• Asian-Australasian Journal of Animal Sciences
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• 제24권9호
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• pp.1199-1203
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• 2011

The objective of this study was to examine the binding potential of sperm to the epithelium of the sperm storage tubules (SST) in vitro and in vivo to assess the functional maturation of fowl sperm. Sperm from the testis, epididymis, as well as the proximal, middle and distal vas deferens were incubated in vitro with either the uterovaginal junction (UVJ)- or infundibular tissue containing SST at $39^{\circ}C$ for 30 min. Aliquots of sperm were also artificially inseminated into the uteri of hens, and the UVJ and infundibulum were collected 24 h post artificial insemination (AI). After incubation and AI, tissues were washed to remove loosely adhered sperm and subjected to fluorescence staining with 4', 6-diamidino-2-phenylindole, dihydrochloride (DAPI) for counting the number of bound sperm per 0.25 mm2 of surface area. Sperm from the testis, epididymis, and the three segments of the vas deferens exhibited their differential (p<0.05) binding capacity, which increased gradually from the testicular to distal vas deferens sperm under both in vitro and in vivo conditions. Existing similar trend, sperm, regardless of their source had a lesser affinity to bind to the epithelium of the infundibular SST than to the UVJ-SST. These experimental results suggested that fowl sperm may undergo gradual changes in the process of functional maturation, whereby they gain the ability to bind to the epithelium of the SST during their passage through the male reproductive tract (MRT).

• 한국발생생물학회지:발생과생식
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• 제16권1호
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• pp.53-58
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• 2012

Circadian rhythmicity (e.g. secretory pattern of hormones) plays an important role in the control of reproductive function. We hypothesized that the alteration of feeding pattern via meal time shift/restriction might disrupt circadian rhythms in energy balance, and induce changes in reproductive activities. To test this hypothesis, we employed simple animal model that not allowing $ad$ $libitum$ feeding but daytime only feeding. The animals of $ad$ $libitum$ feeding group (Control) have free access to food for 4 weeks. The day feeding (=reverse feeding, RF) animals (RF group) have restricted access to food during daytime (0900-1800) for 4 weeks. After completing the feeding schedules, body weights, testis and epididymis weights of animals from both group were not significantly different. However, the weights of seminal vesicle (control : RF group = $0.233{\pm}0.014g$ : $0.188{\pm}0.009g$, $p$<0.01) and prostate (control : RF group = $0.358{\pm}0.015g$ : $0.259{\pm}0.015g$, $p$<0.001) were significantly lower in RF group animals. The mRNA levels of pituitary common alpha subunit ($C{\alpha}$; control : RF group = $1.0{\pm}0.0699$ AU : $0.1923{\pm}0.0270$ AU, $p$<0.001) and $FSH{\beta}$ (control : RF group = $1.0{\pm}0.1489$ AU : $0.5237{\pm}0.1088$ AU, $p$<0.05) were significantly decreased in RF group. The mRNA levels of ACTH were not significantly different. We were unable to find any prominent difference in the microstructures of epididymis, and there were slight alterations in those of seminal vesicles after 4 weeks of reversed feeding when compared to control samples. The present study demonstrates that the shift and/or restriction of feeding time could alter the pituitary gonadotropin expression and the weights of seminal vesicle and prostate in rats. These data suggest the lowered gonadotropin inputs may decrease androgen secretion form testis, and consequently results in poor response of androgen-dependent tissues such as seminal vesicle and prostate.

