Volume 57, Issue 8 pp. 912-918

ORIGINAL ARTICLE

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Evaluation of sperm membrane functionality during epididymal transit in red-rumped agouti (Dasyprocta leporina)

Maiko Roberto Tavares Dantas,

Maiko Roberto Tavares Dantas

Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido – UFERSA, Mossoró, Brazil

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Nayra Rachel Nascimento Luz,

Nayra Rachel Nascimento Luz

Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido – UFERSA, Mossoró, Brazil

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Luana Grasiele Pereira Bezerra,

Luana Grasiele Pereira Bezerra

Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido – UFERSA, Mossoró, Brazil

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Samara Sandy Jerônimo Moreira,

Samara Sandy Jerônimo Moreira

Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido – UFERSA, Mossoró, Brazil

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Moacir Franco de Oliveira,

Moacir Franco de Oliveira

Department of Animal Sciences, Universidade Federal Rural do Semi-Árido – UFERSA, Mossoró, Brazil

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Alexandre Rodrigues Silva,

Corresponding Author

Alexandre Rodrigues Silva

[email protected]

Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido – UFERSA, Mossoró, Brazil

Correspondence

Alexandre Rodrigues Silva, Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido – UFERSA, BR 110, Km 47, Costa and Silva, 59625-900 Mossoró, RN, Brazil.

Email: [email protected]

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Maiko Roberto Tavares Dantas,

Maiko Roberto Tavares Dantas

Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido – UFERSA, Mossoró, Brazil

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Nayra Rachel Nascimento Luz,

Nayra Rachel Nascimento Luz

Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido – UFERSA, Mossoró, Brazil

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Luana Grasiele Pereira Bezerra,

Luana Grasiele Pereira Bezerra

Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido – UFERSA, Mossoró, Brazil

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Samara Sandy Jerônimo Moreira,

Samara Sandy Jerônimo Moreira

Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido – UFERSA, Mossoró, Brazil

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Moacir Franco de Oliveira,

Moacir Franco de Oliveira

Department of Animal Sciences, Universidade Federal Rural do Semi-Árido – UFERSA, Mossoró, Brazil

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Alexandre Rodrigues Silva,

Corresponding Author

Alexandre Rodrigues Silva

[email protected]

Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido – UFERSA, Mossoró, Brazil

Correspondence

Alexandre Rodrigues Silva, Laboratory on Animal Germplasm Conservation, Universidade Federal Rural do Semi-Árido – UFERSA, BR 110, Km 47, Costa and Silva, 59625-900 Mossoró, RN, Brazil.

Email: [email protected]

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First published: 02 May 2022

https://doi.org/10.1111/rda.14140

Funding information

This study was supported, in part, by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, Brasil (CAPES, Financial Code 001). AR Silva and MF Oliveira are CNPq investigators

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Abstract

We studied the sperm membrane functionality through the epididymal transit by comparing different hypoosmotic solutions and verifying possible associations among osmotic response and functional parameters of sperm in red-rumped agouti (Dasyprocta leporina). For this purpose, epididymal sperm from six sexually mature male agoutis were collected via flotation. Then, analyses of sperm parameters and hypoosmotic swelling test using different hypoosmotic solutions (0, 50 and 200 mOsm/L) in different regions of the epididymis (caput, corpus and cauda) were performed. There was an increase (p < .05) in the values for sperm concentration, the total number of sperm recovered, total and progressive motility, average path velocity, straight-line velocity, curvilinear velocity, and rapid and medium subpopulations following the caput-corpus-cauda direction. Regardless of the hypoosmotic solution, the agouti sperm membrane presented similar functional integrity in all the epididymal regions. Moreover, the highest (p < .05) osmotic responses were reached with the use of 50 mOsm/L solution in comparison to 0 and 200 mOsm/L for all the regions. Significant correlations among osmotic response and some sperm kinetic parameters were observed, especially in epididymal caput, while no correlations were found in the region of the cauda. In summary, red-rumped agouti sperm present similar membrane functionality during epididymal transit, but there are evident correlations among such functionality and sperm kinetic parameters, especially in the caput region. Moreover, we indicate the use of a 50 mOsm/L hypoosmotic solution for the analysis of this parameter through the hypoosmotic swelling test.

