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Characterization and Expression of Trypsinogen and Trypsin in Medaka Testis
- Source :
- Zoological Science. 31:840-848
- Publication Year :
- 2014
- Publisher :
- Zoological Society of Japan, 2014.
-
Abstract
- Previously, we reported that the medaka testis abundantly expresses the mRNA for trypsinogen, which is a well-known pancreatic proenzyme that is secreted into and activated in the intestine. Currently, we report our characterization of the medaka trypsin using a recombinant enzyme and show that this protein is a serine protease that shares properties with trypsins from other species. Two polypeptides (28- and 26-kDa) were detected in the testis extracts by Western blot analysis using antibodies that are specific for medaka trypsinogen. The 28-kDa polypeptide was shown to be trypsinogen (inactive precursor), and the 26-kDa polypeptide was shown to be trypsin (active protease). We did not detect enteropeptidase, which is the specific activator of trypsinogen, in the testis extract. Immunohistochemical analyses using the same trypsinogen-specific antibody produced a strong signal in the spermatogonia and spermatozoa of the mature medaka testis. Substantial staining was found with spermatocytes, whereas extremely weak signals were observed with spermatids. In vitro incubation of testis fragments with the trypsinogen antibody strongly inhibited the release of sperm from the testis into the medium. Trypsin activity was detected in sperm extracts using gelatin zymographic analysis. Immunocytochemistry showed that trypsinogen and trypsin were localized to the cell membranes surrounding the sperm head. Collectively, these results suggest that trypsin plays an important role in the testis function of the medaka.
- Subjects :
- Male
Enteropeptidase
Trypsinogen
medicine.medical_treatment
Immunocytochemistry
Oryzias
digestive system
chemistry.chemical_compound
Western blot
Testis
medicine
Animals
Trypsin
reproductive and urinary physiology
Serine protease
Protease
biology
medicine.diagnostic_test
Immunohistochemistry
Spermatozoa
Molecular biology
Sperm
digestive system diseases
Gene Expression Regulation
chemistry
Biochemistry
biology.protein
Animal Science and Zoology
medicine.drug
Subjects
Details
- ISSN :
- 02890003
- Volume :
- 31
- Database :
- OpenAIRE
- Journal :
- Zoological Science
- Accession number :
- edsair.doi.dedup.....26d9d300dd07d1ae3d206d42046fbebe