Sperm analysis by flow cytometry using the fluorescent thiol labeling agent monobromobimane

Seligman, Judith; Shalgi, Ruth; Oschry, Yitzchak; Kosower, Nechama S.

doi:10.1002/mrd.1080290310

Cited by 29 publications

(18 citation statements)

References 19 publications

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“…Spermatozoa were collected from cauda epididymis as described by Seligman et al (1991). Sperm count in cauda epididymis was determined by our laboratory procedure (Krishnamoorthy et al , 2007).…”

Section: Methodsmentioning

confidence: 99%

Lycopene supplementation prevents reactive oxygen species mediated apoptosis in Sertoli cells of adult albino rats exposed to polychlorinated biphenyls

Giribabu¹,

Selvakumar²,

Venkataraman³

et al. 2013

Interdisciplinary Toxicology

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Sertoli cell proliferation is attenuated before attaining puberty and the number is fixed in adult testes. Sertoli cells determine both testis size and daily sperm production by providing physical and metabolic support to spermatogenic cells. Polychlorinated biphenyls (PCBs) exposure disrupts functions of Sertoli cells causing infertility with decreased sperm count. On the other hand, lycopene is improving sperm count and motility by reducing oxidative stress in humans and animals. Hence we hypothesized that PCBs-induced infertility might be due to Sertoli cell apoptosis mediated by oxidative stress and lycopene might prevent PCBs-induced apoptosis by acting against oxidative stress. To test this hypothesis, animals were treated with vehicle control, lycopene, PCBs and PCBs + lycopene for 30 days. After the experimental period, the testes and cauda epididymidis were removed for isolation of Sertoli cells and sperm, respectively. We observed increased levels of oxidative stress markers (H2O2 and LPO) levels, increased expression of apoptotic molecules (caspase-8, Bad, Bid, Bax, cytochrome C and caspase-3), decreased anti-apoptotic (Bcl2) molecule and elevated apoptotic marker activity (caspase-3) in Sertoli cells of PCBs-exposed animals. These results were associated with decreased sperm count and motility in PCBs exposed animals. On the other hand, lycopene prevented the elevation of Sertoli cellular apoptotic parameters and prevented the reduction of sperm parameters (count and motility). The data confirmed that lycopene as an antioxidant scavenged reactive oxygen substances, prevented apoptosis, maintained normal function in Sertoli cells and helped to provide physical and metabolic support for sperm production, thereby treating infertility in men.

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Section: Methodsmentioning

confidence: 99%

Lycopene supplementation prevents reactive oxygen species mediated apoptosis in Sertoli cells of adult albino rats exposed to polychlorinated biphenyls

Giribabu¹,

Selvakumar²,

Venkataraman³

et al. 2013

Interdisciplinary Toxicology

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“…Although human spermatozoa can exhibit a dense, homogeneous chromatin structure, a considerable proportion may have a coarse, granular appearance with vacuoles of varying number and size (Bedford et al, 1973;Evenson et al, 1978;Jager, 1990). This coarse, granular appearance can be found within morphologically normal spermatozoa (Seligman et al, 1991) and is similar to that seen in late spermatids of other species (Fawcett, 1958). Therefore, the presence of this type of chromatin may be indicative of immaturity due to inadequate time in the epididymis (Seligman et al, 1994).…”

Section: Human Sperm Chromatinmentioning

confidence: 99%

Sperm Chromatin Structure Assay: Its Clinical Use for Detecting Sperm DNA Fragmentation in Male Infertility and Comparisons With Other Techniques

