• Animal Bioscience
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• 제36권6호
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• pp.829-839
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• 2023

Objective: The aim of this study was to clone the mRNA sequence of the Acyl-CoA dehydrogenase long chain (ACADL) gene of goats and explore the effect of ACADL on the differentiation of subcutaneous fat cells on this basis. Methods: We obtained the ACADL gene of goats by cloning and used quantitative real-time polymerase chain reaction (qPCR) to detect the ACADL expression patterns of different goat tissues and subcutaneous fat cells at different lipid induction stages. In addition, we transfect intramuscular and subcutaneous adipocytes separately by constructing overexpressed ACADL vectors and synthesizing Si-ACADL; finally, we observed the changes in oil red stained cell levels under the microscope, and qPCR detected changes in mRNA levels. Results: The results showed goat ACADL gene expressed in sebum fat. During adipocyte differentiation, ACADL gradually increased from 0 to 24 h of culture, and decreased. Overexpression of ACADL promoted differentiation of subcutaneous adipocytes in goat and inhibited their differentiation after interference. Conclusion: So, we infer ACADL may have an important role in positive regulating the differentiation process in goat subcutaneous adipocytes. This study will provide basic data for further study of the role of ACADL in goat subcutaneous adipocyte differentiation and lays the foundation for final elucidating of its molecular mechanisms in regulating subcutaneous fat deposition in goats.

• Asian-Australasian Journal of Animal Sciences
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• 제13권2호
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• pp.155-160
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• 2000

In order to understand the effects of sex or age on cellular characteristics of adipocytes from Hanwoo and sheep, samples were obtained from omental, subcutaneous, intermuscular and intramuscular adipose tissue depots of bulls, steers, heifers and cows in Hanwoo, and perirenal, omental and subcutaneous adipose tissues of fetal lambs, suckling lambs and wethers in sheep. In case of Hanwoo, mean diameter, surface area and volume of adipocytes from each depot were obtained by multisizer II (Coulter Co., UK). Osmium-fixed adipocytes were sized and counted using $560{\mu}m$ aperture. For samples obtained from sheep, cellularity was measured by using microscope and MCV program of Texas Instrument. Bulls had less subcutaneous and kidney fat than steers even though their slaughter and carcass weight were heavier. The amounts of fat from cows were greater in subcutaneous, kidney and internal organs than heifers. Steers had larger adipocytes in subcutaneous, intermuscular and intramuscular adipose tissues than bulls, although the differences were significant only for the subcutaneous adipose tissue depots. Adipocytes appeared to be largest in omental and smallest in intramuscular adipose tissue, although there were no significant differences among tissues. In a comparison of heifers and cows, significant site effects (p<0.05) were shown in adipocyte diameter, surface area and volume, and adipocyte appeared to be largest in omental tissue. Statistical difference (p<0.05) was only shown in cell volume of intramuscular tissue which was higher in cow than heifer. Intramuscular adipose tissue tended to have relatively greater numbers of cells per gram tissue and reflect lesser maturity of intramuscular adipose tissue relative to other adipose tissues. In sheep, regardless of adipose tissue depots, wethers had the greater adipocyte diameters than those at any other growth stage of sheep. Within adipose depots, the ranking of cell size was the greatest in the omental tissue of wether and the lowest in the renal and subcutaneous adipose tissue depots of fetal lamb. The cell size of adipocyte became larger with age, especially from fetal to suckling lamb due to a rapid hypertrophy of both perirenal and subcutaneous adipocytes during the suckling period.

• Asian-Australasian Journal of Animal Sciences
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• 제17권5호
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• pp.588-593
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• 2004

The gene expression of porcine adiponectin and stearoyl coenzyme A desaturase (SCD) was investigated in this study. The partial gene sequences for adiponectin and SCD were amplified by RT-PCR from subcutaneous adipose tissue and cloned by TA cloning techniques. Sequences of these genes were determined and found to be highly homologous to that of other species, suggesting similar function of these genes as in other species. The transcripts of these adipocyte-related genes in pig tissues were measured by Northern analysis. The transcripts for adiponectin and SCD were highly expressed in porcine subcutaneous adipose tissue; the transcripts for SCD were also barely detected in the liver, but the greatest concentrations were in the adipose tissue. In porcine stromalvascular cells (S/V cells) cultured in vitro, transcripts for adiponectin and SCD increased gradually during adipocyte differentiation. The level of adipocyte adiponectin mRNA was associated with late adipocyte differentiation, indicating the gene may not be involved in adipocyte differentiation but has great importance in porcine adipocyte functions. The SCD transcripts were not detectable until 2 d after induction of adipocyte differentiation. It was highly expressed in differentiating porcine adipocytes (2 to 10 d after the induction of adipocyte differentiation), indicating a significant role of SCD in adipocytes.

