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Rapid responses to reverse T₃ hormone in immature rat Sertoli cells: calcium uptake and exocytosis mediated by integrin.

Zanatta AP, Zanatta L, Gonçalves R, Zamoner A, Silva FR - PLoS ONE (2013)

Bottom Line: The implication of T-type voltage-dependent calcium channels and chloride channels in the effect of rT₃ was evidenced using flunarizine and 9-anthracene, respectively.Furthermore, the outcomes indicate that rT₃ also stimulates cellular secretion since the cells manifested a loss of fluorescence after 4 min incubation, indicating an exocytic quinacrine release that seems to be mediated by the integrin receptor.These findings indicate that rT₃ modulates the calcium entry and cellular secretion, which might play a role in the regulation of a plethora of intracellular processes involved in male reproductive physiology.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Florianópolis-Santa Catarina, Brazil.

ABSTRACT
There is increasing experimental evidence of the nongenomic action of thyroid hormones mediated by receptors located in the plasma membrane or inside cells. The aim of this work was to characterize the reverse T₃ (rT₃) action on calcium uptake and its involvement in immature rat Sertoli cell secretion. The results presented herein show that very low concentrations of rT₃ are able to increase calcium uptake after 1 min of exposure. The implication of T-type voltage-dependent calcium channels and chloride channels in the effect of rT₃ was evidenced using flunarizine and 9-anthracene, respectively. Also, the rT₃-induced calcium uptake was blocked in the presence of the RGD peptide (an inhibitor of integrin-ligand interactions). Therefore, our findings suggest that calcium uptake stimulated by rT₃ may be mediated by integrin αvβ₃. In addition, it was demonstrated that calcium uptake stimulated by rT₃ is PKC and ERK-dependent. Furthermore, the outcomes indicate that rT₃ also stimulates cellular secretion since the cells manifested a loss of fluorescence after 4 min incubation, indicating an exocytic quinacrine release that seems to be mediated by the integrin receptor. These findings indicate that rT₃ modulates the calcium entry and cellular secretion, which might play a role in the regulation of a plethora of intracellular processes involved in male reproductive physiology.

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Involvement of ionic channels and intracellular calcium on stimulatory effect of rT3 on 45Ca2+ uptake.(A) Influence of flunarizine, (B) BAPTA-AM and (C) 9-AC on stimulatory effect of rT3 on 45Ca2+ uptake in Sertoli cells. Pre-incubation: 15 min in KRb, additional pre-incubation: 60 min with 0.1 µCi/mL of 45Ca2+ and incubation time: 60 s with 0.1 µCi/mL of 45Ca2+ in the presence or absence of flunarizine (1 µM), BAPTA-AM (50 µM) and 9-AC (1 µM) with/without rT3 (10-17 M). Means ± S.E.M. For control, n=10; rT3, n=7; flunarizine, n=8; rT3 + flunarizine, n=8; BAPTA-AM, n=8; rT3 + BAPTA-AM, n=6; 9-AC, n=6; rT3 + 9-AC, n=6. ***P < 0.001 and **p < 0.01 compared with control group; ###p < 0.001; ##p < 0.01 and #p < 0.05 compared with rT3 group.
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pone-0077176-g003: Involvement of ionic channels and intracellular calcium on stimulatory effect of rT3 on 45Ca2+ uptake.(A) Influence of flunarizine, (B) BAPTA-AM and (C) 9-AC on stimulatory effect of rT3 on 45Ca2+ uptake in Sertoli cells. Pre-incubation: 15 min in KRb, additional pre-incubation: 60 min with 0.1 µCi/mL of 45Ca2+ and incubation time: 60 s with 0.1 µCi/mL of 45Ca2+ in the presence or absence of flunarizine (1 µM), BAPTA-AM (50 µM) and 9-AC (1 µM) with/without rT3 (10-17 M). Means ± S.E.M. For control, n=10; rT3, n=7; flunarizine, n=8; rT3 + flunarizine, n=8; BAPTA-AM, n=8; rT3 + BAPTA-AM, n=6; 9-AC, n=6; rT3 + 9-AC, n=6. ***P < 0.001 and **p < 0.01 compared with control group; ###p < 0.001; ##p < 0.01 and #p < 0.05 compared with rT3 group.

Mentions: We also investigated whether T-type voltage-dependent calcium channels (T-VDCC) could be involved in the rT3 stimulatory action on 45Ca2+ uptake. To this aim, Sertoli cells were incubated in the presence of rT3 with/without flunarizine (1 µM) which blocks T-VDCCs [18]. In Figure 3A it can be observed that flunarizine ified the rT3 stimulatory effect indicating the involvement of T-type VDCC in the calcium uptake in Sertoli cells.


