Toll-like receptor 4 (TLR4) is known to contribute to the modulation of insulin resistance and systemic inflammation seen in obesity and the metabolic syndrome. The present study was performed to investigate the fertility competence of TLR4 knock out male mice (TLR4 mice) on a high-fat diet (HFD), compared to a normal-chow diet (NCD). The controls included wildtype (WT) mice fed on a HFD or NCD. Six-week-old male mice were fed with either a NCD or HFD for 20 weeks. Body and organ weights, serum levels of glucose, triglycerides and hepatoxicity, sperm quality and spermatogenesis were observed after the sacrifice. Also, randomly selected male mice were mated with virgin female mice after feeding of 19 weeks. The weight of the body and organs increased in WT and TLR4 mice on a HFD compared to those of mice on a NCD. The weights of the reproductive organs did not vary among the treatment groups. The motility and concentration of the epididymal spermatozoa decreased in both WT and TLR4 mice fed a HFD. The pregnancy rate and litter size declined in the HFD-fed WT mice compared to the HFD-fed TLR4 mice. In conclusion, the HFD alters energy and steroid metabolism in mice, which may lead to male reproductive disorders. However, fertility competence was somewhat restored in HFD-fed TLR4 male mice, suggesting that the TLR4 is involved in testis dysfunction due to metabolic imbalance.

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 (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.

Iron is required for cell viability but is toxic in excess. While the iron-mediated malfunction of testicular cells is well appreciated, the underlying mechanism(s) of this effect and its relationship with fertility are poorly understood. Ferritin is a ubiquitous intracellular protein that controls iron storage, ferroxidase activity, immune response, and stress response in cells. Ferritin light chain protein (FTL) is the light subunit of the Ferritin. Previously, we had identified the FTL in bovine spermatozoa following capacitation. In present study, to investigate the role of Ferritin in sperm function, mice spermatozoa were incubated with multiple doses (1, 10 and 100 μM) of sodium nitroprusside (SNP), an iron donor. SNP was increased Ferritin levels in a dose-dependent manner. The Ferritin was detected on the acrosome in spermatozoa by immunocytochemistry. Short-term exposure of spermatozoa to SNP increased tyrosine phosphorylation and the acrosome reaction (AR). Finally, SNP affected a significant decrease in the rate of fertilization as well as blastocyst formation during early embryonic development. On the basis of these results, we propose that the effects of Ferritin on the AR may reduce overall sperm function leads to poor fertility in males and compromised embryonic development.

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.

Hybrid sterility is one of the major barrier to the application of wide crosses in plant breeding and is commonly encountered in crosses between indica and japonica rice varieties. Ten mapping populations comprised of two reciprocal F2 and eight BC1F1 populations generated from the cross between Ilpumbyeo (japonica) and Dasanbyeo (indica) were used to identify QTLs and to interpret the gametophytic factors involved in hybrid fertility or sterility between two subspecies. Frame maps were constructed using a total of 107 and 144 STS markers covering 12 rice chromosomes in two reciprocal F2 and eight BC1F1 populations, respectively. A total of 15 main-effect QTLs and 17 significant digenic- epistatic interactions controlling spikelet fertility (SF) were resolved in the the entire genome map of F2 BC1F1 populations . Among detected QTLs responsible for hybrid ferility, four QTLs, qSF5.1 and and qS F5.2 on chromosome 5, qSF6.2 on chromosome 6, and qSF12.2 on chromosome 12 were identified as major QTLs since they were located at corresponding position in at least three mapping populations. Loci qSF5.1, qSF6.1 and qSF6.2 were responsible for both female and male sterility, whereas qSF3.1, qSF7 and qSF 12.2 affected the spikelet fertility only through embryosac factors, and qSF9.1 did through pollen factors. Five new QTLs identified in this study will be helpful for understanding the hybrid sterility and for breeding programs via inter-subspecific hybridization.

Hybrid sterility is one of the major barrier to the application of wide crosses in plant breeding and is commonly encountered in crosses between indica and japonica rice varieties. Ten mapping populations comprised of two reciprocal F2 and eight BC1F1 populations generated from the cross between Ilpumbyeo (japonica) and Dasanbyeo (indica) were used to identify QTLs and to interpret the gametophytic factors involved in hybrid fertility or sterility between two subspecies. Frame maps were constructed using a total of 107 and 144 STS markers covering 12 rice chromosomes in two reciprocal F2 and eight BC1F1 populations, respectively. A total of 15 main-effect QTLs and 17 significant digenic- epistatic interactions controlling spikelet fertility (SF) were resolved in the the entire genome map of F2 BC1F1 populations . Among detected QTLs responsible for hybrid ferility, four QTLs, qSF5.1 and and qS F5.2 on chromosome 5, qSF6.2 on chromosome 6, and qSF12.2 on chromosome 12 were identified as major QTLs since they were located at corresponding position in at least three mapping populations. Loci qSF5.1, qSF6.1 and qSF6.2 were responsible for both female and male sterility, whereas qSF3.1, qSF7 and qSF 12.2 affected the spikelet fertility only through embryosac factors, and qSF9.1 did through pollen factors. Five new QTLs identified in this study will be helpful for understanding the hybrid sterility and for breeding programs via inter-subspecific hybridization.

