title: Peptidylglycine α-amidating monooxygenase heterozygosity alters brain copper handling with region specificity.
abstract: Copper (Cu), an essential trace element present throughout the mammalian nervous system, is crucial for normal synaptic function. Neuronal handling of Cu is poorly understood. We studied the localization and expression of Atp7a, the major intracellular Cu transporter in the brain, and its relation to peptidylglycine α-amidating monooxygenase (PAM), an essential cuproenzyme and regulator of Cu homeostasis in neuroendocrine cells. Based on biochemical fractionation and immunostaining of dissociated neurons, Atp7a was enriched in post-synaptic vesicular fractions. Cu followed a similar pattern, with ~ 20% of total Cu in synaptosomes. A mouse model heterozygous for the Pam gene (PAM+/−) was selectively Cu deficient in the amygdala. As in cortex and hippocampus, Atp7a and PAM expression overlap in the amygdala, with highest expression in interneurons. Messenger RNA levels of Atox-1 and Atp7a, which deliver Cu to the secretory pathway, were reduced in the amygdala but not in the hippocampus in PAM+/− mice, GABAB receptor mRNA levels were similarly affected. Consistent with Cu deficiency, dopamine β-monooxygenase function was impaired as evidenced by elevated dopamine metabolites in the amygdala, but not in the hippocampus, of PAM+/− mice. These alterations in Cu delivery to the secretory pathway in the PAM+/− amygdala may contribute to the physiological and behavioral deficits observed. Atp7a, a Cu-transporting P-type ATPase, is localized to the trans-Golgi network and to vesicles distributed throughout the dendritic arbor. Tissue-specific alterations in Atp7a expression were found in mice heterozygous for peptidylglycine α-amidating monooxygenase (PAM), an essential neuropeptide-synthesizing cuproenzyme. Atp7a and PAM are highly expressed in amygdalar interneurons. Reduced amygdalar expression of Atox-1 and Atp7a in PAM heterozygous mice may lead to reduced synaptic Cu levels, contributing to the behavioral and neurochemical alterations seen in these mice.
authors.affiliation: Department of Neuroscience, University of Connecticut Health Center, Farmington, Connecticut, USA.
authors.forename: Eric D
authors.initials: ED
authors.lastname: Gaier
authors.forename: Megan B
authors.initials: MB
authors.lastname: Miller
authors.forename: Martina
authors.initials: M
authors.lastname: Ralle
authors.forename: Dipendra
authors.initials: D
authors.lastname: Aryal
authors.forename: William C
authors.initials: WC
authors.lastname: Wetsel
authors.forename: Richard E
authors.initials: RE
authors.lastname: Mains
authors.forename: Betty A
authors.initials: BA
authors.lastname: Eipper
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D000679:Amygdala / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D002395:Catecholamines / Q000378:metabolism; D027682:Cation Transport Proteins / Q000378:metabolism; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000378:metabolism; D003300:Copper / Q000378:metabolism*; D000073840:Copper-Transporting ATPases; D005260:Female; D006579:Heterozygote; D006624:Hippocampus / Q000378:metabolism; D007395:Interneurons / Q000166:cytology / Q000201:enzymology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000235:genetics* / Q000378:metabolism*; D011247:Pregnancy; D013347:Subcellular Fractions / Q000378:metabolism; D021601:trans-Golgi Network / Q000378:metabolism
chemical_list: C486201:Atp7a protein, mouse; D002395:Catecholamines; D027682:Cation Transport Proteins; D009097:Multienzyme Complexes; D003300:Copper; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase; D000251:Adenosine Triphosphatases; D000073840:Copper-Transporting ATPases
keywords: Atp7a; PSD; amygdala; dopamine β-monooxygenase; neuropeptide; subcellular fractionation
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