With a novel antibody against the rat Na⁺-d-glucose cotransporter SGLT2 (rSGLT2-Ab), which does not cross-react with rSGLT1 or rSGLT3, the ∼75-kDa rSGLT2 protein was localized to the brush-border membrane (BBM) of the renal proximal tubule S1 and S2 segments (S1 > S2) with female-dominant expression in adult rats, whereas rSglt2 mRNA expression was similar in both sexes. Castration of adult males increased the abundance of rSGLT2 protein; this increase was further enhanced by estradiol and prevented by testosterone treatment. In the renal BBM vesicles, the rSGLT1-independent uptake of [¹⁴C]-α-methyl-d-glucopyranoside was similar in females and males, suggesting functional contribution of another Na⁺-d-glucose cotransporter to glucose reabsorption. Since immunoreactivity of rSGLT2-Ab could not be detected with certainty in rat extrarenal organs, the SGLT2 protein was immunocharacterized with the same antibody in wild-type (WT) mice, with SGLT2-deficient ( Sglt2 knockout) mice as negative control. In WT mice, renal localization of mSGLT2 protein was similar to that in rats, whereas in extrarenal organs neither mSGLT2 protein nor mSglt2 mRNA expression was detected. At variance to the findings in rats, the abundance of mSGLT2 protein in the mouse kidneys was male dominant, whereas the expression of mSglt2 mRNA was female dominant. Our results indicate that in rodents the expression of SGLT2 is kidney-specific and point to distinct sex and species differences in SGLT2 protein expression that cannot be explained by differences in mRNA.

Various metals of unknown function in the body (Cd, Cr, Hg, Pb, U), trace elements in excessive concentrations (Co, Cu, Fe, Zn), or metals used in cancer therapy (Pt, V), accumulate in the mammalian kidney, largely in the proximal tubule (PT) cells, and cause functional and structural damage that results in reabsorptive and secretory defects. The intracellular mechanisms of their toxicity in the PT cells are not well known. Recent studies have indicated an oxidative stress with associated lipid peroxidation, apoptosis, and necrosis as common phenomena in the course of nephrotoxicity of these metals. However, a number of other phenomena, such as the selective inhibition and/or loss of various membrane transporters, enhancement of ion conductances, increased cytoplasmic concentration of calcium, deranged cytoskeleton and cell polarity, impaired endocytosis, swelling and fragmentation of mitochondria, increased expression of metallothionein, heat-shock and multidrug resistance proteins, loss of cell membrane integrity, as well as the damage of mitochondrial and genomic DNAs have been fragmentarily demonstrated for the action of some toxic metals, but their importance for the course of nephrotoxicity and the sequence of events in relation to oxidative stress, apoptosis, and necrosis have not been clearly established. Recent studies of metal toxicity in various tissues and cells of non-renal and renal origin enable us to estimate ‘causes and consequences’ of various phenomena in the metal-induced nephrotoxicity, and to assemble them in a possible common, time-related sequence.

In rats, the secretion of p-aminohippurate (PAH) by the kidney is higher in males (M) than in females (F). The role of the major renal PAH transporters, OAT1 and OAT3, in the generation of these gender differences, as well as the responsible hormones and mechanisms, has not been clarified. Here we used various immunocytochemical methods to study effects of gender, gonadectomy, and treatment with sex hormones on localization and abundance of OAT1 and OAT3 along the rat nephron. Both transporters were localized to the basolateral membrane: OAT1 was strong in proximal tubule S2 and weak in the S3 segments, whereas OAT3 was stained in proximal tubule S1 and S2 segments, thick ascending limb, distal tubule, and in principal cells along the collecting duct. Gender differences in the expression of both transporters in adult rats (M > F) were observed only in the cortical tubules. OAT1 in the cortex was strongly reduced by castration in adult M, whereas the treatment of castrated M with testosterone, estradiol, or progesterone resulted in its complete restitution, further depression, or partial restitution, respectively. In adult F, ovariectomy weakly increased, whereas estradiol treatment of ovariectomized F strongly decreased, the expression of OAT1. The expression of OAT3 in the M and F cortex largely followed a similar pattern, except that ovariectomy and progesterone treatment showed no effect, whereas in other tissue zones gender differences were not observed. In prepubertal rats, the expression of OAT1 and OAT3 in the kidney cortex was low and showed no gender differences. Our data indicate that gender differences in the rat renal cortical OAT1 and OAT3 (M > F) appear after puberty and are determined by both a stimulatory effect of androgens (and progesterone in the case of OAT1) and an inhibitory effect of estrogens.