Summary of Study ST003748

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002333. The data can be accessed directly via it's Project DOI: 10.21228/M82K0C This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Study ID	ST003748
Study Title	Phosphatidylinositol remodeling by lysophospholipid acyltransferase 11 ensures embryonic radial glial cell integrity - part 2
Study Summary	Since fragmentation of the Golgi apparatus is associated with defects in phosphoinositide (PIP) metabolisms, we measured PIPs in E11.5-E13.5 cortices by supercritical fluid chromatography-mass spectrometry (SFC-MS). In Mboat7 KO mice, PI4P with arachidonic acid (38:4 PI4P), which is the major species in Mboat7 heterozygous mice, significantly decreased, and instead, PI4P with monounsaturated fatty acid (34:1, 34:2, 36:1, 36:2 PI) and PI4P with docosapentaenoic acid (DPA) or docosahexaenoic acid (DHA) (38:6, 40:5, 40:6, 40:7 PI4P) increased. PI(4,5)P2 also showed changes in the molecular species similar to those observed in PI4P. Interestingly, the total amount of PI(4,5)P2 started to decrease at E12.5 in Mboat7 KO mice, whereas the total amount of PI4P was unchanged. This is because the decrease of arachidonic acid-containing PI(4,5)P2 species was not fully compensated by the increase of other PI(4,5)P2 species. To rule out the possibility that changes in systemic lipid metabolism in global Mboat7 KO mice affect radial glial cells in a non-cell-autonomous manner, we generated neural-specific Mboat7 KO mice (hereafter, Mboat7-Sox1-KO) by crossing Mboat7fl/fl mice with Sox1-Cre transgenic mice, which drives the expression of Cre recombinase before E8.5 in the neuroepithelium. Detachment of RGCs from the ventricular wall was not observed in the E12.5 cortices of Mboat7-Sox1-KO. On the other hand, E13.5 cortices of Mboat7-Sox1-KO showed detachment from the ventricular wall and apoptosis to the same extent as those of Mboat7 KO mice. Also, Mboat7-Sox1-KO showed rounding of the Golgi apparatus at E12.5 when the detachment of RGCs from the ventricular wall was not observed, which is consistent with the phenotype observed in E12.0 cortices of Mboat7 KO mice. Moreover, E-cadherin overexpression failed to rescue the detachment of RGCs from the apical ventricular wall. In the Mboat7-Sox1-KO cortices, a significant change in the fatty acyl composition of PI and PIPs was observed. The total amount of PI(4,5)P2 started to decrease at E12.5 in the Mboat7-Sox1-KO cortices as well as in the Mboat7 KO cortices. Taken together, these results indicate that Mboat7-Sox1-KO mice mimicked almost all the phenotypes observed in the brain of Mboat7 KO mice.
Institute	University of Tokyo
Last Name	Kono
First Name	Nozomu
Address	Hongo 7-3-1,, Bunyo-ku, Tokyo, 113-0033, Japan
Email	nozomu@mol.f.u-tokyo.ac.jp
Phone	81-3-5841-4723
Submit Date	2025-01-25
Raw Data Available	Yes
Raw Data File Type(s)	mzML
Analysis Type Detail	LC-MS
Release Date	2025-10-28
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR002333
Project DOI:	doi: 10.21228/M82K0C
Project Title:	Phosphatidylinositol remodeling by lysophospholipid acyltransferase 11 ensures embryonic radial glial cell integrity
Project Summary:	Arachidonic acid, a vital polyunsaturated fatty acid in brain development, is enriched in phosphatidylinositol (PI). The arachidonic acyl chain in PI is introduced by lysophospholipid acyltransferase 11 (LPLAT11)/membrane-bound O-acyltransferase 7 (MBOAT7), the loss of which causes microcephaly in humans and mice. Here, we show that LPLAT11 deficiency impaired indirect neurogenesis in the developing neocortex, resulting in fewer layer II-V neurons. LPLAT11-deficient radial glial cells had defects in differentiation into intermediate progenitor cells and increased apoptosis. Prior to these anomalies, LPLAT11 deficiency caused a rounding of the Golgi apparatus, accompanied by impaired apical trafficking of E-cadherin, and deregulated apical detachment. Moreover, impaired PI acyl chain remodeling led to a decreased amount of PI(4,5)P2, leading to Golgi apparatus rounding. Thus, these results clarify the underlying mechanism of microcephaly by LPLAT11 deficiency and highlight the critical role of arachidonic acid in PI in the integrity of radial glial cells.
Institute:	University of Tokyo
Department:	Graduate School of Pharmaceutical Sciences
Laboratory:	Department of Health Chemistry
Last Name:	Kono
First Name:	Nozomu
Address:	Hongo 7-3-1,, Bunyo-ku, Tokyo, 113-0033, Japan
Email:	nozomu@mol.f.u-tokyo.ac.jp
Phone:	81-3-5841-4723