Rapid responses to reverse T₃ hormone in immature rat Sertoli cells: calcium uptake and exocytosis mediated by integrin.

Zanatta AP, Zanatta L, Gonçalves R, Zamoner A, Silva FR - PLoS ONE (2013)

Involvement of ionic channels and intracellular calcium on stimulatory effect of rT3 on 45Ca2+ uptake.(A) Influence of flunarizine, (B) BAPTA-AM and (C) 9-AC on stimulatory effect of rT3 on 45Ca2+ uptake in Sertoli cells. Pre-incubation: 15 min in KRb, additional pre-incubation: 60 min with 0.1 µCi/mL of 45Ca2+ and incubation time: 60 s with 0.1 µCi/mL of 45Ca2+ in the presence or absence of flunarizine (1 µM), BAPTA-AM (50 µM) and 9-AC (1 µM) with/without rT3 (10-17 M). Means ± S.E.M. For control, n=10; rT3, n=7; flunarizine, n=8; rT3 + flunarizine, n=8; BAPTA-AM, n=8; rT3 + BAPTA-AM, n=6; 9-AC, n=6; rT3 + 9-AC, n=6. ***P < 0.001 and **p < 0.01 compared with control group; ###p < 0.001; ##p < 0.01 and #p < 0.05 compared with rT3 group.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3795021&req=5

pone-0077176-g003: Involvement of ionic channels and intracellular calcium on stimulatory effect of rT3 on 45Ca2+ uptake.(A) Influence of flunarizine, (B) BAPTA-AM and (C) 9-AC on stimulatory effect of rT3 on 45Ca2+ uptake in Sertoli cells. Pre-incubation: 15 min in KRb, additional pre-incubation: 60 min with 0.1 µCi/mL of 45Ca2+ and incubation time: 60 s with 0.1 µCi/mL of 45Ca2+ in the presence or absence of flunarizine (1 µM), BAPTA-AM (50 µM) and 9-AC (1 µM) with/without rT3 (10-17 M). Means ± S.E.M. For control, n=10; rT3, n=7; flunarizine, n=8; rT3 + flunarizine, n=8; BAPTA-AM, n=8; rT3 + BAPTA-AM, n=6; 9-AC, n=6; rT3 + 9-AC, n=6. ***P < 0.001 and **p < 0.01 compared with control group; ###p < 0.001; ##p < 0.01 and #p < 0.05 compared with rT3 group.
Mentions: We also investigated whether T-type voltage-dependent calcium channels (T-VDCC) could be involved in the rT3 stimulatory action on 45Ca2+ uptake. To this aim, Sertoli cells were incubated in the presence of rT3 with/without flunarizine (1 µM) which blocks T-VDCCs [18]. In Figure 3A it can be observed that flunarizine ified the rT3 stimulatory effect indicating the involvement of T-type VDCC in the calcium uptake in Sertoli cells.

Bottom Line: The implication of T-type voltage-dependent calcium channels and chloride channels in the effect of rT₃ was evidenced using flunarizine and 9-anthracene, respectively.Furthermore, the outcomes indicate that rT₃ also stimulates cellular secretion since the cells manifested a loss of fluorescence after 4 min incubation, indicating an exocytic quinacrine release that seems to be mediated by the integrin receptor.These findings indicate that rT₃ modulates the calcium entry and cellular secretion, which might play a role in the regulation of a plethora of intracellular processes involved in male reproductive physiology.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Florianópolis-Santa Catarina, Brazil.

ABSTRACT
There is increasing experimental evidence of the nongenomic action of thyroid hormones mediated by receptors located in the plasma membrane or inside cells. The aim of this work was to characterize the reverse T₃ (rT₃) action on calcium uptake and its involvement in immature rat Sertoli cell secretion. The results presented herein show that very low concentrations of rT₃ are able to increase calcium uptake after 1 min of exposure. The implication of T-type voltage-dependent calcium channels and chloride channels in the effect of rT₃ was evidenced using flunarizine and 9-anthracene, respectively. Also, the rT₃-induced calcium uptake was blocked in the presence of the RGD peptide (an inhibitor of integrin-ligand interactions). Therefore, our findings suggest that calcium uptake stimulated by rT₃ may be mediated by integrin αvβ₃. In addition, it was demonstrated that calcium uptake stimulated by rT₃ is PKC and ERK-dependent. Furthermore, the outcomes indicate that rT₃ also stimulates cellular secretion since the cells manifested a loss of fluorescence after 4 min incubation, indicating an exocytic quinacrine release that seems to be mediated by the integrin receptor. These findings indicate that rT₃ modulates the calcium entry and cellular secretion, which might play a role in the regulation of a plethora of intracellular processes involved in male reproductive physiology.

Show MeSH
Related in: MedlinePlus