Validation of non-fluorescent methods to reliably detect acrosomal and plasma membrane integrity of common marmoset (Callithrix jacchus) sperm

Valle, Rodrigo del Rio do; Valle, C. M. R; Nichi, Marcílio; Muniz, José Augusto Pereira Carneiro; Nayudu, P.L.; Guimarães, Marco Aurélio

doi:10.1016/j.theriogenology.2008.03.011

Cited by 24 publications

(35 citation statements)

References 12 publications

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“…The percentage of sperm with intact plasma membrane was determined after evaluation of 200 sperm in a smear of semen stained with eosin-nigrosin, with the microscope at ×1,000 magnification [Valle et al, 2008]. For acrosome integrity assessment, 2 different stains were used.…”

Section: Semen Evaluationmentioning

confidence: 99%

“…To perform fast-green/bengal-rose staining [Pope et al, 1991;Valle et al, 2008], 5 μL of semen was mixed with 5 μL of stain, the solution was incubated for 70 s, and a smear was prepared. For the commercial kit Spermac ® staining (Stain Enterprises, Onderstepoort, South Africa) [Valle et al, 2008], an additional smear was prepared with diluted semen, and the slide was stained following the manufacturer's protocol. In both cases, 200 sperm were assessed, at ×1,000 magnification.…”

Section: Semen Evaluationmentioning

confidence: 99%

“…Although there are only a few studies available on the semen characteristics in Neotropical primates to date, in species from the genus Callithrix , Saimiri , Ateles , and Saguinus , as well as the species A. caraya , Cebus ( S. ) apella , and L. chrysomelas [Sankai et al, 1997;Barnabe et al, 2002;Vidal et al, 2007;Hernandéz-Lópes et al, 2008;Massarotto et al, 2010;Poches et al, 2013;Silva et al, 2013a, b;Carvalho et al, 2014;Valle et al, 2014;Leão et al, 2015;Sampaio et al, 2017], major differences are known between the groups [Valle et al, 2008]. These differences are found in several seminal parameters, ranging from volume and concentration to plasma membrane integrity and sperm DNA fragmentation.…”

Section: Introductionmentioning

confidence: 99%

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Collection, Evaluation, and Coagulum Dissolution of Semen from Goeldi's Monkey, Callimico goeldii

Arakaki¹,

Carvalho²,

Castro³

et al. 2017

IJFP

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Research on Neotropical primates' reproduction is necessary due to the lack of available information and the increasing threat to these species. Callimico goeldii is listed as Vulnerable on the IUCN Red List of Threatened Species. This study aimed to test rectal electrostimulation for semen collection and evaluate seminal characteristics. Therefore, semen from 6 captive Goeldi's monkeys was collected and, for the first time, seminal characteristics are described. Coagulum formation was noted in all ejaculates, and we obtained partial or complete liquefaction of the samples. Results were (means ± SD): volume = 26.9 ± 11.87 μL; pH = 7.61 ± 0.28; concentration = 143.18 ± 174.96 × 10⁶ spermatozoa/mL; total sperm motility = 83.33 ± 5.16%; linear progressive motility = 46 ± 24.08%; plasma membrane integrity = 36.38 ± 16.11%; acrosome integrity using fast-green/bengal-rose staining = 63.41 ± 11.72%, and kit Spermac® = 69.36 ± 11.81%; abnormal sperm = 72.5 ± 17.7%, with 16.2 ± 7.7% major defects and 56.3 ± 10% minor defects; sperm with high mitochondrial activity class I = 16.45 ± 22.25%. Rectal electrostimulation was an efficient method for semen collection in this species. Investigations are required to improve semen collection and handling, including cryopreservation methods.