• 한국수정란이식학회지
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• 제24권2호
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• pp.77-87
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• 2009

To understand molecular and cellular mechanisms of many gene products in the female reproductive organs including the ovary and uterine endometrium as well as during embryo development, researchers have developed and utilized many effective methodologies to analyze gene expression in cells, tissues and animals over the last several decades. For example, blotting techniques have helped to understand molecular functions at DNA, RNA and protein levels, and the reverse transcription-polymerase chain reaction (RT-PCR) method has been widely used in gene expression analysis. However, some conventional methods are not sufficient to understand regulation and function of genes expressed in very complex patterns in many organs. Thus, it is required to adopt more high-throughput and reliable techniques. Here, we describe several techniques used widely recently to analyze gene expression, including annealing control based-PCR, differential display-PCR, expressed sequence tag, suppression subtractive hybridization and microarray techniques. Use of these techniques will help to analyze expression pattern of many genes from small scale to large scale and to compare expression patterns of genes in one sample to another. In this review, we described principles of these methodologies and summarized examples of comparative analysis of gene expression in female reproductive organs with help of those methodologies.

• 한국동물생명공학회지
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• 제37권2호
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• pp.96-105
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• 2022

The ovary undergoes substantial physiological changes along with estrus phase to mediate negative/positive feedback to the upstream reproductive tissues and to play a role in producing a fertilizable oocyte in the developing follicles. However, the disorder of estrus cycle in female can lead to diseases, such as cystic ovary which is directly associated with decline of overall reproductive performance. In gene expression studies of ovaries, quantitative reverse transcription polymerase chain reaction (qPCR) assay has been widely applied. During this assay, although normalization of target genes against reference genes (RGs) has been indispensably conducted, the expression of RGs is also variable in each experimental condition which can result in false conclusion. Because the understanding for stable RG in porcine ovaries was still limited, we attempted to assess the stability of RGs from the pool of ten commonly used RGs (18S, B2M, PPIA, RPL4, SDHA, ACTB, GAPDH, HPRT1, YWHAZ, and TBP) in the porcine ovaries under different estrus phase (follicular and luteal phase) and cystic condition, using stable RG-finding programs (geNorm, Normfinder, and BestKeeper). The significant (p < 0.01) differences in Ct values of RGs in the porcine ovaries under different conditions were identified. In assessing the stability of RGs, three programs comprehensively agreed that TBP and YWHAZ were suitable RGs to study porcine ovaries under different conditions but ACTB and GAPDH were inappropriate RGs in this experimental condition. We hope that these results contribute to plan the experiment design in the field of reproductive physiology in pigs as reference data.

• 한국동물생명공학회지
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• 제37권4호
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• pp.276-284
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• 2022

Endocrine-disrupting chemicals found in many commercial products may interfere with the normal functioning of the endocrine system and are unsafe because of their cumulative effect on the human body. However, little is known about the effects of combinations of endocrine-disrupting chemicals in humans. Methoxychlor and bisphenol A are toxic to male reproductive organs. Therefore, we studied the effects of methoxychlor and bisphenol A on male reproductive function. Male mice were divided into four treatment groups: control, 400 mg methoxychlor, 1 mg bisphenol A, and 400 mg methoxychlor + 1 mg bisphenol A/kg/day. Methoxychlor and bisphenol A were dissolved in sesame oil and acetone and administered orally for 4 weeks. After administration, the weight and histological changes in the testicles and epididymis, sperm count and health were observed biochemical tests and whole blood counts were performed. The results showed that the mice in the bisphenol A and methoxychlor + bisphenol A groups gained more weight than those in the control and methoxychlor group. The weights of the testes and epididymis were higher in the experimental groups than in the control. Sperm motility and progression were significantly reduced in the bisphenol A and methoxychlor + bisphenol A groups. Histological observation showed a reduced number of sperm, smaller seminiferous tubules, and destroyed lumen in the methoxychlor + bisphenol A group compared to the other groups. In conclusion, our study showed that methoxychlor and bisphenol A destroy male reproductive tissues and decrease sperm quality.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.123-126
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• 2010

