When looking at this paragraph in Midorikawa's work for a fluorescence review I was confused, specifically around a time constant of ~4s being compared to ~300 ms.
- It is not explicitly mentioned so I assume the 300 ms is a time constant correct?
- How is the conclusion drawn that it is the step after tethering, priming, by the discrepancies in time constant? I am not hoping any readers know why biologically it is this way but rather how I can understand a 13x larger time constant can lead to conclusions that it is not rate-limiting? (emphasis mine)
By comparing the time course of fusion events and tethering events, it was suggested that tethering of new vesicles could refill the empty release sites with a time constant of ∼4 s. This is also surprising since the replenishment of the readily releasable vesicles (RRVs) at the calyx of Held terminal was estimated to be ∼300 ms by the electrophysiological experiments (Neher and Sakaba, 2008; Hallermann and Silver, 2013). Given that a depolarization elicit fusions of only a small fraction of already-tethered vesicles in TIRFM imaging, the discrepancy suggests that the replenishment time course was determined by the step after the tethering, i.e. priming.
Reference Midorikawa, M. (2018) Real-time imaging of synaptic vesicle exocytosis by total internal reflection fluorescence (TIRF) microscopy. Neuroscience Research, Volume 136, (2018), pp. 1-5, ISSN 0168-0102, https://doi.org/10.1016/j.neures.2018.01.008. (https://www.sciencedirect.com/science/article/pii/S0168010217307551)
Thank you for the help!