Radioimmunoassay of total and free corticosterone in rat plasma: Measurement of the effect of different doses of corticosterone

doi:10.1016/0039-128X(88)90056-6

Steroids

Volume 51, Issues 5–6, May–June 1988, Pages 609-622

https://doi.org/10.1016/0039-128X(88)90056-6 Get rights and content

Abstract

A radioimmunological method was developed for determining total and free corticosterone in rat plasma. This method was used to determine the dose-response curve of corticosterone and to measure the elimination and study the half-lives of total and free corticosterone in rat plasma with a dose of 5 mg/kg. The elimination with a dose of 5 mg/kg, when drawn on the half-logarithmic scale, formed a straight line. The half-lives for total and free corticosterone were 25 and 15 min, respectively.

References (34)

S Chattopadhyay et al.
Rat serum corticosterone assay: use of radiosteroid assay method
Steroids
(1971)
SJ Henning
A sensitive and convenient method for measurement of corticosterone in rat serum
Steroids
(1980)
HA Gross et al.
A radio-immunoassay for plasma corticosterone
Steroids
(1972)
CJ Nabors et al.
Radio-immunoassay of human plasma corticosterone: method, measurement of episodic secretion and adrenal suppression and stimulation
Steroids
(1974)
RJ Etches
A radioimmunoassay for corticosterone and its application to the measurement of stress in poultry
Steroids
(1976)
M Schoneshofer
Simultaneous radioimmunoassay for corticosterone and deoxycortisol in human serum: Sex differences in the mean serum concentrations
J Steroid Biochem
(1977)
HK Kley et al.
Evaluation of adrenal function in rats by the measurement of urinary free corticosterone, free aldosterone and free 11-deoxycorticosterone
Steroids
(1978)
JK Mathew et al.
An improved method for the extraction of corticosterone from cell homogenate and subcellular fractions of the rat adrenal cortex
Steroids
(1981)
W Shaw et al.
The carrier effect of gamma globulin when a polyethylene glycol separation technique is used in digoxin radioimmunoassay
Clin Chim Acta
(1975)
LE McDonald
Hormones influencing metabolism

Z Acs et al.

The role of transcortin in the distribution of corticosterone in the rat

Horm Metab Res

(1973)

WR Slaunwhite et al.

Inactivity in vivo of transcortin-bound cortisol

Science

(1962)

R. Ekins

Free hormones in blood: Their physiological significance and measurement

ML Sweat

Sulfuric acid-induced fluorescence of corticosteroids

Anal Chem

(1954)

JH Solem et al.

An evaluation of a method for determination of free corticosteroids in minute quantities of mouse plasma

Scand J Clin Lab Invest

(1965)

HH Newsome et al.

The simultaneous assay of cortisol, corticosterone, 11-deoxycortisol, and corticone in human plasma

J Clin Endocrinol Metab

(1972)

EH Cameron et al.

Determination of corticosterone in rat plasma by competitive protein binding assay and its use in assessing the effects of C-B154 and perphenazine on adrenal function

J Endocrinol

(1973)

Cited by (51)

