Reprinted from Mammalian Hibernation III edited by Kenneth C. Fisher, Albert R. Dawe, Charles P.

Lyman, Eduard Schdnbaum, Frank E. South, Jr. Published by Oliver & Boyd Ltd W, and by American Elsevier, New York, in the U-S.A.

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pf

Atq-

2zft;ufq,,it:& 6;btic:3 gpatt-909

CESIUM.137 METABOLISM IN

ACTIVE AI\D HIBERNATING CITELLUS LATERALIS*

MenvIN L. RrrpnsEr,, Yu-CHoNG LIN, Jevrns J. YouNc AND GoRDoN H. Bnvext .iq67" D ep ar tment d u"If{,:r#;I:'fr':^t.'

Iry:i #f*ioy ., * r TUalura I A^rrrrrrrr(,' tt,, H ; b ev^ na-fi on @ sy n',poiu vn, L( t; u' fDronfrl Aaatacl<) t3 -,b 5e/f
INTRODUCTION

tqbs.

Electrolyte metabolism during mammalian hibernation might be expected to involve changes in the circulatory and excretory systems as well as changes in cellular mechanisms concerned with transport and retention of electrolytes in tissues. A favourable homeostasis must exist during hibernation because anirnals hibernate one to forty days or longer. On the other hand, changes in the rate and effectiveness of the circulatory system and chemical processes involved in electrolyte transport across cell membranes can be eipected to accompany hibernation and lowering of body temperature by 3o-32'C. Studies in this field have been primarily limited to analyses of body fluids, excreta, and one or two tissues for a given electrolyte (Calkin" it ol. 1954, Eliassen 1963, Riedesel 196o, Willis ry64a). Standard chemical procedures are laborious and permit analyses of a limited number of samples per experiment. In addition when'an increase (or decrease) of an electrolyte is observed in a given tissue or fluid, the source (or fate) of the electrolyte is unknown. Radio-isotope techniques provide an opportunity to examine changes in an electrolyte concentration in a number of tissues. The present study represents an attempt to describe cesium-r37 metabolism in active, hypothermic and hibernating conditions at the whole animal and tissue levels. Cesium-r37 is a useful tool in studying hibernation because it is an intracellular electrolyte, is metabolized by all tissues, is a gamma emitter, and has a long biological half life. On the other hand, hibernation is a useful tool for studying cesium metabolism because * Supported by United States Atomic Energy Commission research
AT(29-z)-$zs.
contract

t National Institutes of Health, Post-Doctorate Fellow, zFz}jD-4,374-?2,464-65' Present Address: School of Pharmacy, Montana State University, Missoula, Montana'

220

RIEDESEL ET AL.

the effect of temperature may be observed on the processes which limit the rate of exchange of cesium between intracellular and extracellular spaces, and also between an animal and its environment. Richmond el a/. $96z) have related turnover of cesium to metabolic rate whereas comar and wasserman (1956) emphasized the role of tissue and cellular compartments which retain cesium. Although skeletal muscle is considered the critical organ following administration of cesium, all tissues metabolize cesium (Ballou and rhompson r95B). Emphasis in cesium research has been on long-term studies; and as a result, little information is available regarding rate and mechanism of uptake of cesium by tissr.res.

whole-body counter permits frequent counting of intact animals and provides a good_ comparison of tissue activity to whole-body activity as the whole animal and tissues are counted undir identical conditions. The standard (o'r58 microcuries, cesium-r37) and background counts _source w-ere made daily. The mean and standard error'of counts 4o "rr"-tinrrte of the standard cesium-r37 source and the background were rrz 136 c.p.g. 176 (s.e.) and r36r + 8 (s.e.), respectively. The term, hypothermia, refers to the condition of animals which have had their body temperatures lowered as a result of cornbined cord exposure and carbon dioxide anaesthesia (Andjus et ar. ry56b). Animals were placed in jars (9oo rttl capacity) which were closed. *it-h ."."* caps and exposed to - rooc. opening the jar at 15- to zo-minute intervals ensured adequate oxygen supply. within 45 to 60 minutes the above procedure resulted in lowering of body temperature to rB" to zooc. subsequent immersion in ice water and exposure to air at 8"c kept rectal temperature at Bo to rooc. Animals injected with cesium-r37 during hypothermia had rectal temperatures of 8" to rooC 3o minutes prior to injiction.

