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Evidence of Alu and B1 expression in dbEST. dbEST中Alu和B1表达的证据。
Pub Date : 2007-07-01 DOI: 10.1080/01485010701426422
Boris Umylny, Gernot Presting, W Steven Ward

Alus and B1s are short interspersed repeat elements (SINEs) derived from the 7SL RNA gene. Alus and B1s exist in the cytoplasm as non-coding RNA indicating that they are actively transcribed, but their function, if any, is unknown. Transcription of individual SINEs is a prerequisite for retroposition, but it is also possible that individual Alu and B1 elements have some cellular functions. Previous studies suggest that transcription of Alu elements depends on the presence of an RNA polymerase-III bipartite promoter and the poly-A tail. Sequencing of small RNAs has demonstrated that the members of the Y and S subfamily are expressed. We analyzed almost one million Alu sequences longer than 200 nucleotides for the presence of RNA polymerase-III bipartite promoter sequences. More than half contained a promoter indicating some potential for expression. We searched 7.7 million human EST sequences in dbEST for the presence of Alu non-coding RNAs and found evidence for the expression of 452. Analysis of mouse spermatogenic dbEST libraries revealed an apparent relationship between the level of differentiation and the level of B1-related sequences in the EST library.

Alus和B1s是源自7SL RNA基因的短穿插重复序列(short interspersed repeat element,简称SINEs)。Alus和B1s以非编码RNA的形式存在于细胞质中,表明它们是积极转录的,但它们的功能(如果有的话)尚不清楚。单个SINEs的转录是逆转录的先决条件,但也可能单个Alu和B1元件具有某些细胞功能。先前的研究表明,Alu元件的转录依赖于RNA聚合酶- iii双部启动子和poly-A尾部的存在。小rna的测序表明,Y和S亚家族的成员表达。我们分析了近100万个长度超过200个核苷酸的Alu序列,以寻找RNA聚合酶- iii双部启动子序列的存在。一半以上的基因含有一个启动子,表明有表达的潜力。我们在dbEST中搜索了770万个人类EST序列,寻找Alu非编码rna的存在,发现了452个表达的证据。对小鼠生精dbEST文库的分析表明,EST文库中b1相关序列的水平与小鼠的分化水平有明显的关系。
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引用次数: 9
Quantitative assessment of transition proteins 1, 2 spermatid-specific linker histone H1-like protein transcripts in spermatozoa from normozoospermic and asthenozoospermic men. 正常精子和弱精子男性精子中过渡蛋白1,2精子特异性连接蛋白h1样蛋白转录物的定量评估。
Pub Date : 2007-07-01 DOI: 10.1080/01485010701426430
Piotr Jedrzejczak, Bartosz Kempisty, Artur Bryja, M Mostowska, Magdalena Depa-Martynow, Leszek Pawelczyk, Pawel Piotr Jagodzinski

Spermatid-specific linker histone H1-like protein (HILS1), transition proteins 1 and 2 (TNP1 and TNP2), and protamines 1 and 2 (PRM1 and PRM2) contribute to considerable dense packing of spermatid chromatin during spermiogenesis. We evaluated the HILS1, TNP1, and TNP2 transcript levels in spermatozoa isolated from normozoospermic and asthenozoospermic men. Human ejaculates from normozoospermic (n = 70) and asthenozoospermic (n = 100) donors were purified by centrifugation through a discontinuous Percoll density gradient. RNA was isolated from spermatozoa according to the Chomczynski and Sacchi method, treated with DNase I and reverse-transcribed into cDNA. Quantitative analysis of HILS1, TNP1, and TNP2 transcripts was performed by real-time quantitative (RQ-PCR) SYBR green I analysis. We found significantly lower levels of HILS1, TNP1, and TNP2 transcripts in spermatozoa from asthenozoospermic men compared to normozoospermic men. Our observations suggest that a reduction in HILS1, TNP1, and TNP2 transcripts may be associated with asthenozoospermia.

