Regulation and Role of Neuron-Derived Hemoglobin in the Mouse Hippocampus

Hemoglobin (Hb) is the oxygen transport protein in erythrocytes. In blood, Hb is a tetramer consisting of two Hb-alpha (Hb-α) chains and two Hb-beta (Hb-β) chains. A number of studies have also shown that Hb-α is also expressed in neurons in both the rodent and human brain. In the current study, we examined for age-related regulation of neuronal Hb-α and hypoxia in the hippocampus and cerebral cortex of intact male and female mice. In addition, to confirm the role and functions of neuronal Hb-α, we also utilized lentivirus CRISPR interference-based Hb-α knockdown (Hb-α CRISPRi KD) in the non-ischemic and ischemic mouse hippocampus and examined the effect on neuronal oxygenation, as well as induction of hypoxia-inducible factor-1α (HIF-1α) and its downstream pro-apoptotic factors, PUMA and NOXA, and on neuronal survival and neurodegeneration. The results of the study revealed an age-related decrease in neuronal Hb-α levels and correlated increase in hypoxia in the hippocampus and cortex of intact male and female mice. Sex differences were observed with males having higher neuronal Hb-α levels than females in all brain regions at all ages. In vivo Hb-α CRISPRi KD in the mouse hippocampus resulted in increased hypoxia and elevated levels of HIF-1α, PUMA and NOXA in the non-ischemic and ischemic mouse hippocampus, effects that were correlated with a significant decrease in neuronal survival and increased neurodegeneration. As a whole, these findings indicate that neuronal Hb-α decreases with age in mice and has an important role in regulating neuronal oxygenation and neuroprotection.

[1]  T. Floyd,et al.  Aged Mouse Hippocampus Exhibits Signs of Chronic Hypoxia and an Impaired HIF-Controlled Response to Acute Hypoxic Exposures , 2022, Cells.

[2]  Yujiao Lu,et al.  Ganglioside GD3 is up‐regulated in microglia and regulates phagocytosis following global cerebral ischemia , 2021, Journal of neurochemistry.

[3]  J. Svensson,et al.  Growth Hormone and Neuronal Hemoglobin in the Brain—Roles in Neuroprotection and Neurodegenerative Diseases , 2021, Frontiers in Endocrinology.

[4]  R. Tekmal,et al.  Astrocyte-Derived Estrogen Regulates Reactive Astrogliosis and is Neuroprotective following Ischemic Brain Injury , 2020, The Journal of Neuroscience.

[5]  Quanguang Zhang,et al.  G-protein-coupled estrogen receptor activation upregulates interleukin-1 receptor antagonist in the hippocampus after global cerebral ischemia: implications for neuronal self-defense , 2019, Journal of Neuroinflammation.

[6]  F. Gage,et al.  CRISPR interference-based specific and efficient gene inactivation in the brain , 2018, Nature Neuroscience.

[7]  E. Barbier,et al.  Transit time mapping in the mouse brain using time‐encoded pCASL , 2018, NMR in biomedicine.

[8]  J. Day,et al.  Applications of CRISPR/Cas9 in the Mammalian Central Nervous System

 , 2017, The Yale journal of biology and medicine.

[9]  D. Gell.  Structure and function of haemoglobins. , 2017, Blood cells, molecules & diseases.

[10]  S. Lanigan,et al.  The Effects of Hypoxia and Inflammation on Synaptic Signaling in the CNS , 2016, Brain sciences.

[11]  Stefan Wiehr,et al.  Longitudinal PET-MRI reveals β-amyloid deposition and rCBF dynamics and connects vascular amyloidosis to quantitative loss of perfusion , 2014, Nature Medicine.

[12]  R. Brekken,et al.  Hypoxia Studies with Pimonidazole in vivo. , 2014, Bio-protocol.

[13]  W. Murphy.  The sex difference in haemoglobin levels in adults - mechanisms, causes, and consequences. , 2014, Blood reviews.

[14]  B. Stoica,et al.  Traumatic brain injury in aged animals increases lesion size and chronically alters microglial/macrophage classical and alternative activation states , 2013, Neurobiology of Aging.

[15]  Dong-Sun Han,et al.  Acetylation of the Pro-Apoptotic Factor, p53 in the Hippocampus following Cerebral Ischemia and Modulation by Estrogen , 2011, PloS one.

[16]  Dong-Sun Han,et al.  C terminus of Hsc70-interacting protein (CHIP)-mediated degradation of hippocampal estrogen receptor-α and the critical period hypothesis of estrogen neuroprotection , 2011, Proceedings of the National Academy of Sciences.

[17]  Isidro Ferrer,et al.  Unexpected expression of α- and β-globin in mesencephalic dopaminergic neurons and glial cells , 2009, Proceedings of the National Academy of Sciences.