• 농업생명과학연구
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• 제46권5호
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• pp.47-55
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• 2012

본 연구는 가축 체지방 감소를 위한 수동면역학적 기법의 기초자료 확보를 위해 흰쥐 지방부위별(복강 및 피하지방) 특이 다클론 항체를 개발하고자 실시되었다. 흰쥐 부위별 지방 조직에서 지방세포를 분리하고 각각 primary 배양시킨 후 개발된 복강지방(AAb) 및 피하지방 특이 다클론 항체(SAb)를 주입한 뒤 media 내 lactate dehydrogenase(LDH) 방출 수준을 조사하였다. 희석배율 1:1,000배를 기준으로 비면역 혈청은 항원-항체 결합 반응이 거의 측정되지 않았고, AAb 및 SAb는 희석배율 1:128,000배까지 각각 항원-항체 반응이 감지되었다. 부위별 지방과 비교했을 때, AAb 및 SAb는 흰쥐 타 장기들과는 특이한 반응을 나타내지 않았다(p<0.001). 본 연구에서 개발한 두 항체들은 모두 항원으로 이용된 부위의 지방세포 PMP와 가장 높은 반응을 나타내었으며, 지방세포 배양을 통한 LDH 수준 검사에서는 AAb 및 SAb 모두 비면역혈청에 비해 유의적으로 높은 세포파괴가 일어났음을 확인할 수 있었다(p<0.01). 이상의 결과를 종합할 때 본 연구에서 개발된 AAb 및 SAb는 높은 역가, 타장기 안전성 및 in vitro 지방 감소 효과가 있었다고 생각된다.

• 한국축산식품학회지
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• 제28권5호
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• pp.651-659
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• 2008

본 연구는 국내 고유종인 한우의 부위별 불가식 체지방 감소를 위한 특이 다클론 항체의 개발 및 타장기 안전성을 확인하고자 실시되었다. Collagenase digestion 방법으로 한우의 복강 및 피하지방세포 원형질막 단백질을 분리하여 면양에 3회에 걸쳐 수동면역 주사하고, 면역 주사 전 및 후에 비면역혈청과 항혈청(항체)을 생산하였다. 생산된 한우 부위별 지방 항체의 역가와 한우의 주요 장기 조직인 심장, 신장, 간장, 폐, 근육 및 비장세포의 원형질막 단백질에 대한 타장기 교차반응성과 한우의 부위별 지방 조직에서 지방세포를 분리하고 각각 배양시킨 후 개발된 한우 지방 항체를 직접 주입한 뒤 LDH 수준을 조사하였다. 복강 및 피하지방세포 원형질막 단백질들은 서로 유사하면서도 특이적인 단백질을 가지고 있는 것으로 SDS-PAGE 분석을 통해 확인할 수 있었다. 희석배율 1:1,000배를 기준으로 비 면역혈청은 항원-항체 결합 반응이 거의 측정되지 않았으나, 복강 및 피하지방 항체는 희석배율 1:128,000배 및 1:64,000배까지 각각 항원-항체 반응이 감지되었으며, 이는 본 연구에서 생산한 부위별 지방 특이 다클론 항체가 지방세포 원형질막 단백질에 대해 매우 강한 역가를 가진 항체임을 시사한다. 또한 복강 및 피하지방 항체는 타 장기들과는 특이한 반응을 나타내지 않았다. 본 연구에서 개발한 두 항체들은 모두 항원으로 이용된 부위의 지방세포 원형질막 단백질과 가장 높은 반응을 나타내었으며, 복강 및 피하지방특이항체는 비 면역혈청에 비해 유의적으로 높은 세포독성 효과가 있음을 확인할 수 있었다. 이상의 결과를 종합할 때 본 연구에서 개발된 복강 및 피하 지방 감소 다클론 항체는 높은 역가, 타 장기 안전성 및 세포 파괴 효과가 있었으며 생체 타장기 안전성 등 향 후 기존의 전체 지방에서 생산한 항체의 단점을 보완할 수 있는 연구가 지속될 경우 불가식 체지방이 감소된 저지방 한우 고급육 생산이 가능하리라고 사료된다.

