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. Author manuscript; available in PMC: 2012 Sep 4.
Published in final edited form as: Cell Rep. 2012 Aug 9;2(2):223–230. doi: 10.1016/j.celrep.2012.07.001

Figure 2. Collagen XVIII/endostatin is necessary for the organization of CF terminals.

Figure 2

A. Neurons that synapse onto Purkinje cells (PC). BC – basket cell; CF – climbing fiber; GC – granule cell; IO – inferior olivary neuron; PF – parallel fiber; SC – stellate cell. B. Nerve terminal markers used to label terminals on PCs. Parentheses indicate Syt2 is present in a small set of PF terminals. C,D,F,G,I,J,L,M. Immunostaining for VGluT1 (C,D), Bassoon (F,G), Syt2 (I,J) and VGluT2 (L,M) in P56 control (Ctl) and col18a1-/- mutant cerebellum. Insets in I,J show high magnification of Syt2-positive axo-somatic terminals on PCs (see arrows). Insets in L,M show high magnification of VGluT2 distribution in CFs: note its diffuse axonal distribution in col18a1-/- mutants (arrows in L,M). E,H,K. Average fluorescent intensity of VGluT1-, Bassoon-, and Syt2-immunoreactivity in P56 control (Ctl) and mutants (KO). Data shown are mean ± SEM; n >4 mice per genotype. N. Quantification of the total number of VGluT2-positive varicosities per image field in P14 and P56 col18a1-/- mutants compared to controls (dashed line). Data shown are mean ± SD; n >4 mice per genotype. *Differ from litter mate controls at p<0.001 by Students t-test. O. Western blots show no difference in VGluT2 levels in P56 Ctl and KO cerebellum (CB). Actin was a loading control. P. qPCR of vglut2 mRNA expression levels in P56 IO and CB of KOs and controls (dashed line). Data are mean ± SEM; n =3. Q. Adult KOs exhibit impaired mobility on a rotating rod. Data are mean ± SEM; n =10 Ctls and 13 KOs. *Differ from Ctls at p<0.05 by Tukey HSD Test for differences between means. R,S. Immunostaining for Calb and VGluT2 in P21 Ctl and KO cerebella. T. Quantification of the percent of ML crossed by VGluT2-containing nerve terminals. Data are mean ± SEM; n = 60 measures from 3 mice. U,V. DiI labeled climbing fibers in the ML of P20 control and col18a1-/- mutant cerebellum. W. CF length in ML of KO and Ctl cerebella. X. Quantification of the extent of ML crossed by the CFs in KO and Ctl cerebella. For W,X data are mean ± SEM; n = 26 control CFs and 23 mutants CFs. Scale bar in C = 30 μm for C,D,F,G,I,J,L,M, in insets = 18μm, in Q= 25 μm for Q,R, and in T = 30 μm for T,U.