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Comp. Biochem. Physiol.,7971, VoL 38A, pp.727 to 727. Pergamon Press.

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RESPIRATORY QUOTIENT AS AN INDEX OF SELECTIVE TISSUE CATABOLISM BY WATER-DEPRIVED LABORATORY RATS AND SPERMOPHILUS LATERALIS
GARY L. BINTZ, LORELEI B. BINTZ and MARVIN L. RIEDESEL*
Department of Biology, Eastern Montana College, Billings, Montana 59101
(Receioed 30

June 1970)

Abstract-1, Respiratory quotients and

observations of dissected animals indicated that during periods of water deprivation laboratory rats selectively catabolized fat tissue whereas ground squirrels in summer and thin ground squirrels in winter selectively catabolized skeletal muscle. Under similar conditions fat squirrels in winter appeared to selectir-ely catabolize fat and muscle
at different times.

2. Catabolism of muscle results in net water gain for an animal during homeothermia. Reduced insensible water loss during hypothermia allows net water gain from catabolism of fat.
INTRODUCTION

al. (1964) and Bintz & Riedesel (1967) noted that several species of ground squirrel (Spermophilu.s) capable of hibernation are more tolerant to drinkingwater deprivation than rnany similar-sized rodents incapable of hibernation. Bintz (1965) reported (i) ground squirrels lost less weight during drinking-water deprivation in fall and winter than in spdng and summer, and (ii) fat ground squirrels lost less weight during water deprivation than thin squirrels. A possible means of coping with reduced water intake is selective tissue catabolism, as tissue catabolism makes free water and metabolic water available. Most tissues have a high content of free water. Fat tissue contains little free water, but yields large amounts of metabolic water. With limited food and no water intake, the amount of water available for essential body processes can vary considerably depending upon which tissue is catabolized. In an earlier study (Bintz, 1968) laboratory rats and Spermophilus lateralis underwent 5-12 days of continuous drinking-lvater deprivation. Dissection demonstrated that water-deprived laboratory rats contained less intraperitoneal white fat tissue, whereas ground squirrels contained less skeletal muscle than
Rrnnnsar, et

control animals. Thus, during water deprivation laboratory rats appeared to preferentially catabolize white fat tissue and S. lateralis appeared to preferentially
catabolize skeletal muscle tissue.

* Department
87106.

of Biology, University of New Mexico, Albuquerque, New

Mexico

t2l

722

Ganv L. BrN:rz, Lonnr-nr B. Brwtz aNo MaRvrN L. Rrnorssr,

The present study employs respiratory quotient (RQ) measurements to substantiate the above observations, namely preferential tissue catabolism by starving, water-deprived laboratory rats and ground squirrels. In addition, it was desirable to study the effects of season and body fat content on the ability of ground squirrels to tolerate negative water balance. According to Hawk & Bergeim (1931) oxidation of carbohydrate, protein, and fat results in RQ's of 1'00, 0'80 and 0'70 respectively. The above values were determined in ztitro where variation in RQ of a given foodstuff would be slight. Respiratory quotients of animals are somewhat more variable. Under control conditions RQ approximates 0'83 because a mammal usually oxidizes a combination of fat and carbohydrate.
MATERIALS AND METHODS

One group (n : 6) of albino laboratory rats (Rattus norv^egicus) and four groups (n : 6 in each group) of golden-mantled ground squirrels (Spermophilus lateralis) were maintained in the laboratory (19-23"C, relative humidity 40-50%, 12 hr hghtlday) at least 2 months prior to experimentation. The four groups of ground squirrels were designated as summer-thin, summer-fat, winter-thin and winter-fat (Table 1). Respiratory quotients of summer squirrels and rats were measured in May and July and RQ's of r",.inter squirrels were measured in December. Thin squirrels were obtained by limiting food whereas fat squirrels were

obtained by allowing squirrels ad lib. food consumption. Animals were fed Wayne Lab Blox (7-8 per cent water content) throughout the study. TesLT 1-MEAN INITIAL BoDY
WEIGHTS AND MEAN peT cent WEIGHT LOSSES DURINC OF WATER DEPRIVATION BY LABORATORY RATS AND GROUND SQUIRRELS

10 dayS

Summer-thin
Rats

Summer-fat
squirrels
263

Winter-thin
squirrels
212 (7.18)
30.7

Winter-fat
squirrels
299 (1s.07)
72-O

squirrels
189

wt.(e) Wt. loss (%)

Initial

body

323* (9's9) 37'0


(0.66)

(8.0s)
45.0 (1.88)

(11.4e)
31.1

(2.4s)

(3.30)

(0.82)