Subject:

Subject ID:	SU004409
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

Factors:

Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id	local_sample_id	Sample source	Gestational ages	Genotype
SA496777	E11.5_Het_2	brain cortex	E11.5	Mboat7+/-
SA496778	E11.5_Het_1	brain cortex	E11.5	Mboat7+/-
SA496779	E11.5_Het_3	brain cortex	E11.5	Mboat7+/-
SA496780	E11.5_KO_1	brain cortex	E11.5	Mboat7-/-
SA496781	E11.5_KO_2	brain cortex	E11.5	Mboat7-/-
SA496782	E11.5_KO_3	brain cortex	E11.5	Mboat7-/-
SA496783	E12.5_Het_1	brain cortex	E12.5	Mboat7+/-
SA496784	E12.5_Het_2	brain cortex	E12.5	Mboat7+/-
SA496785	E12.5_Het_3	brain cortex	E12.5	Mboat7+/-
SA496791	E12.5_KO_1	brain cortex	E12.5	Mboat7-/-
SA496792	E12.5_KO_2	brain cortex	E12.5	Mboat7-/-
SA496793	E12.5_KO_3	brain cortex	E12.5	Mboat7-/-
SA496794	E12.5_Sox1_KO1	brain cortex	E12.5	Mboat7f/f Sox1-Cre
SA496795	E12.5_Sox1_KO4	brain cortex	E12.5	Mboat7f/f Sox1-Cre
SA496796	E12.5_Sox1_KO2	brain cortex	E12.5	Mboat7f/f Sox1-Cre
SA496797	E12.5_Sox1_KO3	brain cortex	E12.5	Mboat7f/f Sox1-Cre
SA496786	E12.5_Sox1-Het_5	brain cortex	E12.5	Mboat7+/f Sox1-Cre
SA496787	E12.5_Sox1-Het_4	brain cortex	E12.5	Mboat7+/f Sox1-Cre
SA496788	E12.5_Sox1-Het_3	brain cortex	E12.5	Mboat7+/f Sox1-Cre
SA496789	E12.5_Sox1-Het_2	brain cortex	E12.5	Mboat7+/f Sox1-Cre
SA496790	E12.5_Sox1-Het_1	brain cortex	E12.5	Mboat7+/f Sox1-Cre
SA496798	E13.5_Het_7	brain cortex	E13.5	Mboat7+/-
SA496799	E13.5_Het_5	brain cortex	E13.5	Mboat7+/-
SA496800	E13.5_Het_4	brain cortex	E13.5	Mboat7+/-
SA496801	E13.5_Het_3	brain cortex	E13.5	Mboat7+/-
SA496802	E13.5_Het_2	brain cortex	E13.5	Mboat7+/-
SA496803	E13.5_Het_1	brain cortex	E13.5	Mboat7+/-
SA496804	E13.5_Het_6	brain cortex	E13.5	Mboat7+/-
SA496808	E13.5_KO_5	brain cortex	E13.5	Mboat7-/-
SA496809	E13.5_KO_6	brain cortex	E13.5	Mboat7-/-
SA496810	E13.5_KO_3	brain cortex	E13.5	Mboat7-/-
SA496811	E13.5_KO_2	brain cortex	E13.5	Mboat7-/-
SA496812	E13.5_KO_1	brain cortex	E13.5	Mboat7-/-
SA496813	E13.5_KO_4	brain cortex	E13.5	Mboat7-/-
SA496814	E13.5_Sox1_KO1	brain cortex	E13.5	Mboat7f/f Sox1-Cre
SA496815	E13.5_Sox1_KO2	brain cortex	E13.5	Mboat7f/f Sox1-Cre
SA496816	E13.5_Sox1_KO3	brain cortex	E13.5	Mboat7f/f Sox1-Cre
SA496817	E13.5_Sox1_KO4	brain cortex	E13.5	Mboat7f/f Sox1-Cre
SA496818	E13.5_Sox1_KO5	brain cortex	E13.5	Mboat7f/f Sox1-Cre
SA496819	E13.5_Sox1_KO6	brain cortex	E13.5	Mboat7f/f Sox1-Cre
SA496805	E13.5_Sox1-Het_1	brain cortex	E13.5	Mboat7+/f Sox1-Cre
SA496806	E13.5_Sox1-Het_2	brain cortex	E13.5	Mboat7+/f Sox1-Cre
SA496807	E13.5_Sox1-Het_3	brain cortex	E13.5	Mboat7+/f Sox1-Cre

Showing results 1 to 43 of 43

Showing results 1 to 43 of 43

Collection:

Collection ID:	CO004402
Collection Summary:	Embryos at different gestational ages were collected from Mboat7 heterozygous intercrosses or from crosses between Mboat7 f/f; Sox1-Cre and Mboat7 +/f; Sox1-Cre, and the cortices were subsequently dissected from the embryos.
Sample Type:	Brain cortex

Treatment:

Treatment ID:	TR004418
Treatment Summary:	Not applicable.

