Workgroup 1: article “Epigenetic programming by maternal behavior”
Also about figure 4.
Figure 1: maternal care alters cytosine methylation of the GR
promotor. About methylation levels.
A: sequence map of the exon GR promotor
- Sequence of the promotor 1,7 area. Each spot where
methylation can take place, is numbered. There are 17
potential areas.
- Area 16 and 17 are crucial for the transcription.
B, C: methylation analysis
- The colored block: pups from low licking mothers
White blocks: pups from high licking mothers.
- We want to know whether there is a difference
between that two groups, in the methylation.
- There is significantly more methylation in the black goup.
- The * means significant
- Is that difference really because of the licking? There are 3 potential reasons for the
methylation:
o Behavior mother: Differential behavior of the mother in licking and grooming
o Germ line methylation. Specific to sperm cells and sperm cells.
o Genetic
▪ You can test the cause due to cross fostering: you put the pups of the high
licking mice to the low licking mice and the other way around.
- In spot 17 there is equal level of methylation → focus on spot 16
- Picture C is focusing on the binding spots
D: about the cross fostering experiment→it is the behavior of the mother, not the other 2.Important!
- First letter is the biologic mother, the second letter is the behavior of the foster mother.
- You also test L-L and H-H, this is a negative control. It is the control of taking away the pup to
another mother. The effect of the experimentation.
- L-L: high level of methylation. H – H: low methylation levels . switching nests has no effect.
- H-L: Expectation if the methylation is genetic: you think it will be the same. This is not the
case, the methylation levels are in line with the behavior of the fosting mother.
E:
- P19 is 19 days after birth: here they are adults.
- You need 10 days of the mouse for 2 years of the human.
- A natural de novo effect.
- The groups that receive high licking for 10 days, at day 16 the methylation is still gone. If
there is no licking the methylation stays present. It is for the rest of their lives!
With a western blot you separate proteins (of the brain cortisol). With immunostaining you can make
the specific protein visible. see the cortical gene. You can also use fluorescence (antibody), this is
expensive.
15
Also about figure 4.
Figure 1: maternal care alters cytosine methylation of the GR
promotor. About methylation levels.
A: sequence map of the exon GR promotor
- Sequence of the promotor 1,7 area. Each spot where
methylation can take place, is numbered. There are 17
potential areas.
- Area 16 and 17 are crucial for the transcription.
B, C: methylation analysis
- The colored block: pups from low licking mothers
White blocks: pups from high licking mothers.
- We want to know whether there is a difference
between that two groups, in the methylation.
- There is significantly more methylation in the black goup.
- The * means significant
- Is that difference really because of the licking? There are 3 potential reasons for the
methylation:
o Behavior mother: Differential behavior of the mother in licking and grooming
o Germ line methylation. Specific to sperm cells and sperm cells.
o Genetic
▪ You can test the cause due to cross fostering: you put the pups of the high
licking mice to the low licking mice and the other way around.
- In spot 17 there is equal level of methylation → focus on spot 16
- Picture C is focusing on the binding spots
D: about the cross fostering experiment→it is the behavior of the mother, not the other 2.Important!
- First letter is the biologic mother, the second letter is the behavior of the foster mother.
- You also test L-L and H-H, this is a negative control. It is the control of taking away the pup to
another mother. The effect of the experimentation.
- L-L: high level of methylation. H – H: low methylation levels . switching nests has no effect.
- H-L: Expectation if the methylation is genetic: you think it will be the same. This is not the
case, the methylation levels are in line with the behavior of the fosting mother.
E:
- P19 is 19 days after birth: here they are adults.
- You need 10 days of the mouse for 2 years of the human.
- A natural de novo effect.
- The groups that receive high licking for 10 days, at day 16 the methylation is still gone. If
there is no licking the methylation stays present. It is for the rest of their lives!
With a western blot you separate proteins (of the brain cortisol). With immunostaining you can make
the specific protein visible. see the cortical gene. You can also use fluorescence (antibody), this is
expensive.
15
