The Transgenerational Significance of Bisphenol‒A on Sperm Function and Fertility of CD-1 Male Mice

Md Saidur Rahman,Won-Ki Pang,Ki-Uk Kim,Ye-Ji Kim,Do-Yeal Ryu,Amena Khatun,Sung-Jae Yoon,Woo-Sung Kwon,Myung-Geol Pang 한국수정란이식학회 2016 한국수정란이식학회 학술대회 Vol.2016 No.10

Bisphenol‒A (BPA) is a known endocrine‒disrupting chemical used extensively to manufacture plastic bottles, canned food linings, thermal receipts, and other commonly used items. BPA is capable of inducing chromosomal alterations in germ cell line, thereby produced transgenerational effects on brain function, social recognition, reproductive diseases, sperm quality, gene expression, and obesity. Here, we aimed to investigate the transgenerational effects of BPA on murine male fertility. Six-week-old male mice (F0) were gavaged with corn oil (control), two different doses of BPA (5 mg, and 50 mg·kg bw-1·day-1),andethinylestradiol(EE,0.4mg·kg bw-1·day-1), dailyfor6weeks. Treated male mice were mated with wild‒type female and sibling pairs were bred up to the third generation (F3) in a similar manner with no further BPA exposure. Testes and spermatozoa were collected from 14-week-old males of all generation (F0 to F3) to evaluate testis weight, sperm function, and fertility. We found that high concentration of BPA significantly increased testicular weight in F2. Although the sperm viability, capacitation status, and intracellular ROS levels were not affected by BPA, however, sperm count, motility, hyperactivated motility, and intracellular ATP levels were significantly altered by BPA, dose dependently. In majority of the cases the effects were prominent in F2 followed by F1 and F0, whereas the effects were diminished in F3 generation. Simultaneously, high concentration of BPA significantly decreased cleavage and blastocyst formation rate in both F1 and F2. Similar inhibitory effects on cleavage and blastocyst were also noted in F1 by low dose of BPA. Depending on these findings we conclude that BPA decreases the fertility potential of exposed males and has an adverse impact on sperm function and fertility in subsequent generations.

Sodium Nitroprusside Induce Phosphorylation of Ferritin Light Chain Protein, Regulates the Acrosome Reaction and Fertility of Mice Spermatozoa

Md Saidur Rahman,Won-Ki Pang,Ye-Ji Kim,Do-Yeol Ryu,Amena Khatun,Sung-Jae Yoon,Woo-Sung Kwon,Myung-Geol Pang 한국동물생명공학회(구 한국동물번식학회) 2015 발생공학 국제심포지엄 및 학술대회 Vol.2015 No.10

Alterations of Male Fertility following Exposure to Bisphenol A

Md Saidur Rahman 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11

Bisphenol A (BPA) is a common industrial chemical that has been used extensively to make certain plastics and resins since the 1960s. As a potential endocrine disruptors, BPA has been investigated for its impact on fertility/reproduction in animals and humans. However, the molecular mechanisms of BPA action and standard method for detecting BPA-related health hazards are unclear. Considering in-vitro experimental model, we investigated the effects of BPA (0.0001 to 100 μM) exposure on mouse spermatozoa. We revealed that BPA affects several sperm functions by triggering the mitogen-activated protein kinase, phosphatidylinositol 3-kinase, and protein kinase-A (PKA) activity. High doses of this chemical was also likely for the activation of protein tyrosine phosphorylation in a PKA-dependent signaling consequently induced a precocious acrosome reaction. Simultaneously, BPA has been found to decrease the rate of fertilization and early embryonic development. In addition, BPA induced differential protein expression in spermatozoa were responsible for the pathogenesis of many diseases. Considering in vivo experimental model, we deliberate the effects of gestational BPA exposure (TDI, NOAEL, and LOAEL doses) on both ejaculated and capacitated spermatozoa in F1 adult mice. We confirmed that BPA affects several sperm function in F1 male. These effects appeared to be caused by reduced numbers of stage VIII seminiferous epithelial cells in testis and decreased PKA activity and tyrosine phosphorylation (non-capacitated) in spermatozoa. We also noticed that BPA decreased average litter size as well as compromise the rates of cleavage and blastocyst formation. Proteins differentially expressed in both capacitated/ejaculated spermatozoa play a critical role in energy metabolism, stress responses, and fertility, finally predispose to the development of several diseases. On the basis of these results, we suggest that BPA alter spermatozoa function and the proteomic profile, ultimately affecting their fertility potential. Therefore, it is of critical public health significance to reevaluate the levels of BPA exposure that are currently deemed to be acceptable.

