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| Status |
Public on Oct 24, 2019 |
| Title |
Lysophosphatidic acid treatment improves brush border maturation and apical SGLT1 activity in Myo5b deficient mice, a model of MVID |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Myosin Vb (Myo5b) is an essential trafficking protein for membrane recycling in gastrointestinal (GI) epithelial cells and its inactivating mutation causes the congenital diarrheal disease, microvillus inclusion disease (MVID). We previously reported that Myo5b deficiency in mice causes mislocalization of SGLT1 and NHE3, but retained apical function of CFTR, resulting in malabsorption and secretory diarrhea. Activation of lysophosphatidic acid (LPA) receptors can improve diarrhea, but the effects of LPA on MVID symptoms is unclear. We therefore tested whether LPA treatment can ameliorate the epithelial deficits in Myo5b knockout (KO) mice. Methods: Adult tamoxifen-induced, intestine-specific, Myo5b KO (VillinCreERT2;Myo5bflox/flox) and littermate control mice were treated with LPA, an LPAR2 agonist (GRI977143), or vehicle for 4 days after a single tamoxifen injection. Apical SGLT1 and CFTR activities were measured in Üssing chambers. The localization of membrane transporters was evaluated by immunostaining in mouse tissues and enteroids. RNA sequencing and enrichment analysis were performed with isolated jejunal epithelial cells. Results: Daily treatment with LPA decreased the frequency of multivesicular bodies and the expression of cathepsins, but did not affect the formation of microvillus inclusions in Myo5b KO mice. LPA partially restored the brush border height and the localization of SGLT1 and NHE3 in Myo5b KO small intestine and enteroids. SGLT1-dependent short-circuit current (Isc) was increased and abnormal CFTR activities were suppressed in LPA-treated jejunum compared to that of vehicle-treated Myo5b KO mice. Conclusions: Cell autonomous LPA signaling may modulate a Myo5b-independent trafficking mechanism and the brush border maturation, demonstrating a therapeutic potential for LPA in the treatment of MVID.
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| Overall design |
Adult male VillinCreERT2;MYO5Bflox/flox mice were given a single dose of 2 mg of tamoxifen (KO, 7 mice) or corn oil (control, 3 mice) by ip injection. Induced KO mice were treated with LPA (3 mice) or vehicle (4 mice) by ip for 4 days. All mice were euthanized on day 4 and epithelial cells were mechanically isolated from jejunum by micro forceps in ice-cold PBS containing 10 mM EDTA and immersed in TRIzol Reagent (Invitrogen 15596018). RNA was extracted according to the manufacture instructions and RNA-seq was performed by Novogene Corporation Inc. (Sacramento, CA) on 3 or 4 independent preparations from each condition.
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| Contributor(s) |
Kaji I, Goldenring JR |
| Citation(s) |
34197342 |
| Submission date |
Oct 23, 2019 |
| Last update date |
Sep 14, 2021 |
| Contact name |
Izumi Kaji |
| E-mail(s) |
izumi.kaji@vumc.org
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| Organization name |
Vanderbilt University Medical Center
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| Department |
Surgery
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| Street address |
Rm 10465H, MRB IV
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| City |
Nashville |
| State/province |
TN |
| ZIP/Postal code |
37232 |
| Country |
USA |
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| Platforms (1) |
| GPL24247 |
Illumina NovaSeq 6000 (Mus musculus) |
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| Samples (10)
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| Relations |
| BioProject |
PRJNA579150 |
| SRA |
SRP226727 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE139302_GEO_seq_spreadsheet_101819IK.xlsx |
48.8 Kb |
(ftp)(http) |
XLSX |
| GSE139302_RNA-seq_M5B_induced_selected_101819.xlsx |
40.2 Kb |
(ftp)(http) |
XLSX |
| GSE139302_Tmx_KOvsNon_KO_all.GOenrich.significant_metabolism.xlsx |
34.0 Kb |
(ftp)(http) |
XLSX |
| GSE139302_Tmx_KOvsNon_KO_down.GOenrich.significant.metabolism.xlsx |
21.2 Kb |
(ftp)(http) |
XLSX |
| GSE139302_Tmx_KOvsNon_KO_down.KEGGenrich.significant.txt.gz |
2.1 Kb |
(ftp)(http) |
TXT |
| GSE139302_Tmx_KOvsNon_KO_up.GOenrich.significant.metabolic.xlsx |
38.3 Kb |
(ftp)(http) |
XLSX |
| GSE139302_Tmx_KOvsNon_KO_up.KEGGenrich.significant.txt.gz |
594 b |
(ftp)(http) |
TXT |
| GSE139302_fpkm_genename.txt.gz |
2.1 Mb |
(ftp)(http) |
TXT |
| GSE139302_ipL_KOvsTmx_KO_DEG_up.txt.gz |
1.4 Kb |
(ftp)(http) |
TXT |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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