Protocols for assessing mitophagy in neuronal cell lines and primary neurons.

Mitochondria are organelles that regulate essential eukaryotic functions including generating energy, sequestering excess calcium, and modulating cell survival. In order for neurons to thrive, mitochondria have to be continuously replenished by maintaining autophagic-lysosomal mediated degradation of mitochondria (mitophagy) and mitochondrial biogenesis. While a plethora of image- and biochemical-based techniques have been developed for measuring autophagy (macroautophagy) in eukaryotic cells, the molecular toolbox for quantifying and assessing mitophagy in neurons continues to evolve.

PINK1 regulates mitochondrial trafficking in dendrites of cortical neurons through mitochondrial PKA.

Mitochondrial Protein Kinase A (PKA) and PTEN-induced kinase 1 (PINK1), which is linked to Parkinson's disease, are two neuroprotective serine/threonine kinases that regulate dendrite remodeling and mitochondrial function. We have previously shown that PINK1 regulates dendrite morphology by enhancing PKA activity. Here, we show the molecular mechanisms by which PINK1 and PKA in the mitochondrion interact to regulate dendrite remodeling, mitochondrial morphology, content, and trafficking in dendrites.

Role of membrane microdomains in compartmentation of cAMP signaling.

Spatially restricting cAMP production to discrete subcellular locations permits selective regulation of specific functional responses. But exactly where and how cAMP signaling is confined is not fully understood. Different receptors and adenylyl cyclase isoforms responsible for cAMP production are not uniformly distributed between lipid raft and non-lipid raft domains of the plasma membrane.

Integrin upregulation and localization to focal adhesion sites in pregnant human myometrium.

Focal adhesions are integrin-rich microdomains that structurally link the cytoskeleton to the extracellular matrix and transmit mechanical signals. In the pregnant uterus, increases in integrin expression and activation are thought to be critical for the formation of the mechanical syncytium required for labor. The aim of this study was to determine which integrins are upregulated and localized to focal adhesions in pregnant human myometrium.