• Asian-Australasian Journal of Animal Sciences
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• 제18권11호
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• pp.1519-1523
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• 2005

By screening a subtracted cDNA library constructed with mRNA obtained from the longissimus dorsi muscles of F1 hybrids Landrace${\times}$Yorkshire and their Yorkshire female parents, we isolated two partial sequences coding for the H3-K4-specific methyltransferase (KIAA1717) and skeletal muscle myosin regulatory light chain (HUMMLC2B) genes. In the present work we investigated two SNPs, one (C1354T) at the 3' untranslated region (UTR) of KIAA1717 and one (A345G) at the SINE (PRE-1) element of HUMMLC2B, in a resource population derived from crossing Chinese Meishan and Large White pig. The selected pigs were genotyped by means of a PCR-RFLP protocol. Significant associations were observed for the KIAA1717 C1354T polymorphic site with thorax-waist backfat thickness (p<0.05), buttock backfat thickness (p<0.05), average backfat thickness (p<0.05), loin eye height (p<0.05), loin eye area (p<0.05), carcass length to 1$^{st}$ spondyle (p<0.01) and carcass length to 1st rib (p<0.01). HUMMLC2B A345G were significantly associated with loin eye width (p<0.05), loin eye area (p<0.05). Further studies are needed to confirm these preliminary results.

• The Korean Journal of Physiology and Pharmacology
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• 제21권1호
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• pp.1-9
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• 2017

Intestinal disorders often co-occur with inflammation and dysmotility. However, drugs which simultaneously improve intestinal inflammation and co-occurring dysmotility are rarely reported. Atractylodin, a widely used herbal medicine, is used to treat digestive disorders. The present study was designed to characterize the effects of atractylodin on amelioration of both jejunal inflammation and the co-occurring dysmotility in both constipation-prominent (CP) and diarrhea-prominent (DP) rats. The results indicated that atractylodin reduced proinflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 in the plasma and inhibited the expression of inflammatory mediators iNOS and NF-kappa B in jejunal segments in both CP and DP rats. The results indicated that atractylodin exerted stimulatory effects and inhibitory effects on the contractility of jejunal segments isolated from CP and DP rats respectively, showing a contractile-state-dependent regulation. Atractylodin-induced contractile-state-dependent regulation was also observed by using rat jejunal segments in low and high contractile states respectively (5 pairs of low/high contractile states). Atractylodin up-regulated the decreased phosphorylation of 20 kDa myosin light chain, protein contents of myosin light chain kinase (MLCK), and MLCK mRNA expression in jejunal segments of CP rats and down-regulated those increased parameters in DP rats. Taken together, atractylodin alleviated rat jejunal inflammation and exerted contractile-state-dependent regulation on the contractility of jejunal segments isolated from CP and DP rats respectively, suggesting the potential clinical implication for ameliorating intestinal inflammation and co-occurring dysmotility.

• Journal of Photoscience
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• 제4권2호
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• pp.35-40
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• 1997

The sensory transduction processes of blue light in guard cells have been suggested the involvement of Ca$^{2+}$/calmodulin-dependent myosin light chain kinase (MLCK) or MLCK-like proteins. The source of Ca$^{2+}$ required for the signal transduction process was investigated in guard cell protoplasts (GCPs). The GCPs showed the typical H$^+$ pumping activity by blue light (200 $\mu$mol m$^{-2}$ s$^{-1}$) and fusicoccin (10 $\mu$M) under background red light (600 $\mu$mol m$^{-2}$ s$^{-1}$). The blue light-dependent H$^+$ pumping was not significantly affected by the externally changed Ca$^{2+}$ concentrations. The addition of 1 mM Ca$^{2+}$ in the bathing medium ratherly inhibited the H$^+$ pumping. In contrast, the blue light-dependent H$^+$ pumping was inhibited by caffeine and 2,5-di-(tert-butyl)-1,4-benzohydroquinone (BHQ), inhibitor of C$^{2+}$-ATPase in endoplasmic reticulum (ER) without inhibiting the H $^+$ pump. The inhibition by caffeine and BHQ was fully reversible. The extent of inhibition by caffeine and BHQ was larger when they were added together than when added separately. The results suggest that Ca$^{2+}$ required for the blue light-dependent H$^+$ pumping may be released from the intracellular Ca$^{2+}$ stores, probably ER in guard cells.

