Myeloid cells are involved in the pruning of terminal fields in control mice and can be activated to reversibly prune terminal fields in E3–E12 diet mice. (A) Photomicrographs of horizontal sections through the intermediate zone of the NST showing IX (green), GSP (red), and CT (blue) terminal fields and the overlaps of all three nerves (MERGE) in control mice that were injected with saline or minocycline. (B) Terminal field volumes of the IX, GSP, and CT nerves in individual control mice (symbols) and their means injected with saline (n = 7) or with minocycline (n = 6) daily from P15 to P40. Minocycline-injected mice showed reduced pruning of GSP and CT terminals. (C) Horizontal sections through the intermediate zone of the NST showing IX, GSP, and CT terminal fields and the overlaps of all three nerves in control mice fed a control chow (control diet) or with PLX5622 added to the control diet (PLX5622 treated) from P18–P40. The photomicrographs are duplicated for the intermediate zone in Fig. S4 A. (D) Terminal field volumes of the IX, GSP, and CT nerves in control mice fed the control diet (n = 7) or the PLX5622-treated diet (n = 7). PLX5622-treated mice showed reduced pruning of GSP and CT terminals. (E) Horizontal sections through the intermediate zone NST showing IX, GSP, and CT terminal fields and the overlaps of all three nerves in E3–E12 diet mice who were injected with saline or m-csf. The photomicrographs are duplicated for the intermediate zone in Fig. S4 D. (F) Terminal field volumes of the IX, GSP, and CT nerves in E3–E12 diet mice and their means injected with saline (n = 10) or with m-csf (n = 8) every third day from P10 to P40. m-csf–injected mice showed a pruning of GSP and CT terminals. (G) Microglia densities and their means in the rostral NST in saline (n = 3) or minocycline-injected control mice (n = 4) and for saline- (n = 6) or m-csf–injected E3–E12 diet mice (n = 8). Refer to the color guide in the lower portion of E for colors shown in the merged images in A, C, and E. Scale bars = 200 µm. R, rostral; L, lateral shown in the lower portions of E. All statistical comparisons were multiple, unpaired t tests using the Holm–Sidak method for multiple comparisons. Data are shown as mean ± SEM. * denotes P < 0.05. (B) GSP, P = 0.00003; CT, P = 0.0004. (D) GSP, P = 0.000001; CT, P = 0.000001. (F) GSP, P = 000001; CT, P = 0.00006. (G) Minocycline, P = 0.002; m-csf, P = 0.007. One observation/animal was used for analyses.
Figure 4.

Myeloid cells are involved in the pruning of terminal fields in control mice and can be activated to reversibly prune terminal fields in E3–E12 diet mice. (A) Photomicrographs of horizontal sections through the intermediate zone of the NST showing IX (green), GSP (red), and CT (blue) terminal fields and the overlaps of all three nerves (MERGE) in control mice that were injected with saline or minocycline. (B) Terminal field volumes of the IX, GSP, and CT nerves in individual control mice (symbols) and their means injected with saline (n = 7) or with minocycline (n = 6) daily from P15 to P40. Minocycline-injected mice showed reduced pruning of GSP and CT terminals. (C) Horizontal sections through the intermediate zone of the NST showing IX, GSP, and CT terminal fields and the overlaps of all three nerves in control mice fed a control chow (control diet) or with PLX5622 added to the control diet (PLX5622 treated) from P18–P40. The photomicrographs are duplicated for the intermediate zone in Fig. S4 A. (D) Terminal field volumes of the IX, GSP, and CT nerves in control mice fed the control diet (n = 7) or the PLX5622-treated diet (n = 7). PLX5622-treated mice showed reduced pruning of GSP and CT terminals. (E) Horizontal sections through the intermediate zone NST showing IX, GSP, and CT terminal fields and the overlaps of all three nerves in E3–E12 diet mice who were injected with saline or m-csf. The photomicrographs are duplicated for the intermediate zone in Fig. S4 D. (F) Terminal field volumes of the IX, GSP, and CT nerves in E3–E12 diet mice and their means injected with saline (n = 10) or with m-csf (n = 8) every third day from P10 to P40. m-csf–injected mice showed a pruning of GSP and CT terminals. (G) Microglia densities and their means in the rostral NST in saline (n = 3) or minocycline-injected control mice (n = 4) and for saline- (n = 6) or m-csf–injected E3–E12 diet mice (n = 8). Refer to the color guide in the lower portion of E for colors shown in the merged images in A, C, and E. Scale bars = 200 µm. R, rostral; L, lateral shown in the lower portions of E. All statistical comparisons were multiple, unpaired t tests using the Holm–Sidak method for multiple comparisons. Data are shown as mean ± SEM. * denotes P < 0.05. (B) GSP, P = 0.00003; CT, P = 0.0004. (D) GSP, P = 0.000001; CT, P = 0.000001. (F) GSP, P = 000001; CT, P = 0.00006. (G) Minocycline, P = 0.002; m-csf, P = 0.007. One observation/animal was used for analyses.

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