Perilipin Gene Research Articles

The somatic genome of Eptatretus okinoseanus reveals the adaptation to deep-sea oligotrophic environment

BackgroundHagfishes are fascinating creatures that typically inhabit the deep sea. The deep sea is characterized by its lack of sunlight, primary productivity, and diminishing biomass with increasing ocean depth. Therefore, hagfishes living in this environment must develop effective survival strategies to adapt to the limited food supply. Deep-sea hagfishes have been observed to survive without food intake for up to one year. In this study, we have assembled a high-quality somatic genome of the deep-sea hagfish (Eptatretus okinoseanus) captured below 1,000 m. We compared the genome of E. okinoseanus with the genomes of inshore hagfish, lampreys, and other related species to investigate the genetic factors underlying the deep-sea hagfish adaptations to the environment.ResultsThe E. okinoseanus somatic genome was estimated to be 1.89 Gb and assembled into 17 pseudochromosomes. Phylogenetic analysis showed that shallow-sea and deep-sea hagfishes diverged approximately 58.8 million years ago. We found Perilipin gene family was significantly expanded in deep sea E. okinoseanus, which promotes triacylglycerol storage. Furthermore, a series of genes involved in fatty acid synthesis and metabolism, blood glucose regulation, and metabolic rate regulation were also expanded, rapid evolution or positive selection, and these changes contribute to their efficiency in energy utilization. Among these genes, the positively selected gene JNK may play an important role in energy metabolism. In addition, the olfactory receptors of the deep-sea hagfish were significantly expanded to 86, and three conserved motifs present only in hagfishes olfactory receptors were identified, which may facilitate the rapid localization of carrion.ConclusionsThis study provides valuable genomic resources for insights into the survival strategies of deep-sea hagfishes in oligotrophic environments.

Cancer radioresistance is characterized by a differential lipid droplet content along the cell cycle

BackgroundCancer radiation treatments have seen substantial advancements, yet the biomolecular mechanisms underlying cancer cell radioresistance continue to elude full understanding. The effectiveness of radiation on cancer is hindered by various factors, such as oxygen concentrations within tumors, cells’ ability to repair DNA damage and metabolic changes. Moreover, the initial and radiation-induced cell cycle profiles can significantly influence radiotherapy responses as radiation sensitivity fluctuates across different cell cycle stages. Given this evidence and our prior studies establishing a correlation between cancer radiation resistance and an increased number of cytoplasmic Lipid Droplets (LDs), we investigated if LD accumulation was modulated along the cell cycle and if this correlated with differential radioresistance in lung and bladder cell lines.ResultsOur findings identified the S phase as the most radioresistant cell cycle phase being characterized by an increase in LDs. Analysis of the expression of perilipin genes (a family of proteins involved in the LD structure and functions) throughout the cell cycle also uncovered a unique gene cell cycle pattern.ConclusionsIn summary, although these results require further molecular studies about the mechanisms of radioresistance, the findings presented here are the first evidence that LD accumulation could participate in cancer cells’ ability to better survive X-Ray radiation when cells are in the S phase. LDs can represent new players in the radioresistance processes associated with cancer metabolism. This could open new therapeutic avenues in which the use of LD-interfering drugs might enhance cancer sensitivity to radiation.

RS2289487 variation in PERILIPIN gene is a predictor of weight loss and protection against impaired glucose metabolism after a meal-replacement diet in postmenopausal obese females.

