A maternal diet supplemented with creatine from mid-pregnancy protects the newborn spiny mouse brain from birth hypoxia

doi:10.1016/j.neuroscience.2011.05.012

Neuroscience

Volume 194, 27 October 2011, Pages 372-379

https://doi.org/10.1016/j.neuroscience.2011.05.012 Get rights and content

Abstract

The creatine-phosphocreatine shuttle is essential for the maintenance of cellular ATP, particularly under hypoxic conditions when respiration may become anaerobic. Using a model of intrapartum hypoxia in the precocial spiny mouse (Acomys cahirinus), the present study assessed the potential for maternal creatine supplementation during pregnancy to protect the developing brain from the effects of birth hypoxia. On day 38 of gestation (term is 39 days), the pregnant uterus was isolated and placed in a saline bath for 7.5 min, inducing global hypoxia. The pups were then removed, resuscitated, and cross-fostered to a nursing dam. Control offspring were delivered by caesarean section and recovered immediately after release from the uterus. At 24 h after birth hypoxia, the brains of offspring from dams fed a normal diet showed significant increases in lipid peroxidation as measured by the amount of malondialdehyde. In the cortical subplate, thalamus and piriform cortex there were significant increases in cellular expression of the pro-apoptotic protein BAX, cytoplasmic cytochrome c and caspase-3. When pregnant dams were fed the creatine supplemented diet, the increase in malondialdehyde, BAX, cytochrome c and caspase 3 were almost completely prevented, such that they were not different from control (caesarean-delivered) neonates. This study provides evidence that the neuroprotective capacity of creatine in the hypoxic perinatal brain involves abrogation of lipid peroxidation and apoptosis, possibly through the maintenance of mitochondrial function. Further investigation into these mechanisms of protection, and the long-term development and behavioural outcomes of such neonates is warranted.

Highlights

▶We model birth hypoxia using the precocial spiny mouse. ▶Hypoxia significantly increased lipid peroxidation in the cerebrum. ▶Hypoxia caused region-dependent increases in BAX, cytochrome c and caspase-3. ▶Maternal creatine supplementation prevented adverse changes in hypoxia brains. ▶Creatine has neuroprotective capacity for newborn.

Section snippets

Animals

All experiments were approved in advance by Monash University School of Biomedical Sciences Animal Ethics Committee, and conducted according to the Australian Code of Practice for the Care and Use of Animals for Scientific Purposes. The spiny mice used in this study were obtained from our own laboratory colony and housed, bred and time-mated as previously described (Dickinson and Walker, 2007).

Diet

Pregnant dams were fed either a control diet of standard rat and mouse pellets throughout pregnancy,

Lipid peroxidation in the brain

At 24 h of age lipid peroxidation (as measured by MDA) was significantly increased in the cerebrum of pups in the birth hypoxia group compared to all other groups (P≤0.05; Fig. 1). No such increase was observed in the birth hypoxia pups whose mothers had received the creatine supplemented diet from day 20 of pregnancy.

Activation of the apoptotic cell death pathway in the brain

Birth hypoxia induced a significant increase in the expression of the pro-apoptotic protein BAX in the cortical subplate and the thalamus (P<0.05; Fig. 2A, C), and a strong trend

Discussion

This study investigated the potential for maternal creatine supplementation from mid-pregnancy to reduce hypoxia-induced lipid peroxidation, maintain intra-mitochondrial stores of cytochrome-c, and prevent activation of the apoptotic pathway in the newborn brain following birth hypoxia. In pups subjected to intrapartum hypoxia, at 24 h after birth we observed an increase in lipid peroxidation, and region-specific increases in cytoplasmic cytochrome c, and increased expression of the

Conclusions

This study shows that maternal dietary creatine supplementation throughout the second half of pregnancy significantly protects fetal cortical and deep gray matter from injury arising from intrapartum hypoxia. It is yet to be shown whether creatine provides neuroprotection beyond the immediate postnatal period, and further investigation into the long-term brain development of these offspring is warranted, with particular reference to cognitive and motor development. It is appropriate that

Acknowledgments

We thank Dr. Lisa Hutton for assistance with immunohistochemical protocols. This project was supported by grants from the National Health & Medical Research Council of Australia to DWW, MCM and RS.

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^†: Present address of Z. Ireland: Queensland Cerebral Palsy & Rehabilitation Research Centre, Royal Children's Hospital, Herston, QLD, Australia, 4029.

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Regeneration, Repair, and Developmental NeuroscienceResearch PaperA maternal diet supplemented with creatine from mid-pregnancy protects the newborn spiny mouse brain from birth hypoxia

Abstract

Highlights

Section snippets

Animals

Diet

Lipid peroxidation in the brain

Activation of the apoptotic cell death pathway in the brain

Discussion

Conclusions

Acknowledgments

Curr Probl Pediatr Adolesc Health Care

Am J Obstet Gynecol

Neurobiol Aging

Am J Obstet Gynecol

Semin Neonatol

Early Hum Dev

Neurobiol Dis

Biochem Biophys Res Commun

Semin Perinatol

Neuroscience

Physiol Behav

Neuroprotection of creatine supplementation in neonatal rats with transient cerebral hypoxia-ischemia

Dev Neurosci

Perinatal asphyxia: MR findings in the first 10 days

Am J Neuroradiol

Creatine protects the immature brain from hypoxic-ischemic injury

J Soc Gynecol Invest

On the mechanisms of neuroprotection by creatine and phosphocreatine

J Neurochem

Male excess among anatomically normal fetuses in spontaneous abortions

Am J Med Genet

Maternal creatine supplementation from mid-pregnancy protects the diaphragm of the newborn spiny mouse from intrapartum hypoxia-induced damage

Pediatr Res

Lipid peroxidation, caspase-3 immunoreactivity, and pyknosis in late-gestation fetal sheep brain after umbilical cord occlusion

Pediatr Res

Changes in the male to female ratio at different stages of life

Br J Obstet Gynaecol

The Acomys cahirinus (spiny mouse) as a new model for biological and neurobehavioral studies

Pol J Pharmacol Pharm

Preterm fetal hypoxia-ischemia causes hypertonia and motor deficits in the neonatal rabbit: a model for human cerebral palsy?

J Neurosci

Managing a colony of spiny mice (Acomys cahirinus) for perinatal research

Australian and New Zealand Council for the Care of Animals in Research and Training (ANZCCART) News

The spiny mouse (Acomys cahirinus) completes nephrogenesis before birth

Am J Physiol Ren Physiol

In vivo development of brain phosphocreatine in normal and creatine-treated rabbit pups

J Neurochem

Creatine increases survival and suppresses seizures in the hypoxic immature rat

Pediatr Res

Regeneration, Repair, and Developmental Neuroscience
Research Paper
A maternal diet supplemented with creatine from mid-pregnancy protects the newborn spiny mouse brain from birth hypoxia