促性腺激素受体在雌性水牛生殖器官的表达定位研究

为研究促性腺激素受体(FSHR、LHR)在广西雌性水牛生殖器官中的分布情况,运用免疫组化SABC法对处于不同发情周期(卵泡期、黄体期)成年水牛的卵巢、子宫、输卵管中FSHR、LHR分别进行染色定位。结果表明,FSHR/LHR阳性细胞在卵巢主要见于卵巢内膜细胞及卵泡颗粒细胞;子宫主要见于子宫内膜上皮细胞和腺体细胞;输卵管主要见于柱状上皮纤毛细胞。其中,随着发情周期不同,FSHR、LHR的表达量也有所差异,卵巢中卵泡期FSHR、LHR的表达量均高于黄体期;子宫中FSHR的表达量卵泡期高于黄体期,LHR的表达量黄体期高于卵泡期;而输卵管并没有显著差异。     

促卵泡素受体和促黄体素受体在猪子宫中的定位研究

为探明2种促性腺激素受体（FSHR、LHR）在猪子宫中的位置分布,本试验运用免疫组化ABC法对卵泡期、黄体期猪的子宫FSHR、LHR分别进行定位染色。结果显示,FSHR阳性细胞主要见于子宫内膜上皮细胞、血管平滑肌细胞。LHR阳性细胞主要见于子宫内膜上皮细胞、子宫肌层平滑肌细胞及血管平滑肌细胞;动情周期不同,FSHR、LHR的表达量也不同。卵泡期子宫内膜上皮细胞FSHR的表达量高于黄体期,血管平滑肌FSHR的表达量低于黄体期;卵泡期子宫血管平滑肌细胞LHR的的表达量高于黄体期,而子宫内膜上皮细胞、平滑肌细胞LHR的表达量均低于黄体期。     

NGF对输卵管黏膜上皮细胞促性腺激素受体表达的影响

本研究以牛输卵管黏膜上皮细胞为研究对象,利用荧光定量PCR方法检测NGF和酪氨酸激酶抑制剂K252α对输卵管粘膜上皮细胞膜促性腺激素受体FSHR和LHRmRNA表达的影响.添加30、60、100μg/L NGF可以显著增加FSHR mRNA的表达,添加30和60 μg/L的NGF同样增加LHR mRNA的表达.添加抑制剂K252α后,LHR和FSHR的表达量均比对照组显著下降.该结果说明NGF可以通过其受体调节输卵管黏膜上皮细胞促性腺激素受体FSHR和LHR的表达.     

性腺外FSH受体和LH受体表达研究进展

雌性动物的性腺是卵巢,性腺外的生殖组织器官主要包括:输卵管、子宫颈、子宫肌层等,其中子宫是内分泌器官,除有局部内分泌功能外,还可能对下丘脑-垂体-卵巢轴有调节作用.促卵泡素(FSH)和促黄体素(LH)的生理功能是通过其FSHR、LHR来介导的,其受体在性腺和性腺外均有表达.本文主要对雌性性腺外生殖组织器官上两种受体的表达加以综述.     

牦牛睾丸发育过程中INHβB、FSHR和LHR的表达差异性分析

为进一步研究抑制素(INH)、促卵泡素(FSH)和促黄体素(LH)在牦牛精子形成过程中发挥的作用,采用实时荧光定量PCR(RT-qPCR)、蛋白质免疫印迹技术(WB)和免疫组织化学法(IHC)检测抑制素受体(INHβB)、促卵泡素受体(FSHR)和促黄体素受体(LHR)在不同年龄牦牛睾丸中的表达及定位。结果显示,INHβB、FSHR和LHR在不同年龄牦牛睾丸组织中均有表达,其相对表达量随年龄增长呈现先上升后下降的趋势;5岁时表达量最高,并显著高于其他3个年龄段(P<0.05)。INHβB、FSHR和LHR蛋白主要分布在睾丸组织的各级生精细胞、支持细胞及间质细胞中。结果表明,牦牛睾丸发育过程中INHβB、FSHR和LHR均可介导相应激素调控精子的生成与成熟,为进一步探索牦牛睾丸的发育及精子的生成提供了研究基础。     