精细胞特异性连接蛋白h1样蛋白(HILS1)、过渡蛋白1和2 (TNP1和TNP2)以及精蛋白1和2 (PRM1和PRM2)在精子发生过程中有助于精细胞染色质的密集堆积。我们评估了正常精子和弱精子男性分离的精子中HILS1、TNP1和TNP2转录物的水平。正常精子(n = 70)和弱精子(n = 100)供体的人类射精通过不连续Percoll密度梯度离心纯化。按照Chomczynski和Sacchi方法从精子中分离RNA,用DNase I处理后反转录为cDNA。通过实时定量(RQ-PCR) SYBR green I分析对HILS1、TNP1和TNP2转录本进行定量分析。我们发现,与正常精子男性相比,弱精子男性精子中HILS1、TNP1和TNP2转录物的水平显著降低。我们的观察表明,HILS1、TNP1和TNP2转录物的减少可能与弱精子症有关。
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引用次数: 42
High resolution light microscopic evaluation of boar semen quality sperm cytoplasmic droplet retention in relationship with boar fertility parameters. 猪精液质量的高分辨光镜评价:精子细胞质液滴保留与猪生育参数的关系。
Pub Date : 2007-07-01 DOI: 10.1080/01485010701426463
K W Lovercamp, T J Safranski, K A Fischer, G Manandhar, M Sutovsky, W Herring, P Sutovsky

The purpose of this study was to investigate the relationship between fertility and quantitative measures of boar semen quality, including various patterns of sperm cytoplasmic droplet (CD) retention, as determined by high power differential interference contrast (DIC) microscopy. A total of 116 ejaculates were collected from a nucleus herd of 18 Large White boars over an eight month period. Semen quality parameters were analyzed for each ejaculate by calculating the percentage of normal spermatozoa, spermatozoa possessing a CD in the proximal, distal, or distal midpiece reflex position, total spermatozoa with an attached cytoplasmic droplet, spermatozoa with non-CD related aberrations and total spermatozoa with abnormalities. Of the 116 ejaculates received, 71 ejaculates from 13 boars had corresponding fertility data from single-sire inseminations of multiparous sows. The fertility data included farrowing rate (FR) and total number born (TNB). The monthly FR encompassed one month before and one month after the date of semen collection. Detection of differences for fertility and semen quality parameters was performed by separating the boars into either an above-average or below-average group based on the mean FR (74.01 +/- 1.43%) or TNB (12.34 +/- 0.17) for the study. For FR, the boars in the below-average group had a significantly lower percentage of normal spermatozoa and significantly higher percentage of spermatozoa possessing distal CDs, total attached CDs and total abnormalities compared to the boars in the above-average group. Conversely, for TNB there were no significant differences between the above- and below-average groups for the semen quality parameters. These data suggest that the attached CD may negatively affect FR, but not TNB. The detection of relationships between the boar fertility parameters and the retention of the sperm CD after ejaculation, document the advantage of high power DIC microscopy in conventional semen evaluation.

本研究的目的是探讨生育能力与公猪精液质量的定量指标之间的关系,包括高倍率差示干涉对比显微镜(DIC)测定的精子细胞质液滴(CD)保留的各种模式。在8个月的时间里,从18头大白公猪的核心群中收集了116次射精。通过计算正常精子的百分比、近端、远端或远端正中反射位置具有CD的精子的百分比、附着细胞质液滴的总精子的百分比、非CD相关畸变的精子的百分比和异常的总精子的百分比来分析每次射精的精液质量参数。在收到的116次射精中,来自13头公猪的71次射精具有相应的多产母猪单父系授精的生育数据。生育数据包括产仔率(FR)和总出生数(TNB)。月FR包括采精前1个月和采精后1个月。根据平均繁殖率(74.01 +/- 1.43%)和总产仔率(12.34 +/- 0.17),将公猪分为高于平均水平组和低于平均水平组,检测育性和精液质量参数的差异。对于FR,低于平均水平组公猪的正常精子比例显著低于平均水平组,而具有远端cd、总附着cd和总异常的精子比例显著高于平均水平组。相反,对于TNB,精液质量参数在平均水平以上和低于平均水平的组之间没有显著差异。这些数据表明,附加的CD可能对FR产生负面影响,但对TNB没有负面影响。对公猪生育参数与射精后精子CD保留之间关系的检测,证明了高倍率DIC显微镜在常规精液评估中的优势。
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引用次数: 25
Clinical and radiological evaluation of ejaculatory duct obstruction. 射精管梗阻的临床及影像学评价。
Pub Date : 2007-07-01 DOI: 10.1080/01485010701426448
Mehmet Ruhi Onur, Irfan Orhan, Yrfan Orhan, Fatih Firdolas, Fatih Fýrdolaş, Rahmi Onur, Ercan Kocakoç

Ejaculatory duct obstruction (EDO) is a rare but surgically correctable cause of male infertility. With the advent and increased use of transrectal ultrasonography and magnetic resonance imaging, abnormalities of the ejaculatory duct (ED) related to infertility have been diagnosed more frequently. Recently, with the increased awareness of functional obstruction of ED, reports have been focusing on the diagnosis of partial or functional EDO. We present 2 review of the ED pathologies, imaging modalities and treatment options.

