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Method for Measuring the Activity of Deubiquitinating Enzymes in Cell Lines and Tissue Samples
Published on: May 10, 2015
Expression of the deubiquitinating enzyme mUBPy in the mouse brain
Federica Bruzzone1, Mauro Vallarino, Giovanna Berruti
1Department of Biology, University of Genova, V.le Benedetto XV, 5 16132 Genova, Italy.
Abstract:
Mouse UBPy (mUBPy) is an ubiquitin-specific protease which belongs to a family of deubiquitinating enzymes (DUBs) implicated in several cellular processes related to both cell growth and differentiation. Previously, Northern blot analysis revealed an important expression of mUBPy in the testis and brain. However, a more comprehensive map of mUBPy localization in the central nervous system (CNS) is still lacking. In this study, we mapped the distribution of mUBPy in the mouse brain using nonradioactive in situ hybridization and immunohistochemical techniques. In general, transcript and protein showed a similar and widespread distribution. In particular, mUBPy was strongly expressed in the hippocampal formation, septal region, ventral pallidum, preoptic nucleus, periventricular nucleus of hypothalamus, compact part of the substantia nigra, ventral tegmental area, cochlear nucleus and granular cell layer of cerebellum. A moderate expression of mUBPy was found in the amygdaloid complex, supraoptic nucleus, arcuate and ventromedial nuclei of hypothalamus, lateral hypothalamic area and lateral and reticular part of the substantia nigra. Double labelling with the mUBPy antiserum and antisera against specific cell markers showed that the enzyme is generally expressed in neurons and, in specific regions, also in oligodendrocytes. Moreover, by using antisera to TH and mUBPy we found that mUBPy is localized in dopaminergic neurons. The different distribution of mUBPy in the distinct regions of the brain suggests that it could be related to different deubiquitinating processes; in particular, in the areas where it is expressed at high levels, mUBPy could exert a specialized function through its interaction with specific protein substrates.
Insights
Mouse UBPy (mUBPy), a deubiquitinating enzyme, is widely distributed in the brain, primarily in neurons. Its specific localization suggests specialized functions in various brain regions, including dopaminergic neurons.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Ubiquitin-specific proteases (USPs) are crucial deubiquitinating enzymes (DUBs) involved in cellular processes.
- Mouse UBPy (mUBPy) expression was previously noted in the testis and brain, but its central nervous system (CNS) distribution was not fully mapped.
Purpose of the Study:
- To comprehensively map the distribution of mUBPy protein and transcript within the mouse brain.
- To identify the specific cell types expressing mUBPy in the CNS.
- To investigate the localization of mUBPy in dopaminergic neurons.
Main Methods:
- Nonradioactive in situ hybridization to map mUBPy transcript distribution.
- Immunohistochemistry using mUBPy antiserum to determine protein localization.
- Double labeling with cell-specific markers and tyrosine hydroxylase (TH) antiserum.
Main Results:
- mUBPy transcript and protein exhibited a widespread and similar distribution across the mouse brain.
- Strong mUBPy expression was observed in the hippocampus, substantia nigra, ventral tegmental area, and cerebellum.
- mUBPy was found in neurons and, in certain areas, oligodendrocytes, and co-localized with dopaminergic neurons.
Conclusions:
- mUBPy displays a distinct regional expression pattern in the mouse brain, suggesting diverse roles in deubiquitination.
- High expression levels in specific brain areas indicate potential specialized functions for mUBPy.
- The localization in dopaminergic neurons suggests a role in regulating dopaminergic pathways.
