Comparison of sperm motility test methods (except computer-assisted sperm analysis) in rats under the condition of alfa-chlorohydrin treatment. Collaborative investigation.

Mineshima, Hiroshi; Fujioka, Michi; Furukawa, Masatoshi; Ikeda, Takashi; Kinoshita, Kentaro; Koida, Masahiro; Kondoh, Kazuhisa; Ozawa, Shogo; Oi, Akihide; Ohyama, Naoki; Takahashi, Hiroki; Tanaka, Ryota

doi:10.2131/jts.25.5_443

Cited by 3 publications

(9 citation statements)

References 26 publications

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“…Video recordings were blindly evaluated for motility (number of motile cells and spermatozoa speed) by 2 independent observers. A semiquantitative score (graded 0 to 4) of sperm motility was assigned to each sample (Mineshima et al, 2000).…”

Section: Methodsmentioning

confidence: 99%

Localization, Distribution, and Function of the Calcium‐Sensing Receptor in Sperm

Mendoza

Marı́n

García‐Marín

et al. 2012

Journal of Andrology

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ABSTRACT:The intracellular movement of calcium, through calcium channels, plays a major role on sperm cell function. The calcium-sensing receptor (CaSR), a molecular mechanism by which many cells detect changes in extracellular calcium concentration, has not been described in spermatozoa. The aim of this study was to investigate the expression of the CaSR in testicular tissue and sperm cells and the functional consequences of spermatozoid CaSR activation by calcimimetics. CaSR mRNA and protein were identified both in rat testicular tissue and in rat spermatozoa using real-time reverse transcription-polymerase chain reaction and immunohistochemistry, respectively. Functionality of CaSR was evaluated by studying the influence of calcimimetic AMG 641 on rat and pig sperm motility. Treatment with AMG 641 100 nM for 1 hour increased rat sperm motility from a score of 1.0 6 0.1 to 3.8 6 0.3 (P , .05). AMG 641 also resulted in a modest but significant increase in the pig sperm motility parameters evaluated by computer-assisted sperm analysis. AMG 641 was effective in a wide range of concentrations but resulted in a more marked effect at 50-100 nM. In addition, AMG 641 did not have any negative effect on sperm viability, which was measured by flow cytometry. In conclusion, our results demonstrate the expression of functional CaSR in testicular tissue and sperm, which can be activated by calcimimetic AMG 641.

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Section: Methodsmentioning

confidence: 99%

Localization, Distribution, and Function of the Calcium‐Sensing Receptor in Sperm

Mendoza

Marı́n

García‐Marín

et al. 2012

Journal of Andrology

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“…Vickery et al (1974), Hoyt et al (1994) and Ban et al (1999) showed a NOAEL of 1 mg/kg, whereas Li et al (2003) reported a NOAEL of 2 mg/kg bw per day based on decreased sperm count. Other studies did not provide a NOAEL, the lowest doses tested representing LOAELs above 1 mg/kg bw per day (Yamada et al, 1995;Mineshima et al, 2000;Kim et al, 2012;Zhang et al, 2012). 3-MCPD fatty acid esters were tested in Barocelli et al (2011) and Onami et al (2014) and revealed histopathological effects in testis and epididymis at doses corrected for free-3-MCPD content above the NOAEL for free 3-MCPD.…”

Section: Discussionmentioning

confidence: 99%

“…These data indicate a NOAEL of 1 mg 3-MCPD/kg bw per day, based on dosedependent decrease in sperm motility and consequential decrease in fecundity observed at 3 and 10 mg 3-MCPD/kg bw per day. Mineshima et al (2000) reports on a ring study involving 11 laboratories, comparing five different rat sperm motility test methods. Using 3-MCPD as a test compound affecting epididymis sperm motility, 10-15-week-old male rats, in most labs of the Crj:CD strain, were orally treated for 1 week (in one lab for 2 weeks) with dose levels of 2.5, 5 and 10 mg/kg bw per day.…”

Section: Assessmentmentioning

confidence: 99%

“…()Rat, Sprague–Dawley3‐MCPD, 90 days, gavage, 22 animals/dose group, 1/dayM0.25, 0.5, 1.0, 2.0, 4.0, 8.0 or 16.0 mg/kg bwReproductive organs weight and histopathology; Epididymis sperm count, viability and morphology; Testicular enzymes (LDH, LDH‐X) levels 2.0 mg/kg bw per day (sperm count) 4.0 mg/kg bw per day (sperm viability) 4.0 mg/kg bw per day (sperm count) 8.0 mg/kg bw per day (sperm viability) Mineshima et al. ()Rat, Sprague–Dawley Crj:CD (mostly)3‐MCPD, 1–2 weeks, gavage, 4–10 animals/dose group, 1/dayM2.5, 5.0 or 10.0 mg/kg bwCauda epididymis/testis sperm motility, count and morphology; Fertility–2.5 mg/kg bw (sperm motility and fertility)Study designed as a multilaboratory comparison of different methods for sperm motility assessment. Data presentation other than for motility is very limitedOnami et al.…”

