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1842326032
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4 column as described (8).
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-) instead of Taq. Genotypes of sickle cell mice were confirmed by cellulose acetate gel electrophoresis of hemoglobins (Helena Laboratories, Beaumont, TX) [J. B. Whitney III, Biochem. Genet. 16, 667 (1978)].
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1842329025
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note
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O//+]. Interestingly, there was a fivefold increase in sickle cell newborn survival when sickle cell males were bred with non-sickle cell females (number obtained/number expected = 32/122) compared with when non-sickle cell males were bred with non-sickle cell females (number obtained/number expected = 4/82). One possible explanation for this is the presence of segregating "survival" alleles in the mixed genetic background (FVB/N, 129, DBA/2, C57BL/6, Black Swiss) of the breeding population. These alleles would be inherited at a higher frequency from sickle cell males than from non-sickle cell males. Another possible explanation involves the presence of fewer healthy non-sickle cell newborns in litters from sickle cell males. This might allow sickle cell newborns to compete more effectively for maternal attention and enable them to survive more readily. The following animal husbandry practices may influence the survival of sickle cell mice: Micro-Barrier Systems cages (17.8 × 27.9 cm) (Allentown Caging, Allentown, NJ), Bed-O'Cobs bedding (size, 0.32 cm) (Andersons Industrial Products, Maumee, OH), Nestlets nesting material (Ancare Corp., Bellmore, NY), and Purina 5008 food (PMI Inc.).
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1842409416
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note
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O//+ β-thalassemic mice (1.38 ± 0.05; n = 3).
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Blood was mixed (1:1) with a 2% sodium bisulfite solution [G. A. Daland and W. B. Castle, J. Lab. Clin. Med. 33, 1082 (1948)] and wet mounts were prepared immediately. After 15 min, 80% of cells were completely sickled (60% of cells had a single domain or a few parallel domains, 20% had multispiculated holly leaf morphology).
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note
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The increases (fold) in organ weights were calculated from organ weights expressed as a percentage of total body weight [sickle cell (n = 4) versus wild type (n = 5): kidneys, 2.5% versus 1.3%; heart, 1.0% versus 0.5%; spleen, 6.5% versus 0.5%].
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24
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1842333725
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note
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Histological analysis was performed on four sickle cell and two wild-type mice, all 5 to 7 months of age.
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1842327856
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note
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We thank P. Cooper, F. Kuypers, B.-C. Lau, M. Sorette, B. Bookchin, B. Mentzer, J. Zhang, P. Donohue, J. Hanneman, and K. Brinkley for their contribution to this work. Supported by National Heart, Lung, and Blood Institute (NHLBI) grants HL31579 and HL20985 to the Northern California Comprehensive Sickle Cell Center, a Red Cell Program Project Grant (DK32094) from the National Institute of Diabetes, Digestive and Kidney Diseases, a NHLBI contract (NO1-HB-07086) to the Sickle Cell Disease Centralized Pathology Unit, and an NIH Shared Instrumentation Grant (VGBioQ mass spectrometer). Care of experimental animals was in accordance with institutional guidelines. Informed consent was obtained for human blood samples.
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