activities are not included in this report. Measurement of skin temperature by placing a thermocouple at the base of the fur near the shouldeis of the animals provided an indication of whether the animal was active or hibernating- skin temperature readings were taken without disturbing the animals and averaged z'C lower than rectal temperatures. All measurements of radioactivity were made with the packard Instrument co. Model 4roA_auto-gamma spectrometer and a Model 44o, smallanimal whole-body scintillation detector. The animals and tissues were centred in the counting chamber, r r cm in diameter, 20 cm in iength. The

MATERIALS AND METEIODS The experimental animals, citellus lateralis, were collected in northern New Mexico and had been in the laboratory two days to six months prior to experimentation. All animals received a single iniraperitoneal injection of o'48 microcuries of cesium- r 37 (in o.r ml of buffered Ringer's soiution) per roo g body weight. Immediately after injection the first whole-body count was taken. urine and feces were separated in metabolism cages. our studies confirm earlier reports (Ballou and rhompson r95g, coirar and wasserman 1956, Richmond 1958) which establisl the uiine as the principal route of excretion of cesium-r37; therefore, urine and feces

CESIUM-I37 iN CITELLUS LATERALIS

1t

G/gEAy

upper 3 to 5 cm of the small intestine, r to r.5 g of abdominal skin and o.5 to r.5 g of blood. Tissues in paper tares were placed in the counting chamber, and the radioactivity of each tissue was determined by either a single ten-minute count or the mean of three one-minute counts. counts per minute per gramme wet tissue were calculated. In order to assess the effect of temperature as well as time (see Lin r964) on the cesium-r37 uptake, distribution and retention by tissues, it seemed most convenient to express the data in the form of the ratio: c.p.m./g wet tissue Thus, since cesium is continuously being excreted

Data on cesium distribution at varying times after injection (up to z4 hours) were obtained on hibernating ground squirrels; these animals were always injected 6 to rz hours after they had entered hibernation. Data on hibernating animals at the first, second and third biological half times involved (a) injection of active animals; (D) sacrificing animals at rhe first, second or third biological half time only if they had been hibernating two days prior to the half time indicated. A standard procedure was followed just prior to and after sacrificing the animals. The body weight was first measured. Background, standard source, and whole body counts were measured for one minute. Then, the animal was sacrificed by decapitation. organs and tissues were removed and weighed to the nearest o'or g. The whole liver (without gall bladder), heart, lung, spleen and stomach were counted; both kidneys were counted together whereas the right and left whole gastrocnernii were weighed and the c.p.m. measured separately; the whole right femur was used, the

ilFt.

so that the total amount

particular tissue, which is greater than the average aflfrnity of all of the tissues in the body. In retrospect it would have been more informative to express these activities as c.p.m. per g of water. unfortunately water concentrations were not determined. The following terms were used: Tissue Index and rissue Retention Index. Tissue Index (TI) indicates the relative tissue concentrations of cesium-r37 (i.e. the above ratio) within z4 hours after injection, when the uptake process predominates. The same ratio will be called the Tissue Retention Index (TRI) when it refers to the relative tissue concentrations (i.e. the above ratio) at the first, second, third and fourth biological half times; at these times retention rather than uptake is the major factor contributing to the cesium-r37 concentration of the tissues. Lowering of TRI of a specific tissue between the first and second half times represents a situation in which the rate of loss of the radio-isotope from the tissue is in excess of the rate of loss from the animal as a whole.
RESULTS Experiments conducted include: (a) measurement

in the whole body is always decreasing, the concentration of the isotope in any tissue is always expressed in terms of the concentration in the whole body at that moment. Hence a value for cesium-r37 of more than r'oo for this ratio would indicate an affinity of a

of

tissue ind.ices on

aoa

RIEDESEL ET AL.

active, hypothermic and hibernating C. lateralis sacrificed within z4 hours after a single intraperitoneal injection of cesium-r37; (b) determination of cesium-r37 retention curves on rats, active ground squirrels and hibernating ground squirrels exposed to temperatures of 2o" and ro'C; and (c) measurement of cesium-r37 tissue retention indices on C. lateralis active and hibernating at various biological half times.