1 INTRODUCTION

The red-rumped agouti (Dasyprocta leporina) is a South American wild hystricognath rodent that has ecological importance as a seed disperser and as prey for carnivores (Hosken & Silveira, 2001; Jones et al., 2019). Despite declining in some biomes, its populations are stable, and the species is globally classified as least concerned (Emmons & Reid, 2016). For this reason, this agouti species has been currently considered a model for the development of conservative strategies to be applied to close-related endangered rodents such as Dasyprocta ruatanique (Schipper et al., 2016), Dasyprocta coibae (Roach & Naylor, 2019) and Dasyprocta mexicana (Vázquez et al., 2008).

Unlike what is commonly observed for laboratory rodents, agoutis have reproductive organs (testes and epididymis) located in the abdomen, which may result in regulatory mechanisms not yet known (Arroyo et al., 2014). In this sense, various efforts have been conducted for the establishment of protocols for the recovery (Castelo, Souza, et al., 2015; Ferraz et al., 2011) and the preservation (Castelo, Silva, et al., 2015; Silva et al., 2012) of agouti male gamete, but some basic aspects of its sperm physiology, as the modifications of the sperm along the epididymal transit, remain unknown. In addition to species-specific osmotic characteristics, membrane functionality undergoes changes during sperm maturation, as a wide variety of functional, structural and biochemical changes in sperm occur during epididymal transit (Dantas, 2022; Olson et al., 2003). Such modifications affect the repositioning of protein, lipid and glycoprotein components in different specific regions of the intracellular membranes and plasma membrane, remodeling it along the epididymal transit, conferring motility and fertilization capacity (Jones, 1998; Soler et al., 2017).

The hypoosmotic swelling test (HOS test) has been proposed for the evaluation of sperm membrane functionality in various domestic species, such as dogs (Kumi-Diaka, 1993), goats (Fonseca et al., 2001), pigs (Lechniak et al., 2002) and cattle (Martins et al., 2011). For wildlife, however, the use of the test is limited, mainly because the osmotic response of the sperm is unknown for various species. Thus, the test was validated for only a few species, such as lion (Malo et al., 2004), six-banded armadillo (Santos et al., 2011), brown bear (Pérez-Garnelo et al., 2006), collared peccary (Santos et al., 2013) and white-lipped peccary (Barros et al., 2019). The HOS test evaluates the capability of the sperm membrane to adapt to the influx of fluids into the cell in order to provide an osmotic balance between extracellular and intracellular fluids (Jeyendran et al., 1984). Therefore, the test is based on a simple physiological concept: living cells swell when exposed to a hypoosmotic environment, and cells with damaged membranes do not (Zubair et al., 2015). Despite the relative simplicity of the HOS test, there are some points that need to be clarified to obtain greater reliability, such as the ideal osmolarity of the hypoosmotic solution (Melo et al., 2003), since each species can present a different sperm osmotic response (Matson et al., 2009).

To the best of our knowledge, the use of the HOS test throughout sperm maturation was validated neither for agoutis nor for any other hystricognath rodent. Thus, the relationship between functional membrane integrity and other sperm morphophysiological parameters of this species during the epididymal transit remains known. Since electroejaculation remains a low effective technique for sperm obtaining in agoutis (Castelo, Souza, et al., 2015), the use of epididymal sperm is proposed as an effective alternative for the establishment of protocols for sperm processing or evaluation for the species (Ferraz et al., 2011). Therefore, we aimed to investigate the osmotic response of agouti sperm along epididymal transit through the comparison of different hypoosmotic solutions based on distilled water combined with sodium citrate, to access sperm membrane functionality and verify the existence of possible associations among membrane functionality and other sperm metrics.

2 MATERIAL AND METHODS

2.1 Animals

Experimental procedures were approved by the Ethics Committee on the Use of Animals at UFERSA (CEUA – Opinion nº 11/2019) and Instituto Chico Mendes de Biodiversidade (ICMBIO – Opinion nº 66618-1). Animals were originated from the Wild Animal Multiplication Center (CEMAS) of the Federal Rural University of the Semi-arid (UFERSA), a scientific breeding center authorized by the Brazilian Institute of Renewable Resources (IBAMA – Opinion nº 66618-1), which conducts scheduled slaughter of animals destined for scientific experimentation as those used in the present study.

Six sexually mature male agoutis, aged approximately 12 months old, were used for the experiment. The animals were isolated from females for six months before the start of the study and maintained under a 12-hr natural photoperiod. They were allocated to a covered paddock (4 × 5 m) for their grouping and maintenance. Feed for the agoutis was a commercial rabbit ration, with 13% crude protein, 35% ether extract, 16% fiber and 13% minerals. Fresh drinking water was available ad libitum.

