Display Settings:

Items per page

Results: 9

1.
Figure 8

Figure 8. From: Ascl1 genetics reveals insights into cerebellum local circuit assembly.

In the absence of Ascl1 granule cell precursors are marked with Ascl1CreER GIFM. (A,A') Xgal staining of cerebellar sections of E17.5 Ascl1CreER/+;R26lacZ/+ mice administered Tm at E11.5 showing marking of only Pcs, in contrast to Ascl1CreER/Δ;R26lacZ/+ mice in which gcps in the egl (dotted outline) are marked in addition to Pcs (B,B'). (A”,B”) Double-labeling immunohistochemistry of Pax6 (green) and ßgal (red) shows that Ascl1CreER/Δ GIFM marked cells within the egl are gcps (Pax6+). Xgal staining of E17.5 cerebellar sections of Ascl1CreER/+;R26lacZ/+ (C,C') and Ascl1CreER/Δ;R26lacZ/+ (D,D') when Tm was administered at E13.5 reveals marking only of cells within the white matter, some of which are positive for Pax2 (C',D'). Xgal staining of Ptf1aCre/+;Ascl1CreER/Δ;R26lacZ/+ shows more gcps marked within the egl (dotted lines [E]) compared to littermate controls (F). Scale bar: A–F 200 μm; A',A”,B'B”,C'D” 100 μm.

Anamaria Sudarov, et al. J Neurosci. ;31(30):11055-11069.
2.
Figure 6

Figure 6. From: Ascl1 genetics reveals insights into cerebellum local circuit assembly.

Temporal fate mapping of cerebellar glia with Ascl1CreER GIFM reveals marking of Bergmann glia only at one time point and other glia during their proliferative phase. Ascl1CreER GIFM marking of Bergmann glia (A–A”), oligodendrocytes (B–B') and astrocytes (C–C') is shown. (A') Double-labeling immunohistochemistry for GFAP (red) and YFP (green) confirms the identity of Bergmann glia. (A”) BrdU was administered 12 hours prior to Tm administration at E13.5. Double-labeling immunohistochemistry for BrdU (green) and ßgal (red) shows Ascl1CreER GIFM and long term BrdU marked Bergmann glia. (B') The identity of oligodendrocytes is shown by double-labeling immunohistochemistry for CAII (red) and YFP (green). (C') Double-labeling immunohistochemistry for GFAP (red) and YFP (green) shows some Ascl1CreER GIFM marked cells are astrocytes. (D) The percentage of each type of cerebellar glia marked with Ascl1CreER GIFM at each time point during development is shown (mean±SEM; n = 4, unpaired t test). wm, white matter; Pc, Purkinje cell; igl, internal granular layer; ml, molecular layer. Scale bar: 20 μm.

Anamaria Sudarov, et al. J Neurosci. ;31(30):11055-11069.
3.
Figure 5

Figure 5. From: Ascl1 genetics reveals insights into cerebellum local circuit assembly.

A population of cerebellar interneurons is born during embryonic development. BrdU was administered 12 hours prior to Tm administration at E13.5, E14.5, P2 and P7. (A) Double-labeling immunohistochemistry in the outer ml for BrdU (red) and Parvalbumin (green) shows some stellate cells are born at E14.5. (A') Double-labeling immunohistochemistry for ßgal (red) and BrdU (green) shows an Ascl1CreER GIFM fate mapped stellate cell born embryonically. (B) Double-labeling immunohistochemistry in the inner ml for BrdU (red) and Parvalbumin (green) shows some basket cells are born at E14.5. (B') Double-labeling immunohistochemistry for ßgal (red) and BrdU (green) shows an Ascl1CreER GIFM fate mapped basket cell born embryonically. (C) Double-labeling immunohistochemistry in the igl for BrdU (green) and Neurogranin (red) shows some Golgi cells are born at E14.5. (C') Double-labeling immunohistochemistry for ßgal (red) and BrdU (green) shows an Ascl1CreER GIFM fate mapped Golgi cell born embryonically. (D) Quantification of the number of BrdU and ßgal double positive interneurons marked at each time point with Ascl1CreER GIFM is shown (mean±SEM; n = 3, unpaired t test). ml, molecular layer; Pcl, Purkinje cell layer; igl, internal granular layer; pia, pial surface. Scale bar: 20 μm.

Anamaria Sudarov, et al. J Neurosci. ;31(30):11055-11069.
4.
Figure 9

Figure 9. From: Ascl1 genetics reveals insights into cerebellum local circuit assembly.

