• BMB Reports
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• 제39권4호
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• pp.371-376
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• 2006

Crustacean hyperglycemic hormone (CHH) plays a major role in controlling glucose level in the haemolymph and also triggers important events during molting and reproductive cycles. In Penaeus monodon, three types of CHH, namely Pem-CHH1, Pem-CHH2 and Pem-CHH3, have been previously characterized. In this study, mouse polyclonal antibody was raised against recombinant Pem-CHH1 that was expressed in Escherichia coli. The anti-Pem-CHH1 antibody recognized all three types of Pem-CHHs but did not cross-react with either related hormone, molt-inhibiting hormone of P. monodon, or unrelated human growth hormone. The hyperglycemic activity in the extract from the eyestalk neural tissues was significantly depleted after incubating with anti-Pem-CHH antibody. Direct injection of the antibody into shrimp caused about 30-50% reduction in the haemolymph glucose level. The result demonstrates the ability of anti-Pem-CHH1 antibody to deplete the activity of CHH in vivo, and thus provides a possibility of using anti-Pem-CHH1 antibody to inhibit the hormone activity as a strategy to modulate growth and reproduction in this species.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.215-219
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• 2011

Acyl-CoA synthetase 4(ACSL4) is an arachidonate-preferring enzyme abundant in steroidogenic tissues and postulated to modulate eicosanoid production. The human and mouse ACSL4 gene are mapped on chromosome X. The female mice heterozygous for ACSL4 deficiency became pregnant less frequent1y and produced small litters, with 40% of embryos surviving gestation. In this study, we examined the regulation of ACS4 by estradiol-$17{\beta}$ and progesterone (P4) in the human endometrial cancer cell line HTB-1B. ACSL4 mRNA was increased in a dose-dependent manner. Also, expression of ACSL4 gene was up-regulated in a time-dependent manner in HTB-1B cells. However, combined treatment with progesterone and estradiol-$17{\beta}$ modestly decreased the levels of ACS4L mRNA as compared with the estradiol-$17{\beta}$ and progesterone respectively. Overall, these results suggest that the ACSL4 gene is regulated by progesterone and estradiol-$17{\beta}$ in the HTB-1B cells.

• Molecules and Cells
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• 제26권6호
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• pp.554-557
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• 2008

Double-stranded RNA (dsRNA) induces gene silencing in a sequence-specific manner by a process known as RNA interference (RNAi). The RNA-induced silencing complex (RISC) is a multi-subunit ribonucleoprotein complex that plays a key role in RNAi. VIG (Vasa intronic gene) has been identified as a component of Drosophila RISC; however, the role VIG plays in regulating RNAi is poorly understood. Here, we examined the spatial and temporal expression patterns of VIG-1, the C. elegans ortholog of Drosophila VIG, using a vig-1::gfp fusion construct. This construct contains the 908-bp region immediately upstream of vig-1 gene translation initiation site. Analysis by confocal microscopy demonstrated GFP-VIG-1 expression in a number of tissues including the pharynx, body wall muscle, hypodermis, intestine, reproductive system, and nervous system at the larval and adult stages. Furthermore, western blot analysis showed that VIG-1 is present in each developmental stage examined. To investigate regulatory sequences for vig-1 gene expression, we generated constructs containing deletions in the upstream region. It was determined that the GFP expression pattern of a deletion construct (${\Delta}-908$ to -597) was generally similar to that of the non-deletion construct. In contrast, removal of a larger segment (${\Delta}-908$ to -191) resulted in the loss of GFP expression in most cell types. Collectively, these results indicate that the 406-bp upstream region (-596 to -191) contains essential regulatory sequences required for VIG-1 expression.