* Mean. Standard errors in parentheses. The technique employed for RQ measurement was modified from Schmidt-Nielsen & Schmidt-Nielsen (1950). Oxygen was circulated by a pump from a spirometer through a closed system including an animal chamber (12 x 10 x 10 cm), a copper tube immersed in an ethanol-dry ice mixture to remove water vapor, a calcium chloride cartridge to remove possible remaining water vapor, and a soda lime cartridge to absorb carbon dioxide. Each test involved simultaneous measurements of oxygen consumption and carbon dioxide evolution by an animal for 30 min. Respiratory quotients of animals were measured prior to water deprivation, as each animal was to serve as its own control. Animals were then deprived of drinking water but were not food deprived. Subsequent measurements of RQ and body weight were made 7 , 3, 6 and 10 days after water deprivation. Data were analyzed statistically using t-tests of significance between two means. Differences at the 95 per cent
level of confidence r/ere considered significant.

RQ or warrn-DEpRrvED

ANTMALS

t23

RESULTS (a) Weight


loss

by raater-deprioed animals

When a rat or ground squirrel is deprived of drinking water, food consumption by the animal is decreased markedly. Consequently, the animal catabolizes body tissues for energy and weight is lost because of tissue catabolism and loss of water. A criterion of tolerance to water deprivation is rate of weight loss after water
deprivation.

Summer-thin squirrels lost weight more rapidly than laboratory rats, whereas the other three groups of squirrels iost weight less rapidly than rats (Table 1). In this study two factors, body fat content and season, have been observed to be important determinants of tolerance to drinking-water deprivation. Fat squirrels are more tolerant than thin squirrels to water deprivation and squirrels are more tolerant in winter than respective groups of squirrels in summer. When water deprived, winter-fat squirrels lose very little weight and their physical appearance indicated that they are capable of undergoing considerable periods of water deprivation without ill effects. The ability of fat squirrels to tolerate water deprivation in winter appears related to the tendency of these animals to become hypothermic under winter-fat

conditions. During hypothermia metabolic processes are slowed and rates of tissue catabolism and water loss are reduced. Water-deprived winter-fat squirrels entered hypothermia either frequently or chronically. Every squirrel was observed to be hypothermic at least once daily during all 10 days of the water-deprivation period and bouts of hypothermia occasionally lasted as long as 3 days. Body temperatures of squirrels in deepest hypothermia were approximately 1-3"C above room temperature (19-23'C). Hypothermia was not observed in summer-thin squirrels, and infrequentJy in summer-fat and winter-thin squirrels. Thus hypothermia, which would result in greater tolerance to drinking-water deprivation, appears to be restricted to winter-fat squirrels.
of I ab or at or y r at s and gr ound s quirr e ls Mean RQ's of all groups of control (day 0) animals were near unity (Table 2). Presumably animals were not oxidizing carbohydrate exclusively, but were oxidizing a combination of substrates while synthesizing some fat. Fat squirrels arrd rats were gaining weight at the time control RQ's were measured. Swift & Fisher (1964) indicated that synthesis of fat in conjunction with a combination of carbohydrate and fat oxidation may result in an RQ equal to or greater than unity. After
1

(b)

Re spir at ory quo t i ent s

day of water deprivation, the mean RQ's of most groups of animals had decreased.

Respiratory quotients (0'75)

of rats after 3 days of water deprivation indi-

cated fat tissue catabolism. Fat catabolism apparently continued until some time after day 6 (RQ : 0.73) of the water-deprivation period. On day 10 the mean RQ (0'82) of rats indicated muscle catabolism. Apparently the rats had consumed all available fat stores and were dependent upon protein oxidation. After RQ measurements on day 10, rats were dissected and there remained no visible fat stores.

12+

Ganv L. Brwrz, Lonrr-sr B. BrNrz aNn MenvrN L. RrconsrrRQ's oF LABoRAToRy


RATs AND GRoUND seUIRRBLS pRroR WATER DEPRIVATION

Tast,p 2-Mrer

To AND DURING

Dav

Rats

Summer-thin Summer-fat
1'06 (0.046) 1'03 0.96 (0.013)

Winter-fat
1.09 (0.110) 0.88 (0.010) 0.82

Winter-fat
7'07 (1.06e)

1.00x
(0.008) 0.87 (0.032) 0.75 (0.028) 0.73 (0.014)
10

0.8s
(0.030)

0'84
(0.028)

(0.0s0)
0.82

0'82
(0.016) 0.83

0'75
(0.021) 0.77

(0'014)
0.81

(0'024)
0.84 (0.014) 0.80 (0.011)

(0'01s)
0.84 (0.017)

(0'010)
0.83

(o.o2s)
0.77

0.82 (0.014)

(o'034)

(0.202)

* Mean. Standard errors in parentheses.