射精管梗阻(EDO)是一种罕见但可手术矫正的男性不育原因。随着经直肠超声检查和磁共振成像的出现和使用的增加,与不孕症相关的射精管异常(ED)被诊断得越来越频繁。近年来,随着人们对ED功能性梗阻的认识不断提高,部分性或功能性EDO的诊断也越来越受到关注。我们对ED的病理、成像方式和治疗方案进行了综述。
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引用次数: 12
Association between age and epididymal and accessory sex gland function and their relation to sperm motility. 年龄与附睾和副性腺功能的关系及其与精子活力的关系。
Pub Date : 2007-05-01 DOI: 10.1080/01485010701225667
Saad Elzanaty

Increased male age has been associated with significant reduction in pregnancy rates. This study investigated the association between age, the function of epididymal and accessory sex glands, and their relation to sperm motility. Ejaculates from 498 men assessed for infertility were analysed according to WHO [1999] guidelines. Seminal markers of epididymal (neutral alpha-glucosidase (NAG)), prostatic (prostate-specific antigen (PSA) and zinc), and seminal vesicle function (fructose) were measured. Four groups according to age were defined: G(21-30) (21-30 years), G(31-40) (31-40 years), G(41-50) (41-50 years), and G(>50) (51-66 years). Percentage progressive motility was significantly lower in G(>50) compared with G(21-30). NAG, PSA, zinc, and fructose were significantly lower in G(>50) compared with G(21-30). In a multiple regression analysis model, NAG and PSA showed positive significant association with percentage progressive motility. The opposite trend was found regarding zinc. No association between fructose and percentage progressive motility was shown. In this cross-sectional study, declined sperm motility observed in men over 50 years of age might be due to age-dependent changes in epididymal and accessory sex gland function.

男性年龄的增加与怀孕率的显著降低有关。本研究探讨了年龄、附睾和副性腺功能及其与精子活力的关系。根据世界卫生组织[1999]的指导方针,对498名男性不孕症患者的射精进行了分析。测定附睾(中性α -葡萄糖苷酶(NAG))、前列腺(前列腺特异性抗原(PSA)和锌)和精囊功能(果糖)的精液标志物。按年龄分为G(21 ~ 30岁)(21 ~ 30岁)、G(31 ~ 40岁)(31 ~ 40岁)、G(41 ~ 50岁)(41 ~ 50岁)、G(>50岁)(51 ~ 66岁)4组。与G(21-30)相比,G(>50)的进行性运动百分率显著降低。G组NAG、PSA、锌和果糖显著低于G组(>50)(21-30)。在多元回归分析模型中,NAG和PSA与进行性运动百分比呈显著正相关。锌的情况正好相反。果糖和进行性运动百分比之间没有关联。在这项横断面研究中,在50岁以上的男性中观察到精子活力下降可能是由于附睾和副性腺功能的年龄依赖性变化。
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引用次数: 35
Pre-pubertal di(2-ethylhexyl) phthalate (DEHP) exposure of young boars did not affect sperm in vitro penetration capacity of homologous oocytes post-puberty. 公猪青春期前接触邻苯二甲酸二(2-乙基己基)酯(DEHP)不影响青春期后同种卵母细胞的精子体外穿透能力。
Pub Date : 2007-05-01 DOI: 10.1080/01485010701271778
Linda Spjuth, M A Gil, I Caballero, C Cuello, C Almiñana, E A Martínez, N Lundeheim, H Rodríguez-Martínez

Di(2-ethylhexyl) phthalate (DEHP), a plastic softener used in polyvinylchloride (PVC) products (e.g., plastic bags and medical equipment), has been reported to have toxic effects on animal reproduction and is considered an environmental hazard based, mostly, on rodent studies. However, the doses used in these studies are often considerably higher than that presumed in human exposure. In the present study we used young boars as model animals to assess the effects of pre-pubertal DEHP exposure on the ability of spermatozoa to penetrate homologous oocytes in vitro. Eight pairs of cross-bred male boar siblings were used. One brother in each pair became, at random, the test animal exposed to DEHP per os, three times a week, from 3 to 7 weeks of age while the other acted as the control, i.e., placebo-exposed. Semen was collected and frozen between 8 and 9 months of age and stored until spermatozoa were evaluated for their ability to in vitro penetrate in vitro-matured homologous oocytes post-thaw. Both the penetration rate and the number of spermatozoa per oocyte were considered within expected ranges for frozen boar semen of good quality. Penetration rate did not significantly differ (p > 0.05) between the groups with DEHP-exposed: 50%; control: 59%, which could be owing to a large variation between boars, and between replicates. The number of spermatozoa in the ooplasm was low and similar (p > 0.05) between the groups with DEHP-exposed: 1.5 and the control: 1.7. Under the conditions of the present experiment, pre-pubertal exposure to DEHP does not seem to cause a deleterious effect on the in vitro fertilizing ability of frozen spermatozoa post-puberty.

