RESEARCH : Capacitation and acrosome reaction differences of bovine, mouse and porcine spermatozoa in responsiveness to estrogenic compounds

( Do Yeal Ryu ),( Ye Ji Kim ),( June Sub Lee ),( Md Saidur Rahman ),( Woo Sung Kwon ),( Sung Jae Yoon ),( Myung Geol Pang ) 한국동물자원과학회(구 한국축산학회) 2014 한국축산학회지 Vol.56 No.26

Background: Endocrine disruptors are exogenous substance, interfere with the endocrine system, and disrupt hormonal functions. However, the effect of endocrine disruptors in different species has not yet been elucidated. Therefore, we investigated the possible effects of 17ß-estradiol (E2), progesterone (P4), genistein (GEN) and 4-tert-octylphenol (OP), on capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. In this in vitro trial, spermatozoa were incubated with 0.001-100 μM of each chemical either 15 or 30 min and then assessed capacitation status using chlortetracycline staining. Results: E2 significantly increased capacitation and the acrosome reaction after 30 min, while the acrosome reaction after 15 min incubation in mouse spermatozoa. Simultaneously, capacitation and the acrosome reaction were induced after 15 and 30 min incubation in porcine spermatozoa, respectively. Capacitation was increased in porcine spermatozoa after 15 min incubation at the lowest concentration, while the acrosome reaction was increased in mouse spermatozoa after 30 min (P <0.05). E2 significantly increased the acrosome reaction in porcine spermatozoa, but only at the highest concentration examined (P <0.05). P4 significantly increased the acrosome reaction in bovine and mouse spermatozoa treated for 15 min (P <0.05). The same treatment significantly increased capacitation in porcine spermatozoa (P <0.05). P4 significantly increased capacitation in mouse spermatozoa treated for 30 min (P <0.05). GEN significantly increased the acrosome reaction in porcine spermatozoa treated for 15 and 30 min and in mouse spermatozoa treated for 30 min (P <0.05). OP significantly increased the acrosome reaction in mouse spermatozoa after 15 min (P <0.05). Besides, when spermatozoa were incubated for 30 min, capacitation and the acrosome reaction were higher than 15 min incubation in E2 or GEN. Furthermore, the responsiveness of bovine, mouse and porcine spermatozoa to each chemical differed. Conclusions: In conclusion, all chemicals studied effectively increased capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. Also we found that both E2 and P4 were more potent than environmental estrogens in altering sperm function. Porcine and mouse spermatozoa were more responsive than bovine spermatozoa.

Capacitation and acrosome reaction differences of bovine, mouse and porcine spermatozoa in responsiveness to estrogenic compounds

Ryu, Do-Yeal,Kim, Ye-Ji,Lee, June-Sub,Rahman, Md. Saidur,Kwon, Woo-Sung,Yoon, Sung-Jae,Pang, Myung-Geol Korean Society of Animal Sciences and Technology 2014 한국축산학회지 Vol.56 No.7

Background: Endocrine disruptors are exogenous substance, interfere with the endocrine system, and disrupt hormonal functions. However, the effect of endocrine disruptors in different species has not yet been elucidated. Therefore, we investigated the possible effects of $17{\beta}$-estradiol (E2), progesterone (P4), genistein (GEN) and 4-tert-octylphenol (OP), on capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. In this in vitro trial, spermatozoa were incubated with $0.001-100{\mu}M$ of each chemical either 15 or 30 min and then assessed capacitation status using chlortetracycline staining. Results: E2 significantly increased capacitation and the acrosome reaction after 30 min, while the acrosome reaction after 15 min incubation in mouse spermatozoa. Simultaneously, capacitation and the acrosome reaction were induced after 15 and 30 min incubation in porcine spermatozoa, respectively. Capacitation was increased in porcine spermatozoa after 15 min incubation at the lowest concentration, while the acrosome reaction was increased in mouse spermatozoa after 30 min (P < 0.05). E2 significantly increased the acrosome reaction in porcine spermatozoa, but only at the highest concentration examined (P < 0.05). P4 significantly increased the acrosome reaction in bovine and mouse spermatozoa treated for 15 min (P < 0.05). The same treatment significantly increased capacitation in porcine spermatozoa (P < 0.05). P4 significantly increased capacitation in mouse spermatozoa treated for 30 min (P < 0.05). GEN significantly increased the acrosome reaction in porcine spermatozoa treated for 15 and 30 min and in mouse spermatozoa treated for 30 min (P < 0.05). OP significantly increased the acrosome reaction in mouse spermatozoa after 15 min (P < 0.05). Besides, when spermatozoa were incubated for 30 min, capacitation and the acrosome reaction were higher than 15 min incubation in E2 or GEN. Furthermore, the responsiveness of bovine, mouse and porcine spermatozoa to each chemical differed. Conclusions: In conclusion, all chemicals studied effectively increased capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. Also we found that both E2 and P4 were more potent than environmental estrogens in altering sperm function. Porcine and mouse spermatozoa were more responsive than bovine spermatozoa.