• Fisheries and Aquatic Sciences
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• 제17권4호
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• pp.479-486
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• 2014

To evaluate their potential utility as an ornamental organism, novel transgenic marine medaka Oryzias dancena strains with a highly vivid fluorescent phenotype were established through transgenesis of a cyan fluorescent protein gene (cfp) driven by the endogenous fast skeletal myosin light chain 2 gene (mlc2f) promoter. The transgenic marine medaka strains possessed multiple copies of transgene integrants and passed their fluorescent transgenes successfully to subsequent generations. Transgenic expression in skeletal muscles at both the mRNA and phenotypic levels was, overall, dependent upon transgene copy numbers. In the external phenotype, an authentic fluorescent color was dominant in the skeletal muscles of the transgenic fish and clearly visible to the unaided eye. The phenotypic fluorescent color presented differentially in response to different light-irradiation sources; the transgenics displayed a yellow-green color under normal daylight or white room light conditions, a strong green-glowing fluorescence under ultraviolet light, and a cyan-like fluorescence under blue light from a light-emitting diode.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.100-100
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• 2003

Pluripotent embryonic stem (ES) cells differentiate spontaneously into beating cardiomyocytes via embryo-like aggregates. We describe the use of mouse embryonic stem (mES03) cells as a reproducible differentiation system for cardiomyocyte. To induce cardiomyocytic differentiation, mES03 cells were dissociated and allowed to aggregate (EB formation) at the presence of 0 75% dimethyl sulfoxide (DMSO) for 4 days and then another 4 days without DMSO (4+/4-). Thus treated EBs were plated onto gelatin-coated dish for differentiation. Spontaneously contracting colonies which appeared in approximately 4-5 days upon differentiation. Expression of cardiac-specific genes were determined by RT-PCR. Rebust expression of myosin light chain (MLC-2V), cardiac myosin heavy chain $\alpha$, cardiac muscle heavy polypeptide 7 $\beta(\beta$-MHC), cardiac transcription factor GATA4 and skeletal muscle-specific ${\alpha}_1$-subunit of the L-type calcium channel (${\alpha}_1 CaCh_{sm}$) were detected as early as 8 days after EB formation, but message of cardiac muscle-specific $\alpha$$_1$-subunit of the L-type calcium channel (${\alpha}_1$CaCh) were revealed at a low level. Strikingly, the expression of atrial natriuretic factor (ANF) was not detected. When spontaneous contracting cell masses were examined their electrophysiological features by patch-clamp technique, it showed ventricle-like action potential 17 days after the EB formation. This study indicates that mES03 cell-derived cardiomyocytes displayed biochemical and electrophysiological properties of cardiomyocytes and DMSO enhanced development of cardiomyocytes in 4+/4- method.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.76-76
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• 2003

Pluripotent embryonic stem cells can differentiate into beating cardiomyocytes with proper culture conditions and stimulants via embryo-like aggregates. We describe here the use of mouse embryonic stem (mES03) cells as a reproducible differentiation system for cardiomyocyte. mES03 cells growing in colonies were dissociated and allowed to re-aggregated in suspension [embryoid body (EB) formation〕. To induce cardiomyocytic differentiation, cells were exposed to 0.75% dimethyl sulfoxide (DMSO) during EB formation for 4 days and then another 4 days without DMSO (4+/4-). Thus treated EB was plated onto gelatin-coated dishes for differentiation. Spontaneously contracting colonies which appeared in approximately 4~5 days upon differentiation were mechanically dissected, enzymatically dispersed, plated onto coverslips, and then incubated for another 48~72 hrs. By RT-PCR, robust expression of cardiac myosin heavy chain $\alpha$, cardiac muscle heavy polypeptide 7 $\beta$($\beta$-MHC), cardiac transcription factor GATA4, and skeletal muscle-specific $\alpha$$_1$-subunit of the L-type calcium channel ($\alpha$$_1$CaC $h_{sm}$ ) were detected as early as 8 days after EB formation, but message of cardiac muscle-specific $\alpha$$_1$-subunit of the L-type calcium channel ($\alpha$$_1$CaCh) were reveled at a low level. In contrast, expression of myosin light chain (MLC-2V) and atrial natriuretic factor (ANF) were not detected during EB formation for 8 days. However, a strong expression of the atrial-specific ANF gene was expressed from day 8 onward, which were remained constant in EB. (cardiac specialization and terminal differentiation stage). Electrophysiological examination of spontaneously contracting cells showed ventricle-like action potential 17 days after the EB formation. This study indicates that mES03 cell-derived cardiomyocytes via 4+/4- protocol displayed biochemical and electrophysiological properties of subpopulation of cardiomyocytes.