• Journal of Ginseng Research
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• 제43권4호
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• pp.589-599
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• 2019

Background: Panax ginseng Meyer is known as a conventional herbal medicine, and ginsenoside Rg1, a steroid glycoside, is one of its components. Although Rg1 has been proved to have an antiobesity effect, the mechanism of this effect and whether it involves adipose browning have not been elucidated. Methods: 3T3-L1 and subcutaneous white adipocytes from mice were used to access the thermogenic effect of Rg1. Adipose mitochondria and uncoupling protein 1 (UCP1) expression were analyzed by immunofluorescence. Protein level and mRNA of UCP1 were also evaluated by Western blotting and realtime polymerase chain reaction, respectively. Results: Rg1 dramatically enhanced expression of brown adipocyte-especific markers, such as UCP1 and fatty acid oxidation genes, including carnitine palmitoyltransferase 1. In addition, it modulated lipid metabolism, activated 5' adenosine monophosphate (AMP)-activated protein kinase, and promoted lipid droplet dispersion. Conclusions: Rg1 increases UCP1 expression and mitochondrial biogenesis in 3T3-L1 and subcutaneous white adipose cells isolated from C57BL/6 mice. We suggest that Rg1 exerts its antiobesity effects by promoting adipocyte browning through activation of the AMP-activated protein kinase pathway.

• 한국축산식품학회지
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• 제30권1호
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• pp.87-94
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• 2010

본 연구는 저지방 고품질 돈육의 생산을 위해 돼지 복강 및 피하지방 감소를 위한 다클론 항체의 개발 및 실제 세포에 미치는 세포독성효과를 확인하고자 실시되었다. 항원으로써 돼지의 복강 및 피하지방세포 PMP를 분리하여 면양에 3회에 걸쳐 수동면역 주사하고, 면역 주사 전 및 후에 비면역혈청과 항체를 생산하였다. 돼지 AAb 및 SAb 항체의 역가와 생체 주요 장기 조직인 심장, 신장, 간장, 폐, 근육 및 비장세포의 PMP에 대한 타장기 교차반응성을 조사하였다. 돼지 부위별 지방 조직에서 지방세포를 분리하고 각각 primary 배양시킨 후 개발된 AAb 및 SAb를 주입한 뒤 media 내 LDH 방출 수준을 조사하였다. 희석배율 1:1,000배를 기준으로 비면역혈청은 항원-항체 결합반응이 거의 측정되지 않았고, AAb 및 SAb는 희석배율 1:16,000배까지 각각 항원-항체 반응이 감지되었으며, 이는 본 연구에서 생산한 부위별 지방 감소 다클론 항체가 지방세포 PMP에 대해 강한 역가를 가진 항체임을 시사한다. 또한 AAb 및 SAb는 타 장기들과는 특이한 반응을 나타내지 않았다. 본 연구에서 개발한 두 항체들은 모두 항원으로 이용된 부위의 지방세포 PMP와 가장 높은 반응을 나타내었으며, 비면역혈청에 비해 유의적으로 높은 세포독성 효과가 있음을 확인할 수 있었다. 이상의 결과를 종합할 때 본 연구에서 개발된 AAb 및 SAb는 높은 역가, 타 장기 안전성 및 in vitro 지방 감소 효과가 있었으며 향후 영양생리 안전성 및 생체 지방 감소 효과 등의 추가 연구가 지속될 경우 저지방 고품질 돈육의 생산이 가능하리라고 판단된다.