• 한국수산과학회지
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• 제29권6호
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• pp.770-778
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• 1996

A rapid and economical method of simultaneous extraction of DNA and RNA from seaweeds has been developed by the use of lithium chloride. Lithium chloride facilitates the softening of cell walls resulting in a decrease in both compressive and tensile modulus of elasticity. The DNA was characterized by high molecular weight larger than 27 kb and a relative lack of carbohydrate and protein contamination. The DNA and RNA extracted by the method from many seaweeds were of sufficient quality to be used as a template for per amplification with a plant intergenic gene primer set, for RAPD analysis with arbitrary primers, and for differential display with arbitrary primers in the morphologically distinct regions of the matured Porphyra thallus. The cDNA polymorphism indicated that the reproductive tissue types (male, female, patch) had a relatively high degree of similarity; the vegetative tissue types (dividing, non-dividing) also showed a similar pattern with respect to each other. Holdfast tissue had very low similarity with the other tissues, but appeared most similar to vegetative non-dividing tissue type.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 1998년도 제4차 학술발표대회 및 정기총회
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• pp.50-51
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• 1998

guanidinoacetate methyltransferase (GAMT) catalyzes the last step of creatine biosynthesis in mammals. Creatine plays an important role in cellular energy metabolism in variety of tissues including brain and male reproductive tract. Congenital deficiency of the enzyme leads to a neurologic disorder in humans. We used an interspecific backcross DNA panel to map Gamt to the central region of mouse Chromosome (Chr) 10 near the locus of hesitant mutation affecting male fertility. We assigned the human GAMT gene to Chr 19 by PCR analysis of a human/rodent somatic hybrid cell line DNA panel, and further localized the human gene to Chr 19 at band p13.3 by PCR analysis of a human radiation hybrid DNA panel. Human chr 19p13.3 is homologous to the central part of mouse Chr 10 where mouse Gamt is located. Furthermore, this part of mouse Chr 10 contains mutant loci the phenotype of which is similar to the GAMT deficiency in human.

• Reproductive and Developmental Biology
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• 제30권4호
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• pp.229-234
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• 2006

We have generated transgenic mice that expressed mouse extracellular superoxide dismutase (EC-SOD) in their skin. In particular, the expression plasmid DNA containing human keratin K14 promoter was used to direct the keratinocyte-specific transcription of the transgene. To compare intron-dependent and intron-independent gene expression, we constructed two vectors. The vector B, which contains the rabbit -globin intron 2, was not effective for mouse EC-SOD overexpression. The EC-SOD transcript was detected in the skin, as determined by Northern blot analysis. Furthermore, EC-SOD protein was detected in the skin tissue, as demonstrated by Western blot analysis. To evaluate the expression levels of EC-SOD in various tissues, we purified EC-SOD from the skin, lungs, brain, kidneys, livers, and spleen of transgenic mice and measured its activities. EC-SOD activities in the transgenic mice skin were approximately 7 fold higher than in wild-type mice. These results suggest that the mouse overexpressing vector not only induces keratinocyte-specific expression of EC-SOD, but also expresses successfully functional EC-SOD. Thus, these transgenic mice appeared to be useful for the expression of the EC-SOD gene and subsequent analysis of various skin changes, such as erythema, inflamation, photoaging, and skin tumors.

• Applied Microscopy
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• 제46권2호
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• pp.83-88
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• 2016

Structural differentiation of the connective stalk in giant duckweed, Spirodela polyrhiza, was examined to reveal the anatomical and ultrastructural characteristics within reduced shoot. The study focuses primarily on structural features of the connective stalk (CT), which connect offspring to their mother fronds. Photoautotrophic offspring fronds remained connected by stalks to mother fronds in the reproductive pockets until separation. The CT originated from the meristematic region of the abaxial frond and joined the fronds laterally with two abscission layers. The most notable features of the CT were polymorphic mitochondria, random occurrences of fibrillar structures in intercellular spaces, and great variability in cell wall thickness. Vascular tissues in CTs were highly reduced, demonstrating only a central vascular strand. Grana with 2 to 4 thylakoids and starch grains were found in the chloroplasts. A chlorophyll assay indicated high chlorophyll concentrations in daughter fronds and low concentrations in CTs. The frond and CT, while physically connected to each other, functioned independently. Despite great reduction in S. polyrhiza, the CT has proven to be very efficient for separating offspring from the mother frond, which lends to its capacity for rapid vegetative reproduction. The ultrastructural aspects of CTs in S. polyrhiza were characterized for the first time in this study.