• 대한전기학회논문지:시스템및제어부문D
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• 제55권10호
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• pp.458-463
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• 2006

The purpose of this work is to investigate the fundamental properties of the gradual muscle force potentiation. We investigated the dependence of force potentiation on both the pulse-amplitude and the pulse-duration with different ramp-up time. The experimental results showed that the force increment ratio (FIR) during constant electrical stimulation decreased with pulse-amplitude and also with pulse-duration. The FIR was greater with short ramp-up time in both the pulse-amplitude and pulse-width modulation. The feasible mechanism might be that the myosin light chain phosphorylation induces the force potentiation and it occurs only in the fast type muscle fibers which are recruited first. These observations indicate that muscle potentiation must be understood well for the accurate control of muscle force.

• Journal of Periodontal and Implant Science
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• 제34권1호
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• pp.205-221
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• 2004

Periodontal ligament(PDL) cells have been known as playing an important roles in periodontal regeneration and gingival fibroblasts are also important to periodontal regeneration by forming connective tissue attachment. There were rare studies about the gene expression patterns of PDL cells and gingival fibroblasts, therefore in this study, we tried cDNA microarray-based gene expression monitoring to explain the functional differences of PDL cells and gingival fibroblasts in vivo and to confirm the characteristics of PDL cells. Total RNA were extracted from PDL cells and gingival fibroblasts of same person and same passages, and mRNA were isolated from the total RNA using Oligotex mRNA midi kit(Qiagen) and then fluorescent cDNA probe were prepared. And microarray hybridization were performed. The gene expression patterns of PDL cells and gingival fibroblasts were quite different. About 400 genes were expressed more highly in the PDL cells than gingival fibroblasts and about 300 genes were more highly expressed in the gingival fibroblasts than PDL cells. Compared growth factor- and growth factor receptor-related gene expression patterns of PDL cells with gingival fibroblasts, IGF-2, IGF-2 associated protein, nerve growth factor, placental bone morphogenic protein, neuron-specific growth- associated protein, FGF receptor, EGF receptor-related gene and PDGF receptor were more highly expressed in the PDL cells than gingival fibroblasts. The results of collagen gene expression patterns showed that collagen type I, type III, type VI and type VII were more highly expressed in the PDL cells than gingival fibroblasts, and in the gingival fibroblasts collagen type V, XII were more highly expressed than PDL cells. The results of osteoblast-related gene expression patterns showed that osteoblast specific cysteine-rich protein were more highly expressed in the PDL cells than gingival fibroblasts. The results of cytoskeletal proteins gene expression patterns showed that a-smooth muscle actin, actin binding protein, smooth muscle myosin heavy chain homolog and myosin light chain were more highly expressed in the PDL cells than gingival fibrobalsts, and ${\beta}-actin$, actin-capping protein(${\beta}$ subunit), actin- related protein Arp3(ARP) and myosin class I(myh-1c) were more highly expressed in the gingival fibroblasts than PDL cells. Osteoprotegerin/osteoclastogenesis inhibitory factor(OPG/OCIF) was more highly expressed in the PDL cells than gingival fibroblasts. According to the results of this study, PDL cells and gingival fibroblasts were quite different gene expression patterns though they are the fibroblast which have similar shape. Therefore PDL cells & gingival fibroblasts are heterogeneous populations which represent distinct characteristics. If more studies about genes that were differently expressed in each PDL cells & gingival fibroblasts would be performed in the future, it would be expected that the characteristics of PDL cells would be more clear.

• The Korean Journal of Physiology and Pharmacology
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• 제5권4호
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• pp.287-297
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• 2001

