QUICK SEARCH:

[advanced]

	Author:		Keyword(s):

Home Help Feedback Subscriptions Archive Search Table of Contents

	Help viewing high resolution images
	Return to article

(Downloading may take up to 30 seconds.
If the slide opens in your browser, select File -> Save As to save it.)
Click on image to view larger version.

Fig. 6. Effect of Ptf1a absence on the generation and formation of different cell types in retinal explants. Phenotypic comparison of retinal explants from Ptf1a^+/Cre⁽^ex1⁾ R26R and Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R mice after 12 days in culture (DIC). Retinae from two siblings were dissected at E18.5 and were maintained in culture as whole retinae without destroying their cellular integrity for 12 days. Retinal explants were cryosectioned and immunolabeled with antibodies to cell-specific markers of all major classes of retinal cells. Immunofluorescence analysis of Ptf1a^+/Cre⁽^ex1⁾ R26R and Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R retinal explants at E18.5+12DIC with Thy1.2 (A,B) Hu/D (C,D), GABA (E,F), Gly-T1 (G,H), syntaxin (I,J), calbindin (K,L), PKC ${alpha}$ (M,N) and recoverin (O,P). (Q,R) Expression of ß-galactosidase. (S,T) Results of the TUNEL assay. (A) In Ptf1a^+/Cre⁽^ex1⁾ R26R retinal explant, Thy1.2 is expressed in the GCL (green, arrows). (B) In Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R retinal explant, Thy1.2-positive cells are located in the innermost zone of the INL and do not form a distinct cell layer (green, arrow). (C) In Ptf1a^+/Cre⁽^ex1⁾ R26R retinal explant, Hu/D is expressed in the GCL (arrowhead) and INL (arrow). (D) In Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R retinal explant, Hu/D positive cells are strongly reduced (arrows). (E) In Ptf1a^+/Cre⁽^ex1⁾ R26R retinal explant, GABA is expressed in amacrine (arrows) and displaced amacrine cells (asterisks). (F) By contrast, GABA-positive cells (asterisk) are virtually missing in Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R retinal explants. (G) Glycinergic amacrine cells express GlyT1 in Ptf1a^+/Cre⁽^ex1⁾ R26R retinal explant (arrows). (H) GlyT1-positive cells are not found in Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R retinal explants. (I) Syntaxin is found in the inner plexiform (IPL) and in amacrine cells (arrows, green) of Ptf1a^+Cre⁽^ex1⁾ R26R retinal explant. (J) By contrast, the IPL is missing in retinal explant from Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R embryos and the number of amacrine cells (arrows) is strongly reduced. (K) Calbindin is expressed in the OPL, horizontal cells (asterisk), amacrine cells (arrows) and displaced amacrine cells (arrows) of the Ptf1a^+/Cre⁽^ex1⁾ R26R retinal explant. (L) In Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R retinal explant only reduced number of calbindin-positive cells (arrows) can be detected. (M) Retinal explant from Ptf1a^+/Cre⁽^ex1⁾ R26R mice expresses PKC- ${alpha}$ in bipolar cells (arrows, green). (N) In retinal explant from Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R, there are few PKC- ${alpha}$ labeled cells (arrows, green), which do not form a cell layer and show a disarrayed structure. Both Ptf1a^+/Cre⁽^ex1⁾ R26R and Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R retinal explants are positive for recoverin in the ONL (O,P, green). Cells expressing ß-galactosidase (arrows) are significantly reduced in retinal explant of Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R (R) in comparison with Ptf1a^+/Cre⁽^ex1⁾ R26R (Q). In Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R retinal explant (S, arrows), compared with Ptf1a^+/Cre⁽^ex1⁾ R26R retinal explant (T, arrows), the number of apoptotic nuclei is significantly increased in both INL and ONL. In A-P and S,T the background was stained by DAPI (blue), showing the location of nuclei. Scale bar: 50 µm. GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer.

Return to article