Cesium-r37 Uptake Following Intraperitoneal Injection The tissue index data collected on animals sacrificed within z4 hours after injection of the radio-isotope are presented in Tables I and II and Figs. r and z. The right and left skeletal muscle data were collected to show the reproducibility of the mean and s.d. values. Tissues of particular interest are blood, kidney, heart, skeletal muscle, liver and lung. Observations which are pertinent to this report include: (a) Kidney was the first tissue to concentrate the cesium and was the tissue with the highest TI in active, hypothermic and hibernating animals. Apparently an active transport system was concentrating the cesium in the kidney; and although the system was slowed down during hibernation and hypothermia, the kidney still had the fastest uptake of cesium-r37. (D) The cesium content of blood and plasma was always very low and 15 minutes post-injection of active anirnals the cesium content of blood was lower than in any tissue (Table I). Tissue systems concentrating cesium were efiective at body temperatures of roo and 27" C. (r) The TI of heart tissue six hours after injection of hypothermic and hibernating animals was higher than the heart index of active animals sacrificed six hours after injection, p values being o.or or less. The blood flow to the heart and the cesium concentrating mechanism in heart tissue were temperature independent relative to blood flow and concentrating mechanisms in other tissues. (d) Heart tissue of animals active at the time of injection took up cesium-r37 faster than skeletal muscle. This
difference may be due to the greater blood flow per gramme of heart tissue; however, cardiac muscle appeared to have a cesium concentrating system which was not present in skeletal muscle, or at least was not as rapid in skeletal muscle. (e) Tissues in the order of decreasing affinity for cesium were kidney, heart and liver (Table III). This order was not changed by lowering the body temperature prior to injecting cesium-r37.

Cesium-r37 Excretion Metabolic rate and water turnover are key factors determining renal activity; if they increase in rate so does renal excretion. Both of these factors are changed during hibernation. Therefore, the effects of cold exposure and limited water intake were examined. The rates of excretion of cesium-r37 by eight rats exposed to temperatures of roo and zooC are presented in Fig. 3. The slope at zooC (m : o.o3o9) compares favourably with the slope (zz : o.ozg7) reported by Richmond (1958). At ro"C the rate of excretion is considerably faster than at zooC. Retention curves obtained from eight ground squirrels exposed to temperatures of rooC, however, were similar to those at zooC

CESIUM-r37 IN }TTELLUS LATERALIS TeeI,n (n

223

Mean tissue indices of cesium-r37 in active ground squirrels

6 at each time designated)

Time After fnjection

I
Blood
s.d. Plasma s.d.

mxn

3 mln

15 mtn

rhr
O'I I

Jhr
o'17 o'05
O.I I

6hr
o'I2
o'o6 o'07

rz hr
o'r
8

z4 hr o'20

ro8
o'30

r't6
o'36

r'64
o'96 3'99
I.IO

r'72
o'63 s'67
5 Jl

Kidney
s.d.

Heart
s.d.

r'53
o'43 o'27

r'9+
o'65

o'27 o'09 o'53 o'25 r6'26 3'o8 z'69 o'50 o'46 o'r 5 o'48

o'o5 o'20

o'ro
o'30 o'24

o'ro
O'I I o'r o

o'r5
8'42 z'84

o'r2
5'83

o'o4

4'r8
I'IO

r'70
3'80 o'50

r'36

3'r8
o'57 2'58 o'26

z'89
o'33

r05

3'$

2'55

I.76
o'17

Right
Gastrocnemius
s.d.

o'4r
o'o8 o'38

o'r9
o'32

o'62 o'30 o'65

o'35

r'oo
o'17
I.II

o'98 o'69 o'77 o'53 z'26

r'29
o'23

r'93
o'73

Left
Gastrocnemius
s.d.

r'30
o'r 8 2'39 o'36 o'65

o'r+
r'92
o'3 r

o't2
r'87
o'54

o'r4
z'88 o'56

Liver
s.d.

3'3r r'o5
r'43 o'64 r'65

o'28 3'42 o'32

r'79
o'34
I

Lung
s.d. Spleen s.d.

r'49
o'42

r'30
o'33

r'oo
o'34

r'54
o'23

I'02 t'20
o'57

t'28
o'r7
2'09 o'50
2'25

'I3

o'2\.

