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Title: Differential expression of murine adult hemoglobins in early ontogeny

Abstract

A hemoglobin mutation is described that permits study of the expression of the two adult ..beta..-globin genes throughout fetal and postnatal development. Mice with a mutation at the Hbb/sup s/, ..beta..-globin locus, were used to study the relative levels of ..beta..-s2major and ..beta..-sminor globins specified by the mutant Hbb/sup s2/ haplotype during development. At 11.5 days of gestation ..beta..-sminor comprised over 80% and ..beta..-s2major under 20% of the adult beta-globin. The relative level of ..beta..-sminor decreased through fetal development; at birth ..beta..-sminor represented 33.7% of the ..beta..-globin. The adult values of 71.0% ..beta..-s2major and 29.0% ..beta..-sminor globin are expressed in mice six days after birth. Because the two ..beta..-globin genes are expressed in mice of the Hbb/sup 2s/ haplotype, both the ..beta..-smajor and ..beta..-sminor genes must be expressed in mice of the Hbb/sup s/ haplotype. Expression of the ..beta..-sminor gene is elevated to 35.6% in Hbb/sup s2/ mice that have been bled repeatedly. Thus, the 5' ..beta..-s2major and 3' ..beta..-sminor genes of the Hbb/sup s2/ haplotype and, presumably the 5' ..beta..-smajor and 3' ..beta..-sminor genes of the Hbb/sup s/ haplotype, are regulated independently and are homologous to the 5' ..beta..-dmajor and 3' ..beta..-dminor genes of the Hbb/sup d/ haplotype. Micemore » of the Hbb/sup s2/ haplotype are better than mice of the Hbb/sup d/ haplotytpe for studying the mechanisms of hemoglobin switching because the Hbb/sup s2/ each of the three embryonic and two adult hemoglobins can be separated by electrophoresis. 17 refs., 3 figs.« less

Authors:
; ;
Publication Date:
Research Org.:
Tennessee Univ., Oak Ridge (USA). Graduate School of Biomedical Sciences; Research Triangle Inst., Research Triangle Park, NC (USA). Chemistry and Life Sciences Div.; Oak Ridge National Lab., TN (USA)
OSTI Identifier:
5642932
Report Number(s):
CONF-8409198-1
ON: DE85005364
DOE Contract Number:
AC05-84OR21400
Resource Type:
Conference
Resource Relation:
Conference: Conference on hemoglobin switching, Airlie, VA, USA, 30 Sep 1984
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; BLOOD FORMATION; ONTOGENESIS; HEMOGLOBIN; GENES; QUANTITY RATIO; FETUSES; MICE; SICKLE CELL ANEMIA; THALASSEMIA; ANEMIAS; ANIMALS; CARBOXYLIC ACIDS; DISEASES; GLOBIN; HEMIC DISEASES; HETEROCYCLIC ACIDS; HETEROCYCLIC COMPOUNDS; MAMMALS; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; PIGMENTS; PORPHYRINS; PROTEINS; RODENTS; SYMPTOMS; VERTEBRATES; 550800* - Morphology

Citation Formats

Wawrzyniak, C.J., Lewis, S.E., and Popp, R.A. Differential expression of murine adult hemoglobins in early ontogeny. United States: N. p., 1985. Web.
Wawrzyniak, C.J., Lewis, S.E., & Popp, R.A. Differential expression of murine adult hemoglobins in early ontogeny. United States.
Wawrzyniak, C.J., Lewis, S.E., and Popp, R.A. Tue . "Differential expression of murine adult hemoglobins in early ontogeny". United States. doi:.
@article{osti_5642932,
title = {Differential expression of murine adult hemoglobins in early ontogeny},
author = {Wawrzyniak, C.J. and Lewis, S.E. and Popp, R.A.},
abstractNote = {A hemoglobin mutation is described that permits study of the expression of the two adult ..beta..-globin genes throughout fetal and postnatal development. Mice with a mutation at the Hbb/sup s/, ..beta..-globin locus, were used to study the relative levels of ..beta..-s2major and ..beta..-sminor globins specified by the mutant Hbb/sup s2/ haplotype during development. At 11.5 days of gestation ..beta..-sminor comprised over 80% and ..beta..-s2major under 20% of the adult beta-globin. The relative level of ..beta..-sminor decreased through fetal development; at birth ..beta..-sminor represented 33.7% of the ..beta..-globin. The adult values of 71.0% ..beta..-s2major and 29.0% ..beta..-sminor globin are expressed in mice six days after birth. Because the two ..beta..-globin genes are expressed in mice of the Hbb/sup 2s/ haplotype, both the ..beta..-smajor and ..beta..-sminor genes must be expressed in mice of the Hbb/sup s/ haplotype. Expression of the ..beta..-sminor gene is elevated to 35.6% in Hbb/sup s2/ mice that have been bled repeatedly. Thus, the 5' ..beta..-s2major and 3' ..beta..-sminor genes of the Hbb/sup s2/ haplotype and, presumably the 5' ..beta..-smajor and 3' ..beta..-sminor genes of the Hbb/sup s/ haplotype, are regulated independently and are homologous to the 5' ..beta..-dmajor and 3' ..beta..-dminor genes of the Hbb/sup d/ haplotype. Mice of the Hbb/sup s2/ haplotype are better than mice of the Hbb/sup d/ haplotytpe for studying the mechanisms of hemoglobin switching because the Hbb/sup s2/ each of the three embryonic and two adult hemoglobins can be separated by electrophoresis. 17 refs., 3 figs.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Jan 01 00:00:00 EST 1985},
month = {Tue Jan 01 00:00:00 EST 1985}
}

