As discussed in the previous section, before deafening, IPG effects were more stable than the absolute eCAP measures, illustrating their suitability as neural health markers (Fig.
7). After deafening, all measures except for Δthreshold changed significantly, thereby largely confirming our previous findings in acutely implanted normal-hearing (NH) 2-week deaf (2WD) and 6-week deaf (6WD) guinea pigs (Ramekers et al.
2014). Abrupt changes were observed for Δamplitude, Δdynamic range, and Δlevel
50% (between day 14 and day 21), while gradual changes over time after deafening (between day 7 and day 28) were observed for Δslope and Δlatency. Specifically, the gradual change in Δlatency (i.e., IPG effect on N
1 latency) that took place in the first 6 weeks after deafening is highly similar to our previous observations. In that study, mean Δlatency was 6 µs for NH, 14 µs for 2WD, and 24 µs for 6WD animal groups (and 27 µs for 6WD animals in Vink et al.
2020), compared to 5.1 µs, 15 µs, and 26 µs for the animals in the present study for those specific time points, respectively. This gradual change in Δlatency over the course of 4 to 6 weeks after deafening coincides with the loss of ~50 % of the SGC population during the same period of time (Versnel et al.
2007; Agterberg et al.
2008; van Loon et al.
2013; Ramekers et al.
2015a), and may therefore be used as a predictor of neural survival. Alternatively, since Δlatency appears to stabilize beyond 4 weeks after deafening – at least up to 14 weeks after deafening (25 µs in Ramekers et al.
2015a) – it may reflect electrophysiological changes at the individual level as a result of deafness-induced degeneration, such as shrinkage (e.g., Limón et al.
2005), loss of myelin (e.g., Resnick and Rubinstein
2021), or changes in membrane ion channel composition (e.g., Luque et al.
2021). Note that although often suggested as a characteristic phase in the degeneration process of SGCs, in our lab, we do not observe retrograde degeneration, being loss of peripheral processes prior to loss of the SGC soma (Ramekers et al.
2020). As we have argued previously, the magnitude of the latency difference between the two IPG conditions (up to 27 µs) closely resembles the IPG increase itself (27.9 µs), suggesting a preference shift in excitability from first phase of the biphasic current pulse to the second – from normal-hearing to deaf (Ramekers et al.
2014). This hypothesis somewhat resembles one often referred to as polarity sensitivity, asserting that healthy SGCs respond to cathodic currents and (retrogradely) degenerating SGCs to anodic currents (Rattay et al.
2001; Undurraga et al.
2010; Joshi et al.
2017; Hughes et al.
2018; Jahn and Arenberg
2019; Brochier et al.
2021), with the crucial difference that in our lab, all guinea pigs (NH and deaf) respond mainly to cathodic currents (unpublished data), and that they do not lose their peripheral processes prior to losing their SGC soma after deafening (Ramekers et al.
2020).
In the section above, based solely on the latency data, we postulate the hypothesis that the cathodic phase in cathodic-leading pulses is the most excitatory in normal-hearing animals, but that after deafening, this preference shifts to the cathodic (i.e., second) phase in the anodic-leading pulse. As a reminder, it should be noted that since alternating polarity was used for artifact reduction, both pulse polarities have been applied, but that the respective responses cannot be individually assessed. The hypothesized shift in excitation from cathodic-first to cathodic-second occurring simultaneously with SGC degeneration is supported by a smaller IPG effect on both AGF slope and level50% – essentially a smaller enhancement of excitation efficacy. If indeed after deafening the second phase of the biphasic current pulse gradually becomes the predominant excitatory one, the influence of the IPG in separating the two phases becomes increasingly irrelevant, since the IPG then no longer delays the onset of the repolarizing anodic phase – abolishing the preceding action potential generation – but merely delays the excitation itself. The IPG effects on maximum amplitude and dynamic range both increase after deafening, which seemingly contradicts this hypothesis. However, these two effects take place near the upper asymptote of the sigmoidal AGF, and therefore arguably do not reflect the overall excitability of the majority of the SGC population. Rather, a larger maximum amplitude brought about by high stimulation levels may reflect increased neural excitation by the less-effective cathodic-first pulse in the deaf animals.
Although all 10 animals were deaf for exactly 7 weeks, SGC survival did vary across animals. Importantly, even in this small homogenous group of guinea pigs did we find trends in the correlations between IPG effects and SGC survival that were largely similar to those observed previously (Ramekers et al.
2014). Because of the homogeneity of this group, we here for the first time unambiguously show that it is specifically the extent of SGC degeneration, and not for instance the associated duration of deafness, that drives the IPG effects. This is relevant for CI research since although it has been demonstrated repeatedly in histopathological studies (e.g., Spoendlin
1975; Nadol
1990; Nadol et al.
2001; Fayad and Linthicum
2006) that deafness is virtually invariably accompanied by SGC degeneration, the extent of this degeneration can vary substantially.