2'4r r'72
z'63

r'56

o'99
o'5 r

r'76
o'45 o'46
5'3 r

t'20
o'83

r8z
o'98
2'13

lo9
r'54
o'67 5'44 3'48 o'50 o'o8 o'37 o'25

Stomach
s.d.

r'73
o'97

r'23 r'74
o'76 7'26
3'83
o'3 r

r'55
a'67 3'09

r'54
5'59 2'36

o'84 o'67
o'6 r

o'r 6
2'65

Intestine
s.d.

5'7r r'85
o'42 o'r o o'48 o'22

r'53
o'43 o'r 6 o'69 o'30

r'37

Skin
s.d.

r'65 r'24
o'28 o'12

o'4r
o'25

o'2+ o'56

Femur
s.d.

o'17 o'48

o'25

o'r9

o'68 o'39

y07
o'24

o'r8
o'69 o'38

(Fig. +). Apparently rats in air temperature of rooC have an increased metabolic rate, whereas rooc did not represent sufficient thermal stress for ground squirrels to effect an increase in metabolic rate; so no change in the percentage retained at rooC could be observed over that at zo"C. The rate of cesium-r37 excretion by four ground squirrels deprived of drinking water, and four animals with water intake limited to five millilitres per z4 hours, was slower than the excretion rate by control animals with water ad lib. (Fig. S). Obviously limited water intake greatly delays excretion and so can be an important factor determining the characteristics of cesium-r37 metabolism during hibernation. At this writing, we do not have sufficient data to describe the overall efiect of hibernation on cesium-r37 excretion. However, the slope of retention curves (log per cent retention against time in days) for four animals in hibernation from four to 14 days ranged from o'oo7 to o'o5r,

22+

RIEDESEL ET AL.

;) t: sl
t:
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H@O

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CESIUM-I37 IN CITELLUS LATERALIS

225

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226

RIEDESEL ET AL.

TesI,n

II

Mean tissue indices of cesium-t37 in ground squirrels sacrificed

during hypother mia and hib ernation

(n

6 at each time designated)

Time After Injection

Animals in Hypothermia

Animals in Ilibernation
IO MLN

ro min s I Blood
s.d.

hr

Itzhrlz+hr
o'ro I o'ro
o'o+ | o'o8 z'o6 I z.sz
3'14 I 3's+ lr8 | r'53
I

3hr 6hr tzhr

o'38 o'43

e4

hr

Kidney
s.d.

Heart
s.d.

Right
Gastrocnemius
s.d.

o'29 lo'2r1o.25 o'ro lo'16 lo.z: o'8o lo'sols's+ o'43 lo'4ol3.tt o'4o lo.zal.5.8o o'3+ lo.o9 jz.to

,'27 |

,'r3

o'65 o'45 3'44 z'16

o'2t
o'r 9

o'r4 o'23
o'o9

o'r8
4'45 o'35

r'97
2'54 o'85
o'93

6'ss

Left
Gastrocnemius
s.d.

o'o4 lo.rrlo.23 o.3r o-42 I o'o3 lo'o7io.r9 o'r -5 I o'r 3

tt

r'r7
o'47
o'25

\'67
4'oo o'99

5'4r r'57
4'36

r'84 o'62

Liver
s.d.

Lung
s.d. Spleen s.d.

Stomach
s.d.

Intestine
s.d.

Skin
s.d.

Femur
s.d.

lo.z4 | r.r5 r'o7 ) r.o2 o'2o lo.tglo.Tz o'4o I o'6r 6'o+ l+'s6)s'+, z'35 | r'8o v76 lt.solz.t+ rr5 o'7o o'r8 I o'zol o.z6 o'25 )l o.32 o'ro lo.r3 lo.r, o'o8 | o.o5 o'o9 lo.z8]o.r7 o'15 o.z8 o'r7 lo.oalo.ra o'o4 || o.17

o'3r

o'52 lo'3zlt.5t o'48 lo.zZlr.+6 ror r.r4 o'r3 lo'r+lo.+q o'23I o.3-5 o'4r lo.sslo.ss o'62 | o'69 o.3o lo.ztlo.ro o'3o o.z7 I