FSHR与LHR的研究进展

本文重点论述了FSHR与LHR的结构及在卵母细胞体外成熟过程中的表达和调控,并阐述了FSHR与LHR的研究现状及存在的问题。     

促卵泡素受体和促黄体素受体在乏情期和怀孕期牦牛卵巢中的表达

应用免疫组织化学技术和图象分析方法对乏情期、怀孕期牦牛卵巢的促卵泡素受体(FSHR)、促黄体素受体(LHR)的表达特点进行了研究。结果表明,牦牛卵巢的皮质、髓质、颗粒层和膜内层、黄体中都分布有FSHR和LHR。且牦牛卵巢皮质、髓质中的FSHR光密度值在乏情期显著大于怀孕期(P0.05);怀孕期无黄体侧卵巢颗粒层和膜内层中FSHR光密度值均大于乏情期和怀孕期有黄体侧(P0.05)。怀孕期有黄体侧卵巢皮质处LHR光密度值最大,乏情期次之,怀孕期无黄体侧最小,各组间差异显著(P0.05);在怀孕期无黄体侧卵巢髓质处LHR光密度值显著小于怀孕期有黄体侧和乏情期(P0.05);在乏情期、怀孕期无黄体侧和怀孕期有黄体卵巢颗粒层和膜内层处LHR光密度值之间差异均不显著(P0.05)。表明牦牛卵巢中FSHR和LHR随着生殖阶段的不同而变化。     

南江黄羊不同组织FSHR和GnRHR mRNA表达水平研究

为进一步了解FSHR和GnRHR基因的功能,揭示其对山羊繁殖性状的影响。本研究采用荧光定量PCR技术分析了南江黄羊不同组织FSHR和GnRHR基因的表达差异。结果表明:FSHR和GnRHR在下丘脑、垂体、卵巢、输卵管、子宫、睾丸组织中均有表达,其中,FSHR和GnRHR在母羊子宫组织中表达水平较高,而在其他组织中表达水平相差不大;FSHR基因和GnRHR基因在子宫中的表达量均极显著高于其他组织(P0.01);GnRHR基因在卵巢组织中的表达量显著高于垂体、下丘脑、输卵管(P0.05);公羊睾丸组织FSHR和GnRHR基因的表达量显著高于下丘脑和垂体(P0.05)。本研究结果将会为进一步研究该基因对山羊繁殖相关功能影响的研究奠定基础。     

促黄体素受体在母畜生殖道的分布与表达

在多种动物包括灵长类动物、猪、牛、火鸡和人,子宫内都发现有促黄体素(LH)或人绒毛膜促性腺激素(hCG)及其mRNA的结合位点.牛、猪在整个发情周期子宫内都存在LH受体(LHR)蛋白及其mRNA,并在黄体期表达最高.LH通过激活腺苷酸环化酶和磷脂酶C信号途径,增加前列腺素合成酶(PGSH),也称环加氧酶(COX)的活性,从而促进前列腺素(PG)的合成.牛、猪、羊在发情周期的15 d左右PGF2α升高,这与子宫内LHR的活化有关.在不同种动物的输卵管、子宫内膜、子宫肌层和子宫颈,LHR的释放呈现出动态特征,表明LH在发情周期和胚胎附植的分子自分泌-旁分泌调节中可能具有重要的作用.     

日粮能量和蛋白水平对动物繁殖相关基因表达的调控

日粮营养水平对动物繁殖调控影响机制的研究已经到分子水平,日粮营养水平对动物繁殖相关基因表达的研究可揭示其对动物繁殖调控的分子机制。文章就日粮能量和蛋白水平对与动物繁殖相关的FSHR、LHR和IGF基因的表达调控进行了综述,为进一步研究营养水平调控动物繁殖的分子机理提供理论指导。     

Signalling Events and Associated Pathways Related to the Mammalian Sperm Capacitation

Capacitation is a biological phenomenon occurring prior to fertilization and is a multiple event process. Many physiological and biochemical changes takes place during the process; these changes are related to lipid composition of membrane, intracellular modulation of ion concentration, protein phosphorylation, sperm movement and membrane permeability. These events occur when the sperm is exposed to the new environment of ion concentration in the female reproductive tract. Ions such as bicarbonate and calcium facilitate capacitation by activating adenylyl cyclase, thus initiating protein kinase A (PKA) signalling cascade. Extracellular‐regulated kinase pathway is activated by ligand binding to the membrane receptors and intracellular activation by reactive oxygen species (ROS). Activation of these pathways leads to the phosphorylation of different proteins, which is associated with events such as capacitation, hyperactivation and acrosome reaction that are essential for successful fertilization. Extensive studies were carried out on protein phosphorylation in relation to capacitation, but its role still remains ambiguous.     