Model for the temporal generation of the cells that form the cerebellar local circuitry. (A) Summary of the time points at which vz-derived cells are preferentially marked using Ascl1CreER GIFM. The thickness of the line indicates the relative number of each cell type marked at different time points. (B) During early embryogenesis (E10.5–E13.5) Pcs and neurons of the cn are born and begin to make contacts. During mid and late embryogenesis (E13.5-P0), a population of Bg and Golgi interneurons are born. Bg migrate to their final position within the Pc layer to guide newly differentiated gcs towards the igl. Golgi interneurons and gcs within the igl then form synaptic contacts within glomeruli. During early postnatal stages (P0–P5) basket interneurons are primarily generated and contact Pc apical dendrites. Finally, during late postnatal stages (P5–P15), stellate interneurons are mainly generated and occupy the outer region of the ml, and start making contacts with Pc dendrites. Astrocytes and oligodendrocytes are generated after birth and contact their target neurons. Thus, target neurons are generated before the neurons or glia that contact them are generated.

Anamaria Sudarov, et al. J Neurosci. ;31(30):11055-11069.
5.
Figure 3

Figure 3. From: Ascl1 genetics reveals insights into cerebellum local circuit assembly.

Purkinje cells marked with Ascl1CreER GIFM on consecutive days settle in distinct M–L and A–P patterns. (A–C) Wholemount Xgal staining of embryos 24 hours post Tm administration. (A) E11.5 wholemount Xgal staining of Pcs marked at E10.5 reveals that Pcs were uniformly labeled in the lateral cerebellar primordium. (B) At E12.5 wholemount Xgal staining of Pcs marked at E11.5 shows labeling of Pcs in the cerebellar primordium excluding the midline. (C) E13.5 wholemount Xgal staining of Pcs marked at E12.5 reveals that Pcs were uniformly labeled throughout the M–L axis of the cerebellum. (D) P0 wholemount Xgal staining of Pcs marked at E10.5 shows that Pcs were found throughout the cerebellum except the midline. (E) P0 wholemount Xgal staining of Pcs marked at E11.5 depicts four pairs of distinct M–L Pc bands. (F) P0 wholemount Xgal staining of Pcs marked at E12.5 shows a single midline Pc band and two bilateral clusters in the paravermis. (G) Summary schematic of A–P distribution pattern of Pcs marked at E11.5 (red) and E12.5 (blue). Arrows in E and F indicate positions analyzed in G. (H) The percentage of marked Pcs residing in each of the cardinal lobes at E18.5 cerebellum. Mb, midbrain; Cb, cerebellum; anb, anterobasal; and, anterodorsal; cen, central; pos, posterior; inf, inferior. Scale bar: A–C 200 μm, D–F 350 μm.

Anamaria Sudarov, et al. J Neurosci. ;31(30):11055-11069.
6.
Figure 2

Figure 2. From: Ascl1 genetics reveals insights into cerebellum local circuit assembly.

GABAergic cn interneurons and Purkinje cells are the first cell types marked by Ascl1CreER GIFM. (A) Coronal section of the adult cerebellum highlighting where images in B–E and I were taken. (B) Double-labeling immunohistochemistry for GABA (green) and ßgal (red) shows that cn interneurons were marked when Tm was administered at E10.5. Anti-Calbindin (green) and anti-ßgal (red) double-labeling immunohistochemistry shows that fate mapped cells are Purkinje cells when Tm was administered at E10.5 (C), E11.5 (D) and E12.5 (E). (F–H) Dorsal views of wholemount Xgal staining of P21 cerebella when Tm was administered at E10.5 (F), E11.5 (G) and E12.5 (H). (I) BrdU was administered 12 hours prior to Tm administration at E10.5, E11.5 or E12.5 and analysis performed at P21. Double-labeling immunohistochemistry for ßgal (red) and BrdU (green) shows that Ascl1CreER GIFM marked Pcs retained BrdU labeling. (J) Quantification of the number of double positive BrdU and βgal Pcs (mean±SEM; n = 3; unpaired t test). ml, molecular layer; Pcl, Purkinje cell layer; igl, internal granular layer. Scale bar: A 100 μm, B,C,D,E,I 20 μm, F,G,H 600 μm.

Anamaria Sudarov, et al. J Neurosci. ;31(30):11055-11069.
7.
Figure 7

Figure 7. From: Ascl1 genetics reveals insights into cerebellum local circuit assembly.