The PERILIPIN1 (PLIN1) gene encodes an adipocyte-associated protein that modulates weight. The objective was to evaluate the role of the rs2289487 genetic variant of the PLIN1 gene on weight loss and glucose metabolism secondary to a partial meal replacement (pMR) hypocaloric diet. We conducted an interventional study in 111 postmenopausal obese females with body mass index (BMI) > 35 kg/m2. The subjects received two intakes per day of a normocaloric hyperproteic formula for 12 weeks. After the pMR diet, body weight, (BMI), fat mass, waist circumference, fasting insulin levels and HOMA-IR decreased in both genotype groups. The improvements in these parameters were higher in C allele carriers than in subjects with TT genotype. The percentage of patients who achieved 7.5% weight loss was higher in the C carriers (57.4% vs. 27.6%), (adjusted Odds Ratio 2.14, 95% CI = 1.33-9.40; p = 0.02). The decrease in the percentage of diabetes mellitus or impaired fasting glucose decrease was statistically significant in C allele carriers (30.2% vs. 18.9%; p = 0.01) (OR 0.54, 95% CI = 0.22-0.78; p = 0.02). The C allele of rs2289487 predicts the magnitude of weight loss resulting from a pMR diet. These adiposity improvements produce a better improvement in insulin resistance and the percentage of impaired glucose metabolism.

Effects of PLIN1 Gene Knockout on the Proliferation, Apoptosis, Differentiation and Lipolysis of Chicken Preadipocytes.

Perilipin 1 (PLIN1) is one of the most abundant lipid droplet-related proteins on the surface of adipocytes. Our previous results showed that PLIN1 plays an important role in chicken lipid metabolism. To further reveal the role of PLIN1 in the growth and development of adipocytes, a chicken preadipocyte line with a PLIN1 gene knockout was established by the CRISPR/Cas9 gene editing technique, and the effects of the PLIN1 gene on the proliferation, apoptosis, differentiation and lipolysis of chicken preadipocytes were detected. The results showed that the CRISPR/Cas9 system effectively mediated knockout of the PLIN1 gene. After the deletion of PLIN1, the differentiation ability and early apoptotic activity of chicken preadipocytes decreased, and their proliferation ability increased. Moreover, knockout of PLIN1 promoted chicken preadipocyte lipolysis under basal conditions and inhibited chicken preadipocyte lipolysis under hormone stimulation. Taken together, our results inferred that PLIN1 plays a regulatory role in the process of proliferation, apoptosis, differentiation and lipolysis of chicken preadipocytes.

Sex-Dependent Mediation of Leptin in the Association of Perilipin Polymorphisms with BMI and Plasma Lipid Levels in Children.

Variations in the perilipin (PLIN) gene have been suggested to be associated with obesity and its related alterations, but a different nutritional status seems to contribute to differences in these associations. In our study, we examined the association of several polymorphisms at the PLIN locus with obesity and lipid profile in children, and then analyzed the mediation of plasma leptin levels on these associations. The single-nucleotide polymorphisms (SNPs) rs894160, rs1052700, and rs2304795 in PLIN1, and rs35568725 in PLIN2, were analyzed by RT-PCR in 1264 children aged 6–8 years. Our results showed a contrasting association of PLIN1 rs1052700 with apolipoprotein (Apo) A-I levels in boys and girls, with genotype TT carriers showing significantly higher Apo A-I levels in boys and significantly lower Apo A-I levels in girls. Significant associations of the SNP PLIN2 rs35568725 with high-density lipoprotein cholesterol (HDL-cholesterol), Apo A-I, and non-esterified fatty acids (NEFA) were observed in boys but not in girls. The associations of the SNPs studied with body mass index (BMI), NEFA, and Apo A-I in boys and girls were different depending on leptin concentration. In conclusion, we describe the mediation of plasma leptin levels in the association of SNPs in PLIN1 and PLIN2 with BMI, Apo A-I, and NEFA. Different leptin levels by sex may contribute to explain the sex-dependent association of the PLIN SNPs with these variables.

