Water transport in seminiferous tubule: Aquaporins identification in Sertoli cells M.G. Alves1*, T.T. Jesus1*, R.L. Bernardino1, A.D. Martins1, R. Sá2, M. Sousa2,3, P.F. Oliveira1 1 CICS
– UBI – Health Sciences Research Centre, University of Beira Interior, Covilhã, Portugal; CICS – Centro2de Investigação em Ciências da Saúde, Faculdade de Ciências da Saúde, Universidade da Beira Interior, Department of Microscopy, Laboratory of Cell Biology, Institute of Biomedical Sciences Abel Salazar (ICBAS), UMIB, University of Porto, Portugal; 3 Centre for Reproductive Genetics Alberto Barros, Porto, Portugal; @ www.fcsaude.ubi.pt/cics
[email protected] https://sites.google.com/site/reproductiongroupubi/
* Both authors contributed equally.
Introduction
Results
The secretion of the seminiferous tubular fluid (STF), as well as the control of the composition of this fluid is crucial for male fertility. Sertoli cells (SCs) are known to express various types of membrane transporters that are directly involved on the movement of substances across the membrane. Aquaporins (AQPs) are essential for the regulation of water homeostasis and for providing a continuous and rapid movement of water across tight junction epithelia. Altered expression of AQPs has been associated with several disorders of the male reproductive system, which in most of the cases results in the infertility of the individual.
Aim Identify the expression of specific isoforms of AQPs in rat Sertoli cells (rSCs) and confirm their gene and protein expression.
RT-PCR
Testicular Tissue
mRNA Expression of Aquaporin Isoforms in Sertoli Cells SC (+)
(-)
Aquaporin 1 Aquaporin 4 Aquaporin 8
Figure 1. RT-PCR evaluation of mRNA expression of specific isoforms of Aquaporin in Sertoli cells. SC: Sertoli cells; (+) positive control (kidney lysate); (-) negative control without reverse transcriptase.
Protein Expression of Aquaporin Isoforms in Sertoli Cells SC
Material and Methods
Rat
6200-506 Covilhã
(+)
Aquaporin 1
Aquaporin mRNA expression
Aquaporin 4 Aquaporin 8
SCs Figure 2. Immunoblot evaluation of protein expression of specific isoforms of Aquaporin in Sertoli cells. SC: Sertoli cells (+) positive control (kidney lysate).
Cultures
Immunoblot
Aquaporin protein expression
Conclusions
Acknowledgments:
The presence of the mRNA transcripts of the AQP isoforms 1, 4 and 8 was detected by RT-PCR in cultured rat SCs.
This work was supported by the Portuguese “Fundação para a Ciência e a Tecnologia”—FCT, co-funded by FEDER via Programa Operacional Factores de Competitividade—COMPETE/QREN (PTDC/QUI-BIQ/121446/2010 and PEstC/SAU/UI0709/2011). M. G. Alves (SFRH/BPD/80451/2011) and A.D. Martins (PTDC/QUI-BIQ/121446/2010) were financed by FCT and European Social Fund. P. F. Oliveira was financed by FCT through FSE and POPH funds (Programa Ciência 2008).
Protein expression of AQP isoforms 1, 4 and 8 was also confirmed in cultured rat SCs The identification of these AQPs isoforms in SCs provide new and crucial information concerning the possible pathways by which these cells control the composition of the luminal seminiferous fluid.
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