Bile duct ligation differently regulates protein expressions of organic cation transporters in intestine, liver and kidney of rats through activation of farnesoid X receptor by cholate and bilirubin
2023, Acta Pharmaceutica Sinica B
Citation Excerpt :
Protein expressions of corresponding transporters were measured using Western blot. It is contrast to previous reports31,32 that protein expressions of intestinal Oct1 are not detected. Several reports have also showed that metformin concentration-time profiles following oral dose in blood of Oct1-knockout mice are comparable to those in wild-type mice8,59.
Body is equipped with organic cation transporters (OCTs). These OCTs mediate drug transport and are also involved in some disease process. We aimed to investigate whether liver failure alters intestinal, hepatic and renal Oct expressions using bile duct ligation (BDL) rats. Pharmacokinetic analysis demonstrates that BDL decreases plasma metformin exposure, associated with decreased intestinal absorption and increased urinary excretion. Western blot shows that BDL significantly downregulates intestinal Oct2 and hepatic Oct1 but upregulates renal and hepatic Oct2. In vitro cell experiments show that chenodeoxycholic acid (CDCA), bilirubin and farnesoid X receptor (FXR) agonist GW4064 increase OCT2/Oct2 but decrease OCT1/Oct1, which are remarkably attenuated by glycine-β-muricholic acid and silencing FXR. Significantly lowered intestinal CDCA and increased plasma bilirubin levels contribute to different Octs regulation by BDL, which are confirmed using CDCA-treated and bilirubin-treated rats. A disease-based physiologically based pharmacokinetic model characterizing intestinal, hepatic and renal Octs was successfully developed to predict metformin pharmacokinetics in rats. In conclusion, BDL remarkably downregulates expressions of intestinal Oct2 and hepatic Oct1 protein while upregulates expressions of renal and hepatic Oct2 protein in rats, finally, decreasing plasma exposure and impairing hypoglycemic effects of metformin. BDL differently regulates Oct expressions via Fxr activation by CDCA and bilirubin.
Impact of strain and duration of thermal stress on carcass yield, metabolic hormones, immunological indices and the expression of HSP90 and Myogenin genes in broilers
2019, Research in Veterinary Science
Citation Excerpt :
The levels of serum triiodothyronine (T3) and thyroid (T4) were measured by enzymatic immunoassay using commercial kits (Diagnostic Product Corporation, Los Angeles, CA, USA). The cortisol concentration was assayed by a solid phase radioimmunoassay (RIA) procedure (Sainio et al., 1988). The serum electrolytes (potassium, sodium, chloride, calcium and phosphorus) were estimated by automatic analyzer (Kodak Ektachem®; Eastman Kodak Company, Rochester, New York).
The aim was to elucidate the effects of strain and the duration of thermal stress (36 °C) on the carcass yield, metabolic hormones, immunological parameters and the relative expression of liver heat shock protein 90 (HSP90) and Myogenin genes. Two hundred and seventy day-old chicks (135 of each breed; Ross 308 and Cobb 500) were used in this trial. The birds were divided into 3 equal groups (CON: thermoneutral condition; S₁ and S₂ groups were subjected to 4 and 6 h of daily thermal stress, respectively. The relative bursa weight in the thermonuteral Ross broilers, but not Cobb broilers was significantly greater than both heat-stressed groups (P = 0.001). The serum Na and phosphorus levels were significantly decreased when both broiler strains subjected to 6 h of thermals stress (P = 0.020 and 0.022, respectively). A linear decrease in the serum T₃ level was recorded in both broiler strains as the duration of thermal stress increases (P = 0.001). In both broiler strains, heat stress up-regulate the expression of liver HSP90 gene compared with the thermoneutral group (P = 0.012). Meanwhile, the S₂ group of Ross broilers showed a significantly lower IgG level (P = 0.027). Moreover, thermal stress for 6 h down-regulate the relative expression of Myogenin gene compared with the S₁ and thermoneutral groups (P = 0.005). In conclusion, thermal stress for 6 h down-regulate the mRNA expression of liver Myogenin, concomitantly with an increase in the expression of HSP90 gene in both broiler strains. These results could be helpful in the markers assisted selection to develop more heat-tolerant broiler strains.
GABAergic control of the activity of the central nucleus of the amygdala in low- and high-anxiety rats
2015, Neuropharmacology
Citation Excerpt :
Another fact which deserves comment is the possible influence of the weekend breaks on the effects of corticosterone. The half life of serum corticosterone is about 25 min (Sainio et al., 1988). However, it has been shown that the oil suspension of corticosterone is absorbed from the injection site at a much slower rate than water-based solution.
