Apical-basal distribution of different subtypes of spiral ganglion neurons in the cochlea and the changes during aging

Sound information is transmitted from the cochlea to the brain mainly by type I spiral ganglion neurons (SGNs), which consist of different subtypes with distinct physiological properties and selective expression of molecular markers. It remains unclear how these SGN subtypes distribute along the tonotopic axis, and whether the distribution pattern changes during aging that might underlie age-related hearing loss (ARHL). We investigated these questions using immunohistochemistry in three age groups of CBA/CaJ mice of either sex, including 2–5 months (young), 17–19 months (middle-age), and 28–32 months (old). Mouse cochleae were cryo-sectioned and triple-stained using antibodies against Tuj1, calretinin (CR) and calbindin (CB), which are reportedly expressed in all type I, subtype I(a), and subtype I(b) SGNs, respectively. Labeled SGNs were classified into four groups based on the expression pattern of stained markers, including CR(+) (subtype I(a)), CB(+) (subtype I(b)), CR(+)CB(+) (dual-labeled I(a)/I(b)), and CR(-)CB(-) (subtype I(c)) neurons. The distribution of these SGN groups was analyzed in the apex, middle, and base regions of the cochleae. It showed that the prevalence of subtype I(a), I(b) and dual-labeled I(a)/I(b) SGNs are high in the apex and low in the base. In contrast, the distribution pattern is reversed in I(c) SGNs. Such frequency-dependent distribution is largely maintained during aging except for a preferential reduction of I(c) SGNs, especially in the base. These findings corroborate the prior study based on RNAscope that SGN subtypes show differential vulnerability during aging. It suggests that sound processing of different frequencies involves distinct combinations of SGN subtypes, and the age-dependent loss of I(c) SGNs in the base may especially impact high-frequency hearing during ARHL.