Neuronal Mitophagy Detection Assay Service

At Creative Biolabs, we offer in vitro neuronal mitophagy assay services aimed at advancing mechanistic research and drug development for the central nervous system (CNS). For further details about our products and services, as well as for project-specific consultations and pricing information, please submit an inquiry here.

Introduction

Neuronal mitochondrial autophagy is an important cellular process that maintains energy homeostasis and normal function of neurons by selectively removing damaged or dysfunctional mitochondria. Mitophagy involves several key proteins, such as PINK1 and Parkin, which play critical roles in recognizing and degrading damaged mitochondria. This process is regulated by several signaling pathways that play a role in pathological conditions such as neurodegenerative diseases and cerebral ischemia. Abnormal regulation of mitophagy may lead to the onset of neurodegenerative diseases.

Mitophagy Assays

• Mitochondrial Function Assays

• Fluorescent Markers for Autophagy

We provide comprehensive and professional autophagy marker detection services, including those in the PINK1-PRKN-dependent pathway and the LC3 phagocytosis system, as well as through p-S65-Ub antibody and TOM20 expression level changes.

PINK1-PRKN

The PINK1-PRKN-dependent pathway is an important neuronal autophagy mechanism that maintains cellular homeostasis mainly by selectively degrading damaged mitochondria. Under normal circumstances, PINK1 is degraded by the mitochondrial proteasome, but in damaged mitochondria, PINK1 accumulates on the mitochondrial membrane, eventually activating PRKN. Activated PRKN enhances mitophagy signals through ubiquitination, thereby promoting the clearance of damaged mitochondria.

Fig.1 Representative images of colocalization of mitochondrial markers DsRed-Mito and parkin in N2a cells.¹

LC3

LC3 (microtubule-associated protein 1 light chain 3) is a key protein in the process of mitophagy, mediating the clearance of damaged mitochondria by directly interacting with a variety of mitophagy receptors such as BNIP3 and Nix. This mechanism is of great significance in neurological diseases, helping neurons to cope with energy demands and stress conditions by maintaining the quality and quantity of mitochondria.

Fig.2 Representative images of colocalization of mitochondrial markers DsRed-Mito and LC3-II in N2a cells.¹

p-S65-Ub

PINK1 specifically phosphorylates the Ser65 site of ubiquitin (Ub) on damaged mitochondria to form p-S65-Ub, which serves as a signal for the degradation of damaged mitochondria. Antibodies against p-S65-Ub have been used to detect PINK1-PRKN-mediated autophagy and have been shown to be effective in various cell types, including primary human fibroblasts and stem cell-derived dopamine neurons.

TOM20

TOM20, a TOM family member, is found in the mitochondria's outer membrane. TOM20 regulates mitophagy by promoting PINK1 activity and recruiting Parkin. In drug screening, activating or inhibiting these interactions may provide new therapeutic strategies to improve mitochondrial function in neurodegenerative diseases.

• Monitor Autophagy Progression Assays

Our experiments are equipped with super-resolution microscopy and state-of-the-art live cell imaging systems to precisely monitor the process of mitophagy. These tools can effectively distinguish between mitochondria in contact with lysosomes and the engulfed state, providing a deep understanding of the autophagy mechanism.

• Custom Assays

At Creative Biolabs, we provide the option to tailor neuronal mitophagy assays to meet your unique requirements. Alongside this, we supply premium materials, data, and insights to enhance the success of your projects. Reach out to us to discuss your project plans.

Neuronal Mitophagy-Related Products

High-quality primary cells are essential for successful in vitro experiments. Our primary neural cells, upheld by strict quality control standards, offer reliable support for your scientific research.

Reference

1. Li, Jie, et al. "PINK1-parkin-mediated neuronal mitophagy deficiency in prion disease." Cell Death & Disease 13.2 (2022): 162. Distributed under Open Access license CC BY 4.0, without modification.