The aim of this study was to examine the role of GABAergic neurotransmission in amygdala nuclei in low- (LR) and high-anxiety (HR) rats after repeated corticosterone administration and acute injection of the benzodiazepine midazolam. The animals were divided into LR and HR groups based on the duration of their conditioned freezing in a contextual fear test (CFT). Repeated daily administration of corticosterone (20 mg/kg s.c.) for 21 injections increased anxiety-like behavior in the open field and reduced body weight in both the LR and HR groups. These effects of corticosterone administration were more pronounced in the HR group. Moreover, in the HR group, chronic corticosterone administration increased the duration of freezing in the CFT test compared with the appropriate control group and treated LR rats. The behavioral effects in HR rats were accompanied by an increase in the expression of c-Fos in the lateral (LA) and central (CeA) nuclei of the amygdala and by a decrease in GABA-A alpha-2 subunit density in the CeA. Acute midazolam administration significantly attenuated the neophobia and conditioned fear responses, decreased c-Fos expression in the LA and CeA, and increased alpha-2 subunit density in the CeA only in the HR group. These studies have shown that HR rats are more susceptible to the anxiogenic effects of chronic corticosterone administration, which are associated with the attenuation of GABAergic control over the amygdala output that controls emotional responses. The current data may increase understanding of the neurobiological mechanisms responsible for individual differences in the psychopathological processes induced by repeated administration of high doses of glucocorticoids or by elevated levels of these hormones, which are associated with chronic stress and affective pathology.
Determination of endogenous corticosterone in rodent's blood, brain and hair with LC-APCI-MS/MS
2015, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Citation Excerpt :
Because the content of hair corticosterone is considerably low at several pg/mg [15], its analysis requires instruments with high sensitivity, good selectivity and adequate limits of detection and quantitation (LOD and LOQ). Analysis methods of corticosterone used in the existing literatures were enzyme-linked immunosorbent assay (ELISA) [19], radioimmunoassay (RIA) [20,21], fluorometric determination[22], high performance liquid chromatography in combination with ultraviolet detector (HPLC–UV) [23,24], gas chromatography mass spectrometry (GC–MS) [3] and high performance liquid chromatography mass spectrometry coupled to electrospray ionization (LC–ESI–MS) [25,26]. Among these methods, LC-ESI–MS method, showing high sensitivity [25,26], is undoubtedly preferable for analyzing hair corticosterone, while the other methods are not suitable because of the high cross-reactivity with other steroids (e.g. ELISA and RIA [7,16,17]), time-consuming pretreatment procedures (e.g. fluorometric determination [27]), inadequate LOQ (e.g. HPLC–UV [23,24]) and requirement of large sample volume (e.g. GC–MS [3]).
Endogenous corticosterone in rodent’s hair would be a potential biomarker to assess the response of the hypothalamus–pituitary–adrenal axis to chronic stress. However, currently unknown is whether hair corticosterone is associated with endogenous corticosterone in blood and brain. The present study aimed to develop an enhanced assay for determination of endogenous corticosterone in blood, brain and hair, and to examine associations of hair corticosterone with blood and brain corticosterone under basal condition and association with blood corticosterone under chronic stressful condition. Hair at the back and blood samples were collected from non-stressed and stressed rodents, and prefrontal lobe and thalamus from non-stressed rodents. Chronic stress exerted on mice was 30-day repeated social defeat. The analyses were done using high performance liquid chromatography tandem mass spectrometry with atmospheric pressure chemical ionization in positive mode. Limits of detection and quantification were 0.2 and 0.5 ng/ml for rat’s blood, and 0.5 and 1.0 pg/mg for rat’s hair and brain, and 1.25 and 2.50 ng/ml (or pg/mg) for mouse’s blood (or hair). Recovery ranged from 84.2 to 108.0%. The intra- and inter-day coefficients of variation were less than 10%. Additionally, correlation of hair corticosterone with blood corticosterone was significant in both mice and rats, but correlations with corticosterone in prefrontal lobe and thalamus were not significant in rats. Both hair and blood corticosterone were significantly higher in stressed mice compared with controls.
Monitoring the stress-level of rats with different types of anesthesia: A tail-artery cannulation protocol
2014, Journal of Pharmacological and Toxicological Methods
Functional MRI in rats under anesthesia can largely minimize motion artifacts and attenuate the stress of the animal. However, two issues remain to be clarified and improved. First, fMRI results obtained with different types of anesthesia during surgical preparation and imaging show a large variability, which could be caused by the variable stress level of the rodents. Second, the most common surgical procedure used for anesthesia, blood gas analysis and mean arterial blood-pressure (MABP) monitoring is the femoral vein and artery catheterization that makes longitudinal studies difficult.