邻苯二甲酸二(2-乙基己基)酯(DEHP)是一种用于聚氯乙烯(PVC)产品(如塑料袋和医疗设备)的塑料软化剂,据报道对动物生殖有毒性作用,主要基于对啮齿动物的研究,被认为是一种环境危害。然而,这些研究中使用的剂量往往大大高于人体暴露的假定剂量。在本研究中,我们以公猪为模型动物,评估青春期前DEHP暴露对体外精子穿透同源卵母细胞能力的影响。使用了8对杂交公猪兄弟姐妹。从3周龄到7周龄,每对中的一个兄弟随机地成为每周三次暴露于DEHP的测试动物,而另一个作为对照,即暴露于安慰剂。收集精液并在8 - 9个月大时冷冻保存,直到解冻后评估精子在体外穿透体外成熟的同源卵母细胞的能力。每卵母细胞的穿透率和精子数量都在高质量冷冻猪精液的预期范围内。dehp暴露组间刺入率差异无统计学意义(p > 0.05): 50%;对照:59%,这可能是由于公猪之间和重复之间的差异很大。dehp暴露组和对照组的卵浆中精子数分别为1.5和1.7,差异无统计学意义(p > 0.05)。在本实验条件下,青春期前接触DEHP似乎不会对青春期后冷冻精子的体外受精能力造成有害影响。
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引用次数: 7
Current concepts of molecular events during bovine and porcine spermatozoa capacitation. 牛和猪精子获能过程中分子事件的最新概念。
Pub Date : 2007-05-01 DOI: 10.1080/01485010701329386
Melissa L Vadnais, Hannah L Galantino-Homer, Gary C Althouse

Spermatozoa are required to undergo the processes of capacitation before they obtain fertilizing ability. The molecular changes of capacitation are still not fully understood. However, it is accepted that capacitation is a sequential process involving numerous physiological changes including destabilization of the plasma membrane, alterations of intracellular ion concentrations and membrane potential, and protein phosphorylation. There are no known morphological changes that occur to the spermatozoon during capacitation. The purpose of this review is to summarize current evidence on the molecular aspects of capacitation both in vivo and in vitro in bovine and porcine spermatozoa. For the purpose of this review, the process of sperm capacitation will encompass maturational events that occur following ejaculation up to binding to the zona pellucida, that triggers acrosomal exocytosis and initiates fertilization.

精子在获得受精能力之前需要经历获能过程。电容化的分子变化仍未完全了解。然而,人们普遍认为,获能是一个连续的过程,涉及许多生理变化,包括质膜的不稳定、细胞内离子浓度和膜电位的改变以及蛋白质磷酸化。在获能过程中,精子没有发生已知的形态变化。本文综述了牛和猪精子在体内和体外获能的分子方面的最新证据。本综述的目的是,精子获能的过程将包括射精后发生的成熟事件,直到与透明带结合,触发顶体胞吐并开始受精。
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引用次数: 40
Genetic risk factors in male infertility. 男性不育的遗传风险因素。
Pub Date : 2007-05-01 DOI: 10.1080/01485010701271786
Csilla Krausz, Claudia Giachini

The etiopathogenesis of testicular failure remains unknown in about half of the cases and is referred to as "idiopathic infertility". "Idiopathic" testicular failure is of probable genetic origin since the number of genes involved in human spermatogenesis is likely thousands and only a small proportion of them have been identified and screened in infertile men. In parallel with studies aimed to identify mutations with a clear cause-effect relationship in spermatogenesis candidate genes, there is an increasing interest towards genetic susceptibility factors to male infertility. Despite many efforts, only a few clinically relevant polymorphisms have been identified. This is mainly related to the multifactorial nature of male infertility and to the inappropriate study design of the majority of the studies. The most promising polymorphisms are in genes involved in the endocrine regulation of spermatogenesis and on the Y chromosome, the "gr/gr" deletions. Polymorphisms are generally considered as co-factors. Their final effect on testis function and fertility is probably modulated by the genetic background of each individual and/or by the presence of certain environmental factors. In this review, recent findings concerning some of the most widely studied polymorphisms and male infertility will be discussed.

