Ects cells was performed. both anti SynDIG1 mAb and anti-HA recognized a single immunoreactive band at 32 kDa, consistent with the calculated molecular weight of HA SynDIG1. Against a SynDIG1 immunoreactive band of slightly lower molecular weight was also detected in extracts of the mouse brain and dissociated rat hippocampal Epothilone A neurons. COS cells with HA SynDIG1 Immunf coloring Transfected showed identical patterns for anti-HA mAb and anti SynDIG1. To begin, the epitope were generated by mAb anti SynDIG1 two constructions HA removal SynDIG1 identify detected. Deletion of 33 amino acids The C-terminal hydrophobic Dom ne including normal of the second had no effect w on anti SynDIG1 mAb recognition During deletion of 75 amino Acids from the N-terminus resulted in a total loss Anti SynDIG1 mAb immunoreactivity t.
Important that thwart HA immunoreactivity was t Similar for all constructs, which marks the presence of the HA protein. SynDIG1 protein expression peaks w During the second week of postnatal development, OSI-930 the biggest e time of synapse formation in rodents. Moreover, the expression of the brain SynDIG1 is limited, in agreement with the distribution of mRNA SynDIG1 UniGene database. SynDIG1 groups with AMPA receptors colocalize SynDIG1 advantage, S expression in the hippocampus, the expression was in dissociated rat hippocampal neurons with anti SynDIG1 mAb and anti-MAP2 antique Investigated body. Determine the subcellular Re localization of SynDIG1, SynDIG1 expression in rat hippocampal neurons in dissociated culture of different ages was Immunf Staining with anti SynDIG1 mAb and anti-MAP2 antique Investigated body.
In young cultures was SynDIG1 immunoreactivity t in cellpar.in the adjust Bodies and neurites in a point- Shaped and diffuse F Demonstrated staining. At DIV 8, further SynDIG1 immunoreactivity t in cellpar.in the adjust Bodies and dendrites by Immunf Staining with anti-MAP2 co antique Demonstrated rpern occur. In mature dendritic seemed two types of immunoreactivity t, which have been developed over time, are: 1 fl chenhaften and point-shaped spots along the shafts of dendrites, and in particular in thick and visible Prim rdendriten F staining in two projections along the dendrites. SynDIG1 clusters at synapses enriched as defined by the overlap with postsynaptic and pr Synaptic marker associated with inhibitory synapses.
At 7 DIV contain 48% of the synapses SynDIG1. A DIV containing 10 and 15 64% and 56% of the synapses or SynDIG1. The H eh SynDIG1 at the synapses is 31%, 47% and 52% of total puncta SynDIG1 7, 10, and 15 DIV, respectively, suggesting that the current trend continues, an increasing percentage of SynDIG1 is localized to synapses . To determine whether SynDIG1 present on the cell surface Acts of excitatory synapses, neurons were transfected with HA SynDIG1 With anti-HA Antique Live image body marked HA surface Chenepitope f Dyeing, and fixed and angef With anti-PSD95 and anti-VGLUT1 Antique rbt Pr to label body Synaptic specializations.