The purpose of present study was to analyze mineral contents in various tissues and investigate theirs relation with bone mineral density (BMD) in rats. Fifteen Sprague-Dawley rats were fed standard diet for 4 weeks. Body weight gain, feed intake, and feed efficiency ratio were 41.00 g/week, 171.15 g/week, and 0.24 respectively. Among 12 minerals in serum, Ca is the highest with 6.86 mg/dl. Serum Mg, Se, and Cu were 2.52 mg/dl, 0.23 mg/dl and 0.22 mg/dl respectively. Mg contents in liver, spleen, and kidney were $246.36\;{\mu}/g$, $105.01\;{\mu}/g$, and $273.38\;{\mu}/g$ respectively. Tibia contents of Ca, Mg, Zn, Fe and V were 194.91 mg/g, 23.10 mg/g, 0.60 mg/g, 0.35 mg/g and 0.14 mg/g respectively. BMDs of right tibia and spine were 122.04 mg/cm and $153.61\;mg/cm^2$. There were significantly positive correlations between tibia BMD and Se (p<0.05), tibia BMD and V (p<0.01), spinal BMD and V(p<0.05), respectively. It's expected that these results are used as a reference data in following study to elucidate physiological function of minerals.

• 한국환경과학회지
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• 제8권4호
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• pp.515-523
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• 1999

Nine trace metals (Zn, Fe, Al, Pb, Mn, Cu, Ni, Cr, and Cd) and Se concentrations were determined in organs and tissues(muscle, bone, kidney, liver and skin) of the striped field mice, Apodemus agrarius collected at Daejeo-dong, Pusan city and the Sorak Mt., Kangwon Province. All the trate elements were detected from all the mice examined. As for the metal concentrations in the mice from Daejeo-dong, Zn, Fe, Al, Mn, Ni and Cr were significantly higher than those in Sorak Mt.(p<0.05), suggesting that pesticides including Zn and factory wastes containing several metals might contaminated the environment of Daejeo-dong. As regards the element concentrations in each organ and tissue, Fe, Al, Pb, Mn, Cu, Cr and Cd concentrations were high in liver or kidney; Zn in skin and bone; Ni in bone, skin and kidney; and Se in muscle, liver and skin in all the mice examined. There were significant differences(p<0.05) between juveniles and adults in average concentration of metals(Zn, Fe, Pb, Mn, Ni, and Cd) in every organs and tissues of mice collected at Daejeo-dong. Much higher average levels of metals in juveniles indicate that considerable burdens fo metals might be transferred through the placenta. However, there were no significant differences between males and females, and between young and old adults in average metal concentrations, which suggests that the mice might accumulate the metals during their life time, although they might excrete the metals not only through reproductive activities, such as parturition, lactation and ejaculation of semen, but also through molting, judging from higher accumulating ratios of most metals in skin of adults than of juveniles.

• Reproductive and Developmental Biology
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• 제33권2호
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• pp.93-98
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• 2009

In this study, bovine Dnmt1 cDNA was sequenced and detected Dnmt1 mRNA level in bovine tissues by northern blot, methylation pattern of genome by southern blot, specific localization of Dnmt1 in mouse and bovine preimplantation embryos by immunocytostaining and Dnmt1 protein level in ovary and testis by western blot. Bovine Dnmt1 cDNA sequence showed more homology with that of human than mouse and rat. The RNA level of Dnmt1 was 10 times higher expression in placenta than other tissues. This indicates that placenta was hypermethylated compared to others organs. The genomic DNA could not be cut by a specific restriction enzyme (HpaII) in placenta, lung and liver of bovine. It suggests that Dnmt1 in some somatic cells was already methylated. Dnmt1, which has the antibody epitope 1316~1616, was distributed in nucleus and cytoplasm including the stage of pronuclear stage and maturation of oocyte and gradually weaken to blastocyst stage compare to negative. In addition, Dnmt1 was strongly expressed in tetraploid embryo and cloned 8-cell than IVF 8-cell. An aberrant pattern of DNA methylation in cloned embryo may be abnormal development of fetus, embryonic lethality and placenta dysfunction. The somatic specific band (190kDa) was appeared in ovary and testis, but oocyte specific band (175kDa) was not. Further investigations are necessary to understand the complex links between the methyltransferases and the transcriptional activity of genes in the cloned bovine tissues.