2.2 Sperm collection

After being restrained with the use of a hand net, agoutis were medicated with 1 mg/kg of ketamine (Ketalar; Pfizer) and xylazine hydrochloride (Rompun; Bayer) administered via intramuscular, followed by an intracardiac administration of 1 mg/kg potassium chloride (Castelo, Souza, et al., 2015). The pairs of epididymis were transported to the laboratory in a beaker containing gauze moistened with phosphate-buffered saline (PBS) (287 mOsm/kg) that was stored in a polystyrene insulated box. Epididymis were sliced to separate three distinct regions, caput, corpus and cauda, that were placed in individual beakers containing 2 ml of PBS solution at 37°C. Epididymal sperm were recovered by the floating method (Silva et al., 2016) in plastic tubes to obtain the volume (μl) collected per animal.

2.3 Sperm evaluations

During analysis, the samples were placed in a water bath at 37°C. Sperm concentration (×10⁶ sperm/ml) was evaluated through a Neubauer counting chamber. The number of sperm collected (sperm × 10⁶) was determined by multiplying the sperm concentration and the total volume (µl) recovered (Silva et al., 2016). For the analysis of sperm morphology, a 10-µl aliquot of the sperm sample was used to produce a smear stained with Bengal rose, counting 200 cells under light microscopy (×1000) (Silva et al., 2011).

For the analysis of the sperm membrane integrity and mitochondrial activity, an aliquot (10 μl) was incubated at 37°C for 10 min in a solution containing a combination of fluorescent probes composed of 2 μl of Propidium Iodide (PI), NaCl 0.5 mg/ml at 0.9%, 5 μl CMXRos (Mito Tracker Red^®, Molecular Probes, F-7512) at 500 nM (50 μg dilution in 94 μl DPBS) and 3 μl Hoechst 342 (H342) (diluted at 25 mg/ml in DMSO) (Celeghini et al., 2007). Then, the samples were analyzed by an epifluorescence microscope (Leica), counting 200 sperm per sample. Sperms presenting heads marked in blue (H342) were classified as presenting an intact membrane, while those marked totally or partially in red (PI) were classified as non-intact. Moreover, the sperm presenting the region of the midpiece marked in red were considered as having a mitochondrial activity (Celeghini et al., 2007).

A 3-μl aliquot was used for the analysis of the sperm kinetic parameters using a computer-assisted sperm analysis system (CASA, IVOS 7.4 G; Hamilton-Thorne Research, Beverly, MA, USA), following the settings previously established for the species: temperature, 37°C; straightness threshold, 30%; minimum contrast, 45; low-speed mid-lane cutoff (VAP), 10 μm/s; and average VAP cutoff, 30 μm/s (Castelo, Souza, et al., 2015). Therefore, five independent and non-consecutive microscopic fields were selected for the scan. The parameters analyzed were total motility (%), progressive motility (%), velocity average pathway (VAP, μm/s), velocity straight line (VSL, μm/s), velocity curvilinear (VCL, μm/s), amplitude lateral head (ALH, μm), beat cross frequency (BCF, Hz), straightness (STR, %), linearity (LIN, %) and elongation (%). The total sperm population was subdivided into four categories: rapid, medium, slow and static (%).

2.4 Hypoosmotic swelling (HOS) test

For the evaluation of sperm membrane functionality, hypoosmotic solutions composed of only distilled water (0 mOsm/L) or a combination of distilled water (0 mOsm/L) with sodium citrate and fructose solutions at different osmolarities (50 and 200 mOsm/L) were used. Aliquots of 5 μl containing epididymal sperm plus 45 μl of hypoosmotic solutions were incubated in a dry bath for 40 min at 37°C. Then, 10 μl of each treatment were aliquoted and evaluated in a phase-contrast light microscope (×400). There were 200 sperms examined; we considered those with a swollen, coiled tail as having a functional membrane (Fonseca et al., 2005).

2.5 Statistical analysis

The data were analyzed by the sigmaplot (Systat Software Inc, Version 14) and expressed as mean ± SEM. Data were checked for normality using the Shapiro-Wilk test and for homoscedasticity using the Brown-Forsythe test. Then, the sperm functional membrane integrity was evaluated by two-way repeated-measures ANOVA (Two Factor Repetition), followed by Bonferroni t-test for the pairwise multiple comparison procedures among the epididymal regions (caput, corpus and cauda) and different hypoosmotic solutions (p < .05), as well as to compare the other sperm parameters among the regions of the epididymis. The relationship between sperm parameters and functional membrane integrity at different hypoosmotic solutions and regions of the epididymis was analyzed by Spearman's correlation test (p < .05).