The Effect of Conoidin A on Spermatozoa

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

Freezability biomarkers in epididymal spermatozoa

Do-Yeal Ryu,Won-HeeSong,Won-KiPang,Sung-JaeYoon,Md Saidur Rahman,Myung-Geol Pang 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11

Sperm cryopreservation is well known as a valuable method to preserve the genetic traits. Although many studies have established semen cryopreservation protocols, lack of studies were conducted to discover the differences of sperm proteome and functions between ejaculated and epididymal spermatozoa following to cryopreservation. Therefore, the objective of this study was to (i) evaluate the effect of cryopreservation on bull epididymal spermatozoa and (ii) discover the potential biomarkers, which have highly tolerance to freezing on bull epididymal spermatozoa. Our preliminary study demonstrated that spermatozoa from each bulls have different resistance on freezing during cryopreservation. We divided spermatozoa into two groups according to sperm motility following to cryopreservation; high freezing-tolerant spermatozoa (HFS) and low freezing-tolerant spermatozoa (LFS). Several sperm functional parameters, i.e. sperm motility/motion kinematics, speed parameters, viability, mitochondrial membrane potential, and capacitation status. Our results showed that all parameters except for motion kinematics and capacitation status had significant differences between HFS and LFS. Subsequently, two dimensional electrophoresis were conducted to compare the expression levels of sperm proteome between both groups. Three proteins {glutathione s-transferase mu 5 (GSTM5), voltage-dependent anion-selective channel protein 2 (VDAC2), and ATP stynthase subunit beta (ATP1B1)} were differentially expressed. Based on these results, we propose that epididymal spermatozoa from individual bull have different freezability upon cryopreservation and three differentially expressed proteins might be selected as a biomarker to predict high freezing-tolerant epididymal spermatozoa.

Determination of Highly Sensitive Biological Cell Model Systems to Screen BPA-Related Health Hazards Using Pathway Studio

Ryu, Do-Yeal,Rahman, Md Saidur,Pang, Myung-Geol MDPI 2017 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.18 No.9

<P>Bisphenol-A (BPA) is a ubiquitous endocrine-disrupting chemical. Recently, many issues have arisen surrounding the disease pathogenesis of BPA. Therefore, several studies have been conducted to investigate the proteomic biomarkers of BPA that are associated with disease processes. However, studies on identifying highly sensitive biological cell model systems in determining BPA health risk are lacking. Here, we determined suitable cell model systems and potential biomarkers for predicting BPA-mediated disease using the bioinformatics tool Pathway Studio. We compiled known BPA-mediated diseases in humans, which were categorized into five major types. Subsequently, we investigated the differentially expressed proteins following BPA exposure in several cell types, and analyzed the efficacy of altered proteins to investigate their associations with BPA-mediated diseases. Our results demonstrated that colon cancer cells (SW480), mammary gland, and Sertoli cells were highly sensitive biological model systems, because of the efficacy of predicting the majority of BPA-mediated diseases. We selected glucose-6-phosphate dehydrogenase (G6PD), cytochrome b-c1 complex subunit 1 (UQCRC1), and voltage-dependent anion-selective channel protein 2 (VDAC2) as highly sensitive biomarkers to predict BPA-mediated diseases. Furthermore, we summarized proteomic studies in spermatozoa following BPA exposure, which have recently been considered as another suitable cell type for predicting BPA-mediated diseases.</P>

