Germline recombination in a novel Cre transgenic line, <i>Prl3b1</i>‐Cre mouse

Al-Soudy, Al-Sayed; Nakanishi, Takashi; Mizuno, Seiya; Hasegawa, Yoshikazu; Shawki, Hossam H.; Katoh, Michio; Basha, Walaa A.; Ibrahim, Abdelazim; El‐Shemy, Hany A.; Iseki, Hiroyoshi; Yoshiki, Atsushi; Hiromori, Youhei; Nagase, Hisamitsu; Takahashi, Satoru; Oishi, Hisashi; Sugiyama, Fumihiro

doi:10.1002/dvg.22944

Cited by 6 publications

(12 citation statements)

References 26 publications

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“…Mice of the Mafb fl/fl ;CAG-CreER ™ genotype are herein designated Mafb -cKO, and age-matched Mafb fl/fl mice were considered controls ( Fig 5A ). To determine the efficiency of the Cre deleter line used, we crossed CAG-CreER ™ driver mice with double-fluorescent Cre reporter mice R26GRR ( ROSA26 CAG-GFP/tdsRed ) that express GFP fluorescence constitutively in all cell types prior to Cre-mediated excision and tdsRED after excision [ 21 , 22 ]. The resulting male offspring were genotyped, and double heterozygous mice carrying GRR and Cre recombinase were then injected with tamoxifen at 6 weeks age.…”

Section: Resultsmentioning

confidence: 99%

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MAFB is dispensable for the fetal testis morphogenesis and the maintenance of spermatogenesis in adult mice

et al. 2018

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The transcription factor MAFB is an important regulator of the development and differentiation of various organs and tissues. Previous studies have shown that MAFB is expressed in embryonic and adult mouse testes and is expected to act as the downstream target of retinoic acid (RA) to initiate spermatogenesis. However, its exact localization and function remain unclear. Here, we localized MAFB expression in embryonic and adult testes and analyzed its gene function using Mafb-deficient mice. We found that MAFB and c-MAF are the only large MAF transcription factors expressed in testes, while MAFA and NRL are not. MAFB was localized in Leydig and Sertoli cells at embryonic day (E) 18.5 but in Leydig cells, Sertoli cells, and pachytene spermatocytes in adults. Mafb-deficient testes at E18.5 showed fully formed seminiferous tubules with no abnormal structure or differences in testicular somatic cell numbers compared with those of control wild-type mice. Additionally, the expression levels of genes related to development and function of testicular cells were unchanged between genotypes. In adults, the expression of MAFB in Sertoli cells was shown to be stage specific and induced by RA. By generating Mafbfl/fl CAG-CreER™ (Mafb-cKO) mice, in which Cre recombinase was activated upon tamoxifen treatment, we found that the neonatal cKO mice died shortly upon Mafb deletion, but adult cKO mice were alive upon deletion. Adult cKO mice were fertile, and spermatogenesis maintenance was normal, as indicated by histological analysis, hormone levels, and germ cell stage-specific markers. Moreover, there were no differences in the proportion of seminiferous stages between cKO mice and controls. However, RNA-Seq analysis of cKO Sertoli cells revealed that the down-regulated genes were related to immune function and phagocytosis activity but not spermatogenesis. In conclusion, we found that MAFB is dispensable for fetal testis morphogenesis and spermatogenesis maintenance in adult mice, despite the significant gene expression in different cell types, but MAFB might be critical for phagocytosis activity of Sertoli cells.

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Section: Resultsmentioning

confidence: 99%

“…R26GRR reporter mice, as previously described [ 21 , 22 ], were mated with the CAG-CreER ™ driver mouse strain obtained from Jackson laboratories. The progeny were genotyped using the Cre and GFP primers listed in Table 1 .…”

Section: Methodsmentioning

confidence: 99%

MAFB is dispensable for the fetal testis morphogenesis and the maintenance of spermatogenesis in adult mice

et al. 2018

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“…EFCAB2 was stained in blue by Stains-all, and this binding induced only the J band. Alternatively, ruthenium red is known to interact with CaBPs and shows red [26]. The positive activity of EFCAB2 by Stains-all and ruthenium red analyses, suggests its ability for the calcium binding.…”