Immunolocalization of nerve growth factor (NGF) and its receptors (TrkA and p75LNGFR) in the reproductive organs of Shiba goats

The objective of this study was to determine the immunolocalization of NGF and its receptors (TrkA and p75LNGFR) in the reproductive tract of the Japanese Shiba goats. Five adult goats were used in this study and sections of ovaries, uteri and oviducts were immunostained by the avidin-biotin-peroxidase complex method (ABC). The results showed that NGF and its receptors (TrkA and p75LNGFR) were expressed in granulosa cells, theca cells, interstitial cells and lutein cells in ovaries. Immunoreactions for NGF, TrkA and p75LNGFR were also detectable in epithelial cells and muscle cells of the ampulla and isthmus of the oviduct, and in epithelial cells and uterine glands of the uterus. These results strongly suggest autocrine and paracrine regulation of reproductive function by NGF in the reproductive tract of female Shiba goats.     

Immunity in the female sheep reproductive tract

Immune surveillance in the female reproductive tract is dependent on the interplay of many factors that include the expression of pattern recognition receptors on epithelial cells, resident leukocyte populations and hormones, none of which are uniform. The lower reproductive tract must accommodate the presence of commensal organisms whereas the upper reproductive tract is sterile. However, the upper female reproductive tract has its own immunological challenge in that it must tolerate the presence of a semi-allogeneic fetus if pregnancy is to succeed. So, immune activation and effector mechanisms to control pathogens may be qualitatively and quantitatively different along the reproductive tract. Our knowledge of innate and adaptive immunity in the sheep is less comprehensive than that of human or mouse. Nevertheless, comparative studies suggest that there are likely to be conserved innate immune sensory mechanisms (e.g. Toll-like receptors) and defence mechanisms (anti-proteases, defensins) that combine to limit infection in its early stages while shaping the adaptive response that leads to immunological memory and long-term protection. There are many pathogens that target the reproductive tract, and in particular the placenta, where specialised immunoregulatory mechanisms are operational. Among such pathogens are bacteria belonging to the genera Chlamydia/Chlamydophila that chronically infect the reproductive tracts of sheep and humans and ultimately cause disease through inflammation and tissue damage. An understanding of the immunological microenvironment of the reproductive tract is important for the design of novel control strategies to control chlamydial disease.     

Expression of aromatase and oestrogen receptors in reproductive tissues of the stallion and a single cryptorchid visualised by means of immunohistochemistry

Androgen metabolism may proceed to amplify the action of testosterone by its aromatisation to oestradiol. Recently, a growing body of evidence suggests a role of oestrogens in the male reproductive tract via their specific oestrogen receptors (ERs). In order to check whether androgens are converted to oestrogens in the testis, epididymis and prostate of the stallion, the expression of aromatase was visualised by means of immunohistochemistry. Moreover, to show the cellular targets for oestrogens the presence of oestrogen receptor alpha (ERalpha) and oestrogen receptor beta (ERbeta) was demonstrated in these tissues. Finally, to show whether naturally occurring cryptorchidism has any influence on the localisation of aromatase and distribution of ERs, the reproductive tissues of a single horse, bilaterally cryptorchid, were also taken for this study. The results demonstrated that aromatase and ERs are ubiquitously distributed throughout the male reproductive tract, what indicates a putative role of oestrogens in modulating the function of the reproductive tissues of the stallion. In the cryptorchid horse the increase in conversion of androgen to oestrogen was observed as manifested by aromatase overexpression. This is the first report showing the cellular site of oestrogen biosynthesis not only in the testis but also in the epididymis and prostate of sexually mature stallion and a single, adult cryptorchid.     

Presence of melatonin‐catabolizing non‐specific enzymes myeloperoxidase and indoleamine 2,3‐dioxygenase in the ram reproductive tract

The melatonin catabolism is very complex and not completely understood. Melatonin can be metabolized by free radical interaction, but also pseudo‐enzymatically or by enzymatic pathways. We have previously detected the existence of melatonin‐synthesizing enzymes and melatonin receptors MT1 and MT2 in the ram reproductive tract; thus, in order to start to elucidate melatonin catabolism in these organs, we have investigated the presence of the melatonin‐catabolizing enzymes indoleamine 2,3‐dioxygenase (IDO, both IDO1 and IDO2 isoforms) and myeloperoxidase (MPO) in testis, epididymis and accessory glands. Gene expression analyses by real‐time PCR showed the presence of MPO, IDO1 and IDO2 in all the organs of the ram reproductive tract and revealed that MPO is the main melatonin‐catabolizing enzyme, which is mainly expressed in the testis and the bulbourethral glands (p < .05). These results were further corroborated by immunohistochemical staining, and by Western blot. Likewise, MPO was also evidenced in epididymal and ejaculated spermatozoa by indirect immunofluorescence and Western blot. In conclusion, melatonin‐catabolizing enzymes MPO, IDO1 and IDO2 are expressed in the ram reproductive tract, and MPO is the most expressed one, mainly in the testis and the bulbourethral glands. The presented results warrant further studies on the function of these enzymes and their melatonin‐metabolizing activity.     

