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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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IMS showing the intensity and distribution of PCs in the STs of OC males at high magnifications.Micrographs from H&E-stained sections (a-c) show areas surrounded by white dashed lines corresponding to the same areas that display ion images (d-o). Groups A and B STs containing mostly developing cells, and intertubular area (IT) (d-e, g-h, j-k, m-n), show higher levels of the signal intensity compared with group C STs, which contains only mature spermatozoa (Sz) (surrounded by orange dashed circle) (f, i, l, o). The areas containing Sz have very low signal intensities in all groups. Sc = spermatocytes; St = spermatids; Scale bars = 200 μm; Relative intensity bar shows the intensity level of the ion images.
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pone.0120412.g005: IMS showing the intensity and distribution of PCs in the STs of OC males at high magnifications.Micrographs from H&E-stained sections (a-c) show areas surrounded by white dashed lines corresponding to the same areas that display ion images (d-o). Groups A and B STs containing mostly developing cells, and intertubular area (IT) (d-e, g-h, j-k, m-n), show higher levels of the signal intensity compared with group C STs, which contains only mature spermatozoa (Sz) (surrounded by orange dashed circle) (f, i, l, o). The areas containing Sz have very low signal intensities in all groups. Sc = spermatocytes; St = spermatids; Scale bars = 200 μm; Relative intensity bar shows the intensity level of the ion images.

Mentions: Ion images indicating high intensity of PCs, including m/z 798.5 [PC (16:0/18:1) + K]+, 808.5 [PC (18:0/18:2) + Na]+, 826.5 [PC (18:2/20:5 (EPA)) + Na]+, and 872.5 [PC (18:0/22:6 (DHA)) + K]+ in the Figs. 4 and 5, and m/z 756.5 [PC (16:0/16:1) + Na]+, 780.5 [PC (16:0/18:2) + Na]+, 800.5 [PC (16:0/18:0) + K]+, 804.5 [PC (18:2/18:2) + Na]+, 806.5 [PC (18:1/18:2) + Na]+, 810.5 [PC (18:0/18:1) + Na]+, 820.5 [PC (16:0/20:4 (ARA)) + K]+, 830.5 [PC (18:1/20:4 (ARA)) + Na]+, 832.5 [PC (18:0/20:4 (ARA)) + Na]+, 844.5 [PC (16:0/22:6 (DHA)) + K]+, 846.5 [PC (18:0/20:5 (EPA)) + K]+, and 870.5 [PC (18:1/22:6 (DHA)) + K]+ in the supplementary data S3 Fig., showed the distributions pattern of PCs in each ST group of OC males, particularly in groups A and B, and the IT Tables 1 and 2. The H&E-stained sections of the same areas confirmed that the identifications of the ST groups were correct S2 Fig.


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)

IMS showing the intensity and distribution of PCs in the STs of OC males at high magnifications.Micrographs from H&E-stained sections (a-c) show areas surrounded by white dashed lines corresponding to the same areas that display ion images (d-o). Groups A and B STs containing mostly developing cells, and intertubular area (IT) (d-e, g-h, j-k, m-n), show higher levels of the signal intensity compared with group C STs, which contains only mature spermatozoa (Sz) (surrounded by orange dashed circle) (f, i, l, o). The areas containing Sz have very low signal intensities in all groups. Sc = spermatocytes; St = spermatids; Scale bars = 200 μm; Relative intensity bar shows the intensity level of the ion images.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0120412.g005: IMS showing the intensity and distribution of PCs in the STs of OC males at high magnifications.Micrographs from H&E-stained sections (a-c) show areas surrounded by white dashed lines corresponding to the same areas that display ion images (d-o). Groups A and B STs containing mostly developing cells, and intertubular area (IT) (d-e, g-h, j-k, m-n), show higher levels of the signal intensity compared with group C STs, which contains only mature spermatozoa (Sz) (surrounded by orange dashed circle) (f, i, l, o). The areas containing Sz have very low signal intensities in all groups. Sc = spermatocytes; St = spermatids; Scale bars = 200 μm; Relative intensity bar shows the intensity level of the ion images.
Mentions: Ion images indicating high intensity of PCs, including m/z 798.5 [PC (16:0/18:1) + K]+, 808.5 [PC (18:0/18:2) + Na]+, 826.5 [PC (18:2/20:5 (EPA)) + Na]+, and 872.5 [PC (18:0/22:6 (DHA)) + K]+ in the Figs. 4 and 5, and m/z 756.5 [PC (16:0/16:1) + Na]+, 780.5 [PC (16:0/18:2) + Na]+, 800.5 [PC (16:0/18:0) + K]+, 804.5 [PC (18:2/18:2) + Na]+, 806.5 [PC (18:1/18:2) + Na]+, 810.5 [PC (18:0/18:1) + Na]+, 820.5 [PC (16:0/20:4 (ARA)) + K]+, 830.5 [PC (18:1/20:4 (ARA)) + Na]+, 832.5 [PC (18:0/20:4 (ARA)) + Na]+, 844.5 [PC (16:0/22:6 (DHA)) + K]+, 846.5 [PC (18:0/20:5 (EPA)) + K]+, and 870.5 [PC (18:1/22:6 (DHA)) + K]+ in the supplementary data S3 Fig., showed the distributions pattern of PCs in each ST group of OC males, particularly in groups A and B, and the IT Tables 1 and 2. The H&E-stained sections of the same areas confirmed that the identifications of the ST groups were correct S2 Fig.

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