Molecular Cloning, Genomic Organization, and Expression of a Testicular Isoform of Hormone-Sensitive Lipase

Holst, Lena Stenson; Langin, Dominique; Mulder, Hindrik; Laurell, Henrik; Grober, Jacques; Bergh, Anders; Mohrenweiser, Harvey W.; Edgren, Gustaf; Holm, Cecilia

doi:10.1006/geno.1996.0383

Genomics

1996

DOI: 10.1006/geno.1996.0383

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Molecular Cloning, Genomic Organization, and Expression of a Testicular Isoform of Hormone-Sensitive Lipase

Lena Stenson Holst

Dominique Langin

Hindrik Mulder

et al.

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Cited by 83 publications

(60 citation statements)

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“…In situ hybridization experiments performed in rat testis showed strong labeling of cells in the adluminal parts of the seminiferous tubules at stages X-XIV and sparsely distributed grains in the basal parts (6). Immunohistochemistry experiments showed an HSL-like immunoreactivity in the adluminal part of the rat seminiferous tubules at stages XIII-VIII (5). The data suggested that rodent HSL mRNA and protein are expressed in haploid germ cells with a lag between mRNA and protein appearance.…”

mentioning

confidence: 84%

“…In adipose tissue, HSL catalyzes the rate-limiting step in lipolysis, the catabolic pathway that mobilizes fatty acids from triacylglycerol stored in the lipid droplet. HSL is also expressed in rodent and human testis (3)(4)(5). In situ hybridization experiments performed in rat testis showed strong labeling of cells in the adluminal parts of the seminiferous tubules at stages X-XIV and sparsely distributed grains in the basal parts (6).…”

mentioning

confidence: 99%

“…The 2.8-kb mRNA encodes a 775-amino acid protein. A specific form of HSL, named HSL tes , is expressed in human and rat testis (5). The 3.9-kb human HSL tes mRNA encodes a 1076-amino acid protein.…”

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confidence: 99%

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Testis Hormone-sensitive Lipase Expression in Spermatids Is Governed by a Short Promoter in Transgenic Mice

Blaise¹,

Guillaudeux²,

Tavernier³

et al. 2001

Journal of Biological Chemistry

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A testicular form of hormone-sensitive lipase (HSL tes ), a triacylglycerol lipase, and cholesterol esterase, is expressed in male germ cells. Northern blot analysis showed HSL tes mRNA expression in early spermatids. Immunolocalization of the protein in human and rodent seminiferous tubules indicated that the highest level of expression occurred in elongated spermatids. We have previously shown that 0.5 kilobase pairs of the human HSL tes promoter directs testis-specific expression of a chloramphenicol acetyltransferase reporter gene in transgenic mice and determined regions binding nuclear proteins expressed in testis but not in liver (Blaise, R., Grober, J., Rouet, P., Tavernier, G., Daegelen, D., and Langin, D. (1999) J. Biol. Chem. 274, 9327-9334). Mutation of a SRY/Sox-binding site in one of the regions did not impair in vivo testis-specific expression of the reporter gene. Further transgenic analyses established that 95 base pairs upstream of the transcription start site were sufficient for correct testis expression. In gel retardation assays using early spermatid nuclear extracts, a germ cell-specific DNA-protein interaction was mapped between ؊46 and ؊29 base pairs. The DNA binding nuclear protein showed properties of zinc finger transcription factors. Mutation of the region abolished reporter gene activity in transgenic mice, showing that it is necessary for testis expression of HSL tes . Hormone-sensitive lipase (HSL)1 hydrolyzes triacylglycerol and, cholesterol and retinyl esters (1, 2). In adipose tissue, HSL catalyzes the rate-limiting step in lipolysis, the catabolic pathway that mobilizes fatty acids from triacylglycerol stored in the lipid droplet. HSL is also expressed in rodent and human testis (3-5). In situ hybridization experiments performed in rat testis showed strong labeling of cells in the adluminal parts of the seminiferous tubules at stages X-XIV and sparsely distributed grains in the basal parts (6). Immunohistochemistry experiments showed an HSL-like immunoreactivity in the adluminal part of the rat seminiferous tubules at stages XIII-VIII (5). The data suggested that rodent HSL mRNA and protein are expressed in haploid germ cells with a lag between mRNA and protein appearance. However, the precise germ cell type(s) expressing HSL was not determined in rat and it was not possible to rule out HSL expression in Sertoli cells. Moreover, the expression of HSL in germ cells of other mammalian species, including man, has not been documented. The effects of HSL gene disruption in mice have recently been reported (7). The most striking feature of the phenotype is male sterility due to oligospermia. Degenerated spermatocytes and spermatids were observed in HSL-deficient testis with a lack of mature spermatozoa. The data clearly demonstrate that HSL is required for spermatogenesis.The human adipose tissue form of HSL is encoded by 9 exons spanning 11 kb (8). The transcription start site was mapped in a short 5Ј-noncoding exon located 1.5 kb upstream of the first coding exon (9). The 2.8-kb mR...

