Question about bidirectional seeding

Hi!

I wanted to obtain the structural connectivity between bilateral motor cortex. To do this, I performed tractography first and then use tck2connectome with the ROI image of bilateral motor cortex as input.

During the revision, I was questioned by the reviewer if the seeding was done in both directions (from left to right and from right to left). After some search, I realized that this needs to be done with a seed image during the tractography. But I was wondering if my method has the same effect?

Also the reviewer asked if the corpus callosum was used as a inclusion region to avoid spurious streamlines. I was wondering if this can be accounted in my pipeline, or I have to perform the tractography again with an inclusion region?

Thank you very much!