Measurement of hemoglobin synthesis rate in vivo using a stable isotope method

Jacqueline M. Hibbert, George B. Sutherland, Luther L. Wright, Luke G. Wolfe, Kimberly A. Wolfe, Shi Ping Gao, Dennis Gore, Anwar S. Abd-Elfattah

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

We developed a method to measure hemoglobin synthesis rate (SynHb) in humans, assuming that free glycine in the red blood cell (RBC) represents free glycine in bone marrow for hemoglobin synthesis. The present rat study examines this assumption of the method and quantifies SynHb in rats. Sprague-Dawley rats (n = 9) were studied, [2-13C]glycine was intravenously infused over 24 h (2.5 mg kg-1 h-1), blood was drawn for glycine and heme isolation, and bone marrow was harvested for glycine isolation. Isotopic enrichments of glycine and heme were measured, fractional hemoglobin synthesis rate (fSynHb% day-1) was calculated, and from this a value for SynHb (mg g-1 day-1) was derived. Mean body weight was 446 ± 10 g (mean ± SE) and hemoglobin concentration was 14 ± 0.5 g dl-1. At 24 h, the mean isotopic enrichment, atom percentage excess (APE), of the RBC free glycine (1.56 ± 0.18 APE) was similar to the bone marrow (1.68 ± 0.15 APE). The rate of incorporation of 13C into heme increased over time from 0.0004 APE/h between 6 and 12 h, to 0.0014 APE/h between 12 and 18 h, and 0.0024 APE/h between 18 and 24 h. Consequently, fSynHb (1.19 ± 0.32, 2.92 ± 0.66, and 4.22 ± 0.56% day-1, respectively) and SynHb (0.11 ± 0.03, 0.28 ± 0.05, and 0.42 ± 0.05 mg g-1 day-1, respectively) showed similar patterns over the 24-h study period. We conclude that (1) enrichment of free glycine in the circulating RBC approximates enrichment of bone marrow free glycine for heme formation and (2) this pattern of hemoglobin synthesis rate is reflecting the characteristic release and gradual maturation of reticulocytes in the circulation.

Original languageEnglish (US)
Pages (from-to)118-123
Number of pages6
JournalAnalytical Biochemistry
Volume291
Issue number1
DOIs
StatePublished - Apr 1 2001
Externally publishedYes

Fingerprint

Isotopes
Glycine
Hemoglobins
Heme
Atoms
Bone
Blood
Bone Marrow
Rats
Erythrocytes
Reticulocytes
Sprague Dawley Rats
Cells
Body Weight

Keywords

  • Anemia
  • Bone marrow
  • Glycine
  • Hemoglobin synthesis
  • Rats

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Hibbert, J. M., Sutherland, G. B., Wright, L. L., Wolfe, L. G., Wolfe, K. A., Ping Gao, S., ... Abd-Elfattah, A. S. (2001). Measurement of hemoglobin synthesis rate in vivo using a stable isotope method. Analytical Biochemistry, 291(1), 118-123. https://doi.org/10.1006/abio.2001.4997

Measurement of hemoglobin synthesis rate in vivo using a stable isotope method. / Hibbert, Jacqueline M.; Sutherland, George B.; Wright, Luther L.; Wolfe, Luke G.; Wolfe, Kimberly A.; Ping Gao, Shi; Gore, Dennis; Abd-Elfattah, Anwar S.

In: Analytical Biochemistry, Vol. 291, No. 1, 01.04.2001, p. 118-123.

Research output: Contribution to journalArticle

Hibbert, JM, Sutherland, GB, Wright, LL, Wolfe, LG, Wolfe, KA, Ping Gao, S, Gore, D & Abd-Elfattah, AS 2001, 'Measurement of hemoglobin synthesis rate in vivo using a stable isotope method', Analytical Biochemistry, vol. 291, no. 1, pp. 118-123. https://doi.org/10.1006/abio.2001.4997
Hibbert JM, Sutherland GB, Wright LL, Wolfe LG, Wolfe KA, Ping Gao S et al. Measurement of hemoglobin synthesis rate in vivo using a stable isotope method. Analytical Biochemistry. 2001 Apr 1;291(1):118-123. https://doi.org/10.1006/abio.2001.4997
Hibbert, Jacqueline M. ; Sutherland, George B. ; Wright, Luther L. ; Wolfe, Luke G. ; Wolfe, Kimberly A. ; Ping Gao, Shi ; Gore, Dennis ; Abd-Elfattah, Anwar S. / Measurement of hemoglobin synthesis rate in vivo using a stable isotope method. In: Analytical Biochemistry. 2001 ; Vol. 291, No. 1. pp. 118-123.
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abstract = "We developed a method to measure hemoglobin synthesis rate (SynHb) in humans, assuming that free glycine in the red blood cell (RBC) represents free glycine in bone marrow for hemoglobin synthesis. The present rat study examines this assumption of the method and quantifies SynHb in rats. Sprague-Dawley rats (n = 9) were studied, [2-13C]glycine was intravenously infused over 24 h (2.5 mg kg-1 h-1), blood was drawn for glycine and heme isolation, and bone marrow was harvested for glycine isolation. Isotopic enrichments of glycine and heme were measured, fractional hemoglobin synthesis rate (fSynHb{\%} day-1) was calculated, and from this a value for SynHb (mg g-1 day-1) was derived. Mean body weight was 446 ± 10 g (mean ± SE) and hemoglobin concentration was 14 ± 0.5 g dl-1. At 24 h, the mean isotopic enrichment, atom percentage excess (APE), of the RBC free glycine (1.56 ± 0.18 APE) was similar to the bone marrow (1.68 ± 0.15 APE). The rate of incorporation of 13C into heme increased over time from 0.0004 APE/h between 6 and 12 h, to 0.0014 APE/h between 12 and 18 h, and 0.0024 APE/h between 18 and 24 h. Consequently, fSynHb (1.19 ± 0.32, 2.92 ± 0.66, and 4.22 ± 0.56{\%} day-1, respectively) and SynHb (0.11 ± 0.03, 0.28 ± 0.05, and 0.42 ± 0.05 mg g-1 day-1, respectively) showed similar patterns over the 24-h study period. We conclude that (1) enrichment of free glycine in the circulating RBC approximates enrichment of bone marrow free glycine for heme formation and (2) this pattern of hemoglobin synthesis rate is reflecting the characteristic release and gradual maturation of reticulocytes in the circulation.",
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