Respiratory quotients of summer-thin, summer-fat and winter-thin squirrels measured on days 3, 6 and 10 of the water-deprivation period indicated muscle

catabolism.

It is doubtful that muscle was exclusively

catabolized, but rather

muscle was preferentially catabolized, andwas accompanied by some fat catabolism.

This supposition was supported by visual observations during dissection of the animals. After 10 days of water deprivation intraperitoneal fat stores contained l*20 g of fat, with the greatest fat stores occurring in the summer-fat squirrels. In
all three groups of water-deprived squirrels skeletal muscle mass visually appeared much reduced compared to that of dissected control squirrels. Mean RQ's of water-deprived winter-fat squirrels were between 0'80 (protein oxidation) and 0'70 (fat oxidation) on days 3, 6 and 10 (Table 2). Respiratory quotients of individual squirrels on any given day indicated that some were catabolizing fat preferentially whereas other squirrels were catabolizing muscle preferentially (Table 3). In addition, the same squirrel did not appear to catabolize only protein or fat throughout the water-deprivation period. It is possible that winter-fat squirrels can catabolize muscle or fat tissue. Perhaps during hypothermia water-deprived squirrels catabolized fat tissue (fat oxidation during

Tesrr 3-RESrIRAToRv euorIENTS oF wINTER-FAT seuIRRELS oN DAys 3, 6 eNn


Squirrel
7

10 arrun

Day
0.67 0.73

Day
0'71
0.80 0.80

Day 10
0.70
0.80 0.77
0.81

2
.)

0'78
0.75

4
5

0.84
0.68 0.80

0'79
0.81

0.82

0'72

RQ on wATER-DErRr!'ED ANTMALS

725

hibernation is well documented) pieferentially, but during times at which body temperature is near 37'C muscle tissue is preferentially catabolized. When winterfat squirrels were disturbed in their cages just prior to measuring RQ, some lr,'ere hypothermic and others were normothermic. Each squirrel was normothermic when its RQ was measured.

Measurements

of

DISCUSSION RQ and observations on dissected animals indicated that

during water deprivation laboratory rats preferentially catabolized white fat tissue. Under similar conditions thin ground squirrels during both summer and winter and fat squirrels in summer appeared to catabolize skeletal muscle tissue preferentially, whereas winter-fat squirrels appeared to preferentially catabolize fat and skeletal muscle at different times. Fat tissue catabolism results in large amounts of metabolic water; however, catabolism of skeletal muscle results in
more water being available to the animal, primarily because skeletal muscle contains more free water than fat tissue. There are additional factors which affect the net amount of water available to an animal from catabolism of any tissue (Table 4). The net amount of water Tasr-r 4-WATER
BALANCE RESULTING FRoM cATABoLTsM oF 100 g

racH oF wHrrE
water produced
(e)
105.25

FAT TIssuE

Free water
(g)

Solids (ml)/solids

Metab.

water Water (ml)/ Total


total

(g)
75 25

(e)

(e)

solids

Fat Muscle

25 75

-ffi

7.O7*

80.25 10.00

85.00

l. 02
Fat
N4[uscle

required/ respiratory g solid HrO loss/I. O,


2.07* 0.95 *
H2O lost
0.941 0.941

Insensible

Insensible respiratory HrO lossltotal


solids

(s)
741.7 22.3

nitrogen

Urinary

Total water lost during


catabolism of solids
(e)
741.7 48.1

from N
excretion
(e)

Net
water
(e)

(e)
4.0

Fat Muscle

25.8

36'45 (lost) 36'90 (eained)

* B. Schmidt-Nielsen (1954).

Laboratory rat (B. Schmidt-Nielsen & K. Schmidt-Nielsen, 1950).

726

Genv L, BrNrz, LonnrBr B. BrNrz aNo ManvrN L. Rrnonssl

available from catabolism of a tissue is decreased by insensible respiratory water loss, which is considerable at 37'C body temperature and low ambient humidity. When body temperature, ambient humidity and depth of breathing are constant, the insensible respiratory water loss varies directly with oxygen consumption. Because large amounts of oxygen are required per mass of fat, oxidation of fat results in considerable insensible respiratory water loss. Consequently, the metabolic water formed from fat oxidation is negated by respiratory water loss. It may be hypothesized that a significant correlation exists between the type of

