Circadian Rhythm and Response to an Acute Stressor of Urinary Corticosterone, Testosterone, and Creatinine in Adult Male Mice

Abstract

In small laboratory species, steroid measures can be obtained more frequently and less invasively from urine than blood. Insofar as urinary levels reflect systemic levels, they could provide advantages particularly for measurement of glucocorticoids, whose blood levels react rapidly to handling and stress. In Experiment 1, urinary samples were collected from male mice every second hour over a 14:10 h light:dark cycle. Samples were analyzed via enzyme immunoassay for corticosterone, testosterone, and creatinine. Corticosterone had peak concentrations 1 h after light offset and a trough 1 h after light onset. Testosterone showed peak concentrations 5–7 h after light onset and lowest concentrations during the dark phase of the cycle. Creatinine showed some variation over the light-dark cycle, but steroid measures showed similar trends with and without adjustment for creatinine. In Experiment 2, mice were stressed via an injection at times close to the determined peak and trough levels of corticosterone. In urinary samples taken 90 min after injection, corticosterone was significantly higher in injected animals at both times relative to levels in control animals, but testosterone was unaffected by injection stress. In Experiment 3, serum and urine samples were collected from mice every sixth hour across the diurnal cycle. Corticosterone peaked in urine and serum immediately after light offset, and urinary measures predicted those in serum. These data indicate that urinary corticosterone reflects systemic levels in mice, document circadian variation in urinary testosterone, and indicate that circadian variation in creatinine is minimal, but potentially relevant in stressed animals.

