Peroxisomal membrane channel Pxmp2 in the mammary fat pad is essential for stromal lipid homeostasis and for development of mammary gland epithelium in mice

AbstractTo understand the functional role of the peroxisomal membrane channel Pxmp2, mice with a targeted disruption of the Pxmp2 gene were generated. These mice were viable, grew and bred normally. However, Pxmp2−/− female mice were unable to nurse their pups. Lactating mammary gland epithelium displayed secretory lipid droplets and milk proteins, but the size of the ductal system was greatly reduced. Examination of mammary gland development revealed that retarded mammary ductal outgrowth was due to reduced proliferation of epithelial cells during puberty. Transplantation experiments established the Pxmp2−/− mammary stroma as a tissue responsible for suppression of epithelial growth. Morphological and biochemical examination confirmed the presence of peroxisomes in the mammary fat pad adipocytes, and functional Pxmp2 was detected in the stroma of wild-type mammary glands. Deletion of Pxmp2 led to an elevation in the expression of peroxisomal proteins in the mammary fat pad but not in liver or kidney of transgenic mice. Lipidomics of Pxmp2−/−mammary fat pad showed a decrease in the content of myristic acid (C14), a principal substrate for protein myristoylation and a potential peroxisomal β-oxidation product. Analysis of complex lipids revealed a reduced concentration of a variety of diacylglycerols and phospholipids containing mostly polyunsaturated fatty acids that may be caused by activation of lipid peroxidation. However, an antioxidant-containing diet did not stimulate mammary epithelial proliferation in Pxmp2−/− mice.The results point to disturbances of lipid metabolism in the mammary fat pad that in turn may result in abnormal epithelial growth. The work reveals impaired mammary gland development as a new category of peroxisomal disorders

Solute traffic across the mammalian peroxisomal membrane—the role of Pxmp2

Abstract Peroxisomes are small oxidative organelles found in all eukaryotes. They contain a matrix which is surrounded by a single membrane and consists mainly of soluble proteins. Peroxisomal enzymes are involved in a broad spectrum of metabolic pathways including conversion of lipids, amino- and hydroxyacids, purines and reactive oxygen species. The carbon fluxes through peroxisomal pathways require a continuous metabolite crossing of the peroxisomal membrane. A long-standing and still unresolved problem of the physiology of mammalian peroxisomes is the role of the membrane of these organelles as a permeability barrier to solute molecules. In this study, we have shown that the peroxisomal membrane represents a type of biomembrane where channel-forming proteins coexist with solute transporters. Disruption of the mouse Pxmp2 gene, encoding the peroxisomal integral membrane protein Pxmp2 also known as PMP22, leads to partial restriction of peroxisomal membrane permeability to solutes in vitro and in vivo. Multiple-channel recording of liver peroxisomal preparations revealed that the channel-forming components with a conductance of 1.3 nS in 1.0 M KCl were lost in Pxmp2-/- mice. The channel-forming properties of Pxmp2 were confirmed with recombinant protein expressed in insect cells and with native Pxmp2 purified from mouse liver. The Pxmp2 channel, with an estimated diameter of 1.4 nm, shows weak cation selectivity and no voltage dependence. The long-lasting open states of the channel indicate its functional role as a protein forming a general diffusion pore in the membrane. Hence, Pxmp2 is the first peroxisomal pore-forming protein identified, and its existence suggests that the mammalian peroxisomal membrane is permeable to small solutes, while transfer of bulky metabolites, e.g., cofactors (NAD/H, NADP/H, and CoA) and ATP, requires specific transporters. In addition, the phenotypic characterisation of Pxmp2-/- mice has revealed a role for Pxmp2 during the development of the epithelia in the mammary glands of female mice. The disruption of Pxmp2 leads to the impairment of ductal outgrowth of mammary glands at puberty, which is followed by the inability of Pxmp2-/- mice to nurse their offspring