3 RESULTS

3.1 Sperm parameters in different epididymal regions

Regarding sperm parameters (Table 1), fresh sperm samples recovered from the agouti epididymis cauda presented the highest values (p < .05) related to volume concentration (1285.2 ± 319.3 × 10⁶ sperm/ml) and the total number of sperm recovered (1375.0 ± 244.1 × 10⁶ sperm) when compared to other epididymal regions. Additionally, various sperm kinetic parameters, such as total and progressive motility, VAP, VSL, VCL, and rapid and medium subpopulations, presented higher values in epididymal cauda in comparison to other regions (p < .05).

TABLE 1. Mean values (± SEM) for the sperm parameters of red-rumped agouti (Dasyprocta leporina; n = 6) recovered from different regions of the epididymis

Sperm parameters	Epididymal region
Sperm parameters	Caput	Corpus	Cauda
Sperm concentration (× 10⁶ sperm/ml)	105.0 ± 23.8^B	116.3 ± 42.8^B	1285.2 ± 319.3^A
Number of sperm recovered (× 10⁶ sperm)	691.7 ± 39.6^B	691.7 ± 20.1^B	1375.0 ± 244.1^A
Normal morphology (%)	84.0 ± 1.2	85.7 ± 1.4	87.3 ± 1.9
Membrane structural integrity (%)	80.7 ± 4.0	75.2 ± 6.8	83.5 ± 3.4
Mitochondrial activity (%)	73.8 ± 7.2	45.0 ± 12.5	85.8 ± 3.1
Total motility (%)	12.2 ± 4.2^C	37.7 ± 8.8^B	76.8 ± 6.2^A
Progressive motility (%)	0.8 ± 0.3^B	5.3 ± 2.2^B	13.7 ± 2.8^A
Average path velocity (VAP; μm/s)	19.9 ± 4.2^B	33.1 ± 3.8^B	38.9 ± 2.6^A
Straight line velocity (VSL; μm/s)	13.2 ± 3.0^B	22.2 ± 2.9^AB	27.2 ± 2.2^A
Curvilinear velocity (VCL; μm/s)	38.9 ± 8.4^B	62.0 ± 5.1^B	69.4 ± 3.9^A
Amplitude of lateral head displacement (ALH; μm)	4.2 ± 1.0	5.0 ± 0.4	5.7 ± 0.3
Beat cross frequency (BCF; Hz)	31.0 ± 6.4	41.9 ± 0.7	39.5 ± 0.7
Straightness (STR; %)	51.7 ± 10.7	60.5 ± 1.2	65.2 ± 1.3
Linearity (LIN; %)	27.0 ± 5.7	33.0 ± 1.2	36.3 ± 1.5
Elongation (%)	40.3 ± 8.3	49.5 ± 1.9	58.8 ± 2.1
Subpopulations
% of rapid sperm	1.0 ± 0.4^B	8.2 ± 3.8^B	19.8 ± 4.0^A
% of medium velocity sperm	11.0 ± 4.0^C	29.3 ± 5.3^B	57.0 ± 2.8^A
% of slow sperm	0.5 ± 0.2	0.7 ± 0.3	0.2 ± 0.2
% of static sperm	87.3 ± 4.3^A	61.7 ± 8.7^B	22.7 ± 6.2^C

Note

^ABCDifferent superscript uppercase letters indicate significant differences among the sperm parameters and the epididymal region (p < .05).

3.2 Sperm functional membrane integrity during epididymal transit

The HOS test revealed significant differences (p < .05) among the hypoosmotic solutions evaluated, being the most significant percentage of reactive sperm found at the use of a 50 mOsm/L solution in all regions of the epididymis assessed (Table 2). Additionally, there were no differences in sperm membrane functionality for any solution evaluated among different epididymal regions (caput, corpus and cauda).

TABLE 2. Values for (means ± SEM) osmotic response of red-rumped agouti (Dasyprocta leporina; n = 6) sperm to different hypoosmotic solutions in different regions of the epididymis

Epididymal region*	Osmolarity of solutions
Epididymal region*	0 mOsm/L	50 mOsm/L	200 mOsm/L
Caput	37.3 ± 10.6^B	68.2 ± 10.3^A	49.7 ± 9.3^AB
Corpus	35.0 ± 11.8^B	75.8 ± 7.8^A	54.8 ± 12.0^AB
Cauda	42.5 ± 3.5^B	83.3 ± 3.9^A	61.8 ± 3.9^AB

Note

^ABCDifferent superscript uppercase letters indicate significant differences among osmolarities within the same region of the epididymis (p < .05).
* There were no significant differences among the different regions of the epididymis with the same osmolarity (p > .05).

