Distribution of Novel Iron Regulatory Protein Hemojuvelin in Murine Tissues
HÄNNINEN, MILLA (2005)
2005
Biokemia - Biochemistry
Lääketieteellinen tiedekunta - Faculty of Medicine
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Hyväksymispäivämäärä
2005-05-30
Julkaisun pysyvä osoite on
https://urn.fi/urn:nbn:fi:uta-1-14744
https://urn.fi/urn:nbn:fi:uta-1-14744
Tiivistelmä
Hakutermit:
iron, hemochromatosis, hemojuvelin, in situ hybridization, immunohistochemistry, localization
Backgroung and Aims: Juvenile hemochromatosis is an early-onset iron overload disease that leads to severe organ damage typically before 30 years of age. In patients with juvenile hemochromatosis, several mutations have been identified in the gene encoding the protein designated hemojuvelin (HJV). Previous reports based on RT-PCR have shown that the expression pattern of HJV is wider in murine tissues than in human. To understand the mechanisms whereby HJV is connected to iron homeostasis, hemojuvelin mRNA and protein expression was studied at cellular level in different mouse tissues.
Methods: In situ hybridization was used to determine the distribution of murine HJV mRNA. A specific riboprobe was constructed based on the murine HJV cDNA sequence. Hemojuvelin protein expression was detected immunohistochemically with a peroxidase-antiperoxidase method.
Results: HJV mRNA and protein expression appeared to be rather restricted. Tissues that were positive with both methods included the kidney, pancreas, brain and liver. In addition, protein expression was studied in the endometrium, smooth muscle of the colon, ovario, submandibular gland and skeletal muscle, of which the endometrium and smooth muscle of the colon was defined as positive. HJV mRNA and protein expression were mainly negative in the heart, spleen, thymus and lung. The gastrointestinal tract showed negligible signal for mRNA, whereas in the stomach and duodenum weak staining for HJV protein was detected.
Conclusions: Expression pattern of HJV suggests that HJV protein may play important roles in various tissues not only in those classically considered the most important ones for regulation of iron homeostasis.
iron, hemochromatosis, hemojuvelin, in situ hybridization, immunohistochemistry, localization
Backgroung and Aims: Juvenile hemochromatosis is an early-onset iron overload disease that leads to severe organ damage typically before 30 years of age. In patients with juvenile hemochromatosis, several mutations have been identified in the gene encoding the protein designated hemojuvelin (HJV). Previous reports based on RT-PCR have shown that the expression pattern of HJV is wider in murine tissues than in human. To understand the mechanisms whereby HJV is connected to iron homeostasis, hemojuvelin mRNA and protein expression was studied at cellular level in different mouse tissues.
Methods: In situ hybridization was used to determine the distribution of murine HJV mRNA. A specific riboprobe was constructed based on the murine HJV cDNA sequence. Hemojuvelin protein expression was detected immunohistochemically with a peroxidase-antiperoxidase method.
Results: HJV mRNA and protein expression appeared to be rather restricted. Tissues that were positive with both methods included the kidney, pancreas, brain and liver. In addition, protein expression was studied in the endometrium, smooth muscle of the colon, ovario, submandibular gland and skeletal muscle, of which the endometrium and smooth muscle of the colon was defined as positive. HJV mRNA and protein expression were mainly negative in the heart, spleen, thymus and lung. The gastrointestinal tract showed negligible signal for mRNA, whereas in the stomach and duodenum weak staining for HJV protein was detected.
Conclusions: Expression pattern of HJV suggests that HJV protein may play important roles in various tissues not only in those classically considered the most important ones for regulation of iron homeostasis.