• Animal Bioscience
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• 제36권1호
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• pp.144-155
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• 2023

Objective: Adipocyte differentiation is regulated by a variety of functional genes and noncoding RNAs. However, the role of miRNAs in lipid deposition of goat white adipose tissue is still unclear. Therefore, this study revealed the miRNA expression profile in goat subcutaneous adipocytes by sRNA-seq. Methods: The miRNA expressed in goat subcutaneous preadipocytes and the mature adipocytes were sequenced by sRNA-seq. The differentially expressed miRNAs (DEm) were screened and gene ontology (GO) and Kyoto encyclopedia for genes and genomes (KEGG) analyses were performed. Gain-of-function and loss-of-function combined with oil red O staining, Bodipy staining, and quantitative reverse-transcription polymerase chain reaction (qPCR) were utilized to determine the effect of miR-133a-3p on adipocyte differentiation. Results: A total of 218 DEm were screened out. The target genes of these DEm were significantly enriched in GO items such as biological regulation and in KEGG terms such as FAK signaling pathway and MAPK signaling pathway. qPCR verified that the expression trend of miRNA was consistent with miRNA-seq. The gain-of-function or loss-of-function of miR-133a-3p showed that it promoted or inhibited the accumulation of lipid droplets, and CCAAT enhancer binding protein α (C/EBPα) and C/EBPβ were extremely significantly up-regulated or down-regulated respectively (p<0.01), the loss-of-function also led to a significant down-regulation of peroxisome proliferator activated receptor gamma (PPARγ) (p<0.01). Conclusion: This study successfully identified miRNAs expression patterns in goat subcutaneous adipocytes, and functional identification indicates that miR-133a-3p is a positive regulator of the differentiation process of goat subcutaneous adipocytes. Our results lay the foundation for the molecular mechanism of lipid deposition in meat-source goats from the perspective of miRNA.

• Animal cells and systems
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• 제13권4호
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• pp.371-379
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• 2009

The endocannabinoid system (ECS) plays a key role in the regulation of appetite, body weight and metabolism. We undertook the present study to further clarify the regulation of the cannabinoid CB1 receptor (CB1, CNR1) in human adipose tissue in obesity. CB1 receptor mRNA expression was ~1.6-fold (p<0.004) and 1.9-fold higher (P<0.05) in subcutaneous adipocytes from obese compared to non-obese subjects in microarray and quantitative real-time PCR studies, respectively. Higher CB1 receptor mRNA expression levels in both adipose tissue (~1.2 fold, P<0.05) and adipocytes (~2 fold, P<0.01) were observed in samples from visceral compared to subcutaneous depots collected from 22 obese individuals. Immunofluorescence confocal microscopy demonstrated the presence of CB1 receptor on adipocytes and also adipose tissue macrophages. These data indicate that adipocyte CB1 receptor is up-regulated in human obesity and visceral adipose tissue and also suggest a potential role for the ECS in modulating immune/inflammation as well as fat metabolism in adipose tissue.

• Journal of Ginseng Research
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• 제40권2호
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• pp.141-150
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• 2016

Background: Adipocyte-macrophage communication plays a critical role regulating white adipose tissue (WAT) inflammatory gene expression. Because WAT inflammation contributes to the development of metabolic diseases, there is significant interest in understanding how exogenous compounds regulate the adipocyte-macrophage crosstalk. An aqueous (AQ) extract of North American (NA) ginseng (Panax quinquefolius) was previously shown to have strong inflammo-regulatory properties in adipocytes. This study examined whether different ginseng extracts influence adipocyte-macrophage crosstalk, as well as WAT inflammatory gene expression. Methods: The effects of AQ and ethanol (EtOH) ginseng extracts ($5{\mu}g/mL$) on adipocyte and macrophage inflammatory gene expression were studied in 3T3-L1 and RAW264.7 cells, respectively, using real-time reverse transcription polymerase chain reaction. Adipose tissue organ culture was also used to examine the effects of ginseng extracts on epididymal WAT (EWAT) and inguinal subcutaneous WAT (SWAT) inflammatory gene expression. Results: The AQ extract caused significant increases in the expression of common inflammatory genes (e.g., Mcp1, Ccl5, Tnf-${\alpha}$, Nos2) in both cell types. Culturing adipocytes in media from macrophages treated with the AQ extract, and vice versa, also induced inflammatory gene expression. Adipocyte Ppar-${\gamma}$ expression was reduced with the AQ extract. The AQ extract strongly induced inflammatory gene expression in EWAT, but not in SWAT. The EtOH extract had no effect on inflammatory gene expression in either both cell types or WAT. Conclusion: These findings provide important new insights into the inflammo-regulatory role of NA ginseng in WAT.