• 대한한방부인과학회지
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• 제18권4호
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• pp.230-241
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• 2005

Uterine myoma is benign tumor formed from muscular and connective tissues, which is one of the most common tumors found in women's reproductive organs. Uterine myoma is the most common pelvic tumor and they are diagnosed in up to15 to 20% of women in pubertal period. Although myoma is generally considered to be a slowly growing tumor, in 20-40% of women at the age of 35 and more have uterine myoma of significant sizes with severe clinical symptoms. The most common symptoms include abdominal pain or discomfort, low abdominal pressure sensation, hypermenorrhea and urinary symptoms. The treatment of uterine myoma is just observation, operation and hormone replacement therapy. We treated 6 outpatients who has uterine myoma and visited Kyungwon InCheon Oriental Hospital with Herb-medication and acupuncture. So we have gotten good result from them. After 6 outpatients had taken Guichulpazing-tang(歸朮破?湯), the symptoms were　improved and the size of the uterine myoma was reduced significantly, It proves　that oriental medical treatment is effective on the patients with uterine myoma, but　continuous studies are needed.

• Reproductive and Developmental Biology
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• 제30권3호
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• pp.163-167
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• 2006

The purpose of the present study was to establish culture conditions for the in vitro study of the neonatal piglet Sertoli cell. Isolation for the culture of Sertoli cell was established using collagenase and pancreatin digestion of testicular tissues. The effects of various culture media, fetal bovine serum(FBS), follicular stimulating hormone(FSH), epidermal growth factor(EGF) and insulin-transferrin-sodium selenite(ITS) on growth of neonatal piglet Sertoli cells were investigated. The mitogenic effects of Dulbecco's modified Eagle's medium+Ham's F-12 medium was higher than other media used in this experiment. The addition of 1% FBS in cultures was necessary for attachment of Sertoli cell clusters. However, except FBS and EGF, FSH and ITS did not stimulate Sertoli cell proliferation. When Sertoli cells isolated from neonatal piglets were cultured in Dulbecco's modified Eagle's medium+Ham's F-12 medium supplemented with 1% FBS, FSH EGF and ITS, the yield and plating efficiency of Sertoli cells were largely increased. Confluency of Sertoli cells was reached as early as 4 days of culture. The method described here reduces or eliminates many of the drawbacks of the conventional procedures used to isolate and culture of Sertoli cells, thus providing a useful tool in studies of growth kinetics and regulation of cell proliferation in vitro.

• 한국동물생명공학회지
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• 제36권1호
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• pp.51-58
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• 2021

Kisspeptin, a neuropeptide and the master controller of reproductive axis upstream to GnRH neurons, and its receptor are also expressed in extra-hypothalamic tissues, such as ovaries. As systemic kisspeptin has been shown to modulate follicular dynamics in cattle, we hypothesized that kisspeptin has direct actions on the ovarian follicular development. We also hypothesized that kisspeptin regulation of primordial follicle development is via modulation of VEGF expression. In order to test these hypotheses, we cultured caprine ovarian cortical strips in vitro for 7 days with supplementation of kisspeptin at 1, 10 and 100 µM concentration and observed the development of primordial follicles into intermediate, primary and secondary follicles. We also studied the alteration in the expression profile of VEGF and VEGF transcript variant 2 mRNA during follicular development in the presence of kisspeptin. We confirmed the presence of GPR54 in goat ovaries in our preliminary studies. Supplementation of kisspeptin at 1 and 10 µM concentration facilitated the development of primordial follicles into intermediate, primary and secondary follicles with less number of degenerated follicles while the same at 100 µM resulted in degeneration of follicles. We observed a drastic increase in the expression profile of VEGF and VEGF transcript variant 2 mRNA upon culture which was independent of kisspeptin treatment. In conclusion, our studies show that kisspeptin facilitates ovarian primordial development in vitro.