o'rr lo'rr o.z6 o'o6 lo'o3 jo.r, ro3 lo.5olz.a8

tt

o'r9 o'r6
o'r 6 o'89 o'66
O.I I

o'ro o'39 o'45 o'5 r o'ro o'23 o'17 o.ro

o'07
O'I I

| | o'r -5 z'ry | 167 o'88 | o'38 o'r

-5

o'38 o.4r

o'32

o'36

o'43

o'r8 o'r4 o't2 I. I+ 3'r7 4'73 4'02

r'33

o'53

I.4I r'40 r'55 I.5I r'37 r'21

o'40

r'20
o'55
o'8 r

o'37 4'38 o'95 o'52 o'44 o'25 o'24

o'46 o'26 o'3 r o'39 o'3 r

o'42 o'90
o'3 r

o'20

I05
o'49
I .IO

r'30
o'+7

r'44 r'2+

r'36
o'49 3'20
o'95

r'45
2'45 o'07 o'o6 o'r 3 o'28

o'54 o'35 3'65 5'gr

r'99
o'r
5

o'35 o'r 8
o'3 3 o'3 5

o'4r o'33
o'09

o'r9 o'29 o'r3 o'r 5

Teslr III
Values and timing of highest tissue indices

(Taken from Tables

Highest Tissue Index Hvbo.

and

II)
Hypo.
IO mln

Time of Highest Tissue fndex

Active
o'65

Hiber.

Blood Kidney
FIeart

r'r6
t6'26

Liver Lung

3'r6
3+-

o'29 5'54 5'80 2'48

6'ss
4'36
a tJ

3 min
15

rhr

min

r'54

r'46

I.5I

3hr 3hr

6hr 6hr 6hr 6hr

ro min

6hr

rz hr rz hr

6hr

CESIUM-I37 IN CITELLUS LATERALIS

= c c)
E.
6)

/1

m:-o.o3o9

(zo"c)

c q) o
oq)

o J

ci)

Doys After lnjection

Fig. 3. Cesium-r37 retention by rats at roo and zooC following a single intraperitoneal injection (n

8 at each air temperature).

whereas the slope of retention curves for ten active animals during the same time period ranged from o'o87 to o'14 which replesents a twenty-fold decrease in excretion rate during hibernation.

Cesium Distribution after rst, 2nd, 3rd, and 4th Biological Half Times The tissue retention indices (TRI) of skeletal muscle and skin increased with time, rst, znd, 3rd, and 4th biological half times, whereas the indices of other tissues were unchanged or decreased with time (Fig. 6). The long ternr retention of cesium-47 by skeletal muscle confirms reports on other animals (Richmond et al. 1962, Comar and Wasserman 1956). The increased activity of skin may be due to contamination from excreta on
cages.

With the exception of heart tissue, the TRI values of active and hibernating animals were similar (Fig. 6). The TRI of heart tissue from hibernating animals was higher than the TRI of active animals at all three biological half times, p values being o'o5 or less. The increase in TRI of heart tissue may result from this tissue picking up cesium-r37 which has been released into the blood by other tissues because, as noted above, heart tissue was very effective in concentrating cesium-r37 when body temperature was ro"C at the time of injection. It is important to recall that these animals were active at the time of injection of the cesium-r37 and for part of the time interval between injection and sacrifice. They had been in continuous hibernation a minimum of two days prior to sacrificing. A gross examination of the data presented here and in the literature suggests that the sequence of events following a single intraperitoneal injection of cesium-r37 in active animals was as follows:
MHQ

228

RIEDESEL ET AL,

.9
o) a)

E.
C
(J
(lJ
@,

o-

2 S.D.

o
--.,

or

Doys After lnjection

Fig. 4. cesium-r37 retentionby citelrus rateraris at roo and zooc following intraperitoneal injection (n : 8 at each air temperature).
a

8rot2

single

No Woter; m'-0.o!Bg
c

.9
(l' E.

ll'

ml Woler/24hrs; rn=-o.o6re

c (D
o
q,

o-

-J

cl,

Wsler od

libl 6=-9.9926

Doys After lnjection

Fig. 5. Cesium-r37 retention by Citellus lateralis varied by water intake (n
each group).