Key words

corticosterone - testosterone - creatinine - urine - circadian rhythm - stress

• References

• 1 Fediuc S, Campbell JE, Riddell MC. Effect of voluntary wheel running on circadian corticosterone release and on HPA axis responsiveness to restraint stress in Sprague-Dawley rats. J Appl Physiol 2006; 100: 1867-1875
• 2 Gartner K, Buttner D, Dohler K, Friedel R, Lindena J, Trautschold I. Stress response of rats to handling and experimental procedures. Lab Anim 1980; 14: 267-274
• 3 Sapolsky RM, Romero LM, Munck AU. How do glucocorticoids influence stress? Integrating permissive, suppressive, stimulatory, and preparative actions. Endocr Rev 2000; 21: 55-89
• 4 Zimmermann E, Critchlow V. Effects of diurnal variation in plasma corticosterone levels on adrenocortical response to stress. P Soc Exp Biol Med 1967; 125: 658-663
• 5 Faiman C, Winter JSD. Diurnal cycles in plasma FSH, testosterone and cortisol in men. J Clin Endocr Metab 1971; 33: 186-192
• 6 Halberg F, Peterson RE, Silber RH. Phase relations of 24-hour periodicities in blood corticosterone, mitoses in cortical adrenal parenchyma, and total body activity. Endocrinology 1959; 64: 222-230
• 7 Kakihana R, Moore JA. Circadian rhythm of corticosterone in mice: The effect of chronic consumption of alcohol. Psychopharmacologia 1976; 46: 301-305
• 8 Leal AMO, Moreira AC. Daily variation of plasma testosterone, androstenedione, and corticosterone in rats under food restriction. Horm Behav 1997; 31: 97-100
• 9 MacAdam WR, Eberhart RJ. Diurnal variation in plasma corticosteroid concentration in dairy cattle. J Dairy Sci 1972; 55: 1792-1795
• 10 Malisch JL, Breuner CW, Gomes FR, Chappell MA, Garland Jr T. Circadian pattern of total and free corticosterone concentrations, corticosteroid-binding globulin, and physical activity in mice selectively bred for high voluntary wheel-running behavior. Gen Comp Endocr 2008; 156: 210-217
• 11 Plant TM. Time courses of concentrations of circulating gonadotropin, prolactin, testosterone, and cortisol in adult male rhesus monkeys (Macaca mulatta) throughout the 24h light-dark cycle. Biol Reprod 1981; 25: 244-252
• 12 Waite E, Kershaw Y, Spiga F, Lightman SL. A glucocorticoid sensitive biphasic rhythm of testosterone secretion. J Neuroendocrinol 2009; 21: 737-741
• 13 Balcombe JP, Barnard ND, Sandusky C. Laboratory routines cause animal stress. J Am Assoc Lab Anim Sci 2004; 43: 42-51
• 14 Tuli JS, Smith JA, Morton DB. Corticosterone, adrenal and spleen weight in mice after tail bleeding, and its effects on nearby animals. Lab Anim 1995; 29: 90-95
• 15 Armario A, Montero JL, Balasch J. Sensitivity of corticosterone and some metabolic variables to graded levels of low intensity stresses in adult male rats. Physiol Behav 1986; 37: 559-561
• 16 Bronson FH. Establishment of social rank among grouped male mice: relative effects on circulating FSH, LH, and corticosterone. Physiol Behav 1973; 10: 947-951
• 17 Seggie JA, Brown GM. Stress response patterns of plasma corticosterone, prolactin, and growth hormone in the rat, following handling or exposure to novel environment. Can J Physiol Pharm 1975; 53: 629-637
• 18 van Herck H, Baumans V, de Boer SF, van der Gugten J, van Woerkom AB, Beynen AC. Endocrine stress response in rats subjected to singular orbital puncture while under diethyl-ether anaesthesia. Lab Anim 1991; 25: 325-329
• 19 Van Loo PLP, Mol JA, Koolhaas JM, Van Zutphen BFM, Baumans V. Modulation of aggression in male mice: Influence of group size and cage size. Physiol Behav 2001; 72: 675-683
• 20 deCatanzaro D, Muir C, Beaton E, Jetha M, Nadella K. Enzymeimmunoassay of oestradiol, testosterone and progesterone in urine samples from female mice before and after insemination. Reproduction 2003; 126: 407-414
• 21 deCatanzaro D, Muir C, Beaton EA, Jetha M. Non-invasive repeated measurement of urinary progesterone, 17β-estradiol, and testosterone in developing, cycling, pregnant, and postpartum female mice. Steroids 2004; 69: 687-696
• 22 Muir C, Vella ES, Pisani N, deCatanzaro D. Enzyme immunoassay of 17β-estradiol, estrone conjugates, and testosterone in urinary and fecal samples from male and female mice. Horm Metab Res 2001; 33: 653-658
• 23 Claustrat B, Valatx JL, Harthé C, Brun J. Effect of constant light on prolactin and corticosterone rhythms evaluated using a noninvasive urine sampling protocol in the rat. Horm Metab Res 2008; 40: 398-403
• 24 Hausler A, Hauser K, Meeker JB. Effects of subchronic administration of psychoactive substances on the circadian rhythm of urinary corticosterone excretion in rats. Psychoneuroendocrinology 1985; 10: 421-429
• 25 Goodman RL, Hotchkiss J, Karsch FJ, Knobil E. Diurnal variations in serum testosterone concentrations in the adult male rhesus monkey. Biol Reprod 1974; 11: 624-630
• 26 Hoffman K, Nieschlag E. Circadian rhythm of plasma testosterone in the male Djungarian hamster (Phodopus sungorus). Acta Endocrinol-Cop 1977; 86: 193-199
• 27 Kalra PS, Kalra SP. Circadian periodicities of serum androgens, progesterone, gonadotropins and luteinizing hormone-releasing hormone in male rats: the effects of hypothalamic deafferentation, castration and adrenalectomy. Endocrinology 1977; 101: 1821-1827
