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Changes of phosphatidylcholine and fatty acids in germ cells during testicular maturation in three developmental male morphotypes of Macrobrachium rosenbergii revealed by imaging mass spectrometry.

Siangcham T, Chansela P, Hayasaka T, Masaki N, Sroyraya M, Poljaroen J, Suwansa-ard S, Engsusophon A, Hanna PJ, Sobhon P, Setou M - PLoS ONE (2015)

Bottom Line: Imaging mass spectrometry showed remarkably high signals corresponding to PC (16:0/18:1), PC (18:0/18:2), PC (18:2/20:5), and PC (16:0/22:6) in STs of groups A and B.Moreover, most signals were detected in the early developing cells and the intertubular area, but not at the area containing spermatozoa.The increasing amounts of FAs in the SM and OC indicate that they are important for spermatogenesis and spermiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Bangkok, Thailand.

ABSTRACT
Testis maturation, germ cell development and function of sperm, are related to lipid composition. Phosphatidylcholines (PCs) play a key role in the structure and function of testes. As well, increases of polyunsaturated fatty acids (PUFA) and highly unsaturated fatty acids (HUFA), especially arachidonic acid (ARA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) are essential for male fertility. This study is the first report to show the composition and distribution of PCs and total fatty acids (FAs) in three groups of seminiferous tubules (STs) classified by cellular associations [i.e., A (STs with mostly early germ cells), B (STs with mostly spermatids), and C (STs with spermatozoa)], in three morphotypes of Macrobrachium rosenbergii, [i.e., small male (SM), orange claw male (OC), and blue claw male (BC)]. Thin layer chromatography exhibited levels of PCs reaching maxima in STs of group B. Imaging mass spectrometry showed remarkably high signals corresponding to PC (16:0/18:1), PC (18:0/18:2), PC (18:2/20:5), and PC (16:0/22:6) in STs of groups A and B. Moreover, most signals were detected in the early developing cells and the intertubular area, but not at the area containing spermatozoa. Finally, gas chromatography-mass spectrometry indicated that the major FAs present in the testes were composed of 14:0, 16:0, 17:0, 18:0, 16:1, 18:1, 18:2, 20:1, 20:2, 20:4, 20:5, and 22:6. The testes of OC contained the greatest amounts of these FAs while the testes of BC contained the least amounts of these FAs, and there was more EPA (20:5) in the testes of SM and OC than those in the BC. The increasing amounts of FAs in the SM and OC indicate that they are important for spermatogenesis and spermiogenesis. This knowledge will be useful in formulating diets containing PUFA and HUFA for prawn broodstocks in order to improve testis development, and lead to increased male fecundity.

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MS/MS analysis showing product ions from the precursor ions at (A) m/z 780.5 and (B) m/z 798.5.The product ions from the precursor ion at m/z 780.5 represent neutral losses of a PC head group [(CH3)3N(CH2)2PO4H] at m/z 597.5 and trimethylamine [(CH3)3N] at m/z 721.5. The minor peaks at m/z 465.3 and 441.3 correspond to neutral losses of FAs (16:0 and 18:2) from a peak at m/z 721.5. Therefore, the molecule was assigned as [PC (16:0/18:2) + Na]+. The product ions from the precursor ion at m/z 798.5 represent neutral losses of the PC head group [(CH2)2PO4H] at m/z 615.5 and trimethylamine [(CH3)3N] at m/z 739.5. The minor peaks at m/z 483.5 and 457.5 correspond to neutral losses of FAs (16:0 and 18:1) from a peak at m/z 739.5. Therefore, the molecule was assigned as [PC (16:0/18:1) + K]+.
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pone.0120412.g003: MS/MS analysis showing product ions from the precursor ions at (A) m/z 780.5 and (B) m/z 798.5.The product ions from the precursor ion at m/z 780.5 represent neutral losses of a PC head group [(CH3)3N(CH2)2PO4H] at m/z 597.5 and trimethylamine [(CH3)3N] at m/z 721.5. The minor peaks at m/z 465.3 and 441.3 correspond to neutral losses of FAs (16:0 and 18:2) from a peak at m/z 721.5. Therefore, the molecule was assigned as [PC (16:0/18:2) + Na]+. The product ions from the precursor ion at m/z 798.5 represent neutral losses of the PC head group [(CH2)2PO4H] at m/z 615.5 and trimethylamine [(CH3)3N] at m/z 739.5. The minor peaks at m/z 483.5 and 457.5 correspond to neutral losses of FAs (16:0 and 18:1) from a peak at m/z 739.5. Therefore, the molecule was assigned as [PC (16:0/18:1) + K]+.

