QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

This Article Summary Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Toresson, H. Articles by Campbell, K. Search for Related Content PubMed PubMed Citation Articles by Toresson, H. Articles by Campbell, K.

A role for Gsh1 in the developing striatum and olfactory bulb of Gsh2 mutant mice

Wallenberg Neuroscience Center, Division of Neurobiology, Lund University, Sölvegatan 17, BMC A11, S-221 84 Lund, Sweden

View larger version (109K): [in a new window]   Fig. 1. The striatal complex at E18.5 in wild types and Gsh2 homozygous mutants. Immunostaining for ISL1 (A,B) delineates the entire striatal complex while SOX1 staining (C,D) selectively marks the olfactory tubercle and portions of the nucleus accumbens (Acc) (i.e. ventral striatum). The Gsh2 mutant displays a significant reduction in size of the striatal complex (B), which is most prominent in the ventral striatum, in particular within the olfactory tubercle (OT) region (B,D). Stm, striatum.

View larger version (83K): [in a new window]   Fig. 2. Patch-matrix organization at E18.5 in wild types and Gsh2 homozygous mutants. DARPP-32 immunohistochemistry outlines the dorsal striatum and shows intense labeling in striatal patches and the subcallosal streak (scs) of the wild type (A). Labeling can still be detected in the Gsh2 mutant but patches are fewer and smaller and labeling of the scs is weaker (B). At this stage, immunohistochemistry for the µ-OR shows patchy distribution and scs labeling in the wild type (C) and to a lesser extent in the Gsh2 mutant (D). Arrowheads in A-D show representative patches labeled with either DARPP-32 or µ-OR. CaBP expression in the Gsh2 mutant striatum (F) appears to be increased as compared to wild type (E). ac, anterior comissure; cc, corpus callosum.

View larger version (77K): [in a new window]   Fig. 3. Olfactory bulb interneurons at E18.5 in wild types and Gsh2 mutants. DLX immunohistochemistry reveals positive cells in the SVZ, granule cell layer (GCL) and glomerular layer (GL) of the wild type (A). In the Gsh2 mutant (B) fewer than normal cells are positive for DLX and these cells are mostly confined to the SVZ. GABAergic interneurons of the granule cell layer and glomerular layer in the wild type (C) are revealed by immunohistochemistry for GAD67. In the Gsh2 mutant (D), there is a dramatic reduction in the number of GAD67 cells and most are found in the granule cell layer. E and F show the medial portion of the olfactory bulbs at higher magnification to visualize the few TH neurons present in the glomerular layer. Interneurons positive for TH are detected in the glomerular layer of wild-type (E) but not Gsh2 mutant (F) olfactory bulbs.

View larger version (101K): [in a new window]   Fig. 4. Gsh1 expression in wild types and Gsh2 mutants. At E16.5, Gsh1 expression in the wild type is confined to the ventralmost VZ corresponding to the former MGE and medialmost LGE (A). In Gsh2-/- embryos at this stage, Gsh1 transcripts are detected at a high level throughout the LGE (B). Arrowheads in A and B indicate the LGE-cortex boundary. At E12.5, wild-type Gsh1 expression is observed in the MGE and medialmost LGE (C). A slight expansion of the Gsh1 expression domain is evident in the Gsh2 mutant at this stage (D).

View larger version (92K): [in a new window]   Fig. 5. The striatal complex at E18.5 in Gsh1 and Gsh1/2 homozygous mutants. No abnormalities can be seen in ISL1 (A) or SOX1 (C) expression in the Gsh1 mutant striatum (Stm). The Gsh1/2 double mutant has only a rudimentary striatum as revealed by ISL1 (B) and nearly absent SOX1 (D) staining. Acc, nucleus accumbens; OT, olfactory tubercle.

View larger version (80K): [in a new window]   Fig. 6. Patch-matrix organization at E18.5 in Gsh1 and Gsh1/2 double homozygous mutants. The Gsh1 mutant striatum appears identical to wild type for DARPP-32 (A), µ-OR (C) and CaBP (E). The Gsh1/2 double mutant striatum is devoid of DARPP-32-positive neurons (C) and no µ-OR patches can be detected (F). Arrowheads in A and C point to patches. Neurons expressing CaBP are detected in the rudimentary double mutant striatum (F). ac, anterior comissure; cc, corpus callosum.