3.3 Correlations among functional membrane integrity and other sperm parameters

There were significant correlations among spermatozoa reactive to the use of different hypoosmotic solutions and various sperm kinetic parameters for the caput and corpus regions, but none for the cauda (Table 3).

TABLE 3. Values for Spearman correlations (ρ) among the osmotic response to different hypoosmotic solutions and the sperm parameters in different regions of the agouti (Dasyprocta leporina; n = 6) epididymis

Kinetic parameters*

Osmolarity of solutions

Epididymal region

Caput

Corpus

Cauda**

Straight line velocity (VSL; μm/s)

0 mOsm/L

0.886

.033

−0.836

.033

–

Subpopulation of slow sperm (%)

0.878

.033

–

Average path velocity (VAP; μm/s)

50 mOsm/L

0.841

.033

0.886

.033

–

Subpopulation of rapid sperm (%)

200 mOsm/L

0.956

.003

–

Elongation (%)

−0.943

.017

–

* Only the sperm parameters that showed significant correlations were expressed in the table (p < .05).
** There were no significant correlations among osmotic responses in the epididymal cauda region and other sperm parameters (p > .05).

4 DISCUSSION

Much more than just an indication of the structural integrity of the sperm membrane, the HOS test evaluates the functional capacity of the sperm. Through this test, we demonstrated that red-rumped agouti spermatozoa have a very functional membrane during the entire epididymal transit. This was especially evidenced with the use of a hypoosmotic solution of 50 mOsm/L, which revealed values around 70% of spermatozoa with functional membrane independent of the epididymal region; however, the number of spermatozoa reactive to the test varied according to the use of different hypoosmotic solutions.

As soon as the spermatozoa are released from the seminiferous tubules, the sperm membrane is subjected to continuous remodeling upon their transit in the epididymis (Kuo et al., 2016). Given that the osmotic pressure of the epididymal fluid, especially in the tail, is considerably higher than that of seminal plasma or blood (Sahin et al., 2009), the approach in which a hypoosmotic challenge is used to monitor sperm membrane function along epididymal transit is very valuable. During the process of sperm maturation, the sperm membrane undergoes physiological changes along the epididymal transit that reflect on its functionality and consequently on the acquisition of motility and fecundating capacity (Jones, 1998). In this sense, Druart et al. (2009) observed that there is a progressive decrease in the hypotonic resistance of boar spermatozoa during their transit from the caput to the cauda of the epididymis, which could be associated with changes in lipid and protein compositions, resulting in different physical properties of the membrane. In agoutis, however, we did not observe differences related to sperm osmotic response during epididymal transit, similarly to what was described for cattle using a hypotonic challenge for sperm from the caput and the cauda of the epididymis (Sahin et al., 2009). Of course, differences in the composition of the plasma membrane of the spermatozoa of agoutis must also occur along the epididymal transit, but the HOS test probably does not have the necessary sensitivity to express such detailed changes (Gervasi & Visconti, 2017).

Independently from the epididymal region, the agouti sperm showed the greatest osmotic response in the presence of an intermediate osmolarity (50 mOsm/L) in comparison to the extreme ones (0 and 200 mOsm/L). However, in murine rodents, such as mice, the use of the ’water test’ at 0 mOsm/L appears to be well established for the analysis of sperm membrane function (Sliwa, 1993). When first described in 1984, the hypoosmotic test was used to assess human semen by using solutions with osmolarities varying from 50 to 300 mOsm/L, and the best sperm reactive rates were achieved at the use of a 150 mOsm/L solution (Jeyendran et al., 1984). Since then, the HOS test has been applied to several species and differences in sperm osmotic response have been noted even between phylogenetically close species (Zubair et al., 2015). Generally speaking, the ideal hypoosmotic solution would be one that exerts osmotic stress large enough to provide an observable increase in volume, but at the same time small enough that membrane lysis does not occur (Jeyendran et al., 1984). Therefore, due to the specific characteristics of the sperm membrane for each species, it is expected that there will be variation in the number of spermatozoa reactive to the HOS test.

