Hearing loss is a sensory impairment caused by genetic and environmental factors (Dror and Avraham, 2009; Roth et al., 2011), whose incidence is increasing according to reports of the World Health Organization. Factors such as noise and ototoxic drugs are well-known effectors of this impairment, but more recently several epidemiological studies have shown the association between the nutritional status and hearing loss. Precisely, inadequate levels of folic acid have been correlated with hearing loss in the presence of reduced vitamin B12 concentrations or hyperhomocysteinemia (Houston et al., 1999; Cadoni et al., 2004; Lasisi et al., 2010). Additional reports have explored the putative association of mutations in genes of the folate cycle and the impact of hearing loss, their results being somewhat inconsistent (Durga et al., 2006; Uchida et al., 2011). In contrast, two animal studies have demonstrated the link between homocysteine (Hcy) metabolism and hearing loss (Cohen-Salmon et al., 2007; Martinez-Vega et al., 2015a) and the effects of a folate-deficient (FD) diet in the auditory system (Martinez-Vega et al., 2015a). Thus, remarkable increases in auditory brainstem response (ABR) thresholds were detected in C57BL/6J mice after 2-months on a FD diet (Martinez-Vega et al., 2015a). These changes correlated with the presence of severe histological damage in the cochlea and with important alterations in cochlear metabolism. Folate deficiency induced a decrease in Hcy remethylation and its flux through trans-sulfuration, together with an increase in adenosine elimination, as deduced from the important changes observed in the enzymes involved. Altogether these changes led to cochlear accumulation of Hcy, resulting in enhanced protein homocysteinylation, and in redox stress in FD cochleae (Martinez-Vega et al., 2015a). C57BL/6J is a strain known to develop premature hearing loss, a phenotypic trait that was accelerated in FD mice. Therefore, we set up a new experiment using the CBA/Ca mouse strain, less prone to hearing loss, to further evaluate the impact of folate deficiency in the development of this impairment.