Conditional deletion of Ascl1 in the cerebellum differentially affects the number of interneurons and glia in the adult. Hematoxylin and eosin (H&E) stained sections show smaller cerebellum in a Ptf1a-Ascl1 cko (B) compared to a littermate control (A). Inset in A and B is immunostaining for Calbindin showing no difference in the overall morphology of Pcs. H&E stained sections of En1-Ascl1 ckos (F) shows a similar cellular phenotype to Ptf1a-Ascl1 ckos (A). Higher magnification images show severe reductions in the number of ml interneurons in both ckos (D,H) compared to littermate controls (C,G). (I) Quantification of the number of Purkinje cells per mm in Ptf1a-Ascl1 and En1-Ascl1 ckos and littermate controls. (J) Quantification of the average cerebellar circumference in Ptf1a-Ascl1 and En1-Ascl1 ckos and littermate controls with controls set as 1 shows significantly smaller cerebellum size in Ptf1a-Ascl1 ckos (p=0.01) but not En1-Ascl1 ckos. (K) Quantification of the GABAergic interneurons and glia in Ptf1a-Ascl1 and En1-Ascl1 ckos and littermate controls with average control numbers set as 1. (mean±SEM; Ptf1a-Ascl1 cko n = 3, Ptf1a-Ascl1 control, n = 3; En1-Ascl1 cko n = 3, En1-Ascl1 control, n = 3, unpaired t test, red star depicts p<0.003, black star p<0.05) Scale bar: A,B,E,F 100 μm; C,D,G,H 20 μm.

Anamaria Sudarov, et al. J Neurosci. ;31(30):11055-11069.
8.
Figure 4

Figure 4. From: Ascl1 genetics reveals insights into cerebellum local circuit assembly.

Temporal fate mapping of cerebellar interneurons reveals inside to outside settling in the cortex. All sections are analyzed at P21. (A–C) An example of fate mapped cell types when Tm was administered at E14.5. (A,A') Distinct morphology of basket cells, found in the ml close to Pcl, is shown. Double-labeling immunohistochemistry for Parvalbumin (red) and YFP (green) identifies some fate mapped cells in inner ml as basket cells. (B,B') Distinct morphology of stellate cells located in the outer ml is shown. Double-labeling immunohistochemistry for Parvalbumin (red) and YFP (green) in the outer ml identifies some fate mapped cells as stellate cells. (C,C') Large size of fate mapped interneurons in the igl indicates Golgi cells. Double-labeling immunohistochemistry for Neurogranin (red) and YFP (green) confirms Golgi cell identity. (D) A chandelier-shaped candelabrum fate mapped (YFP+) cell in the inner ml is shown. (E–J) Mosaic images of YFP immunohistochemistry in Ascl1CreER GIFM animals when Tm was administered at E13.5 (E), E14.5 (F), E17.5 (G), P0 (H), P1 (I), P4 (J). (K) A comparison of the percentage of each cell type marked at different time points during development is shown (mean±SEM; n = 3, unpaired t test). White dotted line indicates pial surface, red dotted line indicates border between Pcl and igl, and yellow dotted line indicates border between igl and wm. ml, molecular layer; Pcl, Purkinje cell layer; wm, white matter; igl, internal granular layer. Scale bar: A–D 20 μm, E–J 100 μm.

Anamaria Sudarov, et al. J Neurosci. ;31(30):11055-11069.
9.
Figure 1

Figure 1. From: Ascl1 genetics reveals insights into cerebellum local circuit assembly.

The Ascl1CreER allele is expressed similar to Ascl1, and primarily marks cells that rapidly stop dividing. CreER expression (A–E) closely resembles endogenous Ascl1 expression (F–J) in the cerebellum (outline) throughout embryogenesis. (K–O) Ascl1 expressing cells have distinct proliferative characteristics at various time points. (P–T”) Ascl1CreER/+;R262RlacZ/+ animals were given Tamoxifen (Tm) at different time points and sacrificed 24 hours later. One hour BrdU pulse was given to Ascl1CreER/+;R262RlacZ/+ animals prior to sacrificing. (P) Tm was administered at E10.5 and sagittal sections of E11.5 embryos analyzed by Xgal staining. Xgal+ cells were found away from the vz. (P') Double-labeling immunohistochemistry for BrdU and ßgal shows BrdU (green) in the vz, whereas Ascl1CreER GIFM marked cells were BrdU negative. (P”) Higher magnification of P'. Same type of analysis was performed for Tm administration at E11.5 (Q–Q”) and E14.5 (R–R”). Few Ascl1CreER GIFM marked cells incorporated BrdU (pink arrows) when Tm was administered at E14.5 (R',R”). (S') When Tm was administered at E17.5 and the cerebellum analyzed at P2, few cells were Ki67+ (pink arrows). (S”) The majority of ßgal+ cells were Pax2+ (pink arrowheads). (T',T”) When Tm was administered at P1, very few of the fate mapped cells were Ki67+ (pink arrow). vz, ventricular zone; rl, rhombic lip; wm, white matter. Scale bar: A–P',Q,Q',S–T” 50µm; P”,Q”,R” 30 μm.

Anamaria Sudarov, et al. J Neurosci. ;31(30):11055-11069.

Display Settings:

Items per page

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk