Na+, K+-ATPase in ram sperm – Its importance for kinematics, localisation and expression on the sperm surface

•Sodium pump is localised in ram sperm middle piece.•Inhibition of sodium pump results in discrete modification in ram sperm kinematic.•Cryopreservation influences the sodium pump expression in ram sperm membrane. The goals of this study were to investigate (a) the localisation of Na+, K+-ATPase in...

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Bibliographic Details
Published in:	Small ruminant research Vol. 176; pp. 31 - 36
Main Authors:	de Oliveira, Aline Saraiva, Câmara, Diogo Ribeiro, Batista, André Mariano, Silva, Robespierre Augusto Joaquim Araújo, Arruda, Lúcia Cristina Pereira, Monteiro, Millena Maria, Costa, Joana Amélia de Senna, Guerra, Maria Madalena Pessoa
Format:	Journal Article
Language:	English
Published:	Elsevier B.V 01-07-2019
Subjects:	Ionic channel Ram Sodium pump Spermatozoa Spermatozoa Ionic channel Sodium pump Ram
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Summary:	•Sodium pump is localised in ram sperm middle piece.•Inhibition of sodium pump results in discrete modification in ram sperm kinematic.•Cryopreservation influences the sodium pump expression in ram sperm membrane. The goals of this study were to investigate (a) the localisation of Na+, K+-ATPase in ram sperm; (b) cryopreservation like-changes on median fluorescence intensity (MFI) and the expression of Na+, K+-ATPase on the sperm surface (labeled sperm); and (c) the influence of Na+, K+-ATPase inhibition on sperm kinematics. Exp. 1: pooled semen samples (n = 7) from four rams were diluted and frozen. The following sperm features were evaluated in fresh-diluted and post-thawed samples: MFI and expression of Na+, K+-ATPase (Bodipy® FL-Ouabain), plasma and acrosomal membrane integrity (IP; FITC-PNA), sperm viability and plasma membrane stability (Yo-Pro and Merocyanine), as well as sperm kinematics (CASA). Exp. 2: pooled semen samples (n = 5) from four rams were diluted and incubated in medium treated with or without Na+, K+-ATPase inhibitor (ouabain, 10−4 M). Sperm kinematics (CASA) were determined prior to (0 h) and after incubation (3 h). In both fresh and thawed sperm, Na+, K+-ATPase was localised in the middle piece and cryopreservation did not influence the MFI in sperm with intact plasma membranes. However, cryopreservation reduced (P < 0.05) the overall percentage of sperm labeled with Bodipy® FL-Ouabain. Compared to the control, the inhibition of Na+, K+-ATPase reduced VAP at 0 h and both VAP and VSL at 3 h (P < 0.05). Thus, Na+, K+-ATPase is localised in the middle piece of fresh diluted and thawed sperm, and cryopreservation reduces the percentage of sperm expressing Na+, K+-ATPase in their surface, with no influence on the MFI of sperm with intact plasma membranes. Despite its localisation, Na+, K+-ATPase inhibition results in discrete kinematic modifications to ram sperm.
ISSN:	0921-4488 1879-0941
DOI:	10.1016/j.smallrumres.2019.05.012