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Primary Microglia Isolation from Mixed Glial Cell Cultures of Neonatal Rat Brain Tissue
Published on: August 15, 2012
Brainstem microglial cell morphology in neonatal rats with necrotizing enterocolitis and the effects of prenatal
Vonita Chawla1, Yomara S Mendez2, Lorraine C Siebold3
1Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, Arkansas, USA.
Background:
Necrotizing enterocolitis (NEC) is associated with increased neurodevelopmental impairment. Gut-brain interactions through the brainstem may be central to NEC-related microglia-driven neuroinflammation. Heparin-binding epidermal growth factor-like growth factor (HB-EGF) has intestinal protective properties and is a potential therapy for NEC. The aim of this study is to test the hypothesis that HB-EGF in pregnant rats reduces both NEC incidence and proinflammatory changes in the brainstem microglia of newborn rats.
Methods:
We compared four experimental groups, HB-EGF+/NEC-, HB-EGF-/NEC-, HB-EGF+/NEC+ and HB-EGF-/NEC+, depending on whether HB-EGF was given prenatally, and whether the newborn rats underwent the NEC induction protocol. We stained brainstem microglia and performed fractal analyses to provide objective measures of morphological changes.
Results:
NEC incidence was lower in the HB-EGF+/NEC+ group (n=64, p<0.005) than in the HB-EGF-/NEC+ group. Brainstem microglia of breastfed rats had a larger cell area, perimeter, roughness, and less circularity compared with smaller, denser, compact cells in the NEC+ pups (p<0.0001, n=320 cells). HB-EGF+/NEC+ and HB-EGF-/NEC+ pups had similar-appearing microglia.
Conclusions:
Prenatal HB-EGF treatment reduces NEC incidence in neonatal rats. NEC-related proinflammatory changes are seen in microglial cells present in crucial centers controlling the gut-brain pathway. HB-EGF has a growth-promoting effect on healthy microglia in the offspring but does not avert microglial activation in the brainstem of newborn rats with NEC.
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