Bisphenol-A suppresses male fertility in vitro

Md Saidur Rahman,Woo-Sung Kwon,June-Sub Lee,Ye-Ji Kim,Do-Yeal Ryu,Sung-Jae Yoon,Myung-Geol Pang 한국발생생물학회 2014 한국발생생물학회 학술발표대회 Vol.2014 No.9

The present study aims to investigate the effect of BPA on sperm functions, fertilization and to evaluate their association with the activity of fertility related proteins in spermatozoa. We used a comprehensive in vitro test system to evaluate the effect of various concentrations of BPA (0.0001, 0.01, 1, and 100 μM) on mouse spermatozoa following 6 h of incubation. Our results showed that high concentration of BPA inhibited sperm motility and motion kinematics by significantly decreasing ATP levels in spermatozoa. Simultaneously, exposure of spermatozoa to high concentrations of BPA increased the tyrosine phosphorylation of sperm proteins involved in PKA-dependent regulation and induced a robust AR, ultimately results in poor fertilization and compromised embryonic development. Finally, BPA effects on selected group of fertility related proteins in spermatozoa, such as it degraded the β-actin, whereas the levels of peroxiredoxin-5, glutathione peroxidase, glyceraldehyde-3-phosphate dehydrogenase, and succinate dehydrogenase were increased. Based on these results, we propose that high concentration of BPA may alter overall sperm functions, fertilization and embryonic development, in association with degradation and/or phosphorylation of fertility related proteins in spermatozoa.

소,쥐,되지 정자의 에스트로겐 물질에 대한 민감도

( Md Saidur Rahman ),류도열 ( Do Yeal Ryu ),김예지 ( Ye Ji Kim ),이준섭 ( June Sub Lee ),권우성 ( Woo Sung Kwon ),윤성재 ( Sung Jae Yoon ),방명걸 ( Myung Geol Pang ) 한국축산학회 2014 축산기술과 산업 Vol.5 No.3

본 연구는 에스트로켄 물질 [17 β-estradiol(E2), progesterone(P4), genistein(GEN), 4-tert-octylphenol(OP)]이포유동물 (소, 돼지, 생쥐) 정자의 수정능력 획득과 첨체반응 민감도에 미치는 영향을 체외에서 확인하기 위해 실시하였다. E2, P4, GEN, OP(0.001-100㎛)를 15분 또는 30분 동안 5% CO₂, 39℃에서 배양 후 수정능력 획득 및 첨체반응 여부를 확인하기 위하여 Hoechst 33258/Chlortetracycline 염색법을 실시하였다. E2는 15분 및 30분 처리시 생쥐 정자에서 농도에 따라 첨체반응을 유의적으로 증가시켰다(p<0.05). 돼지 정자의 경우는 15분 처리시 가장 낮은 농도(0. 001㎛)의 처리구에서만 수정능력 획득을 유의적으로 증가 시켰고, 30분 처리시 농도에 따라 첨체반응을 유의적으로 증가시켰다(p<0.05). P4는 15분 처리시 소와 생쥐의 정자에서 농도에 따라 첨체반응을 유의적으로 증가시켰다고, 돼지의 경우 농도에 따라 수정능력 획득을 유의적으로 즌가시켰다(p<0.05). 또한, 30분 동안 처리한 생쥐 정자의 농도에 따라 수정능력 획득을 유의적으로 증가시켰다(p<0.05). GEN 은 15분과 30분동안 배양한 돼지 정자와 30분 동안 배양한 생쥐정자의 쳄체반응을 농도에 따라 유의적으로 증가시켰다(p<0.05). OP의 경우 15분 처리시 생쥐정자의 첨체반응을 농도에 따라 유의적으로 증가시켰다(p<0.05). 특히 이들 물질 중 E2 와 P4에 대한 민감도 더 높아 효과적으로 정자의 기능에 영향을 주는 것으로 확인되었다. 본 연구를 종합해 보면 에스트로겐 물질은 소, 돼지, 생쥐의 정자에서 수정능력 획득과 첨체반응을 효과적으로 야기시켰으며 이들 물질에 대한 민감도 차이는 종간에 유의적 차이가 있었다. 이러한 민감도 민감도 차이를 이용하여 정자를 이용한 저농도의 내분비계교란문질 탐색이 가능할 것으로 기대된다.

