Gene_locus Report for: mouse-1plrp

Pancreatic triglyceride lipase (PTL) and its cofactor, colipase, are required for efficient dietary triglyceride digestion. In addition to PTL, pancreatic acinar cells synthesize two pancreatic lipase-related proteins (PLRP1 and PLRP2), which have a high degree of sequence and structural homology with PTL. The lipase activity of PLRP2 has been confirmed, whereas no known triglyceride lipase activity has been detected with PLRP1 up to now. To explore the biological functions of PLRP1 in vivo, we generated Plrp1 knockout (KO) mice in our laboratory. Here we show that the Plrp1 KO mice displayed mature-onset obesity with increased fat mass, impaired glucose clearance and the resultant insulin resistance. When fed on high-fat (HF) diet, the Plrp1 KO mice exhibited an increased weight gain, fat mass and severe insulin resistance compared with wild-type mice. Pancreatic juice extracted from Plrp1 KO mice had greater ability to hydrolyze triglyceride than that from the wild-type littermates. We propose that PLRP1 may function as a metabolic inhibitor in vivo of PLT-colipase-mediated dietary triglyceride digestion and provides potential anti-obesity targets for developing new drugs.

PURPOSE: Differential display analysis was used to look for gender differences in lacrimal gland gene expression. The expression of a female-specific mouse lacrimal gland mRNA that encoded pancreatic lipase-related protein 1 (PLRP1) was identified and characterized.
METHODS: Differential display analysis of the exorbital lacrimal glands of male and female Swiss Webster mice detected a potential female-specific cDNA, designated Y2. Using the technique of rapid amplification of cDNA ends, a full-length cDNA of Y2 was obtained and the nucleotide sequence determined. To assess tissue-specific expression, a labeled Y2 cDNA probe was hybridized to RNA blots of male and female mouse lacrimal, harderian, parotid, mandibular, sublingual, and pancreas glands and liver. Y2 cDNA was also hybridized to RNA blots of male and female rat lacrimal gland and male rat pancreas. To determine subcellular localization, Y2 sense and antisense RNA probes were hybridized to female mouse lacrimal gland frozen sections.
RESULTS: GenBank database sequence comparisons indicated that Y2 encoded mouse PLRP1. RNA blots documented that PLRP1 was expressed in female, but not in male, mouse lacrimal gland. PLRP1 mRNA was also expressed in male and female mouse sublingual gland and pancreas. Expression of PLRP1 was not detected in male or female rat lacrimal gland. In situ hybridization showed that PLRP1 was expressed in the acinar cells of the female mouse lacrimal gland.
CONCLUSIONS: Lacrimal gland expression of PLRP1 mRNA was gender and species specific. Female, but not male, mouse lacrimal gland expressed PLRPI mRNA. Neither female nor male rat lacrimal gland expressed PLRP1 mRNA. PLRP1 protein may be secreted in mouse tears, where it may function as a lipolytic enzyme, modifying tear film lipids.

Pancreatic triglyceride lipase (PTL) and its cofactor, colipase, are required for efficient dietary triglyceride digestion. In addition to PTL, pancreatic acinar cells synthesize two pancreatic lipase-related proteins (PLRP1 and PLRP2), which have a high degree of sequence and structural homology with PTL. The lipase activity of PLRP2 has been confirmed, whereas no known triglyceride lipase activity has been detected with PLRP1 up to now. To explore the biological functions of PLRP1 in vivo, we generated Plrp1 knockout (KO) mice in our laboratory. Here we show that the Plrp1 KO mice displayed mature-onset obesity with increased fat mass, impaired glucose clearance and the resultant insulin resistance. When fed on high-fat (HF) diet, the Plrp1 KO mice exhibited an increased weight gain, fat mass and severe insulin resistance compared with wild-type mice. Pancreatic juice extracted from Plrp1 KO mice had greater ability to hydrolyze triglyceride than that from the wild-type littermates. We propose that PLRP1 may function as a metabolic inhibitor in vivo of PLT-colipase-mediated dietary triglyceride digestion and provides potential anti-obesity targets for developing new drugs.

Only a small proportion of the mouse genome is transcribed into mature messenger RNA transcripts. There is an international collaborative effort to identify all full-length mRNA transcripts from the mouse, and to ensure that each is represented in a physical collection of clones. Here we report the manual annotation of 60,770 full-length mouse complementary DNA sequences. These are clustered into 33,409 'transcriptional units', contributing 90.1% of a newly established mouse transcriptome database. Of these transcriptional units, 4,258 are new protein-coding and 11,665 are new non-coding messages, indicating that non-coding RNA is a major component of the transcriptome. 41% of all transcriptional units showed evidence of alternative splicing. In protein-coding transcripts, 79% of splice variations altered the protein product. Whole-transcriptome analyses resulted in the identification of 2,431 sense-antisense pairs. The present work, completely supported by physical clones, provides the most comprehensive survey of a mammalian transcriptome so far, and is a valuable resource for functional genomics.

