Elsevier

Life Sciences

Volume 161, 15 September 2016, Pages 27-36
Life Sciences

A highly reproducible mice model of chronic kidney disease: Evidences of behavioural abnormalities and blood-brain barrier disruption

https://doi.org/10.1016/j.lfs.2016.07.020Get rights and content

Abstract

Aims

In the present study, a novel mice model of chronic kidney disease (CKD) was developed, and psycho-motor behavioural abnormalities, blood-brain barrier (BBB) integrity and brain histology were studied.

Main methods

Swiss albino female mice were given high adenine diet (0.3% w/w mixed with feed) for 4 weeks. Serum urea and creatinine levels and renal histological studies were performed to validate the model. Psycho-motor behavioural abnormalities and neurological severity were studied. BBB integrity was assessed using Evans blue extravasation method. Nissl staining was performed to see possible morphological aberrations in brain.

Key findings

There was a significant increase in serum urea and creatinine levels in mice given high adenine diet, and the mice had abnormal kidney morphology. Deposition of adenine and 2,8-dihydroxyadenine crystals, and increased collagen deposits in the renal tissues were found, which validate induction of CKD in the mice. Motor behavioural abnormalities, depression-like and anxiolytic behaviour and increase in neurological severity were prevalent in mice with CKD. Evans Blue dye extravasation was found to occur in the brain, which signifies disruption of BBB. However, Nissl staining did not reveal any morphological aberration in brain tissue.

Significance

The present study puts forward a highly reproducible mice model of CKD validated with serum parameters and renal histopathological changes. The mice showed psycho-motor behavioural abnormalities and BBB disruption. It is a convenient model to study the disease pathology, and understanding the associated disorders, and their therapeutic interventions.

Introduction

Chronic kidney disease (CKD) refers to a wide range of disease condition that affect the structure and function of the kidney, lasting for > 3 months, and leads to albuminuria and reduction in glomerular filtration rate (GFR) [7], [40]. The major causes for the development of CKD are diabetes, hypertension, glomerulonephritis and interstitial nephritis [7], [10]. There is an increase in the global prevalence of CKD, affecting > 15% of the population in developed nations [14], [17], [27]. With reduction in the GFR there occurs retention of uremic toxins and electrolyte imbalance [40], which necessitates, renal replacement therapies in the form of dialysis and kidney transplantation [44]. Due to retention of uremic toxins and electrolyte imbalance, CKD leads to cardiovascular disease, stroke [48], [49] and neurological complications (reviewed in Ref. [36]). In CKD patients, anxiety, depression, cognitive dysfunction, restless leg syndrome and reduced intelligence quotient are common diagnosis which hamper the quality of life [23], [28], [34], [36], [38], [39], [41], [60], [65]. In animal models, depression, anxiety and reduced exploratory and locomotor activities are reported [5], [16], [66].

One of the most widely used animal models of CKD is the surgical model [2], [21], [26], [37], [50]. However, owing to high mortality and need of sophisticated surgical methods, which alter the morphology of the animals as well, alternative models are sought. Yokozawa et al. [74] introduced rat model of CKD by administration of adenine with feed. Adenine and its metabolite (2,8-dihydroxyadenine) get deposited in renal tissues as crystals, as shown by Ikeda et al. [30] in rat model, and cause renal damages [5], [58], which leads to elevation in serum urea, creatinine and renal histological damage [9], [45], [62].

In rats with adenine-induced CKD, cardio-vascular damages have been investigated [22], while Ali et al. [5] reported on psycho-motor behavioural abnormalities. However, no behavioural study has been undertaken yet in mice model, and female mice have not yet been used as adenine-induced CKD model, to the best of our knowledge. In mice model of acute renal injury, disruption of blood-brain barrier (BBB) has been reported [42]. However, any such study is lacking in adenine-induced CKD model. In the present study, we induced CKD in adult female Swiss albino mice by administering high adenine diet. In this novel model, we investigated for renal fibrosis, psycho-motor behavioural abnormalities, neurological severity, BBB disruption, and histopathological changes in discrete brain regions.

Section snippets

Animals

Adult Swiss albino female mice of weight between 25 and 27 g (10 weeks old) were obtained from Pasteur Institute, Shillong, Meghalaya (India). They were housed in standard conditions of 12 h light/dark cycle, 24 ± 2 °C temperature and 60 ± 5% humidity; and were given standard feed and water ad libitum. An acclimatization time of 10 days was given prior to start of the experiment. The experimental protocols met the National Guidelines and were approved by the Institutional Animal Ethics Committee.

Chemicals and others

High adenine diet leads to decrease in body weight

Adenine, when given at 0.3% w/w mixed with feed for 4 weeks, leads to a significant reduction in body weight by 12.59% (compared to the initial body weight), while control mice (given standard feed) gained weight by 13.92% (Fig. 1A, B).

High adenine diet elevates serum urea and creatinine level

The level of serum urea has been found to be significantly higher in the adenine fed animals as compared to control (Fig. 1D). The treated mice had a mean urea level of 111.93 ± 2.40 mg/dL, while the control had 36.8 ± 0.56 mg/dL, thereby an increase of > 3-fold. The

Discussion

CKD is a global health issue with significant morbidity and mortality, and affects all levels of the nervous system (see Ref. [36]). However, detail study of the same are lacking in animal models, except the study by Ali et al. [5] in rat. In the present study, we established CKD in female Swiss albino mice by administering high adenine, and report on behavioural abnormalities (motor, psychometric and neurological severity) and BBB disruption.