Section: Appendix a – Studies Selected For Full Text Appraisalmentioning

confidence: 99%

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Update of the risk assessment on 3‐monochloropropane diol and its fatty acid esters

Knutsen¹,

Alexander²,

Barregård³

et al. 2018

EFS2

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The CONTAM Panel updated the assessment of the risks for human health related to the presence of 3-monochloropropane diol (3-MCPD) and its fatty acid esters in food published in 2016 in view of the scientific divergence identified in the establishment of the tolerable daily intake (TDI) in the Joint FAO/WHO Expert Committee on Food Additives and Contaminants (FAO/WHO) report published in 2017. In this update, dose-response analysis was performed following the recent EFSA Scientific Committee guidance on the use of benchmark dose (BMD) approach in risk assessment, and a review of available data on developmental and reproduction toxicity was included. The outcome of this review indicates that in rats short-term exposure to 3-MCPD above 1 mg/kg body weight (bw) per day can induce reduced sperm motility associated with reduced male fecundity. Decreased sperm count and histopathological changes in the testis and epididymis were observed following longer treatment periods at higher doses. Regarding increased incidence kidney tubular hyperplasia, BMD analysis using model averaging resulted in a BMDL 10 of 0.20 mg/kg bw per day in male rats, which was selected as the new Reference Point (RP) for renal effects. For the effects on male fertility, decreased sperm motility was selected as the most sensitive relevant endpoint and a BMDL 05 of 0.44 mg/kg bw per day was calculated. The RP for renal effects was considered to derive an updated group TDI of 2 lg/kg bw per day for 3-MCPD and its fatty acid esters and was considered protective also for effects on male fertility. The established TDI of 2 lg/kg bw per day is not exceeded in the adult population. A slight exceedance of the TDI was observed in the high consumers of the younger age groups and in particular for the scenarios on infants receiving formula only.

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“…21, 1997). In some end-points of sperm analysis, measure-ment of the sperm motility has been used as an indicator for assessment of male fertility, because some investigators have reported that impaired sperm motion affects fertility index and reproductive status (Mineshima et al, 2000;Ban et al, 2001). Establishment of a technique to discriminate viable and dead spermatozoa is needed for accurate evaluation of spermatozoa which have reduced mobility or have died, when measurement of the sperm motility is decreased by some component.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Mitochondrial Function and Membrane Integrity by Dual Fluorescent Staining for Assessment of Sperm Status in Rats

Kato¹,

Makino²,

Kimura³

et al. 2002

J. Toxicol. Sci.

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Dual fluorescent staining (DFS) with calcein acetoxy methyl ester (CAM), which labels the cellular esterase activity that is a major component of energy metabolism in cellular mitochondria, and with ethidium homodimer-1 (EthD-1) was used to evaluate mitochondrial function and membrane integrity in rat spermatozoa. The spermatozoa stained by DFS could be classified into three different populations microscopically when excited at 490 nm after 60 min incubation. 1) Spermatozoa, which were stained with CAM alone and had maintained either mitochondrial function or membrane integrity, were identified as live during incubation. 2) Spermatozoa, which were stained with EthD-1 alone and had lost either mitochondrial function or membrane integrity, were identified as already dead at the beginning of incubation. 3) Spermatozoa, which were stained with both CAM and EthD-1 and had maintained mitochondrial function with membrane breached, were identified as having died during incubation. Two toxicological tests, an in vitro triton X-100 experiment and an in vivo nitrobenzene experiment, were done. All spermatozoa were immobilized and lost either mitochondrial function or membrane integrity by 1.0% triton X-100 treatment. Almost no motile sperm were found at 0.1% in the triton X-100 group and in the groups treated with 60 and 40 mg/kg/day of nitrobenzene, and these spermatozoa maintained their mitochondrial function but had their membrane breached. In conclusion, the DFS procedure, which uses CAM and EthD-1, can clearly and visually identify the population of viable and dead spermatozoa simultaneously by fluorescence microscopy in rats. This is a useful technique to characterize sperm status, which is determined by the mitochondrial function assessed by CAM and membrane integrity evaluated by EthD-1.

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Comparison of sperm motility test methods (except computer-assisted sperm analysis) in rats under the condition of alfa-chlorohydrin treatment. Collaborative investigation.

Cited by 3 publications

References 26 publications

Localization, Distribution, and Function of the Calcium‐Sensing Receptor in Sperm

Localization, Distribution, and Function of the Calcium‐Sensing Receptor in Sperm

Update of the risk assessment on 3‐monochloropropane diol and its fatty acid esters

Evaluation of Mitochondrial Function and Membrane Integrity by Dual Fluorescent Staining for Assessment of Sperm Status in Rats

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