ABCMdb

PMID: 21625623

Xu WM, Chen J, Chen H, Diao RY, Fok KL, Dong JD, Sun TT, Chen WY, Yu MK, Zhang XH, Tsang LL, Lau A, Shi QX, Shi QH, Huang PB, Chan HC
Defective CFTR-dependent CREB activation results in impaired spermatogenesis and azoospermia.
PLoS One. 2011;6(5):e19120. Epub 2011 May 9., [PubMed]

Sentences

No. Mutations Sentence Comment

29 ABCC7 p.Ser489* Results Impaired spermatogenesis in CF mice models To investigate possible involvement of CFTR in spermatogenesis, we used a CFTR knockout (Cftrtm1Unc , also referred as S489X) mice. Login to comment 30 ABCC7 p.Ser489* Since most homozygous S489X CF mice die at a young age or less frequently available, heterozygous mice were used for quantitative measurement. Login to comment 31 ABCC7 p.Ser489* Morphological study showed that testis tissue size and weight were significantly lower in heterozygous S489X CF mice than that of wild-type control (Fig. 1A). Login to comment 32 ABCC7 p.Ser489* Daily sperm production (DSP), which is often used to evaluate the spemiogenesis in the testis [21], was significantly reduced in heterozygous S489X CF mice compared with wide-type control (Fig. 1B). Login to comment 33 ABCC7 p.Ser489* The sperm number recovered from the epididymis of the heterozygous S489X CF mice was also significantly lower than that of wide-type control (Fig. 1C). Login to comment 34 ABCC7 p.Ser489* H&E staining showed slight decrease in diameter of seminiferous tubules (Fig. 1D,E) and slight cytoplasmic shrinkage of spermatocytes and round spermatids (Fig. S1) in S489X CF mice. Login to comment 35 ABCC7 p.Ser489* Realtime PCR results showed significant reduction of Protamine-2, a specific marker of spermatid and spermatozoa, in heterozygous and homozygous S489X CF mice at mRNA level (Fig. 1F). Login to comment 36 ABCC7 p.Ser489* Immunofluorescence results further demonstrated the down-regulation of Protamine-2 in S489X CF mice at protein level (Fig. 1G). Login to comment 37 ABCC7 p.Ser489* CREM, a spermatid-specific transcription factor, was also down-regulated in the homozygous S489X CF mice compared to wild-type as indicated by Western blot and immunofluorescent staining respectively (Fig. 1H&J), suggesting defect in spermatogenesis in CF mice. Login to comment 38 ABCC7 p.Ser489* Interestingly, immunofluorescence result showed that cAMP-responsive element binding protein (CREB), an important transcription factor in Sertoli cells during spermatogenesis, was decreased in the Sertoli cells of homozygous S489X CF mice, as compared to heterozygotes and wild-type (Fig. 1I). Login to comment 39 ABCC7 p.Ser489* Western blot result also showed downregulation of phosphorylated and total CREB in homozygous S489X CF mice compared to wild-type and heterozygotes (Fig.1K). Login to comment 70 ABCC7 p.Ser489* (A) Significantly reduced testis size and weight seen in heterozygous S489X mice compared with wild-type mice (n = 3; *:p,0.05). Login to comment 71 ABCC7 p.Ser489* (B) Reduced daily sperm production (DSP) values retrieved from heterozygous S489X testes (n = 4; *:p,0.05). Login to comment 72 ABCC7 p.Ser489* (C) Reduced sperm numbers recovered from the epididymis of heterozygous S489X mice (+/+ n = 6, +/2 n = 4; *:p,0.05). Login to comment 73 ABCC7 p.Ser489* (D) H&E staining of the cross sections of wild2type, heterozygous and homozygous S489X CF mice testes. Login to comment 78 ABCC7 p.Ser489* (F) Realtime PCR of protamine-2 in S489X CF mice testes. Login to comment 79 ABCC7 p.Ser489* Protamine-2 mRNA level is significantly lowered in heterozygous and homozygous S489X CF mice compared to their wildtype littermates. Login to comment 80 ABCC7 p.Ser489* (G) Immunofluorescent staining of Protamine-2 in +/+, +/2 and 2/2 S489X CF mice testes (stage VII-VIII). Login to comment 111 ABCC7 p.Ser489* (H) Immunofluorescent staining of CREM in S489X CF mice testes (stage VIII-I). Login to comment 115 ABCC7 p.Ser489* (I) Immunofluorescent staining of CREB in S489X CF mice testes shows stronger immunoactivity in +/+ and +/2 mice Sertoli cell compared to that of 2/2. Login to comment 117 ABCC7 p.Ser489* (J) Western blot analysis of CREM expression in S489X CF testes. Login to comment 119 ABCC7 p.Ser489* (K) Western blot analysis of CREB expression in S489X CF testes. Login to comment 126 ABCC7 p.Ser489* Materials and Methods Animal and cell culture cftrtm1Unc (S489X) mice [45] were from Jackson`s laboratory and maintained in LASEC of CUHK. Login to comment