Characterization of a splice variant of perilipin 5

Lipid storage provides energy stores for the body in times of increased oxidative requirements, such as fasting or exercise, but when the balance of lipid storage and mobilization is disrupted human health suffers. Implications for increased lipid storage include risk factors for developing type II diabetes mellitus and non‐alcoholic fatty liver disease. The perilipin family of proteins associate with the surface of lipid droplets and are essential to regulation of stored triacylglycerols. Perilipin 5 functions to maintain a balance between the storage of triacylglycerols in lipid droplets or their mobilization when energetically required. Several members of the perilipin gene family exhibit differential splicing, most notably perilipin 1 but reports exist of alternative messages for shortened forms of perilipins 2 and 3. We have identified a shortened form of perilipin 5 we have termed perilipin 5b. Western blots indicate that the protein, of approximately 35 kDa, is present in C2C12 cultured myoblasts and in heart and liver tissue from fed and fasted mice. Analysis of murine genomic DNA sequences reveals stop codons found in the introns between exons 6 and 7 and 7 and 8, either of which could give rise to a protein of approximately 35 kDa, indicating the likelihood of the observed protein being a splice variant, rather than a post‐translationally modified protein. Using RT‐PCR we have identified multiple amplicons suggesting that perilipin‐5b contains a read through of intron 6 of the murine perilipin 5 gene. Mining transcriptome databases reveals several transcripts with retained introns. Analysis of the amino acid sequence of perilipin 5b indicates that this truncation would lack the reported C‐terminal domain essential for interactions with ATGL and CGI‐58 but would retain the PKA site found at Ser155. The function of perilipin 5b is yet unknown, but the fact that it is found primarily in heart tissue seems to suggest that it plays a role in the oxidation of the lipid droplet. These data are consistent with what is currently known about Perilipin‐5 and the other PAT family proteins. Determination of perilipin 5b function will give a more accurate picture of the overall mechanism of lipid storage and metabolism.

The effect of the PLIN1 gene on the metabolism and mitochondria of porcine skeletal muscle satellite cells.

Perilipin 1 (PLIN1) is a lipid droplet scaffolding protein that plays a regulatory role in fat decomposition and mitochondrial function. In this study, the effects of PLIN1 gene knockout (PLIN1-KO) and PLIN1 gene overexpression (PLIN1-EX) on cell metabolism and mitochondrial function in porcine skeletal muscle satellite cells were assessed. Porcine skeletal muscle satellite cells were used as the control group (NC). The expression of mitochondrial function-related proteins was detected by western blot. Apoptosis, cell cycle, mitochondrial function-related indices, mitochondrial structure, and morphology were measured by flow cytometry. Our results demonstrated that stable expression of the PLIN1 gene in skeletal muscle satellite cells is critical to maintaining cell metabolism and mitochondrial function. After knockout and overexpression of the PLIN1 gene, the anti-apoptotic ability of cells was enhanced, and the metabolic activity of the cells was accelerated, but at the cost of mitochondrial structural damage, reduction in the number of mitochondria, and decreased mitochondrial function. This study explored the effect of the PLIN1 gene on the mitochondria and metabolism of porcine skeletal muscle satellite cells and provided a theoretical basis for the subsequent study of the effects of PLIN1 on muscle tissue development and meat quality.

Perilipin Genetic Variation Correlated with Obesity and Lipid Profile in Metabolic Syndrome

BACKGROUND: Perilipin is very important for the regulation of the deposition and mobilization of fats. The human perilipin gene (PLIN) is near the locus for risk of obesity and hypertriglyceridemia. The PLIN gene is thought to be involved in the occurrence of metabolic syndrome. AIM: The aim of this research is to determine the role of variations of the PLIN gene (PLN4 11482 G>A) as a risk factor for component of metabolic syndrome. METHODS: This study involved a total of 160 subjects consisting of 80 with metabolic syndrome and 80 controls. Genotype analysis was done with the polymerase chain reaction-restriction fragment length polymorphism method. The data were analyzed with t-tests to compare the subjects’ characteristics between metabolic syndrome groups and controls. Risk factors of PLIN genotypes were calculated with odds ratio and multivariate regression analysis was used to determine the role of the PLIN gene with each biochemical characteristic. RESULTS: The result was significant differences between the characteristics of the metabolic syndrome subjects with controls (p < 0.05). There was no difference in genotypes between patients with metabolic syndrome and controls. The multivariate analysis of the genetic role with biochemical components showed the PLIN gene in AA carriers as a risk factor for metabolic syndrome compare GA+GG, risk of obesity, and hypercholesterolemia with p < 0.05. CONCLUSION: It can be concluded that PLIN variation has a role in the incidence of metabolic syndrome, especially in relation to obesity and hypercholesterolemia. Further study is needed to determine the role of other gene variations as a risk factor for metabolic syndrome.

