Fig. 1: Effective separation of synaptic proteins from trisynaptic circuit synapses using laser-capture microdissection and biochemical processing of hippocampal layers. | Nature Communications

Fig. 1: Effective separation of synaptic proteins from trisynaptic circuit synapses using laser-capture microdissection and biochemical processing of hippocampal layers.

From: Synaptic proteome diversity is shaped by the levels of glutamate receptors and their regulatory proteins

Fig. 1: Effective separation of synaptic proteins from trisynaptic circuit synapses using laser-capture microdissection and biochemical processing of hippocampal layers.

a Representative image of a coronal section showing the hippocampus, as used for laser-capture microdissection (LCM). CA1/CA3 pyramidal layers and dDG granular layer are visible. b Representative image of CA1 before microdissection. The Pyramidal Layer is shown in purple, the green line marks collected neuropile fragments from the Stratum Radiatum (width ~100 µm). c CA1 subfield (in b) after LCM. d Representative image of the Dentate Gyrus after LCM, fragments (width ~100 µm) collected from the Molecular Layer neuropile. Granular layer in orange. e Representative image of the CA3 subfield after LCM, fragments (width ~100 µm) collected from Stratum Lucidum. The pyramidal layer is green. f Total area (mm2) microdissected and number of brain sections (as shown in Fig. 1d) collected for the 3 biological replicas used in proteomics of dDG. g Total area (mm2) microdissected and number of brain sections (as shown in Fig. 1e) collected for the 3 biological replicas used in proteomics of CA3. h Total area (mm2) microdissected and number of brain sections (as shown in Fig. 1c) collected for the 3 biological replicas used in proteomics of CA1. i Outline of synaptic protein enrichment from LCM samples. j Immunoblot of 1% Triton X-100 insoluble (Pellet) and soluble (Supern.) fractions from CA1, CA3 and dDG proved for Psd95 and Synaptophysin (Syp). k Relative Psd95 abundance in 1% Triton soluble (Supern.) and insoluble (Pellet) fractions. IU: intensity units. Bars = mean ± SEM. Statistics, unpaired two-sided Student’s T-test, sample size (n) = 3. ** p < 0.01. l Relative Syp abundance in 1% Triton soluble (Supern.) and insoluble (Pellet) fractions. IU: intensity units. Bars = mean ± SEM. Statistics unpaired two-sided Student’s T-test, sample size (n) = 3. ** p < 0.01. m Protein yield in Triton X-100 pellets per microdissected area. To recover ~20 μg, ~100 mm2 neuropile must be microdissected. Error bars show SEM. Sample size (n, biological replicates) 5. Scale Bar in a 1000 μm, in b–e 300 μm. Source data are provided as a Source Data file. Parts of this figure were created with BioRender. Roher, N. (2025) https://BioRender.com/pin1gen.

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