Molecular cloning and tissue‐specific expression of the mouse homologue of the rat brain 14‐3‐3 θ protein: Characterization of its cellular and developmental pattern of expression in the male germ line

Abstract

The highly conserved 14-3-3 family of proteins, originally reported as brain-specific and then found in various somatic cells and oocytes, interacts with several important signal transduction kinases so that actually the 14-3-3 protein are considered as modulators of multiple signal transduction pathways. Here we show that a 14-3-3 protein is also expressed in the male germ cells, thus extending the protein cellular distribution to a cell line never reported to express 14-3-3 proteins. Screening of a mouse spermatogenic cells lambda gt11 cDNA library with affinity-purified polyclonal antibodies to the tyrosine kinase SP42 allowed the isolation of several positive clones. Sequencing of a positive cDNA clone revealed a 735-nucleotide open reading frame encoding a protein of 245 amino acids (27,778 Da). The predicted protein was found to be identical to the most recently discovered 14-3-3 isoform, the theta subtype from a rat brain. Here we demonstrate that 14-3-3 theta mRNA is highly expressed in testis and brain only. Western immunoblot analyses confirm the Northern blot data. Developmental Northern and Western blot analyses are consistent with an expression and translation of the 14-3-3 theta gene throughout spermatogenesis. However, analysis of RNA from purified populations of spermatogenic cells at different developmental stages and immunohistochemistry on adult testis sections reveal that within the testis the 14-3-3 theta gene products are most abundant in meiotic prophase spermatocytes, and, above all, in differentiating spermatids. Both testicular and epididymal spermatozoa are negative. The present study is the first report on the presence and molecular characterization of the 14-3-3 theta gene product in the male germ line. Our observations suggest that this specific member of the 14-3-3 protein family could play distinct modulatory roles in the complex development of the mammalian male germ cell lineage.

The adaptor protein 14-3-3 binds to the calcium-sensing receptor and attenuates receptor-mediated Rho kinase signalling

“…The 14-3-3 isoforms are differentