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1.
Figure 3

Figure 3. Western blots show that GLE dose-dependently decreased retinal TH expression. From: Gestational Lead Exposure Selectively Decreases Retinal Dopamine Amacrine Cells and Dopamine Content in Adult Mice.

LS, MD and HD GLE significantly decreased TH expression by 21.9 ± 2.4%, 37.1 ± 4.1% and 53.0 ± 5.9%, respectively. The expression of choline acetyltransferase (ChAT), that co-labels with calretinin in IPL S2 and S4 (Haverkamp and Wässle, 2000), showed no significant treatment-related differences. GAPDH was used as a loading control. Representative Western blots are from four to five retinas from four to five different litters per treatment group. Values represent the mean ± SEM and were considered significant from controls if p < 0.05.

Donald A. Fox, et al. Toxicol Appl Pharmacol. ;256(3):258-267.
2.
Figure 4

Figure 4. GLE did not change the number of inner nuclear layer (INL) choline acetyltransferase (ChAT-IR) ACs. From: Gestational Lead Exposure Selectively Decreases Retinal Dopamine Amacrine Cells and Dopamine Content in Adult Mice.

Confocal images of the dorsal region of retinal wholemounts from control, LD, MD and HD GLE mice reveal that the number of ChAT-IR ACs in the INL was not different between treatment groups. Similar results were seen in the ventral region and with fixed-frozen vertical sections (Table 3). Images are representative of four to seven different retinas per treatment group. Scale bar = 40 µM.

Donald A. Fox, et al. Toxicol Appl Pharmacol. ;256(3):258-267.
3.
Figure 1

Figure 1. Gestational lead exposure (GLE) produced a dose- and region-dependent decrease in the number of tyrosine hydroxylase immunoreactive (TH-IR) dopaminergic amacrine cells (DA AC). From: Gestational Lead Exposure Selectively Decreases Retinal Dopamine Amacrine Cells and Dopamine Content in Adult Mice.

Confocal images of control, low- (LD), moderate- (MD) and high-dose (HD) GLE wholemounted retinas show a significant dose-dependent decrease in the number of TH-IR DA ACs. The images are from dorsal retina, which had a greater loss of DA ACs than the ventral retina (Table 3). Images are representative of four to seven different retinas per treatment group. Scale bar = 80 µM.

Donald A. Fox, et al. Toxicol Appl Pharmacol. ;256(3):258-267.
4.
Figure 6

Figure 6. GLE decreased DA metabolism in light- and dark-adapted retinas. From: Gestational Lead Exposure Selectively Decreases Retinal Dopamine Amacrine Cells and Dopamine Content in Adult Mice.

(A–C) *p<0.05 and **p<0.01 compared to corresponding controls. Values that share the same superscript are significantly different at p<0.05. Retinal DA, DOPAC and HVA content were determined by HPLC. (A) In light-adapted retinas, GLE significantly and dose-dependently decreased retinal DA, DOPAC and HVA content. (B) In light-adapted retinas, the [DOPAC]/[DA] and [HVA]/[DA] ratios were decreased significantly in retinas from GLE mice. (C) In controls, the retinal DA content was similar in dark- and light-adapted retinas and the retinal DOPAC content increased during light adaptation. In MD GLE retinas, the retinal DA content was decreased significantly relative to controls, however, it was similar in dark- and light-adapted retinas. Similarly, in MD GLE retinas, the retinal DOPAC content was decreased significantly, but it increased during light adaptation. [DOPAC]/[DA] was decreased significantly for each GLE treatment compared to control, while [HVA]/[DA] was decreased significantly in LD and MD GLE, but not in HD GLE compared to controls. In light-adapted retinas [DA] was unchanged in MD GLE mice, while [DOPAC] was decreased significantly compared to controls.

Donald A. Fox, et al. Toxicol Appl Pharmacol. ;256(3):258-267.
5.
Figure 5

Figure 5. GLE did not change the pattern or labeling intensity of monoamine oxidase A (MAO-A). From: Gestational Lead Exposure Selectively Decreases Retinal Dopamine Amacrine Cells and Dopamine Content in Adult Mice.

Fixed-frozen vertical sections show that MAO-A intensely labeled the entire retina, except for the outer nuclear layer (ONL). Since no previous retinal MAO-A confocal studies are published, double-label confocal experiments characterized its colocalization with the mitochondrial marker, COX IV. MAO-A and COX IV are colocalized and highly expressed in the inner segments (IS) and outer plexiform layer (OPL), while they are colocalized and moderately expressed in the IPL. In the INL, MAO-A-IR was greater than that of COX IV-IR. The location and expression level of MAO-A was not changed by MD GLE. Images are representative of four to five different retinas per treatment group. Scale bar = 20 µm.

Donald A. Fox, et al. Toxicol Appl Pharmacol. ;256(3):258-267.
6.
Figure 2

Figure 2. Moderate-dose GLE decreased the number of DOPA decarboxylase (DDC)-IR and TH-RFP-IR DA ACs and their associated inner plexiform layer (IPL) sublamina 1 (S1) plexus. From: Gestational Lead Exposure Selectively Decreases Retinal Dopamine Amacrine Cells and Dopamine Content in Adult Mice.

(A–B) Confocal images of the dorsal region of retinal wholemounts show that MD GLE significantly decreased the number of (A) DDC-IR and TH-IR co-labeled DA ACs and (B) TH::RFP-IR and TH-IR co-labeled DA ACs. All TH-IR DA ACs are either DDC-IR or TH::RFP-IR. The number and density of TH-IR S1 processes was decreased in GLE retinas. Images are representative of four to seven different retinas per treatment group. Scale bar = 80 µM. (C) Confocal images from fixed-frozen vertical sections show that TH-IR processes in the S1 sublamina (arrows) are markedly decreased in MD GLE retinas compared to controls. Double-label confocal experiments with calretinin, which labels ACs, retinal ganglion cells (RGCs) and IPL sublaminae S2, S3 and S4, reveal no co-labeling with TH and no significant differences between treatment groups. Images are representative of four to five different retinas per treatment group. Scale bar = 20 µm.

Donald A. Fox, et al. Toxicol Appl Pharmacol. ;256(3):258-267.

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