Kubke Lab:Research/CND/Records/HK023

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EmbryoName HK023- chicken embryo- Stage 19.5 - 8.12.14


Experiment

Objective Dyes of different colours on Trigeminal (DiO) vs 7/8th nerve complex (DiI) to see interactions between efferents of one nerve and afferents of other. Also to practice the injection on a younger chicken (slowly moving down to stage 18-17)

Procedure

  • Trigeminal nerve bilaterally DiO crystals
  • 7th/8th nerve bilaterally DiI crystals (Big chunk on 7th nerve on L side and big chunk on 8th on R side)

Notes

No obvious contamination, purposelly put a greater chunk of dye on the 7th nerve on the L side and the 8th nerve on the R side so will see if there is any visible difference between the amount of neurons labelled in different positions within the rhombomere.

Files

  • HK023 folder under Helen Ker in Anatomy All

Results

Facial and Vestibulocochlear

(Orange)

  • Afferents

Shorter than other ages that I am used to and seem to stop suddenly. Interact with the trigeminal and seem to join the ganglion?

  • Efferents

There are some efferents I notice turn before making contact with an afferent, but most do eventually it's just that they turn before actually making physical contact with the afferent. \ Although I know that I put different amounts of dye on thte 7th and 8th on the left and right side, I didn't see any obvious larger dye diffusion in any particular region (I would have thought that the nerve that I dyed the Facial very heavily would have a different amount of efferents labelled when compared to the nerve that I labelled 8th heavily, but after more thought perhaps because it is so upstream it would really not make a difference - and it didn't appear to)

Trigeminal

(Green)

I have found DiO a lot fainter in general and harder to see, is this just the nature of the dye or the imaging techniques we have available? Trigeminal afferents seemed not to align with the VII/VIII afferents, and seemed to join more medially. There are some afferents which look to me as though they join with the ganglion of the VIII nerve? but these are artifact? or just so faint that I'm actually looking at 8th efferents?

Summary

My clearest injection yet. Possibly due to the young age, not the fact that my injection was the right size? I see some efferents turning before making contact with any afferents and that interests me. how do I know that I just haven't labelled the afferent? I guess through repetition which I have done today so hopefully we'll see tomorrow.

Questions - is DiO always that faint? is there a way of changing filters without changing the dimensions of the image? that's a problem when trying to combine the same image with two different filters to map interactions between different nerves. is there any way that the afferents of trigeminal are getting into a VIII nerve?

Future Try and repeat to confirm that efferents turn without afferent's contact. and to see location of the trigeminal and where its axons go

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