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Free keywords:
Mouse testis; CREM; Gene-expression; SSH; Microarray
Abstract:
The transcription factors CREM, CREB, and ATF-1 constitute a subfamily of -Zip transcription factors. Several different kinase cascades regulate the activity of these proteins. The activator splice–isoform CREM is specifically and highly expressed in post-meiotic germ cells during mouse spermatogenesis. Male mice lacking CREM expression are sterile because of stage-specific arrest of sperm maturation as the spermatids undergo apoptosis.
In order to characterize the genes that are controlled by CREM during post-meiotic differentiation of round spermatids, we compared the expression levels of mRNA prepared from testes of wild-type and CREM-deficient mice by suppression subtractive hybridization (SSH) and affymetrix oligonucleotide arrays.
A set of 956 unique sequences found in the CREM SSH library was further characterized by generating stage-specific expression profiles during spermatogenesis by hybridization with cDNAfrom pre-pubertal mice at defined stages of spermatogenesis using nylonDNAarrays. The resulting expression profiles were arranged in a linear order according to similarity in their profile shapes to find co-regulation of functionally related genes.
Our data shows that a large number of genes are transcriptionally activated in round spermatids when CREM activity is maximal, including functional groups like transcription factors, proteins involved in signal transduction, and metabolic enzymes, therefore providing novel information of post-meiotic expression of many known as well as novel genes that are either directly or indirectly influenced by CREM expression.