4 in

CESIUM-I37 IN CITELLUS LATERALIS

229

diffusion from the site of injection into capillaries and general circulation; z. rapid uptake of cesium by kidney tissue, highest TI 15 minutes after injection; high rate of excretion of cesium by kidney, particularly during the 3. first z4 hours after injection (Fig. 3, 4 and 5) (Richmond et al. ry62); concentration of cesium by other body tissues with highest TI 4. occurring at different times (heart, one hour; intestine, one hour; liver, three hours; stomach, three hours; etc.) (Tables I, II and III); 5. the affinity of cells for cesium keeps the concentration in the blood very low with the highest TI for blood occurring within three minutes after injection ; 6. loss of cesium from tissues via blood and urine is very slow, and the rate of loss by muscle tissue and bone is slowest as the TRI values keep increasing past the rst, znd, and 3rd biological half times.
These observations suggest that uptake and retention of cesium-r37 are dependent upon effective circulation as well as cellular processes concerned with active transport and binding of cesium. The maintenance of homeostasis during hibernation can be expected to be dependent upon the
same systems.

r.

DISCUSSION

The TI changes which occur during hypothermia and hibernation, particularly the TI values for blood, kidney, cardiac and skeletal muscles, liver, and lung. Cesium-r37 appears to be present in blood as a free (unbound) electrolyte. This is indicated by (a) the rapid uptake of cesium from blood by kidney and other tissues, and (D) the blood having consistently lower TI and TRI values than any other tissue except during the first few minutes after injection. A comparison of the highest TI values in the blood of active animals (at three minutes) and in that of hypothermic and hibernating animals (at ten minutes), indicates that the rate of blood flow to the site of injection of cesium (i.e., peritoneal cavity) is decreased by a factor of at least three during hypothermia and hibernation. The circulatory adjustments during hypothermia and hibernation appear to be similar judging from the blood data and the time of appearance of the peak radioactivity in tissues. With the exception of the intestine, which is probably contaminated,
the kidney appears to have the highest TI values of any tissue under nearly all conditions, suggesting that the kidney is concentrating cesium by some mechanism, e.g., active transport. If the magnitude of the peak TI values can be used to indicate the rate of activity of this active transport system, then comparing the active to hypothermic and to hibernating animals the 9ro of the process is r'5 and r'2, respectively. On the other hand, the small urine production characteristic of hibernation implies reduced blood

Interpretation of TI Data data have been used to interpret circulatory

230

RIEDESEL ET AL.

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RIEDESEL ET AL,

during hypothermia and hibernation. A favourable electrochemical gradient in heart tissue as described by Ling (1962, 1965) would represenr
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kidney. The authors are of the opinion that both a cellular active transport system and circulation of blood are determinants in the cesium-r37 upiake by kidney tissue. - There are two points regarding metabolism of cesium-r 37 by heart tissue: (a) the TI of the heart is always higher than the Ti of .k.l"tul Tyt"l" during the fi.rst rz hours after injectiin (Tables I and II; Fig. r). (6) The TI of heart tissue reached a higher value when the cesium was injected into hypothermic and hibernating animals (Tables I and II; Fis. r) than when injected into active animals. The higher tissue indices of heart tissue than of other tissues may simply be a result of greater blood flow, and the higher values in hypothermia and. hibernation than in the active animal may result from the fact that the decrease in blood flow to the heart is less than the decrease in blood flow to other tissues during hypothermia and hibernation. In other words, the blood flow to the heart is reduced in hypothermia and hibernation; but the decrease to heart tissue is less than the decrease to other tissues. It takes approximately six hours for the heart TI to reach its peak in hypothermra u"d hibernation, whereas it takes three hours in active animals. These data indicate a twofold decrease in circulation time from the peritoneal cavity to the heart. However, the blood rI data demonstrated a three-fold decrease in circulation to the peritoneal cavity. Therefore heart tissue must have a favourable.electrochemical gradient for concentrating cesium-r37 which may be facilitated by cooling. At least the mechanism taking ,rp ce.inm-r3i in heart tissue is less sensitive to lowering temperature than the mechanism operative in other tissues thereby accounting for higher TI of heart tissue

flow to the kidney, and supports the hypothesis that the lower tissue indices of kidney tissues during hypothermia and hibernation are the result of the effect of temperature on effective blood flow to the kidney. The time required to reach a peak rI in the kidney in the active animal (r5 min, Tib,l: I) indicates a twenty-fold increase over that in the hypothermic and hibernating animal (6 hr, Table II) in the time required to circulate cesium from the peritoneal cavity to the kidney. The blood data indicated a three-fold decrease in circulation to the peritoneal cavity; therefore, there was a six- to seven-fold decrease in thelcirculation to the