Lubabegron fumarate acts as a β-adrenergic receptor antagonist in cultured bovine intramuscular and subcutaneous adipocytes.

We hypothesized that lubabegron fumarate (LUB) (Experior, Elanco Animal Health, Greenfield, IN) would act as an antagonist to β-adrenergic receptor (β-AR) subtypes in primary bovine subcutaneous (s.c.) and intramuscular (i.m.) adipocytes differentiated in culture. This study employed LUB, dobutamine (DOB, a selective β1-agonist), salbutamol (SAL, a selective β2-agonist), and propranolol (PRO, a non-selective β-AR antagonist). Preadipocytes were isolated by standard techniques from bovine longissimus muscle and overlying s.c. adipose tissue and differentiated to adipocytes for 14 d. The adipocyte source x stage of differentiation interaction was significant for β-adrenergic receptors-1 (ADRB1) (P = 0.001) and ADRB2 (P = 0.01) in that expression of ADRB1 and ADRB2 was greater in s.c. adipocytes than in s.c. preadipocytes; expression of the ADRB1-3 did not change after differentiation of i.m. adipocytes. CCATT/enhancer-binding protein alpha (CEBPA) expression increased upon differentiation in both s.c. and i.m. adipocytes (P = 0.006). The source x stage of differentiation interaction was significant for peroxisome proliferator-activated receptor gamma (PPARG) (P ≤ 0.001) and fatty acid binding protein-4 (FABP4) (P = 0.004). Expression of PPARG increased after differentiation of s.c. preadipocytes to adipocytes, but PPARG expression did not change with differentiation of i.m. preadipocytes to adipocytes. FABP4 expression increased after differentiation of both s.c. and i.m. adipocytes, but FABP4 expression increased to a greater extent in s.c. adipocytes. In s.c. adipocytes, DOB elevated cAMP and glycerol production and protein kinase A (PKA) activity, and SAL increased PKA activity; these effects were abolished by LUB and PRO (P < 0.001). Incubation of i.m. adipocytes with SAL increased cAMP production and PKA activity, which was attenuated by LUB and PRO (P ≤ 0.006). In s.c. adipocytes, SAL, LUB + SAL, and LUB + DOB upregulated hormone sensitive lipase (HSL) (P < 0.001) and perilipin (P = 0.002) gene expression. In i.m. adipocytes, DOB and LUB + DOB increased HSL gene expression (P = 0.001) and LUB + SAL depressed adipose triglyceride lipase expression below control levels (P = 0.001). These results demonstrate that LUB is a β-AR antagonist at the β1-AR and β2-AR subtypes in s.c. adipocytes, and that s.c. and i.m. exhibit different responses to β-AA and LUB.

Carcass and fatness traits of Central Javanese local ducks based on Lipoprotein Lipase (LPL) and Perilipin (PLIN) genes

This research is an exploratory study that determines the genotypes of carcass and fatness traits in Central Javanese local ducks based on the lipoprotein lipase (LPL) and perilipin (PLIN) genes. A 35 ducks were sampled from seven local ducks in Central Java. The duck’s DNA was isolated from its feathers. PLP and PLIN genes were amplified by PCR method using specific primers. In the RFLP analysis, PCR products were digested with the restriction enzymes MvaI (BstNI) (5’CC/WGG) and Bsp1286. (SduI) (GDGCH/C), for LPL and PLIN gene, respectively. The results of PCR-RFLP analysis on local ducks in this study showed a monomorphic in exon 2 of the PLIN gene and polymorphic in exon 5 of the LPL gene. Based on the LPL gene, as many as 29 ducks (82.85%) had GG genotypes, 3 ducks (8.57%) had AG types and 3 ducks (8.47%) had AA types. Both PLIN and LPL genotypes were clustered the Central Javanese local ducks into 3 haplotypes (A-C). The duck samples were dominated by haplotype A (82.85%) followed by haplotype B (8.57%) and haplotype C (8.57%). Results showed that 82.85% of Central Javanese local ducks genetically had a good of fatness traits but a low of carcass traits.