The Effects of Peroxiredoxins on Sperm Function and Male Fertility

Do-Yeal Ryu,Woo-Sung Kwon,Md Saidur Rahman,Amena Khatun,Myung-Geol Pang 한국동물생명공학회(구 한국동물번식학회) 2017 발생공학 국제심포지엄 및 학술대회 Vol.2017 No.10

Peroxiredoxins (PRDXs), which are essential antioxidant enzymes for physiological activity, have reported that influence sperm function and consequently male fertility. Although PRDXs are well known as important factor to affect male fertility, there has been a lack of attempt to discover the accurate role and mechanism of PRDXs in male fertility. Therefore, the principal objective of this study is to elucidate the role of PRDXs in sperm function and fertilization. We treated mouse spermatozoa with different dose of specific inhibitor of PRDXs, conoidin A (1, 10, or 100 μM). Our results revealed that conoidin A significantly decreased oxidized form of PRDXs (PRDXs-SO3) in mouse spermatozoa. Inhibited PRDXs activity are then resulted in a significant decrease in sperm motility/motion kinematics, viability, mitochondrial membrane potential, and intracellular ATP levels. On the other hand, intracellular levels of ROS and DNA fragmentation index were significantly increased following exposure to conodin A. Next, we evaluated capacitation and acrosome reaction status, and subsequently tyrosine phosphorylation and protein kinase-A activity to investigated underlying mechanism of PRDXs on sperm capacitation status. Capacitation and acrosome reaction were significantly decreased perhaps due to reduction of tyrosine phosphorylation and protein kinase- A activity. Finally, we investigated the effect of PRDXs on fertilization and early embryonic development. Our results described that decreased PRDXs activity significantly decreased fertilization and early embryonic development. Consequently, we demonstrated that inhibition of PRDXs activity has a direct impact on male fertility via decreased important physiological sperm function, eventually resulted in poor fertilisation and embryonic development.

脊髓 感覺神經節 細胞에 대한 重金屬 및 酸素遊離氣의 細胞損傷에 미치는 韓藥劑의 神經細胞損傷 回復에 미치는 影響에 對한 硏究

柳道坤,李廷憲,成疆慶,李建穆,辛民敎,鄭遇悅,李星根,洪起年,尹向錫,李康昌,宋昊俊,禹元洪,朴承澤,田炳薰 대한동의병리학회 2000 동의생리병리학회지 Vol.14 No.1

중금속의 독성을 산화적 손상 측면에서 규명하며 동시에 중금속의 산화적 손상에 의한 한약추출물의 효과를 항산화적측면에서 조사하기 위하여 생쥐의 척수 감각신경절세포에 여러 농도의 FeSO₄를 처리한 후 이의 독성효과를 조사하였다. FeSO₄는 생쥐의 배양 척수 감각신경질세포에 처리한 농도와 시간에 비례하여 세포생존율을 유의하게 감소시켰으며, 15μM FeSO₄의 농도에서 척수 감각신경절세포를 4시간 동안 배양한 결과 MTT50 과 NR50 값을 나타냈으며 이는 고독성인 것으로 나타났다. [고독성; 100μM> MTT50, NR50]. 한편, 한약추출물인 원두충(EU)과 음양곽(EK) 및 감초(GC)는 FeSO₄에 의해서 유도된 신경독성에 대하여 세포생존율을 현저히 증가시킴으로서 유의한 방어효과를 보였다. 이상에서 FeSO₄는 생쥐의 배양 척수 감각신경절세포에 고독성으로 나타났으며, 또한 음양곽이나 익지인 및 감초와 같은 선택적인 한약추출물이 FeSO₄에 의해서 유도된 신경독성을 효과적으로 방어하였다. To clarify the mechanism between neurotoxity of metal and oxidative stress, toxic effect of FeSO₄was examined by MTT assay and NR assay in cultured spinal dorsal root ganglion(DRG) neurons from mouse. And also neuroprotective effects of antioxidant and several herb extracts on FeSO₄decreased cell viability in dose and time dependentry in cultured mouse spinal DRG neurons. MTT50 and NR50 values were determined at 15μM FeSO₄for 4 hours in these cultures, and this value was heighly toxic; MTT50, NR50 < 100μM). Glutathione markedly increasd cell viability after preincubation of DRG neros for 2 hours in the media containing cocentrations of 2-6mM glutathione. In neuroprotective of herb extracts, Eucommia lmoides Oliver(EU), Epimedium koreanum Nakai(EK) and Glycyrrhiza uralensis Fisch(GC) were very effective in blocking the neurotoxicity induced by FeSO₄in cultured mouse spinal DRG neurons. These results suggested that FeSO₄was heighly toxic in cultured mouse spinal DRG neurons, and selective herb extracts such as Eucommia ulmoides Oliver(EU), Epimedium koreanum Nakai(EK) and Glycyrrhiza uralensis Fisch(GC) were effective in blocking the FeSO₄-induced neurotoxicity in cultured mouse spinal DRG neurons.

