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Distinct expression and localization of the type II diacylglycerol kinase isozymes δ, η and κ in the mouse reproductive organs.

Shionoya T, Usuki T, Komenoi S, Isozaki T, Sakai H, Sakane F - BMC Dev. Biol. (2015)

Bottom Line: However, these enzymes' functions in the reproductive organs remain unknown.In this study, we first identified the expression sites of type II DGKs in the mouse reproductive organs in detail.These results indicate that the expression patterns of the type II DGK isoforms δ2, η1, η3 and κ differ from each other, suggesting that these DGK isoforms play specific roles in distinct compartments and developmental stages of the reproductive organs, especially in the processes of spermatogenesis and oocyte maturation.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, Graduate School of Science, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba, 263-8522, Japan. aeka3227@chiba-u.jp.

ABSTRACT

Background: We have revealed that the type II diacylglycerol kinases (DGKs) δ, η and κ were expressed in the testis and ovary. However, these enzymes' functions in the reproductive organs remain unknown.

Results: In this study, we first identified the expression sites of type II DGKs in the mouse reproductive organs in detail. Reverse transcription-polymerase chain reaction and Western blotting confirmed that DGKδ2 (splicing variant 2) but not DGKδ1 (splicing variant 1) and DGKκ were expressed in the testis, ovary and uterus. DGKη1 (splicing variant 1) but not DGKη2 (splicing variant 2) was strongly detected in the ovary and uterus. Interestingly, we found that a new alternative splicing product of the DGKη gene, DGKη3, which lacks exon 26 encoding 31 amino acid residues, was expressed only in the testis. Moreover, we investigated the distribution of type II DGKs in the testis, ovary and uterus through in situ hybridization. DGKδ2 was distributed in the primary spermatocytes of the testis and ovarian follicles. DGKη1 was distributed in the oviductal epithelium of the ovary and the luminal epithelium of the uterus. Intriguingly, DGKη3 was strongly expressed in the secondary spermatocytes and round spermatids of the testis. DGKκ was distributed in the primary and secondary spermatocyte of the testis.

Conclusion: These results indicate that the expression patterns of the type II DGK isoforms δ2, η1, η3 and κ differ from each other, suggesting that these DGK isoforms play specific roles in distinct compartments and developmental stages of the reproductive organs, especially in the processes of spermatogenesis and oocyte maturation.

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In situhybridization of the type II DGK mRNAs in the ovary of a 12-week-old female mouse. (A) The type II DGK mRNAs were hybridized and detected with the antisense probes of the type II DGK mRNAs. The ovary sections were also hybridized with the sense probes as controls. (B) High-magnification image of the ovarian follicles (secondary follicles). Although we observed histological specimens from five female mice in different estrous cycles, essentially the same results were obtained. Representatives of five independent experiments with five female mice are shown. Pf, primary follicle; Sf, secondary follicle; Mf, mature follicle; Cl, corpus luteum; Gc, granulosa cell; M, medulla. The scale bars in (A) represent 200 μm, and the scale bars in (B) represent 40 μm.
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Fig4: In situhybridization of the type II DGK mRNAs in the ovary of a 12-week-old female mouse. (A) The type II DGK mRNAs were hybridized and detected with the antisense probes of the type II DGK mRNAs. The ovary sections were also hybridized with the sense probes as controls. (B) High-magnification image of the ovarian follicles (secondary follicles). Although we observed histological specimens from five female mice in different estrous cycles, essentially the same results were obtained. Representatives of five independent experiments with five female mice are shown. Pf, primary follicle; Sf, secondary follicle; Mf, mature follicle; Cl, corpus luteum; Gc, granulosa cell; M, medulla. The scale bars in (A) represent 200 μm, and the scale bars in (B) represent 40 μm.

Mentions: DGKδ mRNA was broadly and modestly expressed in the primary, secondary and mature follicles and the corpus lutea (Figure 4 and Table 2). In the medulla, weak staining was observed. DGKδ mRNA was weakly detected in the oviductal epithelium (Figure 5). Although the ovary and oviduct sections were also hybridized with the sense probe as a control, no distinct staining was detected (Figures 4 and 5).Figure 4


Distinct expression and localization of the type II diacylglycerol kinase isozymes δ, η and κ in the mouse reproductive organs.

Shionoya T, Usuki T, Komenoi S, Isozaki T, Sakai H, Sakane F - BMC Dev. Biol. (2015)

In situhybridization of the type II DGK mRNAs in the ovary of a 12-week-old female mouse. (A) The type II DGK mRNAs were hybridized and detected with the antisense probes of the type II DGK mRNAs. The ovary sections were also hybridized with the sense probes as controls. (B) High-magnification image of the ovarian follicles (secondary follicles). Although we observed histological specimens from five female mice in different estrous cycles, essentially the same results were obtained. Representatives of five independent experiments with five female mice are shown. Pf, primary follicle; Sf, secondary follicle; Mf, mature follicle; Cl, corpus luteum; Gc, granulosa cell; M, medulla. The scale bars in (A) represent 200 μm, and the scale bars in (B) represent 40 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4308931&req=5

Fig4: In situhybridization of the type II DGK mRNAs in the ovary of a 12-week-old female mouse. (A) The type II DGK mRNAs were hybridized and detected with the antisense probes of the type II DGK mRNAs. The ovary sections were also hybridized with the sense probes as controls. (B) High-magnification image of the ovarian follicles (secondary follicles). Although we observed histological specimens from five female mice in different estrous cycles, essentially the same results were obtained. Representatives of five independent experiments with five female mice are shown. Pf, primary follicle; Sf, secondary follicle; Mf, mature follicle; Cl, corpus luteum; Gc, granulosa cell; M, medulla. The scale bars in (A) represent 200 μm, and the scale bars in (B) represent 40 μm.
Mentions: DGKδ mRNA was broadly and modestly expressed in the primary, secondary and mature follicles and the corpus lutea (Figure 4 and Table 2). In the medulla, weak staining was observed. DGKδ mRNA was weakly detected in the oviductal epithelium (Figure 5). Although the ovary and oviduct sections were also hybridized with the sense probe as a control, no distinct staining was detected (Figures 4 and 5).Figure 4

Bottom Line: However, these enzymes' functions in the reproductive organs remain unknown.In this study, we first identified the expression sites of type II DGKs in the mouse reproductive organs in detail.These results indicate that the expression patterns of the type II DGK isoforms δ2, η1, η3 and κ differ from each other, suggesting that these DGK isoforms play specific roles in distinct compartments and developmental stages of the reproductive organs, especially in the processes of spermatogenesis and oocyte maturation.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, Graduate School of Science, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba, 263-8522, Japan. aeka3227@chiba-u.jp.

ABSTRACT

Background: We have revealed that the type II diacylglycerol kinases (DGKs) δ, η and κ were expressed in the testis and ovary. However, these enzymes' functions in the reproductive organs remain unknown.

Results: In this study, we first identified the expression sites of type II DGKs in the mouse reproductive organs in detail. Reverse transcription-polymerase chain reaction and Western blotting confirmed that DGKδ2 (splicing variant 2) but not DGKδ1 (splicing variant 1) and DGKκ were expressed in the testis, ovary and uterus. DGKη1 (splicing variant 1) but not DGKη2 (splicing variant 2) was strongly detected in the ovary and uterus. Interestingly, we found that a new alternative splicing product of the DGKη gene, DGKη3, which lacks exon 26 encoding 31 amino acid residues, was expressed only in the testis. Moreover, we investigated the distribution of type II DGKs in the testis, ovary and uterus through in situ hybridization. DGKδ2 was distributed in the primary spermatocytes of the testis and ovarian follicles. DGKη1 was distributed in the oviductal epithelium of the ovary and the luminal epithelium of the uterus. Intriguingly, DGKη3 was strongly expressed in the secondary spermatocytes and round spermatids of the testis. DGKκ was distributed in the primary and secondary spermatocyte of the testis.

Conclusion: These results indicate that the expression patterns of the type II DGK isoforms δ2, η1, η3 and κ differ from each other, suggesting that these DGK isoforms play specific roles in distinct compartments and developmental stages of the reproductive organs, especially in the processes of spermatogenesis and oocyte maturation.

Show MeSH
Related in: MedlinePlus