Overexpression of PLIN1 Promotes Lipid Metabolism in Bovine Adipocytes.

Simple SummaryIn bovine preadipocytes, the overexpression of the PLIN1 gene promotes the expression of FASN, PPARγ, ACC, LPL, FABP4, DGAT2, and C/EBPβ at the mRNA level, inhibiting the expression of fat metabolism genes such as PLIN2 and ATGL. Furthermore, after overexpression of PLIN1, the oil red O-staining results showed that the number of lipid droplets (LDs) in fat cells, their volume and triglyceride content were increased. The elevation in triglyceride content indicates that PLIN1 can promote the accumulation of triglyceride in bovine preadipocytes and has an important regulatory role in fat metabolism. After transfection with the adenovirues Ad-PLIN1 and Ad-NC (empty virus), RNA-seq was used to analyze the differentially expressed genes in bovine preadipocytes. A total of 1923 differentially expressed genes were detected, and GO and KEGG signaling pathways were analyzed for differentially expressed genes.Perilipin 1 (PLIN1) is a protein encoded by the PLIN1 gene in eukaryotes. PLIN1 is a member of the PAT protein family, a family of proteins related to lipid droplet (LD) surface proteins. PLIN1 phosphorylation plays a vital role during fat metabolism of adipose tissue lipolysis and fat storage in adipocytes. However, to further explore the regulation of the PLIN1 gene on the proliferation, differentiation and lipid metabolism of bovine adipocytes. In this study, the mRNA expression of PLIN1, at day six, was the highest during bovine adipocyte differentiation. Moreover, PLIN1 can promote the proliferation and differentiation of preadipocytes in cattle. On the sixth day, after transfection with, and overexpression of, the PLIN1 gene in bovine preadipocytes via adenovirus, cell samples were collected, and transcriptome sequencing was performed. A total of 1923 differentially expressed genes were detected. Through GO and KEGG pathway analysis, the differentially expressed genes were established to be mainly enriched in the AMPK, Wnt, and PPAR signaling pathways related to fat proliferation and differentiation. In conclusion, at the transcriptional level, PLIN1 plays an important role in regulating fat proliferation and metabolism. Additionally, the sequencing results screened new differentially expressed genes related to fat metabolism, providing theoretical support for molecular breeding of Qinchuan beef cattle.

RXRα Positively Regulates Expression of the Chicken PLIN1 Gene in a PPARγ-Independent Manner and Promotes Adipogenesis.

Perilipin1 (PLIN1), the most abundant lipid droplet (LD)-associated protein, plays a vital role in regulating lipid storage and breakdown in adipocytes. Recently, we found that the overexpression of PLIN1 promotes chicken preadipocyte lipid accumulation. However, the mechanisms by which transcription of the chicken PLIN1 gene is regulated remain unknown. In this study, we investigated the role of retinoid X receptor α (RXRα) in transcription of the chicken PLIN1 gene. Notably, reporter gene and expression assays showed that RXRα activates transcription of the chicken PLIN1 gene in a PPARγ-independent manner. Furthermore, promoter deletion and electrophoretic mobility shift assay (EMSA) analysis revealed that the chicken PLIN1 gene promoter region (-774/-785) contains an RXRα-binding site. Further study demonstrated that RXRα overexpression promotes differentiation of an immortalized chicken preadipocyte cell line (ICP1), causing a concomitant increase in PLIN1 transcripts. Taken together, our results show for the first time that RXRα activates transcription of the chicken PLIN1 gene in a PPARγ-independent manner, which might be at least in part responsible for RXRα-induced adipogenesis.