Contraction of smooth muscle is initiated by an increase in cytosolic $Ca^{2+}$ leading to activation of $Ca^{2+}$/ calmodulin-dependnet myosin light chain (MLC) kinase and phosphorylation of MLC. The types of contraction and signaling mechanisms mediating contraction differ depending on the region. The involvement of these different mechanisms varies depending on the source of $Ca^{2+}$ and the kinetic of $Ca^{2+}$ mobilization. $Ca^{2+}$ mobilizing agonists stimulate different phospholipases $(PLC-{\beta},\;PLD\;and\;PLA_2)$ to generate one or more $Ca^{2+}$ mobilizing messengers $(IP_3\;and\;AA),$ and diacylglycerol (DAG), an activator of protein kinase C (PKC). The relative contributions of $PLC-{\beta},\;PLA_2$ and PLD to generate second messengers vary greatly between cells and types of contraction. In smooth muscle cell derived form the circular muscle layer of the intestine, preferential hydrolysis of $PIP_2$ and generation of $IP_3$ and $IP_3-dependent\;Ca^{2+}$ release initiate the contraction. In smooth muscle cells derived from longitudinal muscle layer of the intestine, preferential hydrolysis of PC by PLA2, generation of AA and AA-mediated $Ca^{2+}$ influx, cADP ribose formation and $Ca^{2+}-induced\;Ca^{2+}$ release initiate the contraction. Sustained contraction, however, in both cell types is mediated by $Ca^{2+}-independent$ mechanism involving activation of $PKC-{\varepsilon}$ by DAG derived form PLD. A functional linkage between $G_{13},$ RhoA, ROCK, $PKC-{\varepsilon},$ CPI-17 and MLC phosphorylation in sustained contraction has been implicated. Contraction of normal esophageal circular muscle (ESO) in response to acetylcholine (ACh) is linked to $M_2$ muscarinic receptors activating at least three intracellular phospholipases, i.e. phosphatidylcholine-specific phospholipase C (PC-PLC), phospholipase D (PLD) and the high molecular weight (85 kDa) cytosolic phospholipase $A_2\;(cPLA_2)$ to induce phosphatidylcholine (PC) metabolism, production of diacylglycerol (DAG) and arachidonic acid (AA), resulting in activation of a protein kinase C (PKC)-dependent pathway. In contrast, lower esophageal sphincter (LES) contraction induced by maximally effective doses of ACh is mediated by muscarinic $M_3$ receptors, linked to pertussis toxin-insensitive GTP-binding proteins of the $G_{q/11}$ type. They activate phospholipase C, which hydrolyzes phosphatidylinositol bisphosphate $(PIP_2),$ producing inositol 1, 4, 5-trisphosphate $(IP_3)$ and DAG. $IP_3$ causes release of intracellular $Ca^{2+}$ and formation of a $Ca^{2+}$-calmodulin complex, resulting in activation of myosin light chain kinase and contraction through a calmodulin-dependent pathway.

• 한국수산과학회지
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• 제14권1호
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• pp.15-23
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• 1981

말쥐치 등육 골낙근을 $0^{\circ}C$로 유지하면서 사후경과중 선도의 변화에 따라 단백질의 조성과 단백질의 아미노산 조성 및 유리아미노산의 조성 등을 분석 검토하였다. 말쥐치의 육은 약 $20\%$의 단백질을 함유하고 있었으며, 이 단백질은 근원질단백질 $31\%$, 근원섬유단백질 $55\%$, 세포내잔사단백질 $10\%$, 그리고 기질단백질 $4\%$로 구성되어 있었다. 근원질단백질과 근원섬유단백질은 사후선도의 변화와 더불어 감소하여 갔고, 세포내잔사단백질은 상대적으로 증가하는 관계를 보였다. 근원섬유단백질과 세포내잔사단백질은 전기영동과 densitometry에 의하여 분석한 결과, 근원섬유단백질의 약 $70\%$가 액틴과 미요신, 약 $20\%$가 조절단백질, 그리고 $10\%$전후의 미지단백질로 구성되어 있었다. 세포내잔사단백질은 그 대부분이 근원섬유단백질을 구성하는 단백질로 이루어져 있었다. 한편, 사후경과와 더불어 근원섬유단백질을 구성하는 단백질중 트로포닌-T, 트로포닌-C 및 미요신의 light chain 2는 각각 감소하는 추세를 보였다. 죽인 직후의 말쥐치 육단백질의 아미노산 조성은 다른 어육의 그것과 비슷하였으나, 트?토판과 함황아미노산은 보다 부족하였다. 휘발성 염기질소양에비추어 부패초기에 해당하는 사후 18일이 경과했을 때는 프롤린과 시스테인이 두드러지게 줄었으며, 로이신과 이소로이신 및 페닐알라닌은 조금 증가하였다. 말쥐치육의 유리아미노산중에는 타우린이 특히 많이 함유되어 있었으며, 알라닌과 글리신 및 리신도 비교적 많았다. 사후 18일이 경과했을 때는 타우린, 히스티딘, 발린 및 메치오닌은 증가하였으나 리신, 아르기닌, 아스파르트산, 스레오닌, 세린, 로이신 및 이소로이신은 반대로 감소하였다.