Publisher Summary This chapter discusses the formal genetics of mouse anemias and their significance for an analysis of patterns of mammalian gene action. It also describes normal hemopoietic development, homeostasis, and hemoglobin polymorphism. Anemias are classified into five categories—(1) anemias of maturation arrest, (2) anemias involving defects in iron utilization, (3) anemias involving greatly reduced erythrocyte life-span, (4) hemoglobinopathies, and (5) anemias almost secondary to genetically induced defects. During the total life of a mouse, including prenatal development, hemopoiesis takes place in four different sites. Although hemopoiesis continues in the liver until birth and for a brief time thereafter, hemopoiesis (largely myelopoiesis) commences in the spleen on fetal day 15 or 16 and in the marrow around the time of birth. Hematologically normal adult mice from many different inbred strains regularly produce more than one kind of hemoglobin. There is a widespread genetic polymorphism at both the Hba locus, determining the α-globin structure and the Hbb locus, determining the β-globin structure, and both the loci involve considerable genetic complexity.

Hereditary Anemias of the Mouse: A Review for Geneticists*

Butyric acid, a potent inducer of erythroid differentiation in cultured erythroleukemic cells

The autofluorescence other than intrinsic protein emission of viable cultured mammalian cells has been investigated. The fluorescence was found to originate in discrete cytoplasmic vesicle-like regions and to be absent from the nucleus. Excitation and emission spectra of viable cells revealed at least two distinct fluorescent species. Comparison of cell spectra with spectra of known cellular metabolites suggested that most, if not all, of the fluorescence arises from intracellular nicotinamide adenine dinucleotide (NADH) and riboflavin and flavin coenzymes. Various changes in culture conditions did not affect the observed autofluorescence intensity. A multiparameter flow system (MACCS) was used to compare the fluorescence intensities of numerous cultured mammalian cells.

Autofluorescence of viable cultured mammalian cells.

Addition of the potent tumor promoter, 12-O-tetradecanoyl-phorbol-13-acetate (TPA), to murine erythroleukemia cell lines in suspension cultures inhibited both spontaneous differentiation and differentiation induced by hexamethylene bisacetamide (HMBA), dimethyl sulfoxide, or butyric acid. Inhibition was unrelated to cytotoxicity and was reversible. When several plant diterpenes were tested, there was a positive correlation between tumor-promoting activity and inhibition of differentiation. TPA inhibited HMBA-induced differentiation only if added prior to the time of commitment to differentiation, as assayed by scoring for differentiation after transfer of cells from HMBA to fresh medium without HMBA. TPA-mediated inhibition of differentiation was associated with a decrease in globin mRNA accumulation.

Tumor promoters inhibit spontaneous and induced differentiation of murine erythroleukemia cells in culture

We have developed a general approach to the cloning of specific segments of the mammalian genome that involves a two-step purification of EcoRI fragments of mammalian DNA and their in vitro insertion into a suitably constructed EK2 derivative of bacteriophage lambda. The combination of fragment purification, exclusion of parental-type recombinants, and simple phage screening techniques permits the isolation of virtually any gene segment for which there is an identifying hybridization probe. We illustrate the approach by describing the cloning of an approximately 7000-base-long segment of mouse DNA containing globin and surrounding gene sequences.

https://www.pnas.org/content/pnas/74/10/4406.full.pdf

Cloning specific segments of the mammalian genome: bacteriophage lambda containing mouse globin and surrounding gene sequences.

The thymidine analogue 5-bromodeoxyuridine (BUdR) preferentially inhibits the synthesis of those proteins which are characteristic of the differentiated state of a cell1–5. The observed effects of BUdR have been suggested to be due to its incorporation into DNA1,4,6 and/or due to its interference with the synthesis of carbohydrates needed for the formation of some cell membrane constituents7,8. There has been no direct evidence, however, in favour of any hypothesis and nothing is known about the actual mechanisms involved.

Action of 5-bromodeoxyuridine on the induction of haemoglobin synthesis in mouse leukaemia cells resistant to 5-BUdR.

WE have described structural data for mouse and rabbit embryonic β-like e chains and the conclusions which can be drawn with respect to the evolution of e globin chain genes in both species1. Here we present evidence for crossover events in the ancestral mouse β and e-type chain genes leading to Hb Lepore-like globin chain genes which are functional in mice of recent evolutionary origin.

Evidence for Hb Lepore-like hybrid globin beta chain genes in mice.

https://febs.onlinelibrary.wiley.com/doi/pdf/10.1016/0014-5793%2873%2980836-1

Globin messenger in mouse leukemic cells: Activity associated with RNA species in the region of 8 to 16 S

Mammalian embryonic hemoglobins.

Wir haben uns in den letzten Jahren mit der Friend-Virus induzierten Erythroleukamie bei der Maus beschaftigt. Bei dieser werden sowohl abnormal differenzierte Zellen der roten, wie der lymphatischen Reihe gefunden. Deshalb lag nahe, diese multiple Differenzierungsentgleisung auf eine abnormale Vermehrung der gemeinsamen Stammzelle nach Transformation dieser Zelle durch das Friend-Virus zuruckzufuhren. In der Folgezeit zeigten aber verschiedene Autoren [1], das zwei Tumorviren, die nur in ihrer Vermehrung voneinander abhangig sind, unabhangig zwei verschiedene Zelltypen transformieren: Das lymphatische Leukamievirus (LLV) transformiert Zellen, die sich lymphatisch und das spleen focus forming-Virus (SFFV) solche, die sich erythroid differenzieren.

Untersuchungen zur Erythropoese bei erythroleukämischen Zellkulturen

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