Duration of Preservation Affect the Quality of Chilled Black Bengal Buck Semen

Md Gulshan Anowar Pradhan,Md Saidur Rahman,Woo-Sung Kwon,Dipendra Mishra,Md Mostofa Kamal,Mohammad Musharraf Uddin Bhuiyan,Mohammed Shamsuddin 韓國受精卵移植學會 2013 한국동물생명공학회지 Vol.28 No.2

The study focuses on the quality assessment of Black Bengal buck semen preserved at chilled condition. In this in vitro trial, collected semen from Black Bengal bucks was preserved at chilling temperature (4▲5줛) in tris-glucosecitrate yolk medium of 1:5 ratios for four days. Artificial Vagina (AV) method was utilized to collect semen from buck. General evaluation of semen includes the color, mass activity and density were measured by direct visual examination. However, computer-assisted sperm analysis (CASA) and phase contrast microscopy were used to figure out the motility (%), hyper-activated (HYP) motility (%) and number of abnormal spermatozoa (%) initially, and at every 24 h intervals. The result revealed that spermatozoa preserved at chilling temperature showed significantly (P<0.05) lower motility and HYP motility with the progression of preservation. The number of phenotypically abnormal spermatozoa significantly (P<0.05) increased following preservation. Although significant positive correlation (r=0.945; P<0.05) was existed between % motile and % HYP motile spermatozoa however, the % of morphologically abnormal spermatozoa was negatively correlated with % motile (r=긏0.997; P<0.05) and % HYP motile spermatozoa (r=긏0.946; P<0.01). Therefore, we concluded that the quality of chilled semen progressively losses its viability and doesn…t remain useable after certain period of preservation with respect to its motility and morphology.

The Transgenerational Significance of Bisphenol‒A on Sperm Function and Fertility of CD-1 Male Mice

Md Saidur Rahman,Won-Ki Pang,Ki-Uk Kim,Ye-Ji Kim,Do-Yeal Ryu,Amena Khatun,Sung-Jae Yoon,Woo-Sung Kwon,Myung-Geol Pang 한국동물생명공학회(구 한국동물번식학회) 2016 발생공학 국제심포지엄 및 학술대회 Vol.2016 No.10

Application of Ultrasonography to Investigate Postpartum Anestrus in Water Buffaloes

Md Saidur Rahman,Abu Said Shohag,Md Mostafa Kamal,Nasreen Parveen,Mohammed Shamsuddin 한국동물생명공학회(구 한국동물번식학회) 2012 Reproductive & Developmental Biology(Supplement) Vol.36 No.2s

Anestrus is one of the most important production limiting disorders in dairy buffaloes and its underlying causes have been a current topic of studies. The objectives of this study were to explore the causes of anestrus in buffaloes with the application of ultrasonography. Two examinations were performed by transrectal ultrasonography at 12 days apart in buffalo cows that were not seen in oestrus at 60 or more days postpartum. As high as 54.5% buffaloes had silent ovulation and 45.5% suffered from the true anestrus with ovarian dysfunction. The duration of anestrus after calving was 60~90, 91~120, 121~180 and 181~365 days in 27%, 32%, 18% and 23% buffalo cows, respectively. Treatment with prostaglandin of cyclic buffalo cows with a corpus luteum (72.7%) resulted in higher estrous rate as compared with close observation of estrus (23.1%) by the farmer (p=0.021). Acyclic buffalo cows without any corpus luteum on ovaries were successfully treated with gonadotropin releasing hormone (70%), resulting in higher estrous detection rate than those treated with a vitamin-mineral mixture (20%) (p=0.035). In conclusion, poor heat detection due to silent ovulation is the most important cause of apparent anoestrus in buffaloes; however the percentage of the true anestrous is also quite high in postpartum buffaloes.