在大约一半的病例中,睾丸功能衰竭的病因不明,被称为“特发性不孕”。“特发性”睾丸功能衰竭可能是遗传原因,因为涉及人类精子发生的基因数量可能有数千个,其中只有一小部分在不育男性中被识别和筛选。与旨在确定精子发生候选基因中具有明确因果关系的突变的研究同时,人们对男性不育的遗传易感性因素越来越感兴趣。尽管做出了许多努力,但只有少数临床相关的多态性被确定。这主要与男性不育症的多因素性质和大多数研究的不适当研究设计有关。最有希望的多态性是与精子发生的内分泌调节有关的基因和Y染色体上的“gr/gr”缺失。多态性通常被认为是辅助因素。它们对睾丸功能和生育能力的最终影响可能受到每个人的遗传背景和/或某些环境因素的调节。在这篇综述中,关于一些最广泛研究的多态性和男性不育的最新发现将进行讨论。
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引用次数: 114
The preparation of human spermatozoal RNA for clinical analysis. 用于临床分析的人精子RNA的制备。
Pub Date : 2007-05-01 DOI: 10.1080/01485010701216526
Robert Goodrich, Graham Johnson, Stephen A Krawetz

The recent identification of RNA as a component of mature spermatozoa necessitated the development of a reliable isolation protocol capable of yielding a high-quality substrate. In addition to the inherent difficulties associated with isolating RNA, the procedure as applied to sperm must overcome the resilient nature and reduced RNA content found within this cell type. Further, the protocol must be suited to the clinical setting. A reliable RNA isolation procedure optimized for this unique cell type is described. Ejaculate is collected, contaminating somatic cells lysed then spermatozoal RNA released by homogenization in a chaotrope. RNA is then purified from the homogenate by chromatography using a commercially available resin. The quality of isolated samples is assessed by PCR and RT-PCR. Once purity is established samples are suitable for numerous applications including amplification and probe synthesis. The reliable and consistent isolation of high-quality RNA from mature spermatozoa will aid in the development of new tools for the clinical assessment of male-factor fertility.

最近鉴定的RNA作为成熟精子的一个组成部分,需要开发一种可靠的分离方案,能够产生高质量的底物。除了与分离RNA相关的固有困难外,应用于精子的过程必须克服这种细胞类型的弹性和减少的RNA含量。此外,该方案必须适合临床环境。可靠的RNA分离程序优化这种独特的细胞类型描述。收集精液,污染裂解的体细胞,然后在混沌中匀浆释放精子RNA。然后用市售树脂从匀浆中通过层析纯化RNA。采用PCR和RT-PCR对分离样品的质量进行评价。一旦纯度确定,样品适用于多种应用,包括扩增和探针合成。从成熟精子中可靠和一致地分离高质量RNA将有助于开发用于临床评估男性因素生育能力的新工具。
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引用次数: 116
Analysis of sperm karyotypes in a patient treated with griseofulvin. 灰黄霉素治疗患者精子核型分析。
Pub Date : 2007-05-01 DOI: 10.1080/01485010701314024
Evelyn M Ko, R Brian Lowry, Renée H Martin

Griseofulvin is known to interfere with chromosome segregation by binding to microtubule-associated proteins. Studies in mouse germ cells have demonstrated that griseofulvin can induce aneuploidy (numerical chromosome abnormalities) at therapeutic concentrations. The aim of this study was to determine if chronic griseofulvin treatment led to an increased frequency of sperm chromosome abnormalities in one male subject. We analyzed 290 full sperm karyotypes using the human sperm-hamster oocyte fusion system. The frequency of X- and Y-bearing sperm was equal. There was no increase in the frequency of numerical (1.7%) or structural (9.3%) abnormalities in the subject compared to unexposed controls. Although reassuring, this is the first report on this subject and future studies are needed to assess the risk of griseofulvin.

灰黄霉素通过与微管相关蛋白结合而干扰染色体分离。对小鼠生殖细胞的研究表明,灰黄霉素在治疗浓度下可诱导非整倍体(数值染色体异常)。本研究的目的是确定慢性灰黄霉素治疗是否会导致一名男性受试者精子染色体异常频率增加。我们利用人类精子-仓鼠卵母细胞融合系统分析了290个完整精子核型。携带X染色体和y染色体的精子出现的频率相等。与未暴露的对照组相比,受试者的数字(1.7%)或结构(9.3%)异常频率没有增加。虽然令人放心,但这是关于该主题的第一份报告,需要进一步的研究来评估灰黄霉素的风险。
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引用次数: 6
期刊
Archives of andrology
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