[18]  M. Chesselet,et al.  Neurons express hemoglobin α‐ and β‐chains in rat and human brains , 2009, The Journal of comparative neurology.

[19]  J. Klein,et al.  Synthesis and secretion of hemoglobin by retinal pigment epithelium. , 2009, Investigative ophthalmology & visual science.

[20]  Armin Schneider,et al.  Expression of Hemoglobin in Rodent Neurons , 2009, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[21]  P. Chan,et al.  Potential Role of PUMA in Delayed Death of Hippocampal CA1 Neurons After Transient Global Cerebral Ischemia , 2009, Stroke.

[22]  R. Ratan,et al.  Antioxidants, HIF prolyl hydroxylase inhibitors or short interfering RNAs to BNIP3 or PUMA, can prevent prodeath effects of the transcriptional activator, HIF-1alpha, in a mouse hippocampal neuronal line. , 2008, Antioxidants & redox signaling.

[23]  R. Kamps,et al.  Haemoglobin expression in human endometrium. , 2008, Human reproduction.

[24]  B. Reeder,et al.  Oxygen‐binding haem proteins , 2008, Experimental physiology.

[25]  T. Yanagihara,et al.  Temporal and topographic profiles of tissue hypoxia following transient focal cerebral ischemia in rats. , 2006, The Journal of veterinary medical science.

[26]  J. Baatz,et al.  Hemoglobin Is Expressed by Alveolar Epithelial Cells* , 2006, Journal of Biological Chemistry.

[27]  Jiankun Cui,et al.  Brain-Specific Knock-Out of Hypoxia-Inducible Factor-1α Reduces Rather Than Increases Hypoxic-Ischemic Damage , 2005, The Journal of Neuroscience.

[28]  S. Kiryu-Seo,et al.  Noxa Is a Critical Mediator of p53-Dependent Motor Neuron Death after Nerve Injury in Adult Mouse , 2005, The Journal of Neuroscience.

[29]  T. Shirasawa,et al.  Presenilin-1-deficient neurons are nitric oxide-dependently killed by hydrogen peroxide in vitro , 2004, Neuroscience.

[30]  P. Chan.  Mitochondria and Neuronal Death/Survival Signaling Pathways in Cerebral Ischemia , 2004, Neurochemical Research.

[31]  K. Suk,et al.  BH3-only Protein Noxa Is a Mediator of Hypoxic Cell Death Induced by Hypoxia-inducible Factor 1α , 2004, The Journal of experimental medicine.

[32]  J. LaManna,et al.  Activation of Hypoxia-Inducible Factor-1 in the Rat Cerebral Cortex after Transient Global Ischemia: Potential Role of Insulin-Like Growth Factor-1 , 2002, The Journal of Neuroscience.

[33]  M. Zeng,et al.  Hemoglobin induction in mouse macrophages. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[34]  M. Gassmann,et al.  Erythropoietin Gene Expression in Human, Monkey and Murine Brain , 1996, The European journal of neuroscience.

[35]  G. Semenza,et al.  Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[36]  V. Gracco,et al.  Speech motor coordination and control: evidence from lip, jaw, and laryngeal movements , 1994, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[37]  C. Koch,et al.  Importance of thiols in the reductive binding of 2-nitroimidazoles to macromolecules. , 1990, Biochemical pharmacology.

[38]  W. Fried,et al.  Renal and extrarenal erythropoietin production in male and female rats of various ages. , 1972, The Journal of laboratory and clinical medicine.

[39]  T. Sugawara,et al.  Neuronal death/survival signaling pathways in cerebral ischemia , 2011, NeuroRX.

[40]  I. Ferrer,et al.  Neuronal hemoglobin is reduced in Alzheimer's disease, argyrophilic grain disease, Parkinson's disease, and dementia with Lewy bodies. , 2011, Journal of Alzheimer's disease : JAD.

[41]  J. LaManna,et al.  Increased sensitivity to transient global ischemia in aging rat brain. , 2007, Advances in experimental medicine and biology.

[42]  G. Semenza.  Expression of hypoxia-inducible factor 1: mechanisms and consequences. , 2000, Biochemical pharmacology.

[43]  B. Wittenberg,et al.  Mechanisms of cytoplasmic hemoglobin and myoglobin function. , 1990, Annual review of biophysics and biophysical chemistry.

[44]  A. Tamura,et al.  Selective vulnerability of the hippocampus to ischemia--reversible and irreversible types of ischemic cell damage. , 1985, Progress in brain research.

[45]  G. K. Ackers,et al.  Quaternary enhancement in binding of oxygen by human hemoglobin. , 1979, Proceedings of the National Academy of Sciences of the United States of America.