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Y Hu, Zoology, The University of Melbourne, Melbourne, Australia
H Yu, Zoology, The University of Melbourne, Melbourne, Australia
A Pask, Zoology, The University of Melbourne, Melbourne, Australia
D O'Brien, Department of Cell & Developmental Biology, University of North Carolina School of Medicine, Chapel Hill, United States
G Shaw, Zoology, The University of Melbourne, Melbourne, Australia
M Renfree, Zoology, The University of Melbourne, Melbourne, 3010, Australia
Correspondence: Marilyn B Renfree, Email: m.renfree{at}unimelb.edu.au
Abstract
A-Kinase Anchor Protein 4 (AKAP4) is an X-linked member of the AKAP family of scaffold proteins that anchor cAMP-dependent protein kinases and play an essential role in fibrous sheath assembly during spermatogenesis and flagellar function in spermatozoa. Marsupial spermatozoa differ in structural organization from those of eutherian mammals but data on the molecular control of their structure and function are limited. We therefore cloned and characterized the AKAP4 gene in a marsupial, the tammar wallaby (Macropus eugenii). The gene structure, sequence and predicted protein of AKAP4 were highly conserved with that of eutherian orthologues and it mapped to the marsupial X-chromosome. There was no AKAP4 expression detected in the developing young. In the adult, AKAP4 expression was limited to the testis with a major transcript of 2.9kb. AKAP4 mRNA was expressed in the cytoplasm of round and elongated spermatids while its protein was found on the principal piece of the flagellum in the sperm tail. This is consistent with its expression in other mammals. Thus AKAP4 appears to have a conserved role in spermatogenesis for at least the last 166 million years of mammalian evolution.
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