In order to examine the variability of the stress level with three different anesthesia protocols using isoflurane (Iso), medetomidine–ketamine (MK) or propofol–remifentanil (PR), we measured the plasma corticosterone (CORT) concentration with ¹²⁵I-radioimmunoassay in blood samples collected prior to, immediately after and 60 min after surgery. Tail-artery and vein catheterization was adapted for long-term monitoring of MABP with periodic blood sampling and is proposed as a less invasive and technically simple alternative to femoral vessel catheterization in fMRI preparation protocols.
We show that the CORT concentration depends on the anesthesia protocol with both alternatives providing more efficient stress reduction than the protocol using Iso. However, only the protocol using PR achieved a significant hormone reduction during surgery. Stress was not reliably manifested in changes in heart-rate and breathing-rate. Anesthesia and strain related changes in these two physiological parameters may be assigned to the pharmacological effects of the premedication and anesthetic agents. The results indicate also that MABP can be monitored over a long period of time (e.g. functional imaging session) through an arterial access point in the rat tail after cannulation with the proposed procedure.
Animals can experience stress during fMRI preparation protocols without obvious signs in commonly monitored physiological parameters. Our results challenge the efficiency of surgical protocols using Iso as mono-anesthetic agent, even when extended with topical analgesia. It was demonstrated that the CORT-based stress-level measurement through tail-artery cannulation can be used for developing anesthesia protocols (i.e. the presented PR protocol) when setting up future fMRI studies. The proposed surgical method for the tail is expected to facilitate longitudinal fMRI studies with permanent arterial access.
The effects of acute stress and acute corticosterone administration on the immobility response in rats
2009, Brain Research Bulletin
Citation Excerpt :
One possibility is derived from the short half-life of the CORT in the rat plasma. Sainio et al. [58] using the same dose (5 mg/kg) found the half-lives for total and free CORT in rat plasma were 25 and 15 min, respectively. Thus the lack of an effect 20 min after the CORT injection could be caused by a reduction in the concentration of this corticosteroid in its target areas in the brain.
The immobility response is an innate antipredatory behavior in a broad variety of species. The immobility response varies in its postural components but in general is characterized by an absence of movement and a relative unresponsiveness to stimuli. Experimentally in rats, clamping the neck followed by body inversion and manual restrain elicits a response called “immobility by clamping the neck”. Stress reactions protect animals against predators and are characterized by activation of the sympathetic and hypothalamic–pituitary–adrenal systems. However, in mammals, the role of acute stress as a modulator of immobility response has been less studied. The aim of our study was to assess the effects of acute stress and the injection of corticosterone (5 mg/kg, ip) on immobility by clamping the neck in rats. We observed that either previous acute stress caused by forced exposure to elevated open platform or application of a heat-pain stimulus to the rat's tail during the immobility increased the duration of the immobility response caused by clamping the neck. Also, the corticosterone produced a rapid increase (15 min after injection) in the duration of this immobility response. Our results show that the acute stress, in rats, is a facilitator of the immobility response and suggest a possible nongenomic rapid action of corticosterone over brain structures that control this behavior.

View all citing articles on Scopus

View full text

Radioimmunoassay of total and free corticosterone in rat plasma: Measurement of the effect of different doses of corticosterone

Abstract

Steroids

Steroids

Steroids

Steroids

Steroids

J Steroid Biochem

Steroids

Steroids

Clin Chim Acta

Hormones influencing metabolism

The role of transcortin in the distribution of corticosterone in the rat

Horm Metab Res

Inactivity in vivo of transcortin-bound cortisol

Science

Free hormones in blood: Their physiological significance and measurement

Sulfuric acid-induced fluorescence of corticosteroids

Anal Chem

An evaluation of a method for determination of free corticosteroids in minute quantities of mouse plasma

Scand J Clin Lab Invest

The simultaneous assay of cortisol, corticosterone, 11-deoxycortisol, and corticone in human plasma

J Clin Endocrinol Metab

Determination of corticosterone in rat plasma by competitive protein binding assay and its use in assessing the effects of C-B154 and perphenazine on adrenal function

J Endocrinol