Developmental Biology: In the article "Spatial and temporal pattern of expression of the cellular retinoic acid-binding protein and the cellular retinol-binding protein during mouse embryogenesis" by Ana V. Perez-Castro, Leslie E. TothRogler, Li-na Wei, and M. Chi Nguyen-Huu, which appeared in number 22, November 1989, ofProc. NatI. Acad. Sci. USA (86,(8813)(8814)(8815)(8816)(8817)
Mutagenesis was studied in cultured F9 embryonal carcinoma cells infected with a variant of Moloney murine leukemia virus. Proviral insertion induced the inactivation of the hypoxanthine phosphoribosyltransferase locus, and the virus was used to isolate the mutated genes rapidly. Mutagenesis by these methods may be useful for the genetic dissection of the various mammalian cell phenotypes.
The MH-3 gene, which contains a homeo box that is expressed specifically in the adult testis, was identified and mapped to mouse chromosome 6. By means of in situ hybridization with adult testis sections and Northern blot hybridization with testis RNA from prepuberal mice and from Sl/Sld mutant mice, it was demonstrated that this gene is expressed in male germ cells during late meiosis. In the embryo, MH-3 transcripts were present at day 11.5 post coitum, a stage in mouse development when gonadal differentiation has not yet occurred. The MH-3 gene may have functions in spermatogenesis and embryogenesis.
P19 embryonal carcinoma (EC) cells can be induced to differentiate in vitro into a variety of cell types by treatment with different concentrations of retinoic acid (RA). A study was conducted to explore the regulation of expression of the genes for cellular retinoic acid-binding protein (CRABP) and cellular retinol-binding protein (CRBP) in P19 cells induced to differentiate by RA. For each retinoid-binding protein, both the level of specific mRNA and of immunoreactive protein were measured, respectively, by RNase protection assay and by a specific RIA. Dramatic increases in CRABP and CRBP were seen, at both the mRNA and protein levels, during the RA-induced differentiation. CRBP induction differed from that of CRABP in several major ways. 1) Induction of CRBP occurred at lower concentrations of RA (10(-9) M) than did that of CRABP (10(-8)-10(-7) M). 2) CRBP induction was an early response (within 3 h) to RA treatment, whereas CRABP induction occurred at a later time (12-24 h). 3) Induction of CRABP mRNA by RA was blocked by the protein synthesis inhibitor cycloheximide, whereas induction of CRBP mRNA was not. 4) Several differentiation inducers were tested for their effects on the expression of CRABP and CRBP in P19 cells. CRBP induction occurred with a wider spectrum of inducers than did that of CRABP. 5) In addition, the induction of CRABP and CRBP mRNAs by RA was examined in six different cell lines, including three EC lines. CRBP induction occurred in a wider spectrum of cell lines than did that of CRABP. The induction of CRABP in EC cells seems, in general, to correlate with their differentiation into neuron-like cells. Taken together, our results suggest that CRBP induction may be a direct response to RA and represent a general event in RA-induced cell differentiation, whereas CRABP induction may be an indirect response and represent a later event restricted to only certain differentiation pathways. CRBP may be an early response gene induced by RA.
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