ESTROGEN AND PROGESTERONE REGULATE + + CNG-A1 AND Na /K - ATPase EXPRESSION IN RENAL CORTEX

Menezes, A.L.; Gracelli, J.B.; Souza-Menezes, J.; Barbosa, C.M.L.; Ornellas, F.S.; Takiya, C.M.; Alves, L.M.; Wengert, M.; Feltran, G.S.; Caruso-Neves, C.; Moyses, M.R.; Prota, L.F.M.; Morales, M.M. Laboratrio de Fisiologia Celular e Molecular, Instituto de Biofsica Carlos Chagas Filho, UFRJ, Rio de Janeiro; RJ
Introduction: Cyclic nucleotide-gated (CNG) channels are nonselective cation channels that have a role in transepithelial transport of calcium and sodium across the colon. It was shown CNG-A1 subunit activity in the renal cortex and expression of its mRNA in the renal medulla, however there were no studies about protein expression and function in the renal tissue. Steroid hormones, estrogen and progesterone, have a role in the body hydroelectrolytic balance, modulating the reabsorption of sodium along the nephron. Objective: This study aimed to report the possible modulation of CNG-A1 by those hormones subjecting ovariectomized rats to a 10-day replacement of 17-estradiol benzoate (2.0 g/kg body weight) or progesterone ( 1.7 mg/kg body weight). Results: Ovariectomized rats showed a decreased CNG-A1 (40%) and 1-Na+/K+-ATPase (74%) cortex protein expression (n=4, p<0.05) as well as the Na+/K+ ATPase activity in cortex (9.52.0 mmol/mg/min; control, 21.02.5 mmol/mg/min) (n=4, p<0.05), compared controls. After 17-estradiol benzoate replacement, both CNG-A1 and 1-Na+/K+-ATPase had their protein levels returned to control levels and Na+/K+-ATPase activity did not change significantly (28.54.5 mmol/mg/min) (n=4, p>0.05). Rats treated with progesterone showed a decrease in cortex protein expression (30% for CNG-A1 and 45% for 1-Na+/K+-ATPase) (n=4, p<0.05), but he Na+/K+ ATPase activity did not differ from the control (15.03.5 mmol/mg/min). Protein level and Na+/K+ATPase activity did not change in renal medulla. Conclusion: This work demonstrated not only the presence of CNG-A1 in renal cortex, but also its modulation by estrogen and progesterone as the Na+/K+-ATPase.

Figure 4. Western blot analysis for CNG-A1 protein in renal cortex and medulla of female control rats. The results are expressed as meansSEM. *P<0.05 vs cortex.

Table 1. Urinary parameters: summary data for urinary flow, GFR, body weight, water intake, FE of K+, Na+, Cl- and urea. Control (C), ovariectomized (OVX), overiectomized with 17-estradiol benzoate (OVE) and progesterone (OVP) replacement. The results are expressed as meansSEM (n=4).*p<0,05 vs to control group

Figure 5. Western blot analysis for CNG-A1 in IRPT cells treated with different concentrations of 17estradiol benzoate. C, control; V, vehicle. The results are expressed as meansSEM (n=3). *P<0.05 vs control group.

Ca+ feedback in rod and cone photoreceptors

Figure 6. Immunolocalization of CNGA1 in distal and proximal tubules of control female rats. Proximal Tubule: A, B, C Figure 1. Analysis of 1-Na+/K+-ATPase protein expression and activity in female rat kidney Control (C), ovariectomized (OVX), overiectomized with 17-estradiol benzoate (OVE) and progesterone (OVP) replacement. Immunoblot for 1Na+/K+-ATPase and Na+/K+-ATPase activity in renal cortex are respectively showed in panels A and B. Immunoblot for 1-Na+/K+-ATPase and Na+/K+-ATPase activity in renal medulla are respectively showed in panels C and D. The results are expressed as meansSEM (n=4).*P<0,05 vs control group CNG-A1 Luminal Pole Nucleus Distal Tubule: D, E, F CNG-A1 Luminal Pole Nucleus

CTRL
2 g/Kg 17-estradiol 17 mg/Kg Progesterone

OVX OVP
Bilateral Ovariectomy Figure 2. Analysis of Na+/K+-ATPase expression and activity in IRPT cells treated with different concentrations of 17-estradiol benzoate. Immunoblot for 1Na+/K+-ATPase in IRPT cells is show in panel A. Na+/K+-ATPase activity in IRPT cells is show in panel B. C, control; V, vehicle. The results are expressed as meansSEM (n=3). *P<0.05 vs control group.

OVE

Presence of CNG-A1 subunit in renal tissue Estrogen and progesterone as physiological modulators of CNG-A1 Na+K+-ATPase expression rats only and activity and activity decreased with in

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ovariectomized replacement;

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Figure 3. Analysis of CNG-A1 protein expression in female rat kidney of Control (C), ovariectomized (OVX), ovariectomized with 17-estradiol benzoate (OVE) and progesterone (OVP) replacement for 10 days. Immunoblots for CNG-A1 sodium channel in renal cortex and medulla is showed respectively in panels A and B. The results are expressed as meansSEM (n=4). *P<0.05 vs control group.