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Daunomycin accumulation and induction of programmed cell death in rat hair follicles

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Abstract

The anthracycline antibiotic daunomycin (DM) is useful for the treatment of leukemia but has side-effects such as alopecia. Using immunocytochemistry, we show that, after a single i.v. injection, DM accumulates in the nuclei of matrix cells and in the outer root sheath of hair follicles. DM-positive matrix cells are detectable up to 48 h after injection and exhibit a characteristic granular morphology, which is not observed in saline-injected controls. TUNEL-staining has revealed that DM injection induces programmed cell death (PCD) in rat hair follicles. Cells undergoing PCD are detectable as late as 5 days postinjection in both the matrix and outer root sheath. Newly developed double-staining has shown that some of the DM-positive matrix cell nuclei are also TUNEL-positive. Staining for activated caspase-3 has demonstrated immunopositive cells following DM administration both in the matrix and in the outer root sheath. Ultrastructural immunocytochemistry has shown the presence of DM-positive cells with two different types of morphology. About half of the immunopositive cells exhibit a morphology typical of classical apoptosis (PCD type 1), whereas the other half show signs of autophagic cell death (PCD type 2). Interestingly, little, if any, DM accumulation or apoptosis has been detected in the dermal hair papillae. This may have a bearing on potential regeneration of the hair follicles. Thus, DM accumulates in a characteristic pattern in hair follicles. This accumulation is associated with the induction of two morphologically distinct forms of PCD.

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Abbreviations

BSA:

Bovine serum albumin

CYP:

Cyclophosphamide

DM:

Daunomycin

DOX:

Doxorubicin (adriamycin)

GA:

Glutaraldehyde

HRP:

Horseradish peroxidase

ICC:

Immunocytochemistry

mAb:

Monoclonal antibody

NGS:

Normal goat serum

pAb:

Polyclonal antibody

PBS:

Phosphate-buffered saline

PCD:

Programmed cell death

PFA:

Paraformaldehyde

RT:

Room temperature

TBS:

TRIS-buffered saline

TBST:

TBS containing 1% Triton X-100

TdT:

Terminal deoxynucleotidyl transferase

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Acknowledgment

We are grateful to K. Ohara, Y. Tamamura and K. Tanaka for their technical assistance throughout this study.

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Authors and Affiliations

  1. Department of Applied Life Science, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, Kumamoto, 860-0082, Japan

    Masashi Shin & Kunio Fujiwara

  2. Department of Cell Biology IBHV, Faculty of Life Sciences, University of Copenhagen, Gronnegaardsvej 7, Frederiksberg C, DK-1870, Denmark

    Lars-Inge Larsson

  3. Department of Clinical Biochemistry, Statens Serum Institut, Copenhagen, Denmark

    David M. Hougaard

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  1. Masashi Shin
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  2. Lars-Inge Larsson
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  4. Kunio Fujiwara
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Correspondence to Kunio Fujiwara.

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Shin, M., Larsson, LI., Hougaard, D.M. et al. Daunomycin accumulation and induction of programmed cell death in rat hair follicles. Cell Tissue Res 337, 429–438 (2009). https://doi.org/10.1007/s00441-009-0840-8

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  • DOI: https://doi.org/10.1007/s00441-009-0840-8

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