Antioxidants Show a Protective Effects against Bisphenol A Induced Stress on Spermatozoa

Md Saidur Rahman,Kyu-Ho Kang,Sarder Arifuzzaman,Won-Ki Pang,Do-Yeal Ryu,Won-Hee Song,Myung-Geol Pang 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11

Bisphenol A (BPA), an endocrine-disrupting chemical, has received tremendous attention in the past few decades because of its detrimental health effects. Growing evidence supports that BPA is capable to alter the reproductive performance of the exposed individual. In spermatozoa, it has been reported that BPA increased oxidative stress by the overproduction of reactive oxygen species (ROS), subsequently affects the sperm function, biochemical properties, and fertility. Since antioxidants minimize cellular oxidative stress, therefore may have protective effects against BPA-induced stress. In the present study, we incubated mice spermatozoa for 6 h in a condition that support in vitro fertilization. The sperm incubation media was additionally supplemented with either BPA or BPA together with antioxidants, such as glutathione, vitamin C, and vitamin E. Our results showed that antioxidant significantly decreased the production of ROS that subsequently supports motility and acrosomal integrity of BPA-exposed spermatozoa. Particularly, glutathione and vitamin E inhibit protein kinase-A dependent phosphorylation of sperm proteins subsequently prevented precocious acrosome reaction. In addition, both antioxidants were found to restore fertilization and early embryo development potentiality of BPA-exposed spermatozoa. Therefore, we conclude that antioxidants minimize oxidative stress in spermatozoa in a BPA containing micro-environment, thus avoiding BPA-mediated harmful consequences. The current finding has both theoretical and clinical significance for developing potential remedies of the BPA toxicity.

Perinatal exposure of female mice to bisphenol-A affects the sperm functions and fertilization of male offspring

Md Saidur Rahman,Woo-Sung Kwon,Ye-Ji Kim,Do-Yeal Ryu,Hyun-Gu Kang,Amena Khatun,Sung-Jae Yoon,Myung-Geol Pang 한국발생생물학회 2015 한국발생생물학회 학술발표대회 Vol.2015 No.9

As an endocrine disruptor, bisphenol-A (BPA) causes several functional and behavioral abnormalities related to reproduction. The current study was design to evaluate the effect of perinatal exposure of female mice to BPA on sperm function of adult F(1) offspring. Pregnant female mice F(0) were gavaged with three different concentration of BPA, such as 50 μg/kg/day (tolerable daily intake value by the European Food Safety Authority), 5 mg/kg/day (no-observed-adverse-effect level; NOAEL), and 50 mg/kg/day (lowest-observed-adverse-effect level; LOAEL) and corn oil (7 mg/kg/day; vehicle control). The functional parameters of F(1) spermatozoa were studied both before and after capacitation, whereas the fertility assessment was evaluated by in vitro and in vivo assay using unexposed females. Our results showed that spermatozoa hyperactivated motility, capacitation, intracellular ATP, Ca2+, and ROS levels after capacitation were significantly affected using NOAEL and LOAEL concentration of BPA. However, the sperm motility was only affected by LOAEL dose after capacitation. All of the tested parameters were potentially unaffected by BPA before capacitation, except intracellular ATP that decreased by all concentrations. Although both NOAEL and LOAEL concentration were effectively reduced the rate of fertilization and embryonic development in vitro, however the average litter size was only affected by LOAEL dose. Our finding suggested that perinatal exposure of 50 μg/kg/day did not produce significant effects; however both NOAEL and LOAEL affects overall sperm function after capacitation, leading to impairments in the fertility of F(1) male offspring.