Characterization of a Splice Variant of Perilipin 5

Lipid storage provides energy stores for the body in times of increased oxidative requirements, such as fasting or exercise, but when the balance of lipid storage and mobilization is disrupted human health suffers. Implications for increased lipid storage include risk factors for developing type II diabetes mellitus and non‐alcoholic fatty liver disease. The perilipin family of proteins associate with the surface of lipid droplets and are essential to regulation of stored triacylglycerols. Perilipin 5 functions to maintain a balance between the storage of triacylglycerols in lipid droplets or their mobilization when energetically required. Several members of the perilipin gene family exhibit differential splicing, most notably perilipin 1 but reports exist of alternative messages for shortened forms of perilipins 2 and 3. We have identified a shortened form of perilipin 5 we have termed perilipin 5b. Western blots indicate that the protein, of approximately 35 kDa, is present in C2C12 cultured myoblasts and in heart and liver tissue from fed and fasted mice. Analysis of murine genomic DNA sequences reveals stop codons found in the introns between exons 6 and 7 and 7 and 8, either of which could give rise to a protein of approximately 35 kDa, indicating the likelihood of the observed protein being a splice variant, rather than a post‐translationally modified protein. Using RT‐PCR we have identified multiple amplicons suggesting that perilipin‐5b contains a read through of intron 6 of the murine perilipin 5 gene. Analysis of the amino acid sequence of perilipin 5b indicates that this truncation would lack the reported C‐terminal domain essential for interactions with ATGL and CGI‐58 but would retain the PKA site found at Ser155. The function of perilipin 5b is yet unknown, but the fact that it is found primarily in heart tissue seems to suggest that it plays a role in the oxidation of the lipid droplet. These data are consistent with what is currently known about Perilipin‐5 and the other PAT family proteins. Determination of perilipin 5b function will give a more accurate picture of the overall mechanism of lipid storage and metabolism.

Function and characterization of the promoter region of perilipin 1 (PLIN1): Roles of E2F1, PLAG1, C/EBPβ, and SMAD3 in bovine adipocytes

Perilipin 1 (PLIN1) protein, also known as lipid droplet-associated protein, is encoded by the PLIN1 gene and is able to anchor itself to the membranes of lipid droplets. The phosphorylation of PLIN1 is critical for the mobilization of fat in adipose tissue and plays an important role in regulating lipolysis and lipid storage in adipocytes. However, research on the synthesis and lipid metabolism of lipid droplets by PLIN1 in bovine adipocytes is limited. In the present study, we found that bovine PLIN1 was highly expressed in subcutaneous adipose tissue. The highest level of PLIN1 mRNA expression in bovine adipocytes was observed on day 6 of differentiation. Moreover, the cytoplasmic subcellular localization of PLIN1 was observed in bovine preadipocytes. To elucidate the molecular mechanism of bovine PLIN1 transcriptional regulation, we cloned eight fragments containing the 5′ regulatory region of the PLIN1 gene. The results showed that the −209/−17 bp region of the bovine PLIN1 gene was the core promoter region. Based on the transcriptional activities of the promoter vector fragments, the luciferase activity of the mutated fragment, the siRNA interference, and the results of the electrophoretic mobility shift assay (EMSA), we identified the binding sites of E2F transcription factor 1 (E2F1), pleiomorphic adenoma gene 1 (PLAG1), CCAAT enhancer binding protein beta (C/EBPβ), and SMAD family member 3 (SMAD3) as the transcriptional activators or repressors of the core promoter region. Further experiments confirmed that the knockdown of the PLIN1 gene affected the ability of these transcription factors to regulate the lipid metabolism in bovine adipocytes. In conclusion, our results reveal a potential mechanism for the transcriptional regulation of PLIN1 in bovine adipocytes.

Childhood Obesity Risk in Relationship to Perilipin 1 (PLIN1) Gene Regulation by Circulating microRNAs.

Childhood obesity is a growing public health burden in many countries. The lipid perilipin 1 (PLIN1) gene is involved in regulation of lipolysis, and thus represents a viable candidate mechanism for obesity genetics research in children. In addition, the regulation of candidate gene expression by circulating microRNAs (miRNAs) offers a new research venue for diagnostic innovation. We report new findings on associations among circulating miRNAs, regulation of the PLIN1 gene, and susceptibility to childhood obesity. In a sample of 135 unrelated subjects, 35 children with obesity (between ages 3 and 13) and 100 healthy controls (between ages 4 and 16), we examined the expression levels of four candidate miRNAs (hsa-miR-4777-3p, hsa-miR-642b-3p, hsa-miR-3671-1, and hsa-miR-551b-2) targeting the PLIN1 as measured by real-time polymerase chain reaction in whole blood samples. We found that the full genetic model, including the four candidate miRNAs and the PLIN1 gene, explained a statistically significant 12.7% of the variance in childhood obesity risk (p = 0.0034). The four miRNAs together explained 10.1% of the risk (p = 0.008). The percentage of variation in childhood obesity risk explained by hsa-miR-642b-3p and age was 19%. In accordance with biological polarity of the observed association, for example, hsa-miR-642b-3p was upregulated, while the PLIN1 expression decreased in obese participants compared to healthy controls. To the best of our knowledge, this is the first clinical association study of these candidate miRNAs targeting the PLIN1 in childhood obesity. These data offer new molecular leads for future clinical biomarker and diagnostic discovery for childhood obesity.

DNA methylation of the PLIN1 promoter downregulates expression in chicken lines.

Evidence suggests that Perilipin-1 (PLIN1) is subject to functional regulation by epigenetic modifications in women with obesity. However, whether chicken PLIN1 expression is regulated by DNA methylation is unknown. Here, Sequenom MassARRAY and real-time polymerase chain reaction (PCR) were conducted to analyze the promoter methylation status and expression of the PLIN1 gene in Northeast Agricultural University broiler lines divergently selected for abdominal fat content. We found that chicken PLIN1 expression was significantly higher in adipose tissue of fat-line broilers than in lean lines at 1–7 weeks of age, and was significantly positively correlated with abdominal fat percentage (AFP) in chicken adipose development (Pearson's , ). The region analyzed for DNA methylation was from to bp upstream of the translation start codon ATG, and had five CpG sites, where only the DNA methylation levels of CpG5 located at position bp were significantly higher in lean compared to fat chickens at 5 and 6 weeks () and were significantly negatively correlated with PLIN1 mRNA levels and AFP (). These results shed new light on the regulation of hypertrophic growth in chicken adipose development.

Changes in liver perilipin gene expression during an oral lipid load in obese OLETF rats

Non‐alcoholic fatty liver disease (NAFLD) is one of the most prevalent liver maladies in the US. NAFLD is associated with obesity, hyperlipidemia, hypertension, insulin resistance, and diabetes; all traits linked to metabolic syndrome, affecting 20% of the US population. Inappropriate hepatic lipid accumulation is associated with metabolic syndrome. Intracellular lipid storage is regulated by lipid droplets (LDs) that are active components of the cell. LDs have surface proteins of which the most studied belong to the PAT family (Perilipin1–5), which participate in storage and lipid metabolism. To evaluate the effect that a high‐lipid load on the expression and dynamics of hepatic perilipins during early onset T2D, an oral fat tolerance test (OFTT) was performed in 28 wk‐old rats (n=5–6/group): (1) healthy, strain control LETO, and (2) obese, diabetic OLETF. Liver and blood were collected before (T0) and after (T1, T2, T4, and T8 hours) the lipid challenge. An acute increase in serum TG was observed at T2 (p= &lt; 0.0001) and was maintained over the next 6 hours in OLETF whereas in LETO it decreased after the T2 mark. The mRNA expression of Plin2 increased 2.5‐fold in OLETF at T2 compared to LETO, and increased 2.5‐fold at T8 in LETO compared to baseline. Plin3 remained higher (p= 0.0077) in LETO compared to OLETF at T8. Plin4 did not change significantly in response to the lipid challenge between groups. Plin5 increased 1.8‐fold in LETO at T8 compared to baseline. These changes help explain the contributions of LDs to the regulation of hepatic lipids during both normal and diabetic conditions in response to a high lipid challenge similar to the consumption of a high‐fat meal. As Plin2 and 3 have been described as structural LD‐associated proteins, being present mostly in nascent LD, suggesting that a high fat load induces an acute induction of hepatocyte lipid storage. The increase in Plin5 suggests that lipid metabolism pathways are being activated, as Plin5 has been related with mitochondrial FA metabolism. The differential expression patterns of perilipins in response to an acute lipid challenge illustrates the timing of the cellular changes during the development of hepatic steatosis, providing useful insights into future development of treatment by targeting the PAT family of proteins.Support or Funding InformationThis work was supported by NIH's MHRT program.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Engineering lipid droplet assembly mechanisms for improved triacylglycerol accumulation in Saccharomyces cerevisiae

