• Title/Summary/Keyword: Fluorescence In situ Hybridization

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Analysis of Telomere Length and Telomerase Activity of Tissues in Korean Native Chicken (한국 재래닭의 주령별 각 조직의 텔로미어 함량과 텔로머레이스 활성도 분석)

  • Jung G.S.;Cho E.J.;Choi D.S.;Lee M.J.;Park C.;Jeon I.S.;Sohn S.H.
    • Korean Journal of Poultry Science
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    • v.33 no.2
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    • pp.97-103
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    • 2006
  • Telomeres are essential for chromosome stability and are related with cell senescence, apoptosis and cancer. Even though telomere length and telomerase activity have been studied extensively, very little is known to analyze the telomere dynamics in chicken cells. This study was carried out to analyze the telomere distribution and telomerase activity of Korean Native Chicken cells along with aging. The cells were collected from brain, heart, liver, kidney and germinal tissues during physiological stages. Telomere distribution was analyzed by Quantitative-Fluorescence in situ Hybridization (Q-FISH) techniques using the chicken telomeric DNA probe. Telomerase activity was performed by Telomeric Repeat Amplification Protocol (TRAP) assay. In results, the telomeres of chicken were found at the ends of all chromosomes with the interstitial telomeres on chromosomes 1, 2 and 3. The amount of telomeres on chicken cells was decreased along with aging in most tissues. Furthermore, the telomere quantity was significantly different among tissues. The relative amount of telomeres in proliferous cells such as testis cells had much more than those of liver, brain, heart, blood and kidney cells. The telomerase activity was down-regulated in cells of brain, heart and liver tissues. Whereas gonadal cells showed a constitutive activity of telomerase during all stage of life. In conclusions, the telomere quantity and telomerase activity in chicken are closely relate to cell proliferation and tissue specificity during developmental stages and aging. There is also closely correlated between the amounts of telomeric DNA and telomerase activity in chicken tissues.

Comparison of Stress Response in Diallel Crossed Korean Domestic Chicken Breeds (토종 종계를 이용한 이면 교배조합 계통 간 스트레스 반응정도 비교 분석)

  • Cho, Eun Jung;Park, Ji Ae;Choi, Eun Sik;Sohn, Sea Hwan
    • Korean Journal of Poultry Science
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    • v.43 no.2
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    • pp.77-88
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    • 2016
  • To establish a new synthetic Korean meat chicken breed, we tested $5{\times}5$ diallel cross mating experiment with domestic chicken breeds. Comparing stress responses among diallel crossed chicken breeds, we analyzed telomere length, DNA damage and expressions of heat shock protein genes (HSPs) as the markers of the stress response. The telomere length was measured by quantitative fluorescence in situ hybridization on the nuclei of lymphocytes. The expression levels of HSP-70, $HSP-90{\alpha}$ and $HSP-90{\beta}$ genes were analyzed by quantitative real-time polymerase chain reaction in lymphocytes. The DNA damage rate of lymphocytes was quantified by the comet assay known as the single cell gel electrophoresis. In results, there were significant differences in the values of the stress markers such as telomere length, HSPs and DNA damage rate, and also were significant differences in viabilities and body weights among the $5{\times}5$ diallel crossed chicken breeds. The telomere shortening rate, expression values of HSPs and DNA damage rate were significant low in W and Y crossed chickens compare to the others, but GG pure breed showed the highest values in the 25 crossed chickens. Estimating correlation coefficient, the survival rate positively correlated to telomere length, but negatively correlated to the expression levels of HSP-70, $HSP-90{\alpha}$, $HSP-90{\beta}$ genes and to the value of % DNA in tail as DNA damage rate. The expression levels of HSP-70, $HSP-90{\alpha}$ and $HSP-90{\beta}$ genes of dead chickens had significantly higher than those of survival chickens. According to the results on the stress marker analysis, it would be considered that the crossed breeds had more stress resistant than the pure breeds, and the crossed chickens with a light strain such as W or Y were relatively resistant to stress, but the crossed chickens with a heavy strain such as G, H, F were susceptible to stress.

Comparison of Stress Response between Korean Native Chickens and Single Comb White Leghorns subjected to a High Stocking Density (닭의 품종 간 스트레스 반응 정도 비교 분석)