time required to reach the highest tissue indic"r indicut".l two-fold

electrochemical gradient which facilitates uptake of cesium and is temperature independent v'ould be consistent with both the TI and TRI data. Liver cells do not appear to have a particular affinity for cesium-r37. The difference in the time required to reach the highest fI in active (th;;e hours), hypothermic (six hours), and hibernating (six hours) indicates that the blood flow to the liver is reduced by a factor of two during hypothermia and hibernation. The TI values obtained on lung tissue suggest there is no particular cesium-concentrating or -retaining system operating within lung iells. The

temperature independent electrolyte concentrating mechanism. An

CESIUM-i37 IN CITELLUS LATERALIS

decrease

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in effective blood flow to lung tissue during hypothermia

and

hibernation.

The data presented on stomach, intestine, spleen, bone and skin are difficult to interpret for several reasons. Residual food material within the digestive tract would have a marked effect on the TI and TRI values. No attempt was made to remove contents of stomach or intestine as such a procedure may wash out cesium-r37. The size of the spleen, o.r to o.3 g, was near the limit of the weighing procedure. Changes in cesium-r37 content of bone are not apparent except on a long-term basis because of the low metabolic rate of bone. Contamination of cages, fur, and skin by excreta complicates the skin data. Future studies should clarify cesium-r37 metabolism in these tissues and in nerve tissue.

Interpretation of TRI Data Initial uptake of cesir.rm is determined by circulation, diffusion and cell
membrane characteristics whereas long term retention (at rst, znd and 3rd biological half times) involves these and possibly other mechanisms which retain cesium within subcellular compartments. Typical subcellular components which may be involved in retaining cesium include: cell organelles, cell proteins or some cell metabolites such as amino acids which have an

aflftnity for electrolytes (cesium). The concentration of potassium in mitochondria is higher than in other celL parts (Price et al. ry56), but there is no evidence that mitochondria or other cell organelles concentrate
cesium.

The TRI of heart tissue was the only TRI value changed (p value of o'o5 or less) by 48 hours of hibernation at the various biological half times

(Fig. 6). Earlier reports on cesium metabolism (Ballou and Thompson

1958, Richmond 1958) have explained skeletal muscle retention of cesium by protein binding. Frotein binding may very well account for the long turnover time of cesium in skeletal muscle, but the differences between the cardiac and skeletal muscle TRI data are most likely related to differences in the characteristics of the cell membranes and of blood flow to these tissues. With the exception of respiratory pigments and enzyme concentrations the proteins within these tissues are similar, therefore the intraceliular protein binding of cesium-r37 in cardiac and skeletal muscle should be similar. The increased cesium-r37 in the hibernating heart may be explained as foltrows: (a) the heart cell systerr for concentrating cesiumr37 is temperature independent relative to the system operating in other tissues, (6) a high blood flow to the heart ensures that heart cells could pick up cesium-r37 released from any tissue. With the exception of heart tissue, hypothermia and hibernation did not change the TRI of the tissues examined at the rst, znd, and 3rd biological half times. These findings support the hypothesis that a temperature independent process, such as protein binding is the primary factor determining retention of cesium-r37 by body tissues. The similarities between the chemical characteristics of potassium and cesium are readily recognized (Finstone and Kinsley 196r); however,

23+

RIEDESEL ET AL.

Comar and Wasserman (1956) have pointed out numerous examples of differences in the biological metabolism of potassium and cesium. Application of findings and hypotheses set forth in this report to potassium metabolism during hypothermia and hibernation may be helpful in the design of experiments regarding potassium, but additional application would be tenuous.

SUMMARY
Water intake and environrriental as well as body temperatures can effect marked changes in cesium-r37 metabolism. The distribution of cesium-q7 among body tissues following a single intraperitoneal injection demonstrates the following: (a) animals made hypothermic by artificial methods had a cesium-r37 metabolism similar to that in animals hibernating during, and immediately following, the time of injection; (6) lowering of body temperature slows down the rate at which cesium-r37 is distributed among body tissues; (c) heart tissue uptake of cesium-r37 is less temperature dependent than uptake by other tissues; (d) the kidney uptake of cesium apparently involves a concentrating system as well as changes in blood flow; and (e) hibernation at the rst, 2nd, and 3rd biological half times did not effect a major change in the distribution of cesium-r37 among body tissues with the exception of an increase in the amount of
radio-isotope in cardiac muscle.

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