Production of triacylglycerols (TAGs) through microbial fermentation is an emerging alternative to plant and animal-derived sources. The yeast Saccharomyces cerevisiae is a preferred organism for industrial use but has natively a very poor capacity of TAG production and storage. Here, we engineered S. cerevisiae for accumulation of high TAG levels through the use of structural and physiological factors that influence assembly and biogenesis of lipid droplets. First, human and fungal perilipin genes were expressed, increasing TAG content by up to 36% when expressing the human perilipin gene PLIN3. Secondly, expression of the FIT2 homologue YFT2 resulted in a 26% increase in TAG content. Lastly, the genes ERD1 and PMR1 were deleted in order to induce an endoplasmic reticulum stress response and stimulate lipid droplet formation, increasing TAG content by 72% for Δerd1. These new approaches were implemented in previously engineered strains that carry high flux of fatty acid biosynthesis and conversion of acyl-CoA into TAGs, resulting in improvements of up to 138% over those high-producing strains without any substantial growth effects or abnormal cell morphology. We find that these approaches not only represent a significant improvement of S. cerevisiae for TAG production, but also highlight the importance of lipid droplet dynamics for high lipid accumulation in yeast.

Adipokine levels and perilipin gene polymorphisms in obese Turkish adolescents with non-alcoholic fatty liver disease

Adipokine levels and perilipin gene polymorphisms in obese Turkish adolescents with non-alcoholic fatty liver disease

Association between perilipin gene polymorphisms and body weight traits in Jinmao Hua chickens

Abstract. The perilipin gene (PLIN) plays a crucial role in lipid metabolism and fat deposition. In order to reveal the genetic effects of PLIN polymorphisms on body weight (BW) traits in chickens, PLIN gene polymorphisms in 322 Jinmao Hua chickens were detected by PCR-SSCP and DNA sequencing methods. For PLIN primer pair 1, five genotypes (AA, AB, BB, JJ and JL) were detected in the Jinmao Hua chicken population and three mutations (g.1889C &gt; T, g.1904T &gt; C and g.1922C &gt; T) were revealed by gene sequencing. For PLIN primer pair 2, three genotypes (CC, CD and DD) were detected in the same population and two mutations (g.2014A &gt; G and g.2020C &gt; T) were revealed by gene sequencing. Least squares analysis showed that individuals with the JJ and CD genotypes performed better than the other Jinmao Hua chicken genotypes. Based on the five SNPs, the frequency distributions of the eight haplotypes were estimated with PHASE2.1 software. C-T-C-G-T was the major haplotype with a frequency of 58.6957 %, while the frequency of C-C-C-A-C was less than 1 %. Fourteen diplotypes were obtained from the eight haplotypes. H1H1 was the dominant diplotype with a frequency of 47.205 %. Least squares analysis indicated that BW with the H3H3 diplotype was the lowest, while the H2H2 diplotype was the highest, suggesting that selecting for the H3H3 diplotype improved the BW traits of Jinmao Hua chickens. The findings of this study should be useful to expand the theoretical basis of the role the PLIN in poultry molecular breeding of poultry.