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. Author manuscript; available in PMC: 2015 Dec 1.
Published in final edited form as: J Comp Neurol. 2014 Aug 25;522(17):3885–3899. doi: 10.1002/cne.23647

Figure 6. Axonal and synaptic pY816 TrkB immunoreactivity is not further reduced within stratum lucidum in ZnT3−/− BDNF−/− double knockouts compared to ZnT3+/+ BDNF−/− single knockout mice.

Figure 6

A&B) Representative low power images of hippocampi stained with pY816, taken from ZnT3+/+ BDNF−/− (A) and ZnT3−/− BDNF−/− (B) mice. Immunoreactivity does not appear further changed in the double knockout (B, arrows) compared to the ZnT3+/+ BDNF−/− animal (A, arrows). Scale bar = 300 µm. C&D) High power micrographs of stratum lucidum taken from pY816 stained sections from the same ZnT3+/+ BDNF−/− (C) and ZnT3−/− BDNF−/− (D) animals as in (A&B), demonstrating that pY816 immunoreactivity within mossy fiber axons, already weak in the absence of BDNF, is not further changed in the absence of both ZnT3 and BDNF (arrows point to pY816 enriched regions shown in Fig. 1 and Supplementary Fig. 1 to correspond anatomically to mossy fiber axons). Scale bar = 50 µm. E) Quantification of axonal pY816 immunoreactivity in ZnT3+/+ BDNF−/− (n=12) and ZnT3−/− BDNF−/− (n=12) mice reveals no change in the double compared to single knockout (p=0.25, Student’s t-test). Quantification is presented as mean pY816 axonal immunoreactivity ± SEM. F) Percentages of synapsin-1 puncta within stratum lucidum found to be pY816+ in ZnT3+/+ BDNF−/− (n=6) and ZnT3−/− BDNF−/− (n=8) mice. No significant difference in synaptic pY816 immunoreactivity was found between the two genotypes (p=0.48, Student’s t- test). A total of 1800 synapsin-1 puncta were analyzed in ZnT3+/+ BDNF−/− mice, and 2401 in ZnT3−/− BDNF−/− animals. Quantification is presented as mean % ± SEM. All data were analyzed by Student’s t-test.