부신기능 저하증을 동반한 전이성 부신암 1례

도주호,류성열,황준영,정재진,이동욱,이상준,권기영,이인규 계명대학교 의과학연구소 2000 계명의대학술지 Vol.19 No.2

Adrenal insufficiency is caused by destruction of the adrenal cortex, deficient pituitary ACTH secretion, or deficient hypothalamic secretion of corticotropin releasing hormone. The prominent symptoms are weakness, fatigue, weight loss, and gastrointestinal complaints, but these symptoms are common to many other diseases. Thus, the diagnosis of adrenal insufficiency needs the exception of other diseases. Primary adernal insufficiency comes from autoimmune disease, granulomatous disease, metastatic malignancies such as lung and breast carcinoma, hemorrhagic infarction associated with anticoagulant therapy or meningococcemia, and rare hereditary diseases. In this paper, we present a case of lung cancer with adrenal metastasis which causes adrenal insufficiency. The patient was treated with chemotherapy and steroid replacement therapy successfully.

津液의 調節過程에 對한 硏究

柳道坤(Ryu Do Gon),鄭遇悅(Jeong Woo Yeal) 대한한의학회 1989 대한한의학회지 Vol.10 No.1

NT5C1B and FH are good markers to predict cryoprotectant tolerance in spermatozoa

Won-HeeSong,Do-Yeal Ryu,Won-KiPang,Sung-JaeYoon,Md Saidur Rahman,Myung-Geol Pang 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11

Sperm cryopreservation preserves genetic resources for animal breeding and for human patients who suffers from permanent testicular damage. Although the sperm cryopreservation has been used for many years, the addition of cryoprotective agent (CPA) during cryopreservation negatively affects sperm function and quality. Our previous study reported that the addition of CPA reduced bull sperm physiological functions. However, the sperm cells collected from individual bulls presented different sensitivity to the damage induced by CPA. In the present study, we examined if CPA affect sperm cells acquired from individual bulls. Individual bull spermatozoa were divided into two groups based on motility parameters; high CPA-tolerant sperm (HCS) and low CPA-tolerant sperm (LCS). Our results showed that the HCS group presented good physiological function after CPA exposure, whereas the LCS group showed a significant decrease in the sperm function. We also found differentially expressed five proteins between the HCS and LCS groups, which refer to cytosolic 5′-nucleotidase 1B (NT5C1B), fumarate hydratase (FH), F-actin-capping protein subunit beta (CAPZB), voltage-dependent anion-selective channel protein 2 (VDAC2), and cytochrome b-c1 complex subunit 1 (UQCRC1). NT5C1B and FH showed abundant expression in the HCS group, while the expression of CAPZB, VDAC2, and UQCRC1 was relatively lower in the HCS group than in the LCS group. The current results suggest that NT5C1B, FH, CAPZB, VDAC2, and UQCRC1 can be used as potential markers to predict CPA-tolerable spermatozoa. Those markers provide a reliable tool to select animals and breeds with CPA tolerance.