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. 2024 Sep 17;7:1165. doi: 10.1038/s42003-024-06844-9

Fig. 2. EphrinB2 in astrocytes in BLA is essential for long-term fear conditioning memory formation.

Fig. 2

A Description of deletion of ephrinB2 in astrocytes of B6.129S7-Efnb2tm2And/J (floxed-ephrinB2) using the hGFAP-mCherry_iCre-WPRE-hGHp(A) AAV (GFAP-Cre). B mCherry expression in the amygdala is colocalized with the GFAP astrocytic marker (n = 5, 96.75 ± 0.35). C Photos that encompass mostly the BLA show that the GFAP-Cre construct is expressed in BLA of wild-type and B6.129S7-Efnb2tm2And/J mice as detected by mCherry. D EphrinB2 is depleted in astrocytes in BLA of floxed-ephrinB2 by the AAV expressing Cre-recombinase. Less mCherry expressing astrocytes in floxed-ephrinB2 mice (n = 11) also express ephrinB2 when compared to wt mice (n = 7) (t(16) = 5.107, p < 0.001). The area of staining of ephrinB2 in mCherry cells is significantly (t(16) = 4.882, p < 0.001) reduced in BLA in floxed ephrinB2 (n = 11) compared to wt mice (n = 7). E Long-term memory experiment: Mice were injected with AAV containing GFAP-Cre. One month later the mice were trained for fear conditioning and tested for contextual fear conditioning memory 24 h after fear conditioning and auditory fear memory 48 h after fear conditioning. F There is no difference in freezing during training between the groups (F(1,22) = 3.397, p = 0.079). G Deleting ephrinB2 leads to impairment of long-term contextual fear conditioning (p = 0.006). H Deleting ephrinB2 in astrocytes in BLA (n = 13) impaired long-term auditory fear memory formation when compared to the wt control group (n = 11) (F(1,22) = 9.243, p = 0.006). There is no treatment × tone trial interaction (F(4,88) = 1.668, p = 0.164). I Short-term memory experiment: Mice were injected with AAV containing GFAP-Cre. One month later the mice were trained for fear conditioning and tested for contextual fear conditioning memory 1 h after fear conditioning and auditory fear memory 2 h after fear conditioning. J There is no difference in freezing during training between the groups (F(1,15) = 2.373, p = 0.144). K There is no difference in contextual fear conditioning STM between the groups (p = 0.236). L EphrinB2 deleted mice (n = 9) were not different from control mice (n = 8) in short-term auditory fear conditioning memory (F(1,15) = 1.134, p = 0.304). Data are presented as mean ± SEM.