Limits...
Molecular cloning, spatial and temporal expression analysis of CatSper genes in the Chinese Meishan pigs.

Song C, Gao B, Wu H, Xie Y, Wang X, Li B, Chen G, Mao J - Reprod. Biol. Endocrinol. (2011)

Bottom Line: Among the four CatSpers, CatSper2, 3, and 4 were more conserved across species, compared with CatSper1.CatSper3 and CatSper4 mRNAs were present in mature sperm cells.Substantial upregulation of CatSper genes was initiated at Day 60 and maintained this marked production until sexual maturity.

View Article: PubMed Central - HTML - PubMed

Affiliation: College of Animal Science & Technology, Yangzhou University, Yangzhou, Jiangsu, 225009, China. chengyilab@hotmail.com

ABSTRACT

Background: Sperm ion channel proteins (CatSpers) are essential for sperm hyperactivated motility, and then penetration through the zona pellucida. The CatSper class of proteins have well been characterized in the mouse and human. However, such data for pigs are not available. In the present study, we cloned the porcine CatSper 1-4 genes, analysed their spatial expression in various organs and temporal expression in the testes from birth until sexual maturity in Meishan boars.

Methods: Rapid amplification of cDNA ends (RACE) was performed to clone the full length cDNAs of porcine CatSper genes and bioinformatics analysis of inferred CatSper proteins was also determined. Various organs were collected from 150 day-old pigs to characterize the spatial expression of CatSper genes by qualitative reverse transcriptase polymerase chain reaction (RT-PCR), and testes from birth to 150 day-old boars were sampled to detect the temporal expression of CatSper genes by quantitative real-time RT-PCR.

Results: The mRNA sequences of CatSper1 (2452 bp), CatSper2 (2038 bp), CatSper3 (1408 bp), and CatSper4 (1799 bp), including full length of cDNAs, 5' and 3' flanks, were obtained. The bioinformatics analysis indicated that coding regions spanning the ion transport domains were conserved for different species analyzed. Among the four CatSpers, CatSper2, 3, and 4 were more conserved across species, compared with CatSper1. In addition, six conservative trans-membrane domains, a pore forming motif, and a coiled-coil motif were also identified. The spatial analysis from different organs showed that CatSper1 was detected in both testes and hypothalamus, CatSper2 was restricted in testes only, CatSper4 was expressed in testes and rete testes; whereas CatSper3 was more ubiquitously. CatSper3 and CatSper4 transcripts were also detected in ejaculated sperm. At Days 1 and 30 of age, CatSper mRNAs exhibited only sparse expression in the testes. However, these transcripts highly expressed at Day 60 and onward till sexual maturity (Day 150 of age).

Conclusions: The spatial and temporal expression profiles of CatSper genes were reported herein for the first time in pigs. CatSper1, CatSper2 and CatSper4 were primarily expressed in testes, while CatSper3 transcript was prevalent in a variety of organs. CatSper3 and CatSper4 mRNAs were present in mature sperm cells. Substantial upregulation of CatSper genes was initiated at Day 60 and maintained this marked production until sexual maturity.

Show MeSH

Related in: MedlinePlus

Temporal expression of porcine CatSpers in testes. Relative expression of CatSper1-4/GAPDH in testes at days 1 (n = 5), 30 (n = 4), 60 (n = 3), 90 (n = 3), and 150 (n = 3) of age. * indicates significant change compared to the former time point (P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3198926&req=5

Figure 7: Temporal expression of porcine CatSpers in testes. Relative expression of CatSper1-4/GAPDH in testes at days 1 (n = 5), 30 (n = 4), 60 (n = 3), 90 (n = 3), and 150 (n = 3) of age. * indicates significant change compared to the former time point (P < 0.05).

Mentions: The temporal expression profile of CatSper mRNAs during testicular development was determined by quantitative real-time RT-PCR. Their expression levels at different stage of development after birth from day 1 to 150 of age were summarized in Figure 7. At Days 1 and 30, CatSper1, CatSper3 and CatSper4 were not detectable, while Catsper2 began to be transcribed and was detected at a low level by Day 1. This low expression level was maintained until Day 30; whereupon, the expression of this gene transcript increased dramatically from Days 30 to 60 (Figure 7). After Day 60, all CatSper genes were abundantly expressed, and maintained at seemingly high constitutive level without alteration until Day 150 (Figure 7), which is the last age that was analyzed.


