I am performing intravitreal injections on postnatal day P2 or P3 neonatal rat pups with an AAV vector and wondering if anyone can comment on how the rats tolerate injections in both eyes as opposed to one? I know typically one is done and the other is left as a control. However I am currently testing different serotypes and perfecting my procedure so would like to inject both eyes. However, I am concerned about whether or not injecting both would worsen the pups chances of survival.
Thanks so much,
Ellen
Though I'm not an eye gene therapy person I can provide perspective on wellbeing. The longer term risks are low - AAV vectors, and expression thereof will have no detriment. The key factor is what type of anaesthesia you are using and how long you are taking per procedure. As you probably know, neonates are a little more tricky than adults regarding anaesthesia and you want to minimise the duration they are unconscious. Tips to optimise their wellbeing - give them painkiller pre-injection, so that it is already acting when they awake. Consider using a small regulated warming device to minimise heat loss, and provide hydration - possibly subcut physiological saline in the scruff of their neck, or i.p.
Typically injections of AAV into the eye are well tolerated so I doubt you'd have any issues there. As mentioned above, the key risks will probably come with your surgery and the post-operative care.
Shaun Wood sir, what is the titer of AAV for Rat tail vein(systemic injection)?
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