALDI anâ L, lI, ZZEH~, Life Sciences ia press (198b) .
6,
A . PELLEGRINO DE IItALDI, L, Y, ZIBHER aad E, DIE ROBERTIS Prog. Hraia Res . 10, sß9 (198x) .
8,
R . J, ~tTYAN, J. AIBLROD and J. E, FISCH88, Scieace 1d3, (1984),
7.
J, AIBLROD, R. J, wIIftTYAN and 8, H, SNYDER, J. Biol. Chea. 949 (198b) .
8.
8, H, SNYDER, J. A~LROD aad Y, ZWEIG, Prac, Natl, Acad,Sci , b~,
9,
aol
(19ßb) .
II, S . Y~ EULBTt snd F, LISHAJSO, Acta Physiol . Scand , 61 348 (1981) .
10,
F, HA7~RG, J, Leaoet T3, 20 (1963),
11 .
R . J, wIIRfYAN and J. A~LROD
Scientific Americaa 213, 60
(198b) . 7.9,
J, IIDSACCHIO and V, IIEISE, P'hattiacologist ~ I68 (1886),