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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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Separation and identification of phosphatidylcholines (PCs) by thin-layer chromatography (TLC) showing bands duplicated bands (upper panels) and histograms of the intensity of PCs (lower panel) in (A) each group of seminiferous tubules (ST), and (B) testes of the three developmental male morphotypes.Group B STs, containing mostly spermatids and some immature spermatozoa, show significant differences of PCs intensities compared with groups A and C (P<0.05; means ± S.D.; n = 5). The data support the IMS results of the PC (16:0/18:1). Moreover, the testes of OC males also contain significantly more PCs than those of SM and BC males (P<0.05). SM = small male; OC = orange claw male; BC = blue claw male. Bar = S.D.; * = significant difference at P<0.05.
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pone.0120412.g002: Separation and identification of phosphatidylcholines (PCs) by thin-layer chromatography (TLC) showing bands duplicated bands (upper panels) and histograms of the intensity of PCs (lower panel) in (A) each group of seminiferous tubules (ST), and (B) testes of the three developmental male morphotypes.Group B STs, containing mostly spermatids and some immature spermatozoa, show significant differences of PCs intensities compared with groups A and C (P<0.05; means ± S.D.; n = 5). The data support the IMS results of the PC (16:0/18:1). Moreover, the testes of OC males also contain significantly more PCs than those of SM and BC males (P<0.05). SM = small male; OC = orange claw male; BC = blue claw male. Bar = S.D.; * = significant difference at P<0.05.

Mentions: The extracted lipids were separated by TLC, and the highest intensity signals were found in PC bands of each group. The PCs bands were expressed as mean ± S.D. which showed different amounts in each of the ST groups Fig. 2A and the male developmental morphotypes Fig. 2B. The STs of group B which contained mostly spermatids and some immature sperms showed significantly higher intensities compared with group A and C (P<0.05) Fig. 2A. The highest amounts of PCs could be observed in the OC males, which is the transitional stage from SM to BC male, and the lowest PC amounts were observed in BC males (with significant difference at P<0.05) Fig. 2B.


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)

Separation and identification of phosphatidylcholines (PCs) by thin-layer chromatography (TLC) showing bands duplicated bands (upper panels) and histograms of the intensity of PCs (lower panel) in (A) each group of seminiferous tubules (ST), and (B) testes of the three developmental male morphotypes.Group B STs, containing mostly spermatids and some immature spermatozoa, show significant differences of PCs intensities compared with groups A and C (P<0.05; means ± S.D.; n = 5). The data support the IMS results of the PC (16:0/18:1). Moreover, the testes of OC males also contain significantly more PCs than those of SM and BC males (P<0.05). SM = small male; OC = orange claw male; BC = blue claw male. Bar = S.D.; * = significant difference at P<0.05.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0120412.g002: Separation and identification of phosphatidylcholines (PCs) by thin-layer chromatography (TLC) showing bands duplicated bands (upper panels) and histograms of the intensity of PCs (lower panel) in (A) each group of seminiferous tubules (ST), and (B) testes of the three developmental male morphotypes.Group B STs, containing mostly spermatids and some immature spermatozoa, show significant differences of PCs intensities compared with groups A and C (P<0.05; means ± S.D.; n = 5). The data support the IMS results of the PC (16:0/18:1). Moreover, the testes of OC males also contain significantly more PCs than those of SM and BC males (P<0.05). SM = small male; OC = orange claw male; BC = blue claw male. Bar = S.D.; * = significant difference at P<0.05.
Mentions: The extracted lipids were separated by TLC, and the highest intensity signals were found in PC bands of each group. The PCs bands were expressed as mean ± S.D. which showed different amounts in each of the ST groups Fig. 2A and the male developmental morphotypes Fig. 2B. The STs of group B which contained mostly spermatids and some immature sperms showed significantly higher intensities compared with group A and C (P<0.05) Fig. 2A. The highest amounts of PCs could be observed in the OC males, which is the transitional stage from SM to BC male, and the lowest PC amounts were observed in BC males (with significant difference at P<0.05) Fig. 2B.

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