A teratoproteomics analysis: heat shock protein 70 is upregulated in mouse forelimb bud by methoxyacetic acid treatment.
BACKGROUND
Methoxyacetic acid (MAA) causes fetal limb abnormalities when the substance is administrated on gestation day (GD) 11 in mice. Limb abnormalities are caused mainly by extensive cell death in the mesoderm of the limb plate. This investigation focused on identifying a protein that is linked with mouse limb teratogenicity.


METHODS
A single dose of MAA at 10 mmol/kg body weight was administered by gavage on GD 11; controls were administered vehicle only. Dams were killed by cervical dislocation 4 hr after treatment and forelimb buds were isolated from both the control and treated embryos. Proteins in forelimb buds GD 11 + 4 hr were precipitated out using 40-60% ammonium sulfate and were then analyzed by 2D SDS-PAGE. Excised protein spots were identified by mass spectrometry and amino acid internal sequence analysis. Identified protein was further confirmed by Western blotting.


RESULTS
Two-dimensional gel analysis indicated that 1 protein spot of 81.7 kDa/pI 7.3 was overexpressed, and the protein matched heat shock protein 70 (HSP70; accession no. P08109, SwissProt).


CONCLUSIONS
The results suggest that MAA, when administered to pregnant mice, upregulates HSP70 in the forelimb buds.
Ruyani
Aceng
A
Program Studi Pendidikan Biologi, Universitas Bengkulu, Jalan Raya Kandang Limun, Bengkulu, Indonesia.
Sudarwati
Sri
S
Sutasurya
Lien A
LA
Sumarsono
Sony H
SH
Kim
Dong-Jo
DJ
Chung
Jun Ho
JH
D000014:Abnormalities, Drug-Induced; D000085:Acetates / Q000633:toxicity*; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D005552:Forelimb; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D017880:Limb Deformities, Congenital / Q000139:chemically induced*; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D011247:Pregnancy; D011270:Pregnancy, Animal; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013723:Teratogens / Q000494:pharmacology*; D015854:Up-Regulation
D000085:Acetates; D018840:HSP70 Heat-Shock Proteins; D013723:Teratogens; C013598:methoxyacetic acid