Overview
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- Prenatal treatment with 0.2ppm MeHg causes ASD-like behavior in young adult mice
- istered 0 and 0.2ppm MeHg (an amount at least 10 times lower than previous in vivo studies40 via drinking
- (Figure 1E). We observed significantly more marbles buried by the 0.2ppm MeHg-treated mice (Figure 1F),
- Figure 1. Prenatal treatment with 0.2ppm MeHg causes ASD-like behaviour in young adult mice
- (A) Schematic of prenatal exposure to MeHg in pregnant mice, created with BioRender.com. 0ppm (control) and 0.2ppm MeHg was administered through
- (C and D) The total number of calls, n = 11 animals/group, Mann–Whitney U test, *p < 0.05, and (D) average call length per call was analyzed,
- n = 10 animals/group, Student’s t test, *p < 0.05.
- 0.2ppm MeHg. (F) Quantitative analysis of number of marbles buried in the cage. n = 9–15 animals/group, Student’s t test, **p < 0.01.
- (H) The time spent in interaction zone for female mice, n = 12 animals/group, Student’s t test, **p < 0.01, ns (non-significant).
- measurement of learning. n = 15–16 animals/group.
- shown in L. (L) The percentage of time spent in the target quadrant between control and 0.2ppm MeHg was analyzed as a measurement of memory.
- n = 15–16 animals/group, Student’s t test, ns (non-significant).
- across 2 days of training with latency to reach the platform as a measurement of learning. n = 15 animals/group.
- in P. (P) The percentage of time spent in the new target quadrant and old target quadrant was analyzed as a measurement of reversal memory. n = 15
- animals/group, Student’s t test, *p < 0.05, ns (non-significant). Error bars indicate the standard error of the mean (SEM). See also Figure S1.
- repetitive behavior. Here, mice prenatally treated with either 0.2ppm MeHg or 0ppm MeHg (control) were
- cate that prenatal 0.2ppm MeHg exposure impairs communication, increases restrictive and repetitive be-
- (A) Schematic of prenatal exposure to MeHg in pregnant mice, created with BioRender.com. 0ppm (control) and 0.2ppm MeHg were administered through
- (B, E) Images of E15 cerebral cortex sections from embryos receiving 0 and 0.2ppm MeHg treatment, immunostained for Pax6 (B, green), Ki67 (B, red) or Tbr2
- progenitors (F) within VZ and SVZ/IZ, determined from sections similar to those shown in (B, E). n = 3 embryos/group, Student’s t test *p < 0.05.
- (G) Images of E15 cerebral cortex sections from embryos receiving 0 and 0.2ppm MeHg treatment, immunostained for DCX (G, green) and counterstained
- (H) Quantitative analysis of the number of DCX+ immature neurons within VZ, determined from sections similar to those shown in (G). n = 4 embryos/group,
- (I) Quantitative analysis of thickness for the CP was determined from coronal sections similar to those shown in (G). n = 3 embryos/group, Student’s t test,
- (J) Images of E15 cerebral cortex sections from embryos receiving 0 and 0.2ppm MeHg treatment, immunostained for Tbr1 (red) and counterstained for
- (K) Quantitative analysis of the number of Tbr1+ deep layer mature neurons within CP, determined from sections similar to those shown in (J). n = 4 embryos/
- group. Error bars indicate the standard error of the mean (SEM). See also Figure S2.
- 0.2ppm MeHg via drinking water to pregnant mice from E0 until E15 and sacrificed the dams to collect
- E15 brain samples from embryos for immunohistochemical analysis (Figure 2A). We found that 0.2ppm
- genitors was not significantly changed by 0.2ppm MeHg exposure (Figures 2E and 2F), the number of DCX+
- rons over total live cells after 0.2ppm MeHg exposure (Figures 2J and 2K). We also reported that 0.2ppm
- exposure of 0.2ppm MeHg promotes premature differentiation of cortical precursors in the VZ to generate
- To ask whether prolonged exposure of 0.2ppm MeHg in pregnant mice through the perinatal stage from E0
- opment, we administered 0 and 0.2ppm MeHg via drinking water to pregnant mice starting from E0 to P7,
- Tbr2+ intermediate progenitor populations within the SVZ in the P7 brain were not altered by 0.2ppm
- was significantly increased after the 0.2ppm MeHg treatment (Figures 3D and 3H). We further examined
- deep and superficial layer cortical neurons and found that 0.2ppm MeHg treatment increased the total
- throughout the cortex (Figures 3L–3N) by 0.2ppm MeHg treatment. In addition, 0.2ppm MeHg treatment
- (A) Schematic of perinatal exposure to MeHg in pregnant mice, created with BioRender.com. 0ppm (control) and 0.2ppm MeHg were administered through
- (B–D) Images of P7 SVZ sections from pups receiving 0 and 0.2ppm MeHg treatment since E0, immunostained for Pax6 (B, green), Ki67 (B, red), Tbr2 (C, red)
- progenitors (G) and DCX+ immature neurons (H) within SVZ of P7 brains, determined from sections similar to those shown in (B, C, D). n = 4 embryos/group,
- (I and L) Images of P7 cerebral cortex sections from pups receiving 0 and 0.2ppm MeHg treatment since E0, immunostained for Tbr1 (I, red) and Satb2 (L, red)
- standard error of the mean (SEM). See also Figure S3.
Methods (brief)
- drinking water to pregnant mice starting at E0 until E15. At E15 brains were collected for immunohistochemistry.
- E15 brain samples from embryos for immunohistochemical analysis (Figure 2A). We found that 0.2ppm
- drinking water to pregnant mice starting at E0 until P7. At P7 brains were collected for immunohistochemistry.
- collected from E13.5 embryos that have been exposed to 0ppm or 0.2ppm MeHg as described above to
- profile dissected and dissociated E13.5 murine cortical cells (Figure 4A). In total, we collected 6 embryonic
- drinking water to pregnant mice starting at E0 until E13.5 at which point the dorsal cortical cells were collected for multiplexing. Two technical replicates
- drinking water to pregnant mice starting at E0 until E15. At E15 brains were collected for immunohistochemistry.
- doi.org/10.1016/S0092-8674(02)00657-8. et al. (2019). MULTI-seq: sample multiplexing
- stem.2012.03.016. Goodman, R.H. (2004). Defining the CREB in the BTBR T + Itpr3 tf/J autistic mouse
- nancy E0 until E15. On E15, pregnant mice were sacrificed, and embryos were collected. The brains of the
- until P7. The P7 pups were sacrificed on the last day of treatment, and their brains were collected. To study
- ment, brains were collected from the E15 embryos for immunohistochemistry. For all immunohistochemical
- Multiplexing samples for scRNA-seq
- cerebral cortices were collected after the removal of meninges from the brains of embryos. The cortical tis-
- was then allowed to settle for 1 min. The supernatant was collected and was centrifuged at 300g for 2 min.
- group (0ppm/0.2ppm MeHg) were duplicated and divided into two samples (250,000 cells/sample). The
- four samples were labeled with four different barcodes. For the barcode labelling, the resulting pellet
- Prof. Zev Gartner from the University of California, San Francisco). Samples were then incubated on ice
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