Mentions: MS/MS analysis showed product ions from precursor ions at m/z 780.5 Fig. 3A, and m/z 798.5 Fig. 3B. These signals were identified as [PC (16:0/18:2) + Na]+ and [PC (16:0/18:1) + K]+, respectively. The product ions from the precursor ion at m/z 780.5 represented neutral losses of a PC head group [(CH3)3N(CH2)2PO4H] at m/z 597.5 and trimethylamine [(CH3)3N] at m/z 721.5. These neutral losses are common for PCs. Another peak at m/z 575.5 indicates the replacement of adduct ion from Na+ to H+. The minor peaks at m/z 465.3 and 441.3 correspond to neutral losses of FAs (16:0 and 18:2) from a peak at m/z 721.5. Therefore, these molecules were assigned as [PC (16:0/18:2) + Na]+Fig. 3A. The product ions from the precursor ion at m/z 798.5 represents neutral losses of a PC head group [(CH2)2PO4H] at m/z 615.5 and trimethylamine [(CH3)3N] at m/z 739.5. The peak at m/z 577.5 indicates the replacement of adduct ion from K+ to H+. The minor peaks at m/z 483.5 and 457.5 correspond to neutral losses of FAs (16:0 and 18:1) from a peak at m/z 739.5. Therefore, these molecules were assigned as [PC (16:0/18:1) + K]+Fig. 3B.


Changes of phosphatidylcholine and fatty acids in germ cells during testicular maturation in three developmental male morphotypes of Macrobrachium rosenbergii revealed by imaging mass spectrometry.

Siangcham T, Chansela P, Hayasaka T, Masaki N, Sroyraya M, Poljaroen J, Suwansa-ard S, Engsusophon A, Hanna PJ, Sobhon P, Setou M - PLoS ONE (2015)

MS/MS analysis showing product ions from the precursor ions at (A) m/z 780.5 and (B) m/z 798.5.The product ions from the precursor ion at m/z 780.5 represent neutral losses of a PC head group [(CH3)3N(CH2)2PO4H] at m/z 597.5 and trimethylamine [(CH3)3N] at m/z 721.5. The minor peaks at m/z 465.3 and 441.3 correspond to neutral losses of FAs (16:0 and 18:2) from a peak at m/z 721.5. Therefore, the molecule was assigned as [PC (16:0/18:2) + Na]+. The product ions from the precursor ion at m/z 798.5 represent neutral losses of the PC head group [(CH2)2PO4H] at m/z 615.5 and trimethylamine [(CH3)3N] at m/z 739.5. The minor peaks at m/z 483.5 and 457.5 correspond to neutral losses of FAs (16:0 and 18:1) from a peak at m/z 739.5. Therefore, the molecule was assigned as [PC (16:0/18:1) + K]+.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4363669&req=5