View larger version (68K): [in a new window]   Fig. 7. Olfactory bulb interneurons at E18.5 in Gsh1 and Gsh1/2 homozygous mutants. DLX (A), GAD67 (C) and TH (E) expression appear normal in Gsh1 mutants. The Gsh1/2 double mutant has very few DLX (B) labeled cells in the SVZ. The double mutants are also completely devoid of GAD67 staining (D) and lack TH-positive cells (F). GL, glomerular layer; GCL, granule cell layer.

View larger version (80K): [in a new window]   Fig. 8. Progenitor cell specification at E12.5 in Gsh2 and Gsh1/2 double homozygous mutants. MASH1 protein is detected in many cells of the MGE and LGE VZ of the wild type (A). In the Gsh2 mutant the LGE level of MASH1 is lower and confined to the ventromedial VZ (B). In the Gsh1/2 double mutant, only a few MASH1-positive cells are seen in the LGE, close to the MGE (C). The majority of VZ and SVZ cells of the wild type are positive for DLX (D). In the Gsh2 mutant, only cells of the medialmost LGE SVZ express DLX proteins (E) while DLX-positive cells in the Gsh1/2 double mutant LGE are very rare (F). In situ hybridization for Raldh3 marks cells in the ventromedial LGE of the wild type (G). Raldh3 expression is present at low levels in the same region of the Gsh2-/- LGE (H) but completely absent in the double mutant (I). High levels of PAX6 protein show a sharp boundary in the dorsal LGE of the wild type (J). Ectopic PAX6 is found in a large region of the LGE VZ in both Gsh2 mutants (K) and Gsh1/2 double mutants (L).

View larger version (85K): [in a new window]   Fig. 9. Progenitor cell specification in Gsh2 and Gsh1/2 double homozygous mutants at E16.5. (A,B) NGN2 expression is detected in many cells of the cortical VZ but not in the LGE of the wild type (A). The Gsh2 mutant looks similar to the wild type at this stage (B). In the Gsh1/2 double mutant (C), high levels of NGN2 are detected only in the cortical VZ but low levels can also be detected in a few cells in the lateral LGE. (D-F) The expression of PAX6 is also similar in wild type (D), Gsh2 single (E) and Gsh1/2 double mutants (F). (G-I) In the wild type, high levels of DLX protein can be detected in the VZ, SVZ and mantle of the ventral telencephalon (G). The Gsh2 mutant shows less labeling in the VZ but otherwise looks similar to the wild type (H). Considerably fewer cells in the Gsh1/2 double mutant LGE SVZ express DLX (I). (J-L) NKX2.1 expression defines the remnant of the MGE (asterisk in J) at this stage and clearly helps to visualize the shortening of the LGE VZ in the Gsh2 mutant (K) and Gsh1/2 double mutant (L) compared to the wild type (J). The NKX2.1-positive region in L is larger than in J and K, this is because the section in L is somewhat more caudal than the others; we do not notice an increase in MGE size in the double mutant. Arrows in I and L shows the boundary between the MGE and LGE. GP, globus pallidus.

View larger version (134K): [in a new window]   Fig. 10. Proliferating cells at E12.5 and E16.5 in Gsh2 and Gsh1/2 double homozygous mutants. Immunohistochemistry for the cell cycle marker Ki67 shows strong labeling in the VZ and SVZ (separated by the dashed line) of the wild-type LGE and MGE (A). Labeling in the Gsh2 mutant is normal in the VZ and MGE SVZ but significantly lower in the LGE SVZ (B). The Gsh1/2 double mutant (C) is similar to the Gsh2 mutant. At E16.5, the SVZ of the Gsh2 mutant LGE (E) has improved, showing numerous Ki67-positive cells which is not the case in the SVZ of the double mutant LGE (E).