Cytoplasmic male sterility caused by DCGMS (Dongbu cytoplasmic and genic male-sterility) cytoplasm and its nuclear restorer-of-fertility locus (Rfd1) with a linked molecular marker (A137) have been reported in radish (Raphanus sativus L.). To construct a linkage map of the Rfd1 locus, linked amplified fragment length polymorphism (AFLP) markers were screened using bulked segregant analysis. A 220-bp linked AFLP fragment sequence from radish showed homology with an Arabidopsis coding sequence. Using this Arabidopsis gene sequence, a simple PCR marker (A220) was developed. The A137 and A220 markers flanked the Rfd1 locus. Two homologous Arabidopsis genes with both marker sequences were positioned on Arabidopsis chromosome 3 with an interval of 2.4 Mb. To integrate the Rfd1 locus into a previously reported expressed sequence tag (EST)-simple sequence repeat (SSR) linkage map, the radish EST sequences located in three syntenic blocks within the 2.4-Mb interval were used to develop single nucleotide polymorphism (SNP) markers for tagging each block. The SNP marker in linkage group 2 co-segregated with male fertility in an F2 population. Using radish ESTs positioned in linkage group 2, five intron length polymorphism (ILP) markers and one cleaved amplified polymorphic sequence (CAPS) marker were developed and used to construct a linkage map of the Rfd1 locus. Two closely-linked markers delimited the Rfd1 locus within a 985-kb interval of Arabidopsis chromosome 3. Synteny between the radish and Arabidopsis genomes in the 985-kbp interval were used to develop three ILP and three CAPS markers. Two ILP markers further delimited the Rfd1 locus to a 220-kb interval of Arabidopsis chromosome 3.

스테로이드 호르몬인 에스트로겐은 여러 조직의 발생 및 분화에 영향을 주는 것으로 알려져 있다. 최근 에스트로겐의 영향을 평가하는데 있어 전통적인 여성 호르몬이라는 인식을 탈피하여 수컷 생식계상의 역할에 관한 연구가 활발하게 이루어지고 있다. 특히 내분비계 교란물질에 관한 다양한 연구 결과가 보고되면서, 수컷에서의 에스트로겐 역할에 대한 관심이 증대되고 있는 실정이다. 따라서 에스트로겐의 정확한 역할을 파악하기 위해서 발생 단계에 따른 에스트로겐 수용체의

유채의 F1 수량이 획기적으로 증수되고 있으나 유질과 유박의 성분이 개량된 F1 육성 보급하는 것이 가장 바람직한 일이라 하겠다. 필자 등은 유질과 유박의 성분이 완전 개량된 세포질 유전자적 웅성불임 계통을 개발한 바 있다. 그러나 화분친으로 이용할 수 있는 성분개량 품종수는 극히 적다. 따라서 국내 육성계통중 성분이 개량된 O-erucic acid, Low-glucosinolate 계통을 화분친으로 이용하는 문제를 검토코저 511육성 계통을 공시하여 F1 의 임성회복력과 F1 의 유용형질의 Heterosis 발현에 대해 조사하였다. 1. MS를 이용한 F1 임성회복력에서 공시 화분친중 81%의 계통들이 완전 임성회복 유전자를 가지고 있었으며 부분 임성회복 품종이 5%, 비회복 계통이 14%에 불와해서 국내 육성계통들의 임성회복능력이 매우 높은 것으로 밝혀졌다. 2. 비회복 육성계통중 11계통은 Fertility index가 1～3으로 웅성불임 계통의 불임성을 유지하는 능력을 가지고 있어 성분개량 유지계통으로 다양하게 활용할 수 있게 됐다. 3. 이들 MS의 Maintainer는 대부분 핵내 유전자가 Sterile인 Isuzu, Bronowski, 유달, Chisaga에 유래하고 있었다. 4. F1 의 Heterosis 발현에서 개화기와 초장은 대부분 중간친보다 늦어지고 긴 경향이며 평균 중간친보다 4일 늦었으며 초장은 45cm나 길었다. 5. 수량구성 형질들은 협장과 1협결실립수만이 중간친보다 짧거나 같은 외는 모든 형질들이 월등히 많고 긴 방향으로 발현하였다. 6. 특히 분지수와 수장 등이 크게 F1 Heterosis를 발현하였을 뿐만 아니라 1 수협수에서도 중간친보다 8개나 더 많았다. 7. 국내 육성계통을 화분친으로 공시한 F1 의 10a당 수량성은 511조합의 F1 중 91%가 다수성 강세를 발현하는데 그 중에서 10a당 450kg이상 560kg나 증수는 조합만도 11조합이 있었다.