TY - JOUR
T1 - Localization and age-dependent expression of the inward rectifier K+ channel subunit Kir 5.1 in a mammalian reproductive system
AU - Salvatore, Lorena
AU - D'Adamo, Maria Cristina
AU - Polishchuk, Roman
AU - Salmona, Mario
AU - Pessia, Mauro
N1 - Funding Information:
We thank Antonio De Blasi and Stephen Tucker for reading and commenting on the manuscript. We are grateful to John Adelman for the generous gifts of Kir 4.1 and Kir 5.1 clones and to Simone Valentino, Cosmo Rossi, Nicola Celli, Antonio Tamburro, Alexander Mironov for fruitful discussion, insightful suggestions and invaluable technical assistance. We thank the Animal Care Unit of CMNS and the financial support of the Italian National Research Council (Convenzione CNR – Mario Negri Sud). M.C. D’Adamo and Roman Polishchuk are fellows of the Alfredo Leonardi Fund and G.L. Pfeiffer Foundation.
PY - 1999/4/23
Y1 - 1999/4/23
N2 - Kir 5.1 is a member of the inward rectifier potassium channel superfamily which does not form functional channels when expressed by itself in Xenopus laevis oocytes. rt-PCR reveals high levels of Kir 5.1 mRNA expression in testis but the function of this channel remains unknown. To determine the cell-specific expression of this channel in the testis we raised a polyclonal antibody against an external epitope of Kir 5.1 and tested its specificity in Xenopus oocytes expressing several cloned Kir subunits. Strong immunoreactivity for Kir 5.1 was found in seminiferous tubules of rat testis and, particularly, in spermatogonia, primary and secondary spermatocytes, spermatids and in the head and body of spermatozoa. The intensity of Kir 5.1 immunofluorescence, quantified using laser scanning microscopy, increased with age at every stage in the development of sperm from spermatogonia and reached a peak in 60-day-old rats. In contrast, the immunofluorescence decreased in 90-day-old animals and was detected mostly in spermatozoa. The results demonstrate that Kir 5.1 expression in the testis is localised to cells involved in spermatogenesis, showing a temporal pattern of expression during sexual maturity. Copyright (C) 1999 Federation of European Biochemical Societies.
AB - Kir 5.1 is a member of the inward rectifier potassium channel superfamily which does not form functional channels when expressed by itself in Xenopus laevis oocytes. rt-PCR reveals high levels of Kir 5.1 mRNA expression in testis but the function of this channel remains unknown. To determine the cell-specific expression of this channel in the testis we raised a polyclonal antibody against an external epitope of Kir 5.1 and tested its specificity in Xenopus oocytes expressing several cloned Kir subunits. Strong immunoreactivity for Kir 5.1 was found in seminiferous tubules of rat testis and, particularly, in spermatogonia, primary and secondary spermatocytes, spermatids and in the head and body of spermatozoa. The intensity of Kir 5.1 immunofluorescence, quantified using laser scanning microscopy, increased with age at every stage in the development of sperm from spermatogonia and reached a peak in 60-day-old rats. In contrast, the immunofluorescence decreased in 90-day-old animals and was detected mostly in spermatozoa. The results demonstrate that Kir 5.1 expression in the testis is localised to cells involved in spermatogenesis, showing a temporal pattern of expression during sexual maturity. Copyright (C) 1999 Federation of European Biochemical Societies.
KW - Confocal microscopy
KW - Immunofluorescence
KW - Immunohistochemistry
KW - Potassium channel
KW - Spermatozoon
KW - Testis
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U2 - 10.1016/S0014-5793(99)00420-2
DO - 10.1016/S0014-5793(99)00420-2
M3 - Article
C2 - 10338121
AN - SCOPUS:0032910451
SN - 0014-5793
VL - 449
SP - 146
EP - 152
JO - FEBS Letters
JF - FEBS Letters
IS - 2-3
ER -