Student University of Minnesota Eden Prairie, Minnesota, United States
Background: Preterm infants are at high risk for phlebotomy-induced anemia (PIA). PIA in neonatal mice causes sex-dependent neurobehavioral deficits and downregulates the expression of CAMKIIa which plays a role in synaptic plasticity and learning and memory. Long term potentiation (LTP) and long term depression (LTD) are forms of synaptic plasticity which help memory consolidation and learning. LTP and LTD require both glutamatergic and GABAergic synaptic transmission. However, whether PIA affects GABA and glutamate receptors as a mechanism of neurodevelopmental impairment or differentially affects males and females is not known. Objective: To determine the effects of PIA on glutamate and GABA receptors in male and female neonatal mice. Design/Methods: Male and female mice were phlebotomized twice daily via facial venipuncture beginning at postnatal day 3 (P3) to reach a target hematocrit level of 18% on P10±1. Pups continued to be phlebotomized once daily to maintain this target hematocrit level until P14. Hippocampal tissue from PIA mice and controls were collected at P14 when RNA and cDNA was prepared. Gene expression of glutamate and GABA receptors were determined by qPCR (n=6/group), assayed in duplicate, and normalized against the reference gene PPIA. Protein levels of Grin2A, Grin2B, Grin1, GABRB1, and GABRB2 will be analyzed using western blotting to be completed by December 2023. Whole brain was frozen and sectioned. Hippocampal sections will be analyzed by NeuN and MAP2 immunohistochemistry January 2024.