Molecular cloning, spatial and temporal expression analysis of CatSper genes in the Chinese Meishan pigs.

Song C, Gao B, Wu H, Xie Y, Wang X, Li B, Chen G, Mao J - Reprod. Biol. Endocrinol. (2011)

Temporal expression of porcine CatSpers in testes. Relative expression of CatSper1-4/GAPDH in testes at days 1 (n = 5), 30 (n = 4), 60 (n = 3), 90 (n = 3), and 150 (n = 3) of age. * indicates significant change compared to the former time point (P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3198926&req=5

Figure 7: Temporal expression of porcine CatSpers in testes. Relative expression of CatSper1-4/GAPDH in testes at days 1 (n = 5), 30 (n = 4), 60 (n = 3), 90 (n = 3), and 150 (n = 3) of age. * indicates significant change compared to the former time point (P < 0.05).
Mentions: The temporal expression profile of CatSper mRNAs during testicular development was determined by quantitative real-time RT-PCR. Their expression levels at different stage of development after birth from day 1 to 150 of age were summarized in Figure 7. At Days 1 and 30, CatSper1, CatSper3 and CatSper4 were not detectable, while Catsper2 began to be transcribed and was detected at a low level by Day 1. This low expression level was maintained until Day 30; whereupon, the expression of this gene transcript increased dramatically from Days 30 to 60 (Figure 7). After Day 60, all CatSper genes were abundantly expressed, and maintained at seemingly high constitutive level without alteration until Day 150 (Figure 7), which is the last age that was analyzed.

Bottom Line: Among the four CatSpers, CatSper2, 3, and 4 were more conserved across species, compared with CatSper1.CatSper3 and CatSper4 mRNAs were present in mature sperm cells.Substantial upregulation of CatSper genes was initiated at Day 60 and maintained this marked production until sexual maturity.

View Article: PubMed Central - HTML - PubMed

Affiliation: College of Animal Science & Technology, Yangzhou University, Yangzhou, Jiangsu, 225009, China. chengyilab@hotmail.com

ABSTRACT

Background: Sperm ion channel proteins (CatSpers) are essential for sperm hyperactivated motility, and then penetration through the zona pellucida. The CatSper class of proteins have well been characterized in the mouse and human. However, such data for pigs are not available. In the present study, we cloned the porcine CatSper 1-4 genes, analysed their spatial expression in various organs and temporal expression in the testes from birth until sexual maturity in Meishan boars.

Methods: Rapid amplification of cDNA ends (RACE) was performed to clone the full length cDNAs of porcine CatSper genes and bioinformatics analysis of inferred CatSper proteins was also determined. Various organs were collected from 150 day-old pigs to characterize the spatial expression of CatSper genes by qualitative reverse transcriptase polymerase chain reaction (RT-PCR), and testes from birth to 150 day-old boars were sampled to detect the temporal expression of CatSper genes by quantitative real-time RT-PCR.

Results: The mRNA sequences of CatSper1 (2452 bp), CatSper2 (2038 bp), CatSper3 (1408 bp), and CatSper4 (1799 bp), including full length of cDNAs, 5' and 3' flanks, were obtained. The bioinformatics analysis indicated that coding regions spanning the ion transport domains were conserved for different species analyzed. Among the four CatSpers, CatSper2, 3, and 4 were more conserved across species, compared with CatSper1. In addition, six conservative trans-membrane domains, a pore forming motif, and a coiled-coil motif were also identified. The spatial analysis from different organs showed that CatSper1 was detected in both testes and hypothalamus, CatSper2 was restricted in testes only, CatSper4 was expressed in testes and rete testes; whereas CatSper3 was more ubiquitously. CatSper3 and CatSper4 transcripts were also detected in ejaculated sperm. At Days 1 and 30 of age, CatSper mRNAs exhibited only sparse expression in the testes. However, these transcripts highly expressed at Day 60 and onward till sexual maturity (Day 150 of age).

Conclusions: The spatial and temporal expression profiles of CatSper genes were reported herein for the first time in pigs. CatSper1, CatSper2 and CatSper4 were primarily expressed in testes, while CatSper3 transcript was prevalent in a variety of organs. CatSper3 and CatSper4 mRNAs were present in mature sperm cells. Substantial upregulation of CatSper genes was initiated at Day 60 and maintained this marked production until sexual maturity.

Show MeSH
Related in: MedlinePlus