Switches in 6-phosphofructo-2-kinase isoenzyme expression during rat sperm maturation.
Here we analyzed Pfkfb3 and Pfkfb4 gene expression in rat testis development, isolated testicular cells and spermatozoa. Real time RT-PCR analysis during testis development showed the maximum expression of Pfkfb3 in pre-puber samples and of Pfkfb4 in adult samples. Western blot analysis showed that uPFK-2 protein, a product of Pfkfb3 gene, was present in all the cell types forming the seminiferous epithelium (Sertoli, interstitial and spermatogenic cells). In contrast, tPFK-2, a product of Pfkfb4 gene, was restricted to spermatogenic cells. Confocal analyses by indirect immunofluorescence also corroborated this expression pattern. Immunoblotting studies of isolated spermatozoa demonstrated the presence of uPFK-2 only in immature sperm and once spermatozoa became fully functional this isozyme was replaced by the testicular isozyme tPFK-2. Moreover, immunostaining confirmed that tPFK-2 was localized mainly in the acrosomal region of the sperm head and in the mid-piece of the flagellum, where other spermatogenic cell-specific glycolytic enzymes have been found.
Gómez
M
M
Unitat Bioquímica i Biologia Molecular, Departament de Ciències Fisiològiques II, Campus de Ciències de la Salut, IDIBELL - Universitat de Barcelona, Feixa Llarga s/n, L'Hospitalet, E-08907 Barcelona, Spain.
Navarro-Sabaté
A
A
Manzano
A
A
Duran
J
J
Obach
M
M
Bartrons
R
R
D000818:Animals; D015971:Gene Expression Regulation, Enzymologic; D007527:Isoenzymes / Q000235:genetics; D008297:Male; D025481:Phosphofructokinase-2 / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013091:Spermatogenesis / Q000235:genetics*; D013094:Spermatozoa / Q000201:enzymology / Q000254:growth & development*; D013737:Testis / Q000201:enzymology / Q000254:growth & development*
D007527:Isoenzymes; C542952:Pfkfb4 protein, rat; C526918:Pfkfb3 protein, rat; D025481:Phosphofructokinase-2