Section: Discussionmentioning

confidence: 99%

“…The coding region of Efcab2 cDNA was amplified by RT-PCR from mouse testes using the sense primer and the antisense primer . The product was digested with NotI/XbaI and ligated into p3xFLAG-CMV-14 expression vector (Sigma, St. Louis, MO) [26]. The 293T cells were transfected with control (empty vector) or Efcab2 -expression plasmid using Fugene 6 transfection reagent (Promega, Madison, WI).…”

Section: Methodsmentioning

confidence: 99%

EFCAB2 is a novel calcium-binding protein in mouse testis and sperm

et al. 2019

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Calcium-binding proteins regulate ion metabolism and the necessary signaling pathways for the maturational events of sperm. Our aim is to identify the novel calcium-binding proteins in testis. The gene EFCAB2 (GenBank NM_026626.3, NP_080902.1) was not previously examined, and its properties and exact mechanisms of action are unknown. In this study, we performed phylogenetic and structure prediction analyses of EFCAB2, which displays definitive structural features. Additionally, the distribution, localization, and calcium binding ability of mouse EFCAB2 were investigated. Results revealed extensive conservation of EFCAB2 among different eukaryotic orthologs. The constructed 3D model predicted that mouse EFCAB2 contains seven α-helices and two EF-hand motifs. The first EF-hand motif is located in N-terminal, while the second is located in C-terminal. By aligning the 3D structure of Ca 2+ -binding loops from EFCAB2 with calmodulin, we predicted six residues that might be involved in Ca 2+ binding. The distribution of the Efcab2 mRNA, as determined by northern blotting, was detected only in the testis among mouse tissues. Native and recombinant EFCAB2 protein were detected by western blotting as one band at 20 kDa. In situ hybridization and immunohistochemical analyses showed its localization specifically in spermatogenic cells from primary spermatocytes to elongate spermatids within the seminiferous epithelium, but neither spermatogonia nor somatic cells were expressed. Moreover, EFCAB2 was specifically localized to the principal piece of cauda epididymal sperm flagellum. Furthermore, the analyses of purified recombinant EFCAB2 by Stains-all, ruthenium red staining, and by applying in vitro autoradiography assay showed that the physiological function of this protein is Ca 2+ binding. These results suggested that EFCAB2 might be involved in the control of sperm flagellar movement. Altogether, here we describe about EFCAB2 as a novel calcium-binding protein in mouse testis and sperm.

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“…Refinements to mouse mutagenesis techniques resulted in trophoblast lineage-specific targeting using Cre/Lox strategies. Regulatory sequences associated with the human Cytochrome P450 family 19 subfamily A member 1 (Cyp19a1) gene, several mouse genes (Trophoblast specific protein alpha, Tpbpa; Prolactin family 3, subfamily d, member 1, Prl3d1; Prolactin family 2, subfamily c, member 2, Prl2c2; PR/SET domain 1, Prdm1; Keratin 5, Krt5; Prolactin family 3, subfamily b, member 1, Prl3b1; Gcm1; Cathepsin Q, Ctsq), and a chimeric Tpbpa/Adenosine deaminase (Ada) enhancer have all been effectively utilized to produce gene disruptions in specific lineages of trophoblast cells [146][147][148][149][150][151][152][153][154][155] (Table 2). Strengths and limitations of Cre mouse lines for placentaspecific gene disruptions have been discussed [156].…”

Section: Targeted Mutagenesis In the Mousementioning

confidence: 99%

Hemochorial placentation: development, function, and adaptations†

Soares

Varberg

Iqbal

2018

Biology of Reproduction

142

158

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Placentation is a reproductive adaptation that permits fetal growth and development within the protected confines of the female reproductive tract. Through this important role, the placenta also determines postnatal health and susceptibility to disease. The hemochorial placenta is a prominent feature in primate and rodent development. This manuscript provides an overview of the basics of hemochorial placental development and function, provides perspectives on major discoveries that have shaped placental research, and thoughts on strategies for future investigation.

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Germline recombination in a novel Cre transgenic line, Prl3b1‐Cre mouse

Cited by 6 publications

References 26 publications

MAFB is dispensable for the fetal testis morphogenesis and the maintenance of spermatogenesis in adult mice

MAFB is dispensable for the fetal testis morphogenesis and the maintenance of spermatogenesis in adult mice

EFCAB2 is a novel calcium-binding protein in mouse testis and sperm

Hemochorial placentation: development, function, and adaptations†

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