• Asian-Australasian Journal of Animal Sciences
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• 제13권7호
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• pp.1026-1034
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• 2000

As described here, most recently developed methods for improving reproduction performance of domesticated animals such as cattle, swine and chicken have been considered to be also usable for restoring some sorts of endangered and/or extinct wild animals in the very near future. Especially, the techniques for in vitro storage of gametes obtained from dead animals shortly after the death, probably 24 h following the sacrifice are also available for obtaining some of experimental specimens. In case of the endangered animals, nobody will be allowed to use any tissues from the living animals, therefore, e.g., the use of skin tissues from these bodies is another possibility of restoring the living animals. Regarding the use of skin tissues, the most highly usable tools must be the cloning techniques for reviving rare cells from the living body. Most possible techniques for cloning cells is nuclear transfer from rare species to highly relative species, and this is the case of germ cells, e.g., primordial germ cells (PGCs) of avian species. One of the possibilities is the nuclear transfer of Crested Ibis (Nipponia nippon) to the PGCs of chicken, resulting in the PGCs with transferred nucleus from the ibis. In mammalian species, the same procedure as in the case of birds would be successful, e.g., the removed nucleus from Giant Pandas will be transferred to the cell, such as somatic cells or germ cells from black bears or lesser pandas, leading to the production of transnucleared cells in the body of female black bears. These two cases are most promising techniques for reviving endangered animals in the world, particularly in Asian countries, mainly in China. As a conclusion, possible production of cloned animals carrying transnucleared cells from endangered animals, such as Giant Pandas and Crested Ibis, may be reproduced gradually in the near future. Scientists are, therefore, required to convert the paradigm from domestic animals to wild animals, including endangered and/or extinct animals on the earth.

• 환경생물
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• 제20권3호
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• pp.224-231
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• 2002

광주기(하루 중 빛의 길이)는 골든 햄스터의 생식을 조절하는 주된 요인이다. 광주기 정보는 멜라토닌을 통하여 생식 내분비계로 전달된다. 따라서 멜라토닌이 생식에 미치는 효과를 여러 광주기에 노출시킨 햄스터에서 조사하였다. 단주기(하루 중 12시간 이하의 조명)에 노출시킨 동물들과 저녁에 멜라토닌을 주사한 동물들의 정소 무게는 현저하게 줄어들었으나, 장주기 (하루 중 12.5시간 이상의 조명)에 유지된 동물과 오전에 멜라토닌을 투여한 동물들의 정소 무게는 줄어들지 않았다. 퇴화된 정소를 조직학적으로 조사한 결과, 세정관 직경이 감소되었고, 세정관내 세포수가 두드러지게 줄어들었다. 또한 생식 능력이 퇴화된 동물의 혈중 여포자극호르몬과 황체호르몬의 수준도 생식 능력을 보유하고 있는 동물에 비해 뚜렷하게 감소하였다. 멜라토닌 수용체가 역전사 polymerase chain reaction으로 동정되었고 조직특이성 또한 조사하였다. 동정된 멜라토닌 수용체는 309염기였으며, 시상하부와 뇌하수체를 포함하는 다양한 장기에서 발현되었다. 생식을 조절하는 핵심 물질인 gonadotropin releasing hormone (GnRH) 유전자의 발현 또한 동정되었다. 그러나 멜라토닌 처리와 광주기 처리는 GnRH유전자 발현에 영향을 미치지 않았다. 종합하면, 광주기의 효과는 멜리토닌을 경유하여 발휘되며, 멜라토닌은 GnRH유전자의 발현보다는, 생성된 GnRH의 분비에 영향을 미쳐 생식내분비계에 간접적으로 작용함을 알 수 있었다.