GBA Mutation Assay

Parkinson's disease (PD) is a common neurodegenerative disease, and its specific pathogenesis mechanism is still not fully elucidated. Plenty of studies indicate that the role of aging and environmental factors leads to protein aggregation, inflammation, mitochondrial dysfunction and other phenomena that lead to the pathogenesis of PD. In addition, the genetic factors of PD cannot be ignored, among which GBA mutation is the largest genetic risk factor. Creative Biolabs now offers professional GBA mutation assay services to our worldwide clients, we hope our services can contribute to advancing your research.

Mechanism of GBA Mutation in PD

Studies found a connection between GBA mutations and PD more than a decade ago. The GBA is a gene located on the first chromosome (1q21), a locus consisting of two pseudogenes and nine genes. The GCase protein encoded by GBA can degrade glucocerebroside into glucose and ceramide, and can also degrade some of the glucosides. The mutations in the GBA gene are often associated with Gaucher disease (GD), a disease that shares similar clinical symptoms and molecular mechanisms with PD, so the pathogenic function of GBA mutation in PD was quickly proposed and confirmed. There are approximately 130 GBA mutations discovered in PD patients, among which C.1448 T > C (L444P) and C.1226A > G (N370S) are the two most common and persistent mutations, and they cover approximately 75% of Parkinson's-associated GBA mutants. Phosphorylated tau levels were higher in GBA mutants, and several key lipids were also higher in plasma. At the same time, the main pathological significance of GBA mutation lies in the decreased activity or loss of function of the encoded GCase, resulting in the accumulation of its metabolic substrate, GlcCer. The excessive presence of GlcCer affects protein trafficking, inhibits the autophagy-lysosomal pathway, and ultimately interferes with the degradation of α-synuclein and causes its aggregation and misfolding which leads to a dysfunction in the neuro system.

Fig 1. Mice hippocampus Wild Type and L144P Mutant neuronal cells, labeled by MitoGreen and MitoRed. (Li, et al., 2018)

Pathology of GBA Mutation in PD

The GBA gene mutations often lead to an increased risk of PD, earlier age of onset, cognitive impairment, and more severe behavioral impairment. Understanding the pathogenic mechanisms and different manifestations of this mutation is critical for geneticists and medical scientists to guide their research and is essential for clinicians to properly implement care.

We provide human neuroblastoma SH-SY5Y cells or mutant mice that have undergone site-directed mutagenesis or GBA knockout with viral plasmids for GBA mutation studies. We have a series of mutation libraries that allow you to choose from a range of heterozygous or homozygous mutations, including commonly used L444P and N370S GBA mutations. Western blot, immunochemical, or mass spectrometry techniques can be used to qualitatively or quantitatively detect the gene product of mutant GBA. In addition, gene sequencing approaches have also been used to characterize GBA mutations in vitro or in vivo.

Fig 2. Western blot assay for GBA, LC311b, Actin to determine the extent of autophagy in PD patient fibroblasts. (Collins, et al., 2018)

If you need any advice for GBA experimentation, assistance for trials or validation, or facile GBA mutation assay, Creative Biolabs can help. We have a professional R&D team to enforce experiment reliability and deliver the most rigorous and relevant GBA mutation assay for your project, please contact us so that we can provide you with more professional and detailed advice and services.

References

1. Li, H.; et al. Mitochondrial dysfunction and mitophagy defect triggered by heterozygous GBA mutations. Autophagy. 2018, 15(1): 113-130.
2. Collins, L.M.; et al. Dermal fibroblasts from patients with Parkinson's disease have normal GCase activity and autophagy compared to patients with PD and GBA mutations. F1000Research. 2018, 6: 1751.