The CKD mice model was generated by giving adenine

Conclusion

The present study reports development of a highly reproducible mice model of CKD induced by adenine feeding at the dose of 0.3% w/w mixed with diet for 4 weeks, having features of elevated serum urea and creatinine, and renal fibrosis. At this dose, deposition of adenine and 2,8-dihydroxyadenine crystals in renal tissues were observed. In this model, BBB disruption and behavioural abnormalities, including motor, psychometric and neurological severity, were found, which are reported hereby in

Conflict of interest

None declared.

Author contribution

MKM contributed by overall design of the experiment, performing the experiments, acquisition and interpretation of data, and drafting of the manuscript. AB contributed by conception of the idea and final approval of the manuscript. AG and SK contributed by revision of the manuscript.

Acknowledgment

We sincerely acknowledge the funding and support provided by the Department of Biotechnology (DBT), Govt. of India (under Rapid Grant for Young Investigator project; Sanction Order No. BT/PR6806/27/480/2012, dated August 05, 2013), and Department of Science and Technology, Govt. of India (Sanction Order No. SB/YS/LS-61/2014 dated March 04, 2015). MKM is recipient of Maulana Azad National Senior Research Fellowship from University Grant Commission, Govt. of India (MANF-2012-13-MUS-ASS-11201). We

References (74)

  • A.B. Fogo

    Animal models of FSGS: lessons for pathogenesis and treatment

    Semin. Nephrol.

    (2003)
  • R. Haobam et al.

    Swim-test as a function of motor impairment in MPTP model of Parkinson's disease: a comparative study in two mouse strains

    Behav. Brain Res.

    (2005)
  • M. Lew-Starowicz et al.

    The sexuality and quality of life of hemodialyzed patients—ASED multicenter study

    J. Sex. Med.

    (2009)
  • Y. Liu et al.

    Metformin attenuates blood-brain barrier disruption in mice following middle cerebral artery occlusion

    J. Neuroinflammation

    (2014)
  • N.P. Mallick et al.

    Haemodialysis

    Lancet

    (1999)
  • I. Matsui et al.

    Fully phosphorylated fetuin-A forms a mineral complex in the serum of rats with adenine-induced renal failure

    Kidney Int.

    (2009)
  • K.P. Mohanakumar et al.

    Neuroprotection by sodium salicylate against 1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine-induced neurotoxicity

    Brain Res.

    (2000)
  • T. Ninomiya et al.

    Chronic kidney disease and cardiovascular disease in a general Japanese population: the Hisayama Study

    Kidney Int.

    (2005)
  • H. Puchtler et al.

    Polarization microscopic studies of connective tissue stained with picro-sirius red FBA

    Beitr. Pathol.

    (1973)
  • T. Sengupta et al.

    2-Phenylethylamine, a constituent of chocolate and wine, causes mitochondrial complex-I inhibition, generation of hydroxyl radicals and depletion of striatal biogenic amines leading to psycho-motor dysfunctions in Balb/c mice

    Neurochem. Int.

    (2010)
  • M. Shuvy et al.

    Histopathology and apoptosis in an animal model of reversible renal injury

    Exp. Toxicol. Pathol.

    (2011)
  • M.J. Smogorzewski et al.

    Altered acetylcholine in uremia: role metabolism of brain of secondary hyperparathyroidism

    J. Ren. Nutr.

    (2008)
  • M.J. Smogorzewski

    Central nervous dysfunction in uremia

    Am. J. Kidney Dis.

    (2001)
  • T. Tanaka et al.

    Urinary L-type fatty acid-binding protein can reflflect renal tubulointerstitial injury

    Am. J. Pathol.

    (2009)
  • F.B. Weihmuller et al.

    Dissociation between biochemical and behavioral recovery in MPTP-treated mice

    Pharmacol. Biochem. Behav.

    (1989)
  • N. Weiss et al.

    The blood-brain barrier in brain homeostasis and neurological diseases

    Biochim. Biophys. Acta

    (2009)
  • B.H. Ali et al.

    Anaemia in adenine-induced chronic renal failure and the influence of treatment with gum acacia thereon

    Physiol. Res.

    (2014)
  • B.H. Ali et al.

    Does treatment with gum Arabic affect experimental chronic renal failure in rats?

    Fundam. Clin. Pharmacol.

    (2004)
  • B.H. Ali et al.

    Motor and behavioral changes in rats with adenine-induced chronic renal failure: influence of acacia gum treatment

    Exp. Biol. Med.

    (2011)
  • ANZDATA

    31st annual report

  • K. Ataka et al.

    Effects of erythropoietin-gene electrotransfer in rats with adenine-induced renal failure

    Am. J. Nephrol.

    (2003)
  • R.S. Barsoum

    Chronic kidney disease in the developing world

    N. Engl. J. Med.

    (2006)
  • M.P.E. Berthelot

    Berthelot's reaction mechanism

    Report Chim. Appl.

    (1859)
  • N. Bhattacharjee et al.

    L-DOPA treatment in MPTP-mouse model of Parkinson's disease potentiates homocysteine accumulation in substantia nigra

    Neurosci. Lett.

    (2016)
  • S.J. Chadban et al.

    Prevalence of kidney damage in Australian adults: the Aus Diab kidney study

    J. Am. Soc. Nephrol.

    (2003)
  • S. Chua et al.

    Inhibition of dipeptidyl peptidase-IV enzyme activity protects against myocardial ischemia-reperfusion injury in rats

    J. Transl. Med.

    (2014)
  • A.D. Costaesilva et al.

    Effect of experimentally induced chronic renal-failure upon the behavior of rats

    Nephron

    (1979)
  • Cited by (0)

    View full text