雄性生殖系统抗氧化研究进展

雄性生殖道代谢活跃,活性氧(reactive oxygen species,ROS)的产生不可避免,低浓度的ROS对哺乳动物生殖具有基础性作用,但ROS的过度累积必然会使精子发生氧化损伤而影响其受精能力。因此精子对抗氧化应激的保护机制非常重要,作者就目前公认的雄性生殖系统中抗氧化酶及氧化还原系统的研究现状进行综述,为更好地开展雄性动物生理和病理学研究提供借鉴。     

Research Progresses on Anti-oxidation in Male Reproductive Systems

It is believed that active metabolism in male reproductive tract is contributed to production of ROS (reactive oxygen species).Low physiological ROS plays a fundamental role in mammalian reproduction, but the excessive accumulation of potentially damaging ROS can affect the function of sperm.Thus, it is important to maintain normal reproductive ability by a protective mechanism against oxidative stress.Here, we review the progress of antioxidant enzymes and redox systems in male reproductive systems, in order to provide references for physiology and pathology researches in male animals.     

Sperm Storage in the Female Reproductive Tract in Birds

The ability to store sperm in the female genital tract is frequently observed in vertebrates as well as in invertebrates. Because of the presence of a system that maintains the ejaculated sperm alive in the female reproductive tract in a variety of animals, this strategy appears to be advantageous for animal reproduction. Although the occurrence and physiological reasons for sperm storage have been reported extensively in many species, the mechanism of sperm storage in the female reproductive tract has been poorly understood until recently. In avian species, the specialized simple tubular invaginations referred to as sperm storage tubules (SSTs) are found in the oviduct as a sperm storage organ. In this review, we summarize the current understanding of the mechanism of sperm uptake into the SSTs, maintenance within it, and controlled release of the sperm from the SSTs. Since sperm storage in avian species occurs at high body temperatures (i.e., 41 C), elucidation of the mechanism for sperm storage may lead to the development of new strategies for sperm preservation at ambient temperatures, and these could be used in a myriad of applications in the field of reproduction.     

Health Assessment of the Reptilian Reproductive Tract

The number of reptiles maintained as pets, along with the captive reproduction of these animals, has increased dramatically over the last 2 decades. Reproductive tract pathology, secondary to an inadequate captive environment, is a common occurrence that can lead to abnormal reproductive activity and resulting disease. Additional factors such as malnutrition, obesity, systemic disease, infection/inflammation, trauma, and dehydration contribute to the development of reproductive tract disorders. The diagnosis of reproductive tract disease requires a thorough history, laboratory testing, and diagnostic imaging. An understanding of a species' biological and environmental requirements is essential for the prevention of reproductive tract disease. The purpose of this article is to review the reproductive tract anatomy and physiology of snakes, turtles, and lizards, and to provide a brief overview of the common disorders affecting the reproductive tract of reptilian species.     

Board-invited review: Estrogen and progesterone signaling: genomic and nongenomic actions in domestic ruminants

Progesterone and estrogens play key roles in regulating various physiological phenomena related to normal growth, development, and reproduction of domestic animals. This review focuses on the mechanisms by which progesterone and estrogens regulate the reproductive processes in these animals. The majority of research on the actions of progesterone and estrogens on the reproductive systems of cattle, sheep, and pigs has been genomic in nature and represents attempts to better understand how these steroids regulate gene expression. Results of recent research suggest that progesterone and estrogens can alter target cell responses nongenomically via membrane receptors. The characteristics of membrane receptors for progesterone and estrogen in various cell types are described and the intracellular signal pathways defined. Estrogens acting via membrane receptors can suppress LH secretion by gonadotropes and stimulate rapid increases in uterine blood flow. Progesterone acting via a membrane receptor has been shown to inhibit binding of oxytocin to oxytocin receptors in isolated endometrial plasma membranes and stimulate capacitation of spermatozoa. Results of research suggest that progesterone and estrogens can act nongenomically to alter target cell responses in domestic animals. The biological implications of this mode of action in these animals are discussed.