  • Sohn, Sea Hwan;Cho, Eun Jung;Park, Dhan Bee;Jang, In Surk;Moon, Yang Soo
    • Korean Journal of Poultry Science
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    • v.41 no.2
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    • pp.115-125
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    • 2014
  • With Single Comb White Leghorn (WL) and Korean Native Chicken (KNC) breeds, we compared the stress response with chicken breeds that were subjected to a high stocking density. Stress response was analyzed by the quantity of telomeric DNA, the rate of DNA damage and the expression levels of heat shock proteins (HSPs) and hydroxyl-3-methyl-glutaryl coenzyme A reductase (HMGCR) genes on tissues and blood. The telomere length and telomere shortening rates were analyzed by quantitative fluorescence in situ hybridization on the nuclei of lymphocytes and tissues. The DNA damage rate of lymphocytes was quantified by the comet assay. The expression levels of HSP70, HSP90-${\alpha}$, HSP90-${\beta}$ and HMGCR genes were measured by quantitative real-time polymerase chain reaction in lymphocytes. There was no significant difference between KNC and WL in body weight, weight gain, telomere shortening rate and DNA damage rate. However, the growth rate significantly decreased in chickens raised under high stocking density conditions, as compared to the control group. The telomere-shortening rate, DNA damage and HSPs expression of the lymphocytes were significantly higher in the high stocking density group than the control. The stress condition and breeds had a significant effect on the expressions of HSP70, HSP90-${\alpha}$ and HSP90-${\beta}$ in lymphocytes, except HMGCR. The stress response of WL was higher than that of KNC, as analyzed to the expression of HSP70 and HSP90-${\alpha}$. Therefore, we concluded that the chickens which were exposed to a high stocking density had increased the individual physiological stress response regardless of breeds, and White Leghorns are more susceptible to stress condition than Korean Native Chickens.

Association between Paternal Telomere Length and Offspring Production Performance in Korean Native Chickens (토종 종계 부계의 텔로미어 길이와 자식의 생산능력 간의 연관성 분석)

  • Kim, Bo Gyeong;Sohn, Sea Hwan
    • Korean Journal of Poultry Science
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    • v.49 no.3
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    • pp.145-156
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    • 2022
  • The effect of paternal telomere length on reproductive performance, relationship between paternal and offspring telomere length, and association between paternal telomere length and offspring production performance were investigated in Korean native chickens. Using 22 paternal chickens and 329 offspring, the paternal reproductive performance such as fertility, embryo mortality, and hatchability, as well as the offspring production performance such as survival rate, body weight, and weight gain were analyzed. Telomere length was analyzed through quantitative fluorescence in situ hybridization using lymphocytes. No significant differences were observed in fertility, embryo mortality, and hatchability between paternal chicken telomere lengths (P<0.05). These results indicate that paternal telomere length had a weak negative correlation with fertility and embryonic death rate but a weak positive correlation with hatchability. The correlation coefficient between paternal telomere length and offspring survival rate was r=0.17 (P>0.05). The group of offspring with long paternal telomeres showed relatively poor growth performance. Moreover, a significant negative correlation was estimated between paternal telomere length and offspring growth performance (P<0.05). The correlation coefficient between paternal and offspring telomere lengths was r=0.075 (P>0.05). In conclusion, there was a weak association between paternal telomere length and reproductive performance, offspring survival rate, and offspring telomere length, respectively. However, paternal telomere length and offspring growth performance showed a negative relationship. Such results could be due to the re-extension of telomere length by telomere reprogramming in the early embryonic stage and the different degree of telomere shortening between individuals with increasing age after birth.

CD45RA+ Depleted Lymphocyte Infusion for Treatment of Refractory Cytomegalovirus Disease in Complete DiGeorge Syndrome: A Case Report

  • HyungJin Chin;Young Hye Ryu;Da Yun Kang;Hyun Jin Park;Kyung Taek Hong ;Jung Yoon Choi;Ki Wook Yun;Bongjin Lee;Hyoung Jin Kang;Eun Hwa Choi
    • Pediatric Infection and Vaccine
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    • v.30 no.3
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    • pp.173-179
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    • 2023
  • Complete DiGeorge syndrome (cDGS) refers to DGS with profound T cell deficiency. Herein, we present the case of an infant with cDGS suffering from refractory cytomegalovirus (CMV) infection and who was treated with CD45RA+ depleted lymphocyte infusion. The patient was diagnosed with cDGS by fluorescence in situ hybridization which verified 22q11.2 deletion and as well as by the observed profound T cell deficiency (CD3+ T cells 69/μL, CD4+ T cells 7/μL). On the 45th day of age, CMV viremia was first detected with a plasma viral load (VL) of 120,000 IU/mL. Ganciclovir treatment effectively reduced VL post 56 days of treatment; however, VL subsequently rebounded. A CMV UL97 phosphotransferase M460V mutation conferring ganciclovir resistance emerged and foscarnet was incorporated. Despite this, high titers of CMV viremia (VL 2,820,000 IU/mL) and CMV retinitis were complicated. To restore T cell immunity and treat refractory CMV infection, CD45RA+ depleted CMV-specific lymphocytes from the patient's father were infused twice on the 196th and 207th days after birth. After receiving the second infusion, a decline in CMV VL was observed, with a decrease to 87,100 IU/mL by the tenth day following infusion, despite the failure in maintaining T cell increase. The patient died of Pneumocystis jirovecii pneumonia and Elizabethkingia meningoseptica sepsis on the 222nd day after birth. CD45RA+ depleted lymphocyte infusion may be a therapeutic option for refractory CMV disease in cDGS patients.