Sample Preparation:

Sampleprep ID:	SP004415
Sampleprep Summary:	"Cortices from E11.5 to E13.5 were isolated in ice-cold DMEM and immediately collected into a safe-lock poly-propylene tube (2ml) with 750 µl of ice-cold quench mix and sonicated. 170 µl of H2O was mixed and stored at -80 ºC before lipid extraction. The single-phase sample (a mixture of 170 µl of an aqueous sample, 10 ng of internal standards [17:0/20:4 PI, 17:0/20:4 PI4P, and 17:0/20:4 PI(4,5)P2], and 750 µl of quench mix) were mixed with 725 µl of CHCl3 and 170 µl of 2 M HCl, followed by vortex-mixing and centrifugation (1,500 g, 5 min at room temperature). The lower organic phase was collected into a new 2 ml safe-lock poly-propylene tube and mixed with 708 µl of pre-derivatization wash solution, followed by vortex-mixing and centrifugation (1,500 g, 3 min at room temperature). The lower phase was collected into another fresh tube and subjected to derivatization. Fifty µl trimethylsilyl diazomethane in hexane (2 M solution) was added to the lipid extracts (~1 ml), and after leaving to stand 10 min at room temperature, the reaction was quenched with 6 µl of acetic acid. Next, 700 µl post-derivatization wash solution was added to the mixture, followed by centrifugation (1,500 g, 3 min). The resultant lower phase was collected and rewashed with a 700 µl post-derivatization wash solution. Samples were temporarily stored at -80 ºC. Ninety µl of methanol and 10 µl of H2O were added to the final collected lower phase. The samples were dried up with a centrifugal evaporator. Samples were dissolved in 80 µl methanol, sonicated for 1 min, and 20 µl H2O was added. After centrifugation at 15,000 g for 30 min at 4 ºC, the supernatants were collected. To avoid degradation, the samples were stored at -80 ºC until use. "

Sampleprep ID:

SP004415

Sampleprep Summary:

"Cortices from E11.5 to E13.5 were isolated in ice-cold DMEM and immediately collected into a safe-lock poly-propylene tube (2ml) with 750 µl of ice-cold quench mix and sonicated. 170 µl of H2O was mixed and stored at -80 ºC before lipid extraction. The single-phase sample (a mixture of 170 µl of an aqueous sample, 10 ng of internal standards [17:0/20:4 PI, 17:0/20:4 PI4P, and 17:0/20:4 PI(4,5)P2], and 750 µl of quench mix) were mixed with 725 µl of CHCl3 and 170 µl of 2 M HCl, followed by vortex-mixing and centrifugation (1,500 g, 5 min at room temperature). The lower organic phase was collected into a new 2 ml safe-lock poly-propylene tube and mixed with 708 µl of pre-derivatization wash solution, followed by vortex-mixing and centrifugation (1,500 g, 3 min at room temperature). The lower phase was collected into another fresh tube and subjected to derivatization. Fifty µl trimethylsilyl diazomethane in hexane (2 M solution) was added to the lipid extracts (~1 ml), and after leaving to stand 10 min at room temperature, the reaction was quenched with 6 µl of acetic acid. Next, 700 µl post-derivatization wash solution was added to the mixture, followed by centrifugation (1,500 g, 3 min). The resultant lower phase was collected and rewashed with a 700 µl post-derivatization wash solution. Samples were temporarily stored at -80 ºC. Ninety µl of methanol and 10 µl of H2O were added to the final collected lower phase. The samples were dried up with a centrifugal evaporator. Samples were dissolved in 80 µl methanol, sonicated for 1 min, and 20 µl H2O was added. After centrifugation at 15,000 g for 30 min at 4 ºC, the supernatants were collected. To avoid degradation, the samples were stored at -80 ºC until use. "

Chromatography:

Chromatography ID:	CH005380
Instrument Name:	Shimadzu Nexera UC/s
Column Name:	ULTRON AF-HILIC-CD (250 x 2.0 mm, 5.0um)
Column Temperature:	10ºC
Flow Gradient:	0-16 min: 5% B to 20% B; 16.01-18 min: 40% B; 18.01-22 min: 5% B
Flow Rate:	1.5 mL/min
Solvent A:	99.99% supercritical carbon dioxide
Solvent B:	2.5% water/97.5% methanol; 0.1% formic acid
Chromatography Type:	Supercritical fluid chromatography

Analysis:

Analysis ID:	AN007083
Analysis Type:	MS
Chromatography ID:	CH005380
Num Factors:	10
Num Metabolites:	36
Units:	pmol/cortex