Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Comparative Analysis of cul5 and rbx2 Expression in the Developing and Adult Murine Brain and Their Essentiality During Mouse Embryogenesis

doi: 10.1002/dvdy.24675

Figure Lengend Snippet: Disruption of cul5 and rbx2 causes embryonic lethality at different developmental stages. Diagram of cul5GT (A) and rbx2GT (B) alleles indicating the LacZ cassette insertion in the first intron in both genes. Notice that whereas cul5GT and rbx2GT generate null alleles, rbx2GT can be converted to a conditional allele (rbx2fl) via FLPe recombination and to a knockout allele (rbx2KO) via CRE recombination. White boxes in exons indicate untranslated regions and gray boxes indicate coding sequences. Genotyping primers and their relative positions are indicated. Detailed explanation of genotyping strategy can be found in the Material and Methods section. C: No homozygous cul5GT embryos were collected from n = 6 litters at E3.5 and n = 5 litters analyzed at P0. On the contrary, homozygous rbx2KO embryos were collected at E3.5 (n = 15 litters) but failed to survive until birth (n = 6 litters) (D). The number of embryos obtained per genotype and age is indicated in each case. Differences from expected Mendelian ratio were tested using the Chi-square test (χ2), and P values are indicated in each case. ß-gal, beta-galactosidase; ß-geo, beta-galactosidase + Neomycin resistance gene; Chr9, chromosome 9; En2 intr1, partial Engrailed 2 Intron 1; En2 SA, Engrail 2 splicing acceptor; NeoR, Neomycin-resistance gene; pA, polyadenylation site; SA, splicing acceptor; T2A, thosea asigna virus 2A peptide.

Article Snippet: Similar to cul5GT , no rbx2 mutant pups (postnatal day P0) were recovered from heterozygous rbx2 KO intercrossing (Chi-square test P = 0.00228) ( ).

Techniques: Disruption, Knock-Out, Virus

Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Comparative Analysis of cul5 and rbx2 Expression in the Developing and Adult Murine Brain and Their Essentiality During Mouse Embryogenesis

doi: 10.1002/dvdy.24675

Figure Lengend Snippet: Co-expression of Cul5 and Rbx2 in the adult brain. Comparative analysis of Cul5 and Rbx2 by immunofluorescence in consecutive wild-type adult brain sections. In the cerebellum, both Cul5 and Rbx2 were detected in the nuclei of Purkinje cells (purple arrowheads) and broadly distributed in the internal granular layer (A). In the hippocampus, Cul5 was detected principally in the cytoplasm of the pyramidal cells of the cornu ammonis and in mossy fibers. Rbx2 was detected in the nuclei of pyramidal cells and in mossy fibers (B). In the neocortical neurons, Cul5 was detected in the cytoplasm and Rbx2 in the nucleus. On the contrary, in the caudate-putamen, both Cul5 and Rbx2 have a nuclear localization. D-D’“: Immunofluorescence against ß-galactosidase and Cul5 in adult rbx2GT tissue shows ubiquitous colocalization, including the neocortex (D), caudate-putamen (D”), hippocampus (D“), and cerebellum (D”’). MF, mossy fibers; sl, stratum lucidum. Scale bars A,B = 100 μm. Scale bar C = 500 μm. Scale bar D = 25 μm.

Article Snippet: Similar to cul5GT , no rbx2 mutant pups (postnatal day P0) were recovered from heterozygous rbx2 KO intercrossing (Chi-square test P = 0.00228) ( ).

Techniques: Expressing, Immunofluorescence

Journal: Developmental dynamics : an official publication of the American Association of Anatomists

Article Title: Comparative Analysis of cul5 and rbx2 Expression in the Developing and Adult Murine Brain and Their Essentiality During Mouse Embryogenesis

doi: 10.1002/dvdy.24675

Figure Lengend Snippet: Expression of rbx2 during development and in the adult brain. rbx2 expression parallels cux5 expression in all the areas analyzed. In the olfactory bulb, rbx2 was strongly expressed in the mitral layer at all ages analyzed, and expression of rbx2 was detected in granule cells at postnatal stages only (A). rbx2 expression in the neocortex was detected in proliferative zones as well as in somatic areas (B) (E16.5 and P0). In the adult neocortex, strong LacZ staining was observed in all cortical layers (AD). Similar to cu/5, rbx2 was detected in proliferative areas of the hippocampus and in the stratum pyramidale at early stages (C) (E16.5). LacZ signal was detected in the dentate gyrus starting at postnatal stages and reaching maximum expression in the adult (P0 and AD). Comparative analysis of rbx2 expression in the forebrain indicated that rbx2 was ubiquitously expressed during development and in the adult (D). In the hindbrain, rbx2 expression was detected principally in Purkinje cells, deep cerebellar nuclei, and medulla (E14.5 and P0). In the adult cerebellum, a strong LacZ staining was observed in Purkinje cells, but especially in granule cells of the internal granular layer, similar to cu/5. th, thalamus; 6N, abducens nucleus; 7N, facial nucleus. Scale bars A,C = 100 μm. Scale bars D,E = 500 μm.

Article Snippet: Similar to cul5GT , no rbx2 mutant pups (postnatal day P0) were recovered from heterozygous rbx2 KO intercrossing (Chi-square test P = 0.00228) ( ).

Techniques: Expressing, Staining