In addition to the functionality of the sperm membrane, we observed an improvement in the morphofunctional parameters during the maturation of agouti sperm, especially those related to kinetics, similarly as described for other rodents as murine (Soler et al., 1994) and hamsters (Olson et al., 2003). Although no differences were found in the osmotic response of spermatozoa between regions of the epididymis, the functionality of the agouti sperm membrane seems to be intrinsically related to the acquisition of sperm movement considering the direction from the caput to the cauda region of the epididymis. This fact is supported by the observation of numerous correlations between the osmotic response and the kinetic parameters of spermatozoa obtained from the caput when compared to those obtained from the epididymal cauda. Although it is known that there are specific biochemical interactions between secretions from the male reproductive tract and the germ cells along the epididymal transit, the cauda and corpus regions are described as mainly responsible for sperm maturation, while the cauda acts mainly as a sperm reservoir (Gervasi & Visconti, 2017; Toshimori, 2003).

In studies with spermatozoa from cats (Comercio et al., 2013), drones (Nur et al., 2011) and humans (Van den Saffele et al., 1992), positive correlations were found between the HOS test and sperm motility. According to the authors, it is expected that there will be correlations between the parameters regarding motility and spermatozoa reactive to the HOS test since it is known that sperm motility depends in part on sperm membrane functionality, as well as biochemical and metabolic processes (Jeyendran et al., 1984). On the other hand, in our study, no correlations were identified between spermatozoa from the epididymal cauda of agouti. These results were similar to those described for spermatozoa ejaculated from goats (Oliveira et al., 2013), horses (Snoeck et al., 2007) and six-banded armadillos (Santos et al., 2011). According to these authors, the reason for this lack of correlations between the HOS test and sperm morphofunctional parameters is a consequence of the different specificities and purposes of these analyses. In fact, while spermatic swelling evidences the functional integrity of sperm membranes in the HOS test, sperm motility and viability depend not only on the transport of substances that pass through the membranes, but also on several other biochemical functions that modulate the sperm metabolism, as well as the microtubular activity of the fibers present in the region of the sperm tail. It is necessary to mention that a small sample number can also influence the identification of significant correlations between the referred parameters; however, this is, unfortunately, a limiting factor in studies conducted with wild species whose availability of individuals to be used in scientific research is scarce.

5 CONCLUSIONS

In summary, we provided novel data related to the function of the sperm membrane along the epididymal transit, which presents significant correlations with some sperm kinetic parameters in the red-rumped agouti (Dasyprocta leporina), especially in the epididymal caput. Moreover, we recommended the use of a 50 mOsm/L hypoosmotic solution for the analysis of this parameter through the hypoosmotic swelling test.

AUTHOR CONTRIBUTIONS

M.R.T. Dantas, N.R.N. Luz, L.G.P. Bezerra, and S.S.J. Moreira executed the experiments and analysis; M.F. Oliveira, A.R. Silva are the main leaders of the team that manage the experiments and the manuscript writing.

CONFLICT OF INTEREST

None of the authors have any conflict of interest to declare.

Open Research

DATA AVAILABILITY

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Volume57, Issue8

August 2022

Pages 912-918

Evaluation of sperm membrane functionality during epididymal transit in red-rumped agouti (Dasyprocta leporina)

Abstract

1 INTRODUCTION

2 MATERIAL AND METHODS

2.1 Animals

2.2 Sperm collection

2.3 Sperm evaluations

2.4 Hypoosmotic swelling (HOS) test

2.5 Statistical analysis

3 RESULTS

3.1 Sperm parameters in different epididymal regions

Note

3.2 Sperm functional membrane integrity during epididymal transit

Note

3.3 Correlations among functional membrane integrity and other sperm parameters

4 DISCUSSION

5 CONCLUSIONS

AUTHOR CONTRIBUTIONS

CONFLICT OF INTEREST

Open Research

DATA AVAILABILITY

REFERENCES

References

Information

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Evaluation of sperm membrane functionality during epididymal transit in red-rumped agouti (Dasyprocta leporina)

Abstract

1 INTRODUCTION

2 MATERIAL AND METHODS

2.1 Animals

2.2 Sperm collection

2.3 Sperm evaluations

2.4 Hypoosmotic swelling (HOS) test

2.5 Statistical analysis

3 RESULTS

3.1 Sperm parameters in different epididymal regions

Note

3.2 Sperm functional membrane integrity during epididymal transit

Note

3.3 Correlations among functional membrane integrity and other sperm parameters

4 DISCUSSION

5 CONCLUSIONS

AUTHOR CONTRIBUTIONS

CONFLICT OF INTEREST

Open Research

DATA AVAILABILITY

REFERENCES

References

Related

Information