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. 2020 Oct 29;21(21):8063.
doi: 10.3390/ijms21218063.

Vasectomy and Photoperiodic Regimen Modify the Protein Profile, Hormonal Content and Antioxidant Enzymes Activity of Ram Seminal Plasma

Melissa Carvajal-Serna  1   2 , Meriem Fatnassi  3 , Felipe Torres-Ruda  2 , Jaime Antonio Cardozo  4 , Henry Grajales-Lombana  2 , Mohamed Hammadi  3 , Jose Alfonso Abecia  1 , Teresa Muiño-Blanco  1 , Rosaura Pérez-Pe  1 , Jose Álvaro Cebrián-Pérez  1 , Adriana Casao  1
Affiliations

Vasectomy and Photoperiodic Regimen Modify the Protein Profile, Hormonal Content and Antioxidant Enzymes Activity of Ram Seminal Plasma

Melissa Carvajal-Serna et al. Int J Mol Sci. .

Abstract

This work aimed to determine the contribution of the testis and epididymis and the effect of the photoperiodic regimen on ram seminal plasma (SP). Semen was collected from 15 mature rams located in an equatorial (Colombian Creole and Romney Marsh, eight intact and two vasectomized) or a temperate climate (Rasa Aragonesa, three intact and two vasectomized). SP proteins were analyzed by Bradford, SDS-PAGE and difference gel electrophoresis (DIGE). Melatonin and testosterone concentrations were quantified by ELISA, and activity of glutathione peroxidase (GPx), glutathione reductase (GRD), and catalase by enzymatic assays. Vasectomy increased protein concentration and the intensity of high molecular weight bands (p < 0.001), with no differences between breeds. DIGE revealed the absence of six proteins in vasectomized rams: angiotensin-converting enzyme, lactotransferrin, phosphoglycerate kinase, sorbitol dehydrogenase, epididymal secretory glutathione peroxidase and epididymal secretory protein E1. Vasectomy also decreased melatonin concentrations in seasonal rams, and testosterone in all of them (p < 0.001), but did not affect antioxidant enzyme activity. Equatorial rams showed lower melatonin and testosterone concentration (p < 0.01) and catalase, but higher GPx activity (p < 0.05). In conclusion, vasectomy modifies the protein profile and hormonal content of ram seminal plasma, whereas the exposure to a constant photoperiod affects hormonal concentration and antioxidant enzymes activity.

Keywords: DIGE; antioxidant enzymes; melatonin; ram; seminal plasma; testosterone; vasectomy.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Representative image of SDS-PAGE of seminal plasma proteins from intact and vasectomized rams subjected to temperate (seasonal rams) or equatorial (non-seasonal rams) photoperiod. 1: intact seasonal, 2: vasectomized seasonal, 3: intact non-seasonal, 4: vasectomized non-seasonal.
Figure 2
Figure 2
Representative image of difference gel electrophoresis (DIGE) analysis of seminal plasma proteins from intact and vasectomized rams. Angiotensin-converting enzyme (a), lactotransferrin (b), sorbitol dehydrogenase (c), phosphoglycerate kinase (d), epididymal secretory glutathione peroxidase (e), epididymal secretory protein E1 (f) and inactive ribonuclease-like protein 9 (g) spots are identified.
Figure 3
Figure 3
(a) Melatonin and (b) testosterone concentration in seminal plasma of intact and vasectomized rams subjected to temperate (seasonal rams) or equatorial (non-seasonal rams) photoperiod. Results are shown as mean ± S.E.M of n = 12 seminal plasma samples for seasonal rams, and n = 24 seminal plasma samples for non-seasonal. ** p < 0.01, *** p < 0.001.
Figure 4
Figure 4
(a) Glutathione peroxidase (GPx), (b) glutathione reductase (GRD) and (c) catalase enzymatic activity in seminal plasma of intact and vasectomized rams subjected to temperate (seasonal) or equatorial (non-seasonal rams) photoperiod. Results are shown as mean ± S.E.M of n = 12 seminal plasma samples for seasonal rams, and n = 24 seminal plasma samples for non-seasonal. * p < 0.05, *** p < 0.001.
See this image and copyright information in PMC

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