Role of Smug1 on the modulation of gene expression in mouse brain during neurodevelopment
1 Purpose
This project aims at understanding the molecular mechanisms played by the DNA glycosylase Smug1 during neurodevelopment. Our previous animal experiments have shown that the Smug1 deficient mice have rewarded-based learning deficit and impulsivity when submitted to behavioural tasks. Recently, we found that Smug1 regulates the expression of olfactory receptors in the olfactory bulb and prefrontal cortex, modulating the sensory perception of smell. Olfaction issues are the first symptoms presented by individuals developing neurodegenerative diseases, but until the moment, little is known about the initiation and progression of this important sign of neurodegeneration. We need more understanding about how the loss of olfaction is triggered to develop prevention and treatment strategies for these patients. We believe that an in-depth characterization of the molecular mechanism associated to the neurodevelopment of Smug1 deficient mice will give us insight into how olfaction deficiency is triggered.
2 Distress:
All mice used in this project will undergo a non-recovery procedure. The animals will be either sacrificed before the procedure or deeply anaesthetised during the procedure and then terminated.
3 Benefits:
Neurological conditions affecting human brains, such as neurodegenerative diseases the body's activities including balance, movement, cognition, and learning. The prevention and treatment of such diseases are still a challenge and therefore require more molecular understanding besides novel therapeutic strategies. The work described here promotes scientific renewal about the brain in development by defining a new principle of gene regulation mediated by the Smug1 enzyme. It represents original research into fundamental mechanisms that may have implications for common brain diseases detected in humans.
4 Animals:
All animals will be bred and provided by FOTS 30099.
Strains: Smug1 deficient mice C57BL/6 (Smug1 KO) and C57BL/6 (WT) as control.
- 212 WT mice: 48 embryos (E14.5), 110 young mice (includes P1, P5, P10, P15, P20), 54 adults (includes P60, 4-6 months
- 212 Smug1KO mice: 48 embryos (E14.5), 110 young mice (includes P1, P5, P10, P15, P20), 54 adults (includes P60, 4-6 months
5 How to adhere to 3R:
In vitro studies using cell lines have been conducted to characterise the molecular mechanisms behind DNA glycosylases. However, cell lines are not able to provide information regarding which brain region is more affected or at which stage of the development alterations in behaviour are compromised by Smug1. We will, therefore, collect data from embryonic, early postnatal mice that are actively developing the brain and compare then with fully developed mice. From the embryonic mice, we will also isolate and propagate neural stem cells and proceed with in vitro experiments to answer our research questions when appropriate, avoiding the use of a higher number of animals. For this, we will breed homozygous Smug1-/- mice that are set up with animals from heterozygous breeding. This strategy guarantees good control of the breeding and avoids the use of further animals.
This project aims at understanding the molecular mechanisms played by the DNA glycosylase Smug1 during neurodevelopment. Our previous animal experiments have shown that the Smug1 deficient mice have rewarded-based learning deficit and impulsivity when submitted to behavioural tasks. Recently, we found that Smug1 regulates the expression of olfactory receptors in the olfactory bulb and prefrontal cortex, modulating the sensory perception of smell. Olfaction issues are the first symptoms presented by individuals developing neurodegenerative diseases, but until the moment, little is known about the initiation and progression of this important sign of neurodegeneration. We need more understanding about how the loss of olfaction is triggered to develop prevention and treatment strategies for these patients. We believe that an in-depth characterization of the molecular mechanism associated to the neurodevelopment of Smug1 deficient mice will give us insight into how olfaction deficiency is triggered.
2 Distress:
All mice used in this project will undergo a non-recovery procedure. The animals will be either sacrificed before the procedure or deeply anaesthetised during the procedure and then terminated.
3 Benefits:
Neurological conditions affecting human brains, such as neurodegenerative diseases the body's activities including balance, movement, cognition, and learning. The prevention and treatment of such diseases are still a challenge and therefore require more molecular understanding besides novel therapeutic strategies. The work described here promotes scientific renewal about the brain in development by defining a new principle of gene regulation mediated by the Smug1 enzyme. It represents original research into fundamental mechanisms that may have implications for common brain diseases detected in humans.
4 Animals:
All animals will be bred and provided by FOTS 30099.
Strains: Smug1 deficient mice C57BL/6 (Smug1 KO) and C57BL/6 (WT) as control.
- 212 WT mice: 48 embryos (E14.5), 110 young mice (includes P1, P5, P10, P15, P20), 54 adults (includes P60, 4-6 months
- 212 Smug1KO mice: 48 embryos (E14.5), 110 young mice (includes P1, P5, P10, P15, P20), 54 adults (includes P60, 4-6 months
5 How to adhere to 3R:
In vitro studies using cell lines have been conducted to characterise the molecular mechanisms behind DNA glycosylases. However, cell lines are not able to provide information regarding which brain region is more affected or at which stage of the development alterations in behaviour are compromised by Smug1. We will, therefore, collect data from embryonic, early postnatal mice that are actively developing the brain and compare then with fully developed mice. From the embryonic mice, we will also isolate and propagate neural stem cells and proceed with in vitro experiments to answer our research questions when appropriate, avoiding the use of a higher number of animals. For this, we will breed homozygous Smug1-/- mice that are set up with animals from heterozygous breeding. This strategy guarantees good control of the breeding and avoids the use of further animals.