Limits...
Expression and localization of aquaporin 1b during oocyte development in the Japanese eel (Anguilla japonica).

Kagawa H, Kishi T, Gen K, Kazeto Y, Tosaka R, Matsubara H, Matsubara T, Sawaguchi S - Reprod. Biol. Endocrinol. (2011)

Bottom Line: Light microscopic immunocytochemical analysis of ovary revealed that the Japanese eel AQP1b was expressed in the cytoplasm around the yolk globules which were located in the peripheral region of oocytes during the primary yolk globule stage; thereafter, the immunoreactivity was observed throughout the cytoplasm of oocyte as vitellogenesis progressed.The immunoreactivity became localized around the large membrane-limited yolk masses which were formed by the fusion of yolk globules during the oocyte maturation phase.These results together indicate that AQP1b, which is synthesized in the oocyte during the process of oocyte growth, is essential for mediating water uptake into eel oocytes.

View Article: PubMed Central - HTML - PubMed

Affiliation: Faculty of Agriculture, University of Miyazaki, Miyazaki 889-2192, Japan. kagawa@cc.miyazaki-u.ac.jp

ABSTRACT
To elucidate the molecular mechanisms underling hydration during oocyte maturation, we characterized the structure of Japanese eel (Anguilla japonica) novel-water selective aquaporin 1 (AQP1b) that thought to be involved in oocyte hydration. The aqp1b cDNA encodes a 263 amino acid protein that includes the six potential transmembrane domains and two Asn-Pro-Ala motifs. Reverse transcription-polymerase chain reaction showed transcription of Japanese eel aqp1b in ovary and testis but not in the other tissues. In situ hybridization studies with the eel aqp1b cRNA probe revealed intense eel aqp1b signal in the oocytes at the perinucleolus stage and the signals became faint during the process of oocyte development. Light microscopic immunocytochemical analysis of ovary revealed that the Japanese eel AQP1b was expressed in the cytoplasm around the yolk globules which were located in the peripheral region of oocytes during the primary yolk globule stage; thereafter, the immunoreactivity was observed throughout the cytoplasm of oocyte as vitellogenesis progressed. The immunoreactivity became localized around the large membrane-limited yolk masses which were formed by the fusion of yolk globules during the oocyte maturation phase. These results together indicate that AQP1b, which is synthesized in the oocyte during the process of oocyte growth, is essential for mediating water uptake into eel oocytes.

Show MeSH

Related in: MedlinePlus

Tissue distribution of aqp1b in Japanese eel. RT-PCR was performed using 1 μg of total RNA prepared from Japanese eel tissues using the Omniscript kit (Qiagen). Amplification products were analyzed on a 2.0% agarose gel and stained by ethidium bromide. Control RT-PCR of β-actin using the same amount of cDNA is also indicated in the lower panel. Lane from the left is brain(1), eye(2), gill(3), esophagus(4), heart(5), liver(6), kidney(7), intestine(8), ovary(9), testis(10), and no template (11), respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3117773&req=5

Figure 3: Tissue distribution of aqp1b in Japanese eel. RT-PCR was performed using 1 μg of total RNA prepared from Japanese eel tissues using the Omniscript kit (Qiagen). Amplification products were analyzed on a 2.0% agarose gel and stained by ethidium bromide. Control RT-PCR of β-actin using the same amount of cDNA is also indicated in the lower panel. Lane from the left is brain(1), eye(2), gill(3), esophagus(4), heart(5), liver(6), kidney(7), intestine(8), ovary(9), testis(10), and no template (11), respectively.

Mentions: Phylogenetic tree of vertebrate AQP1 proteins. The unrooted phylogenetic tree was constructed by neighbor-joining method after alignment of deduced amino acid sequences of AQP1 protein. Values at interior nodes are bootstrap percentages derived from 1000 replications. The scale bar indicates an evolutionary distance of 0.1 amino acid substitution per position in the sequence.