of the squirrel. Only winter-fat squirrels frequently became hypothermic, and only these squirrels had RQ's indicative of fat oxidation. Fat tissue catabolism would result in neutral or positive water balance only during hypothermia when insensible respiratory water loss is reduced. In fact, if insensible water loss were reduced from 0'94 (assumed) to less than 0'70 ml water per liter of oxygen consumed, the animal would realize a net gain in water from fat catabolism. At 37"C, however, ventilation is accompanied by high respiratory water loss and protein catabolism is essential to maintain water balance. Summer-thin, summer-fat, and winter-thin squirrels entered hypothermia infrequently or not at all, and RQ data indicated these squirrels were preferentially catabolizing muscle. Questions raised from the above discussion include (i) what controls the switch between fat catabolism and muscle catabolism ? and (ii) how rapidly can the switch from fat to muscle to fat catabolism occur ? In addition to body temperature, degree of tissue hydration may have an important effect in determining which tissue is selectively catabolized. In an earlier study during which laboratory rats and ground squirrels were water deprived until 30 per cent of initial body weight was lost, it was observed that all rat tissues analyzed were dehydrated, whereas only blood of squirrels was dehydrated (Bintz & Riedesel, 1967). Maintenance of proper tissue hydration may be critical, and therefore the degree of tissue hydration may serve as a stimulus for selective tissue catabolism. Such stimuli may operate through hormonal pathways such as glucocorticoids or growth hormone. The rapidity with which a switch from preferential catabolism of one tissue to another could occur is unknown and warrants further investigation. However, it is rn'ell known that induction of certain enzyme systems in some organisms may occur from within several minutes to a few hours. A comparable rate of induction of protein- and fat-catabolizing enzyme systems in squirrels would certainly be rapid enough to accompany daily variations between hypothermia and normothermia in ground squirrels.
tissue catabolized and the body temperature SUMMARY 1. Employing weight loss as a criterion, ground squirrels were more tolerant than laboratory rats to drinking-water deprivation. 2. Tolerance to water deprivation by squirrels was greatr in winter than in summer and varied directly with body fat content. Tolerance may be related to the ability of the animal to enter hypothermia.

RQ op

wATER-DEeRrVED

ANTMALS

127

3. Respiratory quotients indicated that water-deprived rats catabolized fat, squirrels in summer and winter-thin squirrels selectively catabolized muscle, and winter-fat squirrels selectively catabolized muscle or fat. 4. Data on respiratory water loss indicated that catabolism of muscle tissue results in more net water for the animal than catabolisrn of fat tissue.
Acknozaledgements-This study was supported by AEC Contract AT-29-2-2566 and NSF Grant GB-5339 granted to the Department of Biology of the University of New

Mexico, Albuquerque. Appreciation is expressed to the University of New Mexico for use
of required equipment.

REFERENCES

BtNrz G. L. (1965) Seasonal tolerance of Citellus tridecemlineatus, Citellus lateralis, and the laboratory rat to deprivation of drinking water. M. S. thesis, University of New Mexico. BrNrz G. L. (1968) Uptake and retention of tritiated water by tissues of laboratory rats and
Citellus lateralis. Ph.D. dissertation, IJniversity of New Mexico. BrNrz G. L. & Rrrorspr, M. L. (1967) Water content of ground squimel and laboratory rat tissues. C omp. Biochem. Physiol. 22, 7 5-80. FrNcrn R. W. and Rrro L. S. (1952) The effect of water deprivation and subsequent satiationupongeneralactivityintherat. J.comp.Physiol.Psychol.45,368-372. Hawx P. B. & Brncstla O. (1931) Practical Physiological Chemistry,lOth edn. P. Blakiston, Philadelphia.
RrroBsBr,

M. L., KI-INnstrvnn L. R. & BBNar,r,v N. R. (1964) Tolerance of Citellus lateralis andC. spilosomaforwater deprivation. Ann. Acad, Sci.fenn. Ser. A, IV,71,375-388. ScnNrror-Nrsr-ssN B. (1954) Water conservation in small desert rodents. In Biology of Deserts (Edited by Cr-ouosr,ry-TnolrpsoN J. L.), pp. 773-781. Institute of Biology,
London.

Scnlrror-NrplspN B. & Scnulor-Nrrr.snN K. (1950) Pulmonary water loss in desert rodents. Am. J. Physiol. t62, 31-36. Swrrr R. W. & FrsnBn K. H. (1964) Energy metabolism. In Nutrition, I: Macronutrients and Nutrient Elements (Edited by Brarox G. H. and McHnNnv E. W.), pp. 781-260.
Academic Press, New York.

Key Word Index-Ground squirrel; Spermophilus lateralis; water metabolism; rvater deprivation; respiratory quotient; tissue catabolism; selective tissue catabolism.