Title: Studying the sex differences in mouse hippocampal astrocytes after in vitro ischemia
Legend: Up-regulation of S100B and GFAP expression in sexed hippocampal astrocyte cultures following OGD-REOX. Sexed cultured hippocampal astrocytes were stained for S100b and GFAP expression under normoxic conditions or after 2 h OGD followed by 5 h of REOX (OGD/REOX). Inset: 60X image from hippocampal astrocytes cultured from GFAP null mice. Scale bar: 30 mm.
Citation: Chanana V, Tumturk A, Kintner D, Udho E, Ferrazzano P, Cengiz P (2016) Sex differences in mouse hippocampal astrocytes after in-vitro ischemia. Journal of Visualized Experiments, 25;(116). doi: 10.3791/53695 PMCID: PMC5092229 PMID: 27805577
Abstract: Astrogliosis following hypoxia-ischemia (HI) related brain injury plays a role in increased morbidity and mortality in neonates. Recent clinical studies indicate that the severity of brain injury appears to be sex-dependent and that male neonates are more susceptible to the effects of HI resulting in worse neurological outcomes compared to females with comparable brain injuries. The development of reliable methods to isolate and maintain highly enriched populations of sexed hippocampal astrocytes is essential to understand the cellular basis of sex-based neuropathology following neonatal HI. In this study, we describe a method for creating sex specific hippocampal astrocyte cultures that are subjected to a model of in-vitro ischemia, oxygen glucose deprivation, followed by reoxygenation. Subsequent reactive astrogliosis was examined by immunostaining for the glial fibrillary acidic protein (GFAP) and S100B. This method provides a useful tool to study separately the role of male and female hippocampal astrocytes following HI.
About the Lab: Cengiz’s research focuses on finding a novel therapy for neonatal encephalopathy subsequent to HI. Hypoxia ischemia-related brain injury is associated with high mortality and morbidity. One of every three neonates with HI develops chronic morbidity including cerebral palsy, intellectual and developmental disabilities, learning disabilities or epilepsy. Moderate hypothermia has been shown to improve the neurological outcome of term neonates. However, term neonates after 6 hours of life and pre-term neonates do not qualify for hypothermia therapy. Cengiz’s research continues to focus on exploring a pharmacological therapy as an adjunct to hypothermia and for those neonates who are not eligible for hypothermia therapy. Cengiz is particularly interested in targeting membrane proteins that are either detrimental or beneficial to neuronal survival after hypoxia and ischemia. She is working both with in vitro and in vivo models of hypoxia and ischemia in order to identify the role of these proteins and their effects on neuronal degeneration. Using Vannucci-Rice’s neonatal HI model in nine-day-old mice, she has shown neuroprotection at 72 hours after HI by pharmacologically inhibiting the membrane protein Na+/H+ exchanger isoform-1. Her goal is to establish the mechanism(s) that leads to hippocampal neuroprotection by characterizing the function of Na+/H+ exchanger protein isoform-1 after HI and to translate these preclinical findings into clinical studies that will benefit patients with neonatal encephalopathy after hypoxia and ischemia.