Ethanol-Induced Alterations in Placental and Fetal Cerebrocortical Annexin-A4 and Cerebral Cavernous Malformation Protein 3 Are Associated With Reductions in Fetal Cortical VEGF Receptor Binding and Microvascular Density

Abstract

Jegou et al. (2012) have reported prenatal alcohol exposure (PAE)-induced reductions of angiogenesis-related proteins in mouse placenta. These effects were associated with striking alterations in microvascular development in neonatal cerebral cortex. Here, we employed a rat model of moderate PAE to search for additional proteins whose placental and fetal cortical expression is altered by PAE, along with a subsequent examination of fetal cerebral cortical alterations associated with altered protein expression. Long-Evans rat dams voluntarily consumed either a 0 or 5% ethanol solution 4 h each day throughout gestation. Daily ethanol consumption, which resulted in a mean peak maternal serum ethanol concentration of 60.8 mg/dL, did not affect maternal weight gain, litter size, or placental or fetal body weight. On gestational day 20, rat placental: fetal units were removed by Caesarian section. Placental protein expression, analyzed by 2D-PAGEtandem mass spectroscopy, identified a total of 1,117 protein spots, 20 of which were significantly altered by PAE. To date, 14 of these PAE-altered proteins have been identified. Western blotting confirmed the alterations of two of these placental proteins, namely, annexin-A4 (ANX-A4) and cerebral cavernous malformation protein 3 (CCM-3). Specifically, PAE elevated ANX-A4 and decreased CCM-3 in placenta. Subsequently, these two proteins were measured in fetal cerebral cortex, along with radiohistochemical studies of VEGF binding and histofluorescence studies of microvascular density in fetal cerebral cortex. PAE elevated ANX-A4 and decreased CCM-3 in fetal cerebral cortex, in a pattern similar to the alterations observed in placenta. Further, both VEGF receptor binding and microvascular density and orientation, measures that are sensitive

Ethanol consumption during gestation promotes placental alterations in IGF-1 deficient mice

“…In this sense, ethanol consumption during gestation also reduces placental efficiency due to reductions in angiogenesis-related proteins. 42,43 However, the results suggest that ethanol consumption during gestation did not gravely affect the development of HZ embryo-placenta units. This could be due to the intricate genetics of the placenta, being noted that it is an organ conformed of a fetal part and a maternal part, which are in constant communication.…”

MicroRNA‐150‐5p is upregulated in the brain microvasculature during prenatal alcohol exposure and inhibits the angiogenic factor Vezf1

“…A variety of animal studies have shown that moderate PAE results in decreased cortical blood flow (Bake et al, 2012), changes in cerebral arterial reactivity including vasodilatory responses as a result of increased oxidative stress (Cananzi & Mayhan, 2019), and decreased oxygen and glucose metabolism (Gleason et al, 1997; Gleason & Hotchkiss, 1992; Richardson et al, 1987). In a rat model of PAE, Annexin‐A4, a calcium‐dependent membrane phospholipid binding protein expressed primarily in epithelial cells was upregulated in the fetal cerebral cortex, and has been associated with neurotoxicity in several neuropathologies (Savage et al, 2020). In addition, PAE decreased the expression of cerebral cavernous malformation protein 3 (CCM‐3) in the ventricular zone of the cerebral cortex (Savage et al, 2020).…”

“…In a rat model of PAE, Annexin‐A4, a calcium‐dependent membrane phospholipid binding protein expressed primarily in epithelial cells was upregulated in the fetal cerebral cortex, and has been associated with neurotoxicity in several neuropathologies (Savage et al, 2020). In addition, PAE decreased the expression of cerebral cavernous malformation protein 3 (CCM‐3) in the ventricular zone of the cerebral cortex (Savage et al, 2020). In the presence of VEGF, CCM‐3 is recruited to cell membranes where it protects VEGFR2 receptors from endocytosis, promoting angiogenesis during development and promoting migration of pyramidal cells and radial glia (Savage et al, 2020).…”

“…In addition, PAE decreased the expression of cerebral cavernous malformation protein 3 (CCM‐3) in the ventricular zone of the cerebral cortex (Savage et al, 2020). In the presence of VEGF, CCM‐3 is recruited to cell membranes where it protects VEGFR2 receptors from endocytosis, promoting angiogenesis during development and promoting migration of pyramidal cells and radial glia (Savage et al, 2020). The authors also found a decrease in VEGF binding to VEGF receptors and a reduction in vascular density at the ventricular zone in PAE embryos (Savage et al, 2020).…”

“…In the presence of VEGF, CCM‐3 is recruited to cell membranes where it protects VEGFR2 receptors from endocytosis, promoting angiogenesis during development and promoting migration of pyramidal cells and radial glia (Savage et al, 2020). The authors also found a decrease in VEGF binding to VEGF receptors and a reduction in vascular density at the ventricular zone in PAE embryos (Savage et al, 2020). Overall, this study also demonstrates that PAE alters the development of the cortical microvasculature network through decreased VEGF signaling (Savage et al, 2020).…”