pone.0120412.g003: MS/MS analysis showing product ions from the precursor ions at (A) m/z 780.5 and (B) m/z 798.5.The product ions from the precursor ion at m/z 780.5 represent neutral losses of a PC head group [(CH3)3N(CH2)2PO4H] at m/z 597.5 and trimethylamine [(CH3)3N] at m/z 721.5. The minor peaks at m/z 465.3 and 441.3 correspond to neutral losses of FAs (16:0 and 18:2) from a peak at m/z 721.5. Therefore, the molecule was assigned as [PC (16:0/18:2) + Na]+. The product ions from the precursor ion at m/z 798.5 represent neutral losses of the PC head group [(CH2)2PO4H] at m/z 615.5 and trimethylamine [(CH3)3N] at m/z 739.5. The minor peaks at m/z 483.5 and 457.5 correspond to neutral losses of FAs (16:0 and 18:1) from a peak at m/z 739.5. Therefore, the molecule was assigned as [PC (16:0/18:1) + K]+.
Mentions: MS/MS analysis showed product ions from precursor ions at m/z 780.5 Fig. 3A, and m/z 798.5 Fig. 3B. These signals were identified as [PC (16:0/18:2) + Na]+ and [PC (16:0/18:1) + K]+, respectively. The product ions from the precursor ion at m/z 780.5 represented neutral losses of a PC head group [(CH3)3N(CH2)2PO4H] at m/z 597.5 and trimethylamine [(CH3)3N] at m/z 721.5. These neutral losses are common for PCs. Another peak at m/z 575.5 indicates the replacement of adduct ion from Na+ to H+. The minor peaks at m/z 465.3 and 441.3 correspond to neutral losses of FAs (16:0 and 18:2) from a peak at m/z 721.5. Therefore, these molecules were assigned as [PC (16:0/18:2) + Na]+Fig. 3A. The product ions from the precursor ion at m/z 798.5 represents neutral losses of a PC head group [(CH2)2PO4H] at m/z 615.5 and trimethylamine [(CH3)3N] at m/z 739.5. The peak at m/z 577.5 indicates the replacement of adduct ion from K+ to H+. The minor peaks at m/z 483.5 and 457.5 correspond to neutral losses of FAs (16:0 and 18:1) from a peak at m/z 739.5. Therefore, these molecules were assigned as [PC (16:0/18:1) + K]+Fig. 3B.

Bottom Line: Imaging mass spectrometry showed remarkably high signals corresponding to PC (16:0/18:1), PC (18:0/18:2), PC (18:2/20:5), and PC (16:0/22:6) in STs of groups A and B.Moreover, most signals were detected in the early developing cells and the intertubular area, but not at the area containing spermatozoa.The increasing amounts of FAs in the SM and OC indicate that they are important for spermatogenesis and spermiogenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Bangkok, Thailand.

ABSTRACT
Testis maturation, germ cell development and function of sperm, are related to lipid composition. Phosphatidylcholines (PCs) play a key role in the structure and function of testes. As well, increases of polyunsaturated fatty acids (PUFA) and highly unsaturated fatty acids (HUFA), especially arachidonic acid (ARA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) are essential for male fertility. This study is the first report to show the composition and distribution of PCs and total fatty acids (FAs) in three groups of seminiferous tubules (STs) classified by cellular associations [i.e., A (STs with mostly early germ cells), B (STs with mostly spermatids), and C (STs with spermatozoa)], in three morphotypes of Macrobrachium rosenbergii, [i.e., small male (SM), orange claw male (OC), and blue claw male (BC)]. Thin layer chromatography exhibited levels of PCs reaching maxima in STs of group B. Imaging mass spectrometry showed remarkably high signals corresponding to PC (16:0/18:1), PC (18:0/18:2), PC (18:2/20:5), and PC (16:0/22:6) in STs of groups A and B. Moreover, most signals were detected in the early developing cells and the intertubular area, but not at the area containing spermatozoa. Finally, gas chromatography-mass spectrometry indicated that the major FAs present in the testes were composed of 14:0, 16:0, 17:0, 18:0, 16:1, 18:1, 18:2, 20:1, 20:2, 20:4, 20:5, and 22:6. The testes of OC contained the greatest amounts of these FAs while the testes of BC contained the least amounts of these FAs, and there was more EPA (20:5) in the testes of SM and OC than those in the BC. The increasing amounts of FAs in the SM and OC indicate that they are important for spermatogenesis and spermiogenesis. This knowledge will be useful in formulating diets containing PUFA and HUFA for prawn broodstocks in order to improve testis development, and lead to increased male fecundity.

Show MeSH
Related in: MedlinePlus