In Ménière's disease, an inner ear disorder related to an endolymphatic hydrops, an alteration of the functioning of the endolymphatic sac has been proposed. The endolymphatic sac is assumed to be involved in the secretion/resorption of endolymph. The epithelial transport systems have been indirectly studied by the recording of the endolymphatic sac transepithelial potential (ESP) in control conditions and after the local injection of drugs such as diuretics that have been proposed in the treatment of Ménière's disease. The ESP was recorded, in vivo, in guinea pigs up to 150 minutes after the perisaccular injection of 5 μL of a 150 mmol/L (mM) NaCl solution containing various drugs known to inhibit ionic transport systems. The initial ESP was +8.4 ± 0.3 mV (mean ± SEM, n = 78). The basolateral injection of 5 μL of 150 mM NaCl induced an ESP decrease of 64% ± 6.0% (n = 12), 5 minutes after the end of the injection. Then ESP increased, returning to its initial value at 60 minutes and surpassing it at 120 minutes. Diuretics such as acetazolamide (10−3 mol/L [M]), an inhibitor of carbonic anhydrase, and amiloride (10−4 M), an inhibitor of Na channel or Na/H exchanger, decreased the ESP recovery. At variance, bumetanide (10−6 M, 10−4 M), the Na-K-Cl cotransport inhibitor, and chlorothiazide (10−4 M), a Na-Cl cotransporter inhibitor, failed to alter the ESP as compared with the control group. Ouabain (10−3 M), the Na+,K+-adenosine triphosphatase (ATPase) inhibitor, prevented the ESP recovery otherwise observed 60 minutes after the NaCl injection. Bafilomycin A1, the inhibitor of the vacuolartype H+-ATPase, prevented the recovery of the ESP with a log-dose/effect (10−5 M, 10−6 M, 10−8 M). Disulfonic acid stilbene (DIDS) (10−4 M), an inhibitor of transporters involving HCO3−, also prevented the ESP recovery. These results suggest that the genesis of the ESP was highly dependent on acid-base transport systems including carbonic anhydrase, a vacuolar-type H+-ATPase, and an anionic transport system blocked by DIDS. Further studies are needed to confirm the alteration of the acid-base balance in this epithelium and its possible involvement in the pathogenesis of Ménière's disease.