Evenson

Larson

Jost

2002

Journal of Andrology

904

768

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The Sperm Chromatin Structure Assay (SCSA) was first described in the December 5, 1980 issue of Science (Evenson et al, 1980a). The data described in that article showed a significant difference between proven fertile and subfertility or infertility in men and bulls and the susceptibility to denaturation of their sperm nuclear DNA. A subsequent study (Evenson et al, 1984) described SCSA data obtained from men with testicular cancer. The data were heterogeneous, and begged the question, what was SCSA measuring? Great effort has been spent over the past 15 years on animal model systems, mostly related to dose-response toxicology experiments and large animal fertility trials. The results from these studies showed that SCSA was highly dose-responsive to toxicants, highly repeatable, and provided meaningful biological information on sperm nuclear DNA defects. The extensive data from nonhumans (Evenson et al, 1985(Evenson et al, , 1989a(Evenson et al, ,b, 1993a(Evenson et al, ,b,c, 1994Ballachey et al, 1987Ballachey et al, , 1988Evenson and Jost, 1993; Sailer et al, 1995a,b;Evenson, 1999a), and human toxicology and fertility studies (Evenson et al, 1978(Evenson et al, , 1980a(Evenson et al, ,b, 1984(Evenson et al, , 1991Evenson and Melamed, 1983;Evenson, 1999b;Larson et al, 1999Larson et al, , 2000Larson et al, , 2001 provide This work was supported in part by grant R827019 from the US Environmental Protection Agency, grants EPS-9720642 and OSR-9452894 from the National Science Foundation, and by SCSA Diagnostics, Inc. This is South Dakota Agricultural Experiment Station publication 3273 of the journal series.Correspondence to: Donald P. Evenson, Olson Biochemistry Laboratories, ASC 136, South Dakota State University, Brookings, SD 57006 (e-mail: donald evenson@sdstate.edu).Received for publication September 20, 2001; accepted for publication September 20, 2001. compelling evidence that SCSA will be useful in clinical human semen analysis (Figure 1).After reviewing current infertility center sites on the World Wide Web and approximately a dozen lay-oriented books on infertility and assisted reproductive technology (ART), it was amazing that not a single reference contained any information about the negative influence of fragmented sperm nuclear DNA on successful pregnancy outcome. Furthermore, few physicians are aware of scientific literature on this subject. Currently, if a spermatozoon has some motility as seen under a light microscope, and reasonable morphology, then many ART clinics assume that the sperm DNA must be fine. However, a number of cases have been documented in which couples have had multiple failed attempts at intracytoplasmic sperm injection (ICSI), even using donor eggs, and we found that the fragmentation of sperm DNA was above our threshold for less than 1% probability of achieving a successful pregnancy. Spermatozoa with defective DNA can fertilize an oocyte, produce high-quality early stage embryos, and then, in relationship to the extent of DNA damage, fail in producing a successful term ...

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“…The paraffin sections were cut to 5-6 µm of thickness. The epididymides were rapidly isolated from the testes and divided into caput, corpus and cauda in order to collect the spermatozoa separately from the distinct regions as described by Seligman et al (1991). In brief, pieces of tissue were shaken in 0.1M phosphate buffered saline (PBS) at pH 7.4 and 37°C for 15 min to allow the release of spermatozoa and the epididymal fluid.…”

Section: Immunohistochemistry In Light Microscopymentioning

confidence: 99%

Immunocytochemical Localisation of Phospholipid Hydroperoxide Glutathione Peroxidase in Bull’s Spermatogenic Cells

Parillo

Sylla

Palombi

et al. 2014

Italian Journal of Animal Science

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Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is a selenoprotein, which protects biomembranes from oxidative damages, and it also accounts for almost the entire selenium content of mammalian testis. The present investigation was performed to localise PHGPx in the testis and in epididymal and ejaculated spermatozoa of the bull by using light and electron immunomicroscopy. The study also aimed to further clarify the possible functions of the protein in bull fertility. In the testis, spermatogenic cells of the adluminal tubular compartment showed cytoplasmatic immunostaining; whereas, in the epididymal and ejaculated spermatozoa immunostaining was specifically localised at the level of the head and mid-piece. Ultrastructural data revealed the presence of signals for PHGPx in different subcellular compartments of maturing and mature sperm (mitochondria, chromatin, nuclear envelope, acrosomes, cytoskeletal structures) suggesting that this enzyme plays versatile and important biological roles during spermatogenesis. The final localisation of the immunostaining at acrosomal level puts forward a new role of the protein which further emphasises its relevance in male reproduction: it is reported to anchor substrate of the sperm acrosome to the oocyte zona pellucida during the fertilisation process.

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Sperm analysis by flow cytometry using the fluorescent thiol labeling agent monobromobimane

Cited by 29 publications

References 19 publications

Lycopene supplementation prevents reactive oxygen species mediated apoptosis in Sertoli cells of adult albino rats exposed to polychlorinated biphenyls

Lycopene supplementation prevents reactive oxygen species mediated apoptosis in Sertoli cells of adult albino rats exposed to polychlorinated biphenyls

Sperm Chromatin Structure Assay: Its Clinical Use for Detecting Sperm DNA Fragmentation in Male Infertility and Comparisons With Other Techniques

Immunocytochemical Localisation of Phospholipid Hydroperoxide Glutathione Peroxidase in Bull’s Spermatogenic Cells

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