The RIKEN Mouse Gene Encyclopaedia Project, a systematic approach to determining the full coding potential of the mouse genome, involves collection and sequencing of full-length complementary DNAs and physical mapping of the corresponding genes to the mouse genome. We organized an international functional annotation meeting (FANTOM) to annotate the first 21,076 cDNAs to be analysed in this project. Here we describe the first RIKEN clone collection, which is one of the largest described for any organism. Analysis of these cDNAs extends known gene families and identifies new ones.

In the effort to prepare the mouse full-length cDNA encyclopedia, we previously developed several techniques to prepare and select full-length cDNAs. To increase the number of different cDNAs, we introduce here a strategy to prepare normalized and subtracted cDNA libraries in a single step. The method is based on hybridization of the first-strand, full-length cDNA with several RNA drivers, including starting mRNA as the normalizing driver and run-off transcripts from minilibraries containing highly expressed genes, rearrayed clones, and previously sequenced cDNAs as subtracting drivers. Our method keeps the proportion of full-length cDNAs in the subtracted/normalized library high. Moreover, our method dramatically enhances the discovery of new genes as compared to results obtained by using standard, full-length cDNA libraries. This procedure can be extended to the preparation of full-length cDNA encyclopedias from other organisms.

The RIKEN high-throughput 384-format sequencing pipeline (RISA system) including a 384-multicapillary sequencer (the so-called RISA sequencer) was developed for the RIKEN mouse encyclopedia project. The RISA system consists of colony picking, template preparation, sequencing reaction, and the sequencing process. A novel high-throughput 384-format capillary sequencer system (RISA sequencer system) was developed for the sequencing process. This system consists of a 384-multicapillary auto sequencer (RISA sequencer), a 384-multicapillary array assembler (CAS), and a 384-multicapillary casting device. The RISA sequencer can simultaneously analyze 384 independent sequencing products. The optical system is a scanning system chosen after careful comparison with an image detection system for the simultaneous detection of the 384-capillary array. This scanning system can be used with any fluorescent-labeled sequencing reaction (chain termination reaction), including transcriptional sequencing based on RNA polymerase, which was originally developed by us, and cycle sequencing based on thermostable DNA polymerase. For long-read sequencing, 380 out of 384 sequences (99.2%) were successfully analyzed and the average read length, with more than 99% accuracy, was 654.4 bp. A single RISA sequencer can analyze 216 kb with >99% accuracy in 2.7 h (90 kb/h). For short-read sequencing to cluster the 3' end and 5' end sequencing by reading 350 bp, 384 samples can be analyzed in 1.5 h. We have also developed a RISA inoculator, RISA filtrator and densitometer, RISA plasmid preparator which can handle throughput of 40,000 samples in 17.5 h, and a high-throughput RISA thermal cycler which has four 384-well sites. The combination of these technologies allowed us to construct the RISA system consisting of 16 RISA sequencers, which can process 50,000 DNA samples per day. One haploid genome shotgun sequence of a higher organism, such as human, mouse, rat, domestic animals, and plants, can be revealed by seven RISA systems within one month.

PURPOSE: Differential display analysis was used to look for gender differences in lacrimal gland gene expression. The expression of a female-specific mouse lacrimal gland mRNA that encoded pancreatic lipase-related protein 1 (PLRP1) was identified and characterized.
METHODS: Differential display analysis of the exorbital lacrimal glands of male and female Swiss Webster mice detected a potential female-specific cDNA, designated Y2. Using the technique of rapid amplification of cDNA ends, a full-length cDNA of Y2 was obtained and the nucleotide sequence determined. To assess tissue-specific expression, a labeled Y2 cDNA probe was hybridized to RNA blots of male and female mouse lacrimal, harderian, parotid, mandibular, sublingual, and pancreas glands and liver. Y2 cDNA was also hybridized to RNA blots of male and female rat lacrimal gland and male rat pancreas. To determine subcellular localization, Y2 sense and antisense RNA probes were hybridized to female mouse lacrimal gland frozen sections.
RESULTS: GenBank database sequence comparisons indicated that Y2 encoded mouse PLRP1. RNA blots documented that PLRP1 was expressed in female, but not in male, mouse lacrimal gland. PLRP1 mRNA was also expressed in male and female mouse sublingual gland and pancreas. Expression of PLRP1 was not detected in male or female rat lacrimal gland. In situ hybridization showed that PLRP1 was expressed in the acinar cells of the female mouse lacrimal gland.
CONCLUSIONS: Lacrimal gland expression of PLRP1 mRNA was gender and species specific. Female, but not male, mouse lacrimal gland expressed PLRPI mRNA. Neither female nor male rat lacrimal gland expressed PLRP1 mRNA. PLRP1 protein may be secreted in mouse tears, where it may function as a lipolytic enzyme, modifying tear film lipids.