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mentioning

confidence: 84%

mentioning

confidence: 99%

See 1 more Smart Citation

Testis Hormone-sensitive Lipase Expression in Spermatids Is Governed by a Short Promoter in Transgenic Mice

Blaise¹,

Guillaudeux²,

Tavernier³

et al. 2001

Journal of Biological Chemistry

Self Cite

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“…The effect of glucose was reversible and was not due to an impairment of the differentiation program, since the mRNA levels of CCAAT/enhancer binding protein-␣ (C/EBP␣), a transcription factor important for maintenance of the terminally differentiated state, and adipocyte triglyceride content were not affected by glucose deprivation. Moreover, it was shown that prolonged exposure of isolated rat adipocytes to glucose in the presence of insulin resulted in an increase of basal and stimulated lipolysis and a maintenance of HSL protein levels (14).The adipocyte form of HSL is encoded by nine exons spanning 11 kb (15,16). The transcriptional start site has been mapped in a short 5Ј-noncoding exon located 1.5 kb upstream of the first coding exon.…”

mentioning

confidence: 99%

“…The adipocyte form of HSL is encoded by nine exons spanning 11 kb (15,16). The transcriptional start site has been mapped in a short 5Ј-noncoding exon located 1.5 kb upstream of the first coding exon.…”

mentioning

confidence: 99%

Transcriptional Regulation of Adipocyte Hormone-Sensitive Lipase by Glucose

et al. 2002

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Hormone-sensitive lipase (HSL) catalyzes the ratelimiting step in the mobilization of fatty acids from adipose tissue, thus determining the supply of energy substrates in the body. HSL mRNA was positively regulated by glucose in human adipocytes. Pools of stably transfected 3T3-F442A adipocytes were generated with human adipocyte HSL promoter fragments from ؊2,400/ ؉38 to ؊31/؉38 bp linked to the luciferase gene. A glucose-responsive region was mapped within the proximal promoter (؊137 bp). Electromobility shift assays showed that upstream stimulatory factor (USF)-1 and USF2 and Sp1 and Sp3 bound to a consensus E-box and two GC-boxes in the ؊137-bp region. Cotransfection of the ؊137/؉38 construct with USF1 and USF2 expression vectors produced enhanced luciferase activity. Moreover, HSL mRNA levels were decreased in USF1-and USF2-deficient mice. Site-directed mutagenesis of the HSL promoter showed that the GC-boxes, although contributing to basal promoter activity, were dispensable for glucose responsiveness. Mutation of the E-box led to decreased promoter activity and suppression of the glucose response. Analogs and metabolites were used to determine the signal metabolite of the glucose response. The signal is generated downstream of glucose-6-phosphate in the glycolytic pathway before the triose phosphate step. Diabetes 51:293-300, 2002 H ormone-sensitive lipase (HSL) is a key enzyme for the hydrolysis of adipose tissue triacylglycerol into fatty acids that are the major source of body energy in the absence of dietary lipids. Catecholamines and insulin are important regulators of lipolysis in adipocytes through a modulation of intracellular cAMP levels and reversible phosphorylation of HSL (1). The lipase is thought to catalyze the rate-limiting step in cAMP-dependent lipolysis. In agreement, a strong linear correlation was found between HSL protein levels and the maximum lipolytic capacity of human subcutaneous adipocytes stimulated by a ␤-adrenergic agonist (2). Moreover, targeted disruption of the HSL gene in the mouse results in blunted ␤-adrenergic agonist-induced lipolysis (3,4). Clinical studies also support a role for HSL as a limiting factor in adipose tissue lipolysis and show that, besides the short-term modulation of activity by phosphorylation, variations in HSL expression are associated with changes in lipolytic capacity. Indeed, obese patients and normal-weight subjects with a family trait for obesity show decreased maximal lipolytic effect of catecholamines and blunted HSL expression (5,6). Furthermore, genetic studies suggest that HSL participates in the polygenic background of obesity and type 2 diabetes (7,8).Increased mobilization of fatty acids from adipose tissue stores in diabetic patients leads to high free fatty acid concentrations that contribute to the development of ketoacidosis, a major acute complication of type 1 diabetes (9). Several mechanisms may account for the enhanced lipolysis. Hypoinsulinemia and an increase in the concentrations of stimulatory hormones stimulate HSL activ...

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The N‐terminal His‐tag affects the triglyceride lipase activity of hormone‐sensitive lipase in testis

Wang

Ren

Chen

et al. 2019

J of Cellular Biochemistry

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The sterility of hormone-sensitive lipase (HSL) knockout mice clearly shows the link between lipid metabolism and spermatogenesis. However, which substrate or product of this multifunctional lipase affects spermatogenesis is unclear. We found that an HSL protein with a His-tag at the N-terminus preserved the normal hydrolase activity of cholesteryl ester (CE) but the triglyceride lipase (TG) activity significantly decreased in vitro. Therefore, mice with this functionally incomplete HSL (His-HSL) were produced on a background of HSL deficiency (HSL −/− h). As a result, HSL −/− h testis has an 8.65-fold higher CE activity than wild-type testis but a twofold higher TG activity than wild-type testis. To compare His-HSL and wild-type HSL in vitro and in vivo, we confirmed that the His-tag significantly suppressed HSL TG activity. From our results, we believe that TG activity was affected by the His-tag insertion, but CE activity was not influenced. Furthermore, the His-tag protected HSL from binding to the inhibitor BAY. From our study, TG activity and BAY binding sites were affected by N-terminal His-tag insertion. K E Y W O R D S cholesterol metabolism, hormone-sensitive lipase, lipid metabolism, male infertility, spermatogenesis

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Molecular Cloning, Genomic Organization, and Expression of a Testicular Isoform of Hormone-Sensitive Lipase

Cited by 83 publications

References 1 publication

Testis Hormone-sensitive Lipase Expression in Spermatids Is Governed by a Short Promoter in Transgenic Mice

Testis Hormone-sensitive Lipase Expression in Spermatids Is Governed by a Short Promoter in Transgenic Mice

Transcriptional Regulation of Adipocyte Hormone-Sensitive Lipase by Glucose

The N‐terminal His‐tag affects the triglyceride lipase activity of hormone‐sensitive lipase in testis

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