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A gene-environment study of cytoglobin in the human and rat hippocampus
Christian Ansgar Hundahl1, Betina Elfving, Heidi Kaastrup Müller
1Centre of Excellence for Translational Medicine, University of Tartu, Tartu, Estonia.
Insights
Cytoglobin (Cygb) and neuronal nitric oxide synthase (nNOS) are co-expressed in the hippocampus and their expression increases with stress. The rat hippocampus models human Cygb expression, suggesting Cygb
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Cytoglobin (Cygb), the fourth vertebrate heme-globin, has an unknown function.
- In vitro studies suggest Cygb scavenges reactive oxygen species and metabolizes nitric oxide.
- In vivo studies indicate Cygb is upregulated by hypoxic stress.
Purpose of the Study:
- To determine if the rat hippocampus is a valid model for human Cygb expression.
- To investigate the co-expression of Cygb and neuronal nitric oxide synthase (nNOS) in the rat hippocampus.
- To assess the impact of chronic restraint stress (CRS) on Cygb and nNOS expression in the hippocampus.
Main Methods:
- Immunohistochemistry to compare Cygb expression in human and rat hippocampi.
- Investigated Cygb and nNOS co-expression in rat hippocampus.
- Quantitative real-time PCR and Western blotting to analyze Cygb and nNOS transcription and translation in response to CRS.
Main Results:
- Rat and human hippocampus exhibit similar Cygb expression patterns.
- High co-expression of Cygb and nNOS was observed in the rat hippocampus.
- CRS significantly upregulated nNOS and Cygb protein levels in the dorsal hippocampus of Flinders (FSL/FRL) rats.
Conclusions:
- The rodent hippocampus serves as a valid model for studying Cygb protein localization in the human hippocampus.
- Co-expression of Cygb and nNOS supports Cygb's role in nitric oxide metabolism.
- CRS induces Cygb and nNOS expression, indicating Cygb is stress-responsive and potentially involved in physiological stress responses.
Background:
Cytoglobin (Cygb) was discovered a decade ago as the fourth vertebrate heme-globin. The function of Cygb is still unknown, but accumulating evidence from in vitro studies point to a putative role in scavenging of reactive oxygen species and nitric oxide metabolism and in vivo studies have shown Cygb to be up regulated by hypoxic stress. This study addresses three main questions related to Cygb expression in the hippocampus: 1) Is the rat hippocampus a valid neuroanatomical model for the human hippocampus; 2) What is the degree of co-expression of Cygb and neuronal nitric oxide synthase (nNOS) in the rat hippocampus; 3) The effect of chronic restraint stress (CRS) on Cygb and nNOS expression.
Methods:
Immunohistochemistry was used to compare Cygb expression in the human and rat hippocampi as well as Cygb and nNOS co-expression in the rat hippocampus. Transcription and translation of Cygb and nNOS were investigated using quantitative real-time polymerase chain reaction (real-time qPCR) and Western blotting on hippocampi from Flinders (FSL/FRL) rats exposed to CRS.
Principal Findings:
Cygb expression pattern in the human and rat hippocampus was found to be similar. A high degree of Cygb and nNOS co-expression was observed in the rat hippocampus. The protein levels of nNOS and Cygb were significantly up-regulated in FSL animals in the dorsal hippocampus. In the ventral hippocampus Cygb protein levels were significantly up-regulated in the FSL compared to the FRL, following CRS.
Significance:
The rodent hippocampus can be used to probe questions related to Cygb protein localization in human hippocampus. The high degree of Cygb and nNOS co-expression gives support for Cygb involvement in nitric oxide metabolism. CRS induced Cygb and nNOS expression indicating that Cygb expression is stress responsive. Cygb and nNOS may be important in physiological response to stress.
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