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  • The earliest cells synthesizing Ig in the mouse can be detected by biosynthesis incorporation of leucine and by lactoperoxidase-catalyzed radioionation of fetal liver cells, followed by serological precipitation and identification as 7-8S IgM by gel electrophoresis from day 10 to day 12 of gestation onward. These early Ig-synthesizing cells are large; they display their Ig on the surface but release it rapidly (t/sub 1///sub 2/ = 45 min). These cells may be identical with pre-B cells identified by adoptive transfer experiments to be precursors of antigen-reactive B cells. Up to day 15 of gestation, such large precursor B cells aremore » the only detectable Ig-synthetizing B cells in fetal liver. Stimulation of these small B cells by mitogens leads to clonal growth and maturation of plasma blasts and plasma cells secreting 19S IgM, 7S IgG, and IgA at a rapid rate. Fetal liver cells from as early as day 12 to day 13 of gestation will develop, in single-cell suspension cultures, into mitogen-reactive B cells. Such fetal liver cell cultures, in the presence of LPS, develop plaque-forming cells with a peak of IgM response at the equivalent of days 5 to 7 after birth. The time of this peak response is independent of the age of fetal liver at which the cultures are initiated. LPS can be added to the cultures as late as the equivalent of day 18 or 19 of gestation without changing the size or the kinetics of the PFC response. This indicates that B cells from fetal liver acquire LPS reactivity at around birth either in vitro or in vivo.« less
  • The authors have developed a mAb (JE-6) that recognizes an Id encoded by the most J[sub H]-proximal human V[sub H] gene segment (V[sub H]6) in or near germ-line configuration. This mAb was used to determine the frequency of Id JE6[sup +] B cells in large collections of monoclonal EBV-transformed and short term B cell lines derived from fetal, neonatal, and adult lymphoid tissues. Moreover, they investigated the presence of Id JE-6[sup +] lg in sera from neonates and adults and determined the (auto)antigen binding properties of V[sub H]6-encoded IgM mAb. They detected a fivefold overrepresentation of V[sub H]6-expression IgM producingmore » B cells in fetal tissues, cord blood, and adult bone marrow relative to adult blood. In cord blood, but not in adult blood sera, germ-line V[sub H]6-encoded IgM molecules were readily detectable. IgM secreted by V[sub H]6-expressing B cell clones displayed highly conserved and virtually identical autoantigen binding properties, independent of the length and composition of the IgH chain CDR3 region and L chain isotype. Collectively, these results suggest that the V[sub H]6 gene and the antibodies it encodes play an important role in early human ontogeny. 31 refs., 3 figs., 2 tabs.« less
  • Clustering of mutant pollen grains in a population of normal pollen due to premeiotic mutational events complicates translating mutation frequencies into rates. Embryo ontogeny in barley will be described and used to illustrate the formation of such mutant clusters. The nature of the statistics for mutation frequency will be described from a study of the reversion frequencies of various waxy mutants in barley. Computer analysis by a jackknife method of the reversion frequencies of a waxy mutant treated with the mutagen sodium azide showed a significantly higher reversion frequency than untreated material. Problems of the computer analysis suggest a bettermore » experimental design for pollen mutation experiments. Preliminary work on computer modeling for pollen development and mutation will be described.« less
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