• 28 Keating RJ, Tcholakian RK. In vivo patterns of circulating steroids in adult male rats. I. Variations in testosterone during 24- and 48-hour standard and reverse light/dark cycles. Endocrinology 1979; 104: 184-188
• 29 Lucas LA, Eleftheriou BE. Circadian variation in concentrations of testosterone in the plasma of male mice: A difference between BALB/cBy and C57BL/6By inbred strains. J Endocrinol 1980; 87: 37-46
• 30 Rowsemitt CN, Berger PJ. Diel plasma testosterone rhythms in male Microtus montanus, the Montane vole, under long and short photoperiods. Gen Comp Endocr 1983; 50: 354-358
• 31 Sharma OP. Diurnal variations of plasma testosterone in stallions. Biol Reprod 1976; 15: 158-162
• 32 Munro CJ, Stabenfeldt GH, Cragun JR, Addiego LA, Overstreet JW, Lasley BL. Relationship of serum estradiol and progesterone concentrations to the excretion profiles of their major urinary metabolites as measured by enzyme immunoassay and radioimmunoassay. Clin Chem 1991; 37: 838-844
• 33 deCatanzaro D, Khan A, Berger RG, Lewis E. Exposure to developing females induces polyuria, polydipsia, and altered urinary levels of creatinine, 17β-estradiol, and testosterone in adult male mice (Mus musculus). Horm Behav 2009; 55: 240-247
• 34 Khan A, Berger RG, deCatanzaro D. The onset of puberty in female mice as reflected in urinary steroids and uterine/ovarian mass: Interactions of exposure to males, phyto-oestrogen content of diet, and ano-genital distance. Reproduction 2008; 135: 99-106
• 35 Meijer MK, Lemmens AG, Van Zutphen BFM, Baumans V. Urinary corticosterone levels in mice in response to intraperitoneal injections with saline. J Appl Anim Welf Sci 2005; 8: 279-283
• 36 Touma C, Sachser N, Möstl E, Palme R. Effects of sex and time of day on metabolism and excretion of corticosterone in urine and feces of mice. Gen Comp Endocr 2003; 130: 267-278
• 37 Van Cauter E, Leproult R, Kupfer DJ. Effects of gender and age on the levels and circadian rhythmicity of plasma cortisol. J Clin Endocr Metab 1996; 81: 2468-2473
• 38 Brennan FX, Ottenweller JE, Seifu Y, Zhu G, Servatius RJ. Persistent stress-induced elevations of urinary corticosterone in rats. Physiol Behav 2000; 71: 441-456
• 39 Plymate SR, Tenover JS, Bremner WJ. Circadian variation in testosterone, sex hormone-binding globulin, and calculated non-sex hormone-binding globulin bound testosterone in healthy young and elderly men. J Androl 1989; 10: 366-370
• 40 Veldhuis JD, King JC, Urban RJ, Rogol AD, Evans WS, Kolp LA, Johnson ML. Operating characteristics of the male hypothalamo-pituitary-gonadal axis: Pulsatile release of testosterone and follicle-stimulating hormone and their temporal coupling with luteinizing hormone. J Clin Endocr Metab 1987; 65: 929-941
• 41 Mock EJ, Norton HW, Frankel AI. Daily rhythmicity of serum testosterone concentration in the male laboratory rat. Endocrinology 1978; 103: 1111-1121
• 42 Kinson GA, Liu CC. Diurnal variation in plasma testosterone of the male laboratory rat. Horm Metab Res 1973; 5: 233-234
• 43 Wilson MJ, McMillin JM, Seal US, Ahmed K. Circadian variation of serum testosterone in the adult male rat with a late morning acrophase. Cell Mol Life Sci 1976; 32: 944-945
• 44 Hostetter MW, Piacsek BE. Patterns of pituitary and gonadal hormone secretion during a 24 h period in the male rat. Biol Reprod 1977; 16: 495-498
• 45 Bartke A, Steele RE, Musto N, Caldwell BV. Fluctuations in plasma testosterone levels in adult male rats and mice. Endocrinology 1973; 92: 1223-1228
• 46 Ellis GB, Desjardins C. Male rats secrete luteinizing hormone and testosterone episodically. Endocrinology 1982; 110: 1618-1627
• 47 Armario A, Castellanos JM. A comparison of corticoadrenal and gonadal responses to acute immobilization stress in rats and mice. Physiol Behav 1984; 32: 517-519
• 48 Bliss EL, Frischat A, Samuels L. Brain and testicular function. Life Sci 1972; 11: 231-238
• 49 Demura R, Suzuki T, Nakamura S, Komatsu H, Odagiri E, Demura H. Effect of immobilization stress on testosterone and inhibin in male rats. J Androl 1989; 10: 210-213
• 50 Charpenet G, Tache Y, Bernier M, Ducharme JR, Collu R. Stress-induced testicular hyposensitivity to gonadotropin in rats. Role of the pituitary gland. Biol Reprod 1982; 27: 616-623
• 51 Marić D, Kostić T, Kovačević R. Effects of acute and chronic immobilization stress on rat leydig cell steroidogenesis. J Steroid Biochem 1996; 58: 351-355
• 52 Beaton EA, Khan A, deCatanzaro D. Urinary sex steroids during sexual development in female mice and in proximate novel males. Horm Metab Res 2006; 38: 501-506
• 53 Jäntti SE, Kiriazis A, Reinilä RR, Kostiainen RK, Ketola RA. Enzyme-assisted synthesis and characterization of glucuronide conjugates of neuroactive steroids. Steroids 2007; 72: 287-296
• 54 Vella ES, deCatanzaro D. Novel male mice show gradual decline in the capacity to disrupt early pregnancy and in urinary excretion of testosterone and 17β-estradiol during the weeks immediately following castration. Horm Metab Res 2001; 33: 681-686