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Section: Semen Evaluationmentioning

confidence: 99%

Section: Semen Evaluationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Collection, Evaluation, and Coagulum Dissolution of Semen from Goeldi's Monkey, Callimico goeldii

Arakaki¹,

Carvalho²,

Castro³

et al. 2017

IJFP

View full text Add to dashboard Cite

show abstract

“…Stimulation was continued with 80 Hz and 1-mm amplitude for 1 min and 70 Hz and 1.5-mm amplitude for 1 min. If ejaculation had not occurred, stimulation intensity was increased to 80 Hz and 1.5 mm, 90 Hz and 1.0 mm, and 90 Hz and 1.5 mm [13], pooling two successive ejaculates from the same animal per day to obtain homogeneous samples. All marmosets were maintained under uniform nutritional and environmental conditions to minimize external factor differences and effects on semen quality.…”

Section: Donors and Semen Collectionmentioning

confidence: 99%

“…Slides were prepared by placing the stained sperm sample on the clear end of a frosted microscope slide and dragging the drop across the slide to create a thin feathered smear (two smears per ejaculate). Duplicated sperm smears from each ejaculate were stained as described by Pope et al [14] adapted to the marmoset [13]. Briefly, 5 L of semen were incubated with 5 L of stain solution in a 0.5 mL microtube in the dark for 90 to 120 sec at room temperature (25°C ).…”

Section: Semen Processing and Sample Stainingmentioning

confidence: 99%

Sperm head morphometry in ejaculates of adult marmosets (Callithrix jacchus): A model for studying sperm subpopulations and among-donor variations

et al. 2012

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Assessment of different sperm functional tests in golden‐headed lion tamarins (Leontopithecus chrysomelas)

Arakaki

Salgado

Losano

et al. 2019

American J Primatol

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The golden‐headed lion tamarin (Leontopithecus chrysomelas) is an endangered species endemic to Brazil's Atlantic Forest, a shrinking biodiversity hotspot. As in other Neotropical primates, its semen characteristics and freezability are poorly studied. Hence, reproductive technologies for callitrichids would greatly benefit from reliable methods of semen analysis. In a bid to promote reproductive research in tamarins, we validated simple and inexpensive sperm function tests that can be used to monitor sperm‐egg binding, plasma membrane and acrosome integrity, mitochondrial activity, and DNA fragmentation. Ejaculates from adult males were individually diluted and divided into control and damage‐induced aliquots, and then samples comprising assorted amounts of damaged spermatozoa were examined by organelle‐specific tests. Our findings showed that sperm‐binding in chicken egg perivitelline membrane (EPM) positively correlated with the number of spermatozoa injured by snap‐freezing. Eosin‐nigrosin (EN) and propidium iodide readings were correlated with each other, and both provided robust measurements of plasma membrane integrity. A high correlation between expected and measured amounts of acrosome‐intact spermatozoa was found using Fast Green‐Rose Bengal (FG‐RB), Coomassie Blue (CB), and FITC‐PSA stains, and all three methods exhibited comparable results. Likewise, different percentages of UV‐irradiated spermatozoa were accurately assessed for DNA integrity by Toluidine Blue (TB) and sperm chromatin dispersion (SCD) tests. Comparisons between 3,3′‐diaminobenzidine (DAB) and JC‐1 stains also indicated the reliability of the former assay to ascertain gradual increases in spermatozoa with greater mitochondrial function. These data confirmed that different parts of the tamarin spermatozoa can be simply and consistently evaluated by EPM, EN, FG‐RB, CB, TB, and DAB protocols.

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Validation of non-fluorescent methods to reliably detect acrosomal and plasma membrane integrity of common marmoset (Callithrix jacchus) sperm

Cited by 24 publications

References 12 publications

Collection, Evaluation, and Coagulum Dissolution of Semen from Goeldi's Monkey, Callimico goeldii

Collection, Evaluation, and Coagulum Dissolution of Semen from Goeldi's Monkey, Callimico goeldii

Sperm head morphometry in ejaculates of adult marmosets (Callithrix jacchus): A model for studying sperm subpopulations and among-donor variations

Assessment of different sperm functional tests in golden‐headed lion tamarins (Leontopithecus chrysomelas)

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