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The use of the green fluorescent protein (GFP) to label specific cell types and track gene expression in animal models, such as mice, has evolved to become an essential tool in biological research. Transgenic animals expressing genes of interest linked to GFP, either as a fusion protein or transcribed from an internal ribosomal entry site (IRES) are widely used. Enhanced GFP (eGFP) is the most common...
The expression of all four fgfr genes was extensively examined throughout early embryogenesis of the zebrafish (Danio rerio). fgfr1 alone was expressed maternally throughout the blastoderm, and then zygotically in the anterior neural plate and presomitic mesoderm. fgfr4 expression was first detected in late blastulae and was gradually restricted to the brain. fgfr2 and fgfr3 expression were initiated...
The expression of the gene for zebrafish fibroblast growth factor receptor 4 (fgfr4) in a 3‐somite stage embryo was stained (blue) together with pax2a expression (red), which was seen mainly at the midbrain‐hindbrain boundary, by two‐color in situ hybridization. At this stage, fgfr4 is expressed in the midbrain, anterior hindbrain, and posterior hindbrain, in addition to the paraxial and lateral mesoderm...
We have generated a transgenic mouse line that expresses improved Cre recombinase (iCre) under the control of the testis‐expressed gene 101 (Tex101) promoter. This transgenic mouse line was named Tex101‐iCre. Using the floxed ROSA reporter mice, we found that robust Cre recombinase activity was detected in postnatal testes with weak or no activity in other tissues. Within the testis, Cre recombinase...
The SCUBE gene family encode secreted, extracellular proteins that share a distinct domain organization of at least five recognizable motifs, including an amino‐terminal signal peptide sequence, multiple EGF‐like domains, a large spacer region containing multiple N‐linked glycosylation sites, three repeated stretches of six‐cysteine residues and a carboxy‐terminal CUB domain. We describe a Scube3tm1Dge/H...
As a putative transcription factor, Af17 may play a role in multiple signaling pathways. However, the Af17 expression profile during development and in adult tissues remains largely uncharacterized. The importance of Af17 function in embryogenesis, hematopoiesis, and animal survival has never been addressed before. Here we report the generation of the first Af17 mutant mouse model and characterization...
The mammary gland is a developmentally dynamic, hormone‐responsive organ that undergoes proliferation and differentiation within the secretory epithelial compartment during pregnancy. The epithelia are maintained by pro‐survival signals (e.g., Stat5, Akt1) during lactation, but undergo apoptosis during involution through inactivation of cell survival pathways and upregulation of pro‐apoptotic proteins...
Early stages of avian development are not well‐suited for in ovo studies. A critical limitation of ex ovo culture is the short period of growth that can be attained for explanted embryos. Here we report a modified and simplified version of an existing ex ovo culture method, the cornish pasty culture. We show that this modified method, referred to as MC culture, can be used to grow chick and quail...
The transcription factors Foxd3 and Pax3 are important early regulators of neural crest (NC) progenitor cell properties. Homozygous mutations of Pax3 or a homozygous NC‐specific deletion of Foxd3 cause marked defects in most NC derivatives, but neither loss of both Foxd3 alleles nor loss of one Pax3 allele alone greatly affects overall development of cardiac NC derivatives. In contrast, compound mutant...
Summary:Irxl1 (Iroquois‐related homeobox like‐1) is a newly identified three amino‐acid loop extension (TALE) homeobox gene, which is expressed in various mesoderm‐derived tissues, particularly in the progenitors of the musculoskeletal system. To analyze the roles of Irxl1 during embryonic development, we generated mice carrying a null allele of Irxl1. Mice homozygous for the targeted allele were...
Cre/loxP‐dependent expression of fluorescent proteins represents a powerful biological tool for cell lineage, fate‐mapping, and genetic analysis. Live tissue imaging has significantly improved with the development of far‐red fluorescent proteins, with optimized spectral characteristics for in vivo applications. Here, we report the generation of the first transgenic mouse line expressing the far‐red...
The Connexin‐40 (Cx40) gene encodes a gap junction protein that plays an important role in cell‐cell communication in cardiomyocytes of the atria and cardiac conduction system and endothelial cells of large arteries. During embryonic development, Cx40 expression is tightly regulated and correlates with progressive ventricular conduction system (VCS) differentiation and vessel function. We have generated...
Transgenesis enables the elucidation of gene function; however, constant transgene expression is not always desired. The tetracycline responsive system was devised to turn on and off transgene expression at will. It has two components: a doxycycline (dox)‐controlled transactivator (TA) and an inducible expression cassette. Integration of these transgenes requires two transfection steps usually accomplished...
Female reproductive decline is one of the first aging phenotypes in humans, manifested in increasing rates of infertility, miscarriage, and birth defects in children of mothers over 35. Recently, Caenorhabditis elegans (C. elegans) has been developed as a model to study reproductive aging, and several studies have advanced our knowledge of reproductive aging regulation in this organism. In this review,...
Confocal images of immunofluorescence showing the specific expression of Cx40‐Cre allele through the expression of RFP (Red) colocalizing with alpha‐actinin in green in Purkinje fibers of the ventricular conduction system and with PECAM (blue) in endothelial cells of coronary vessels. See the article by Beyer at al. in this issue.
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