Expression and localization of aquaporin 1b during oocyte development in the Japanese eel (Anguilla japonica).

Kagawa H, Kishi T, Gen K, Kazeto Y, Tosaka R, Matsubara H, Matsubara T, Sawaguchi S - Reprod. Biol. Endocrinol. (2011)

Tissue distribution of aqp1b in Japanese eel. RT-PCR was performed using 1 μg of total RNA prepared from Japanese eel tissues using the Omniscript kit (Qiagen). Amplification products were analyzed on a 2.0% agarose gel and stained by ethidium bromide. Control RT-PCR of β-actin using the same amount of cDNA is also indicated in the lower panel. Lane from the left is brain(1), eye(2), gill(3), esophagus(4), heart(5), liver(6), kidney(7), intestine(8), ovary(9), testis(10), and no template (11), respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3117773&req=5

Figure 3: Tissue distribution of aqp1b in Japanese eel. RT-PCR was performed using 1 μg of total RNA prepared from Japanese eel tissues using the Omniscript kit (Qiagen). Amplification products were analyzed on a 2.0% agarose gel and stained by ethidium bromide. Control RT-PCR of β-actin using the same amount of cDNA is also indicated in the lower panel. Lane from the left is brain(1), eye(2), gill(3), esophagus(4), heart(5), liver(6), kidney(7), intestine(8), ovary(9), testis(10), and no template (11), respectively.
Mentions: Phylogenetic tree of vertebrate AQP1 proteins. The unrooted phylogenetic tree was constructed by neighbor-joining method after alignment of deduced amino acid sequences of AQP1 protein. Values at interior nodes are bootstrap percentages derived from 1000 replications. The scale bar indicates an evolutionary distance of 0.1 amino acid substitution per position in the sequence.

Bottom Line: Light microscopic immunocytochemical analysis of ovary revealed that the Japanese eel AQP1b was expressed in the cytoplasm around the yolk globules which were located in the peripheral region of oocytes during the primary yolk globule stage; thereafter, the immunoreactivity was observed throughout the cytoplasm of oocyte as vitellogenesis progressed.The immunoreactivity became localized around the large membrane-limited yolk masses which were formed by the fusion of yolk globules during the oocyte maturation phase.These results together indicate that AQP1b, which is synthesized in the oocyte during the process of oocyte growth, is essential for mediating water uptake into eel oocytes.

View Article: PubMed Central - HTML - PubMed

Affiliation: Faculty of Agriculture, University of Miyazaki, Miyazaki 889-2192, Japan. kagawa@cc.miyazaki-u.ac.jp

ABSTRACT
To elucidate the molecular mechanisms underling hydration during oocyte maturation, we characterized the structure of Japanese eel (Anguilla japonica) novel-water selective aquaporin 1 (AQP1b) that thought to be involved in oocyte hydration. The aqp1b cDNA encodes a 263 amino acid protein that includes the six potential transmembrane domains and two Asn-Pro-Ala motifs. Reverse transcription-polymerase chain reaction showed transcription of Japanese eel aqp1b in ovary and testis but not in the other tissues. In situ hybridization studies with the eel aqp1b cRNA probe revealed intense eel aqp1b signal in the oocytes at the perinucleolus stage and the signals became faint during the process of oocyte development. Light microscopic immunocytochemical analysis of ovary revealed that the Japanese eel AQP1b was expressed in the cytoplasm around the yolk globules which were located in the peripheral region of oocytes during the primary yolk globule stage; thereafter, the immunoreactivity was observed throughout the cytoplasm of oocyte as vitellogenesis progressed. The immunoreactivity became localized around the large membrane-limited yolk masses which were formed by the fusion of yolk globules during the oocyte maturation phase. These results together indicate that AQP1b, which is synthesized in the oocyte during the process of oocyte growth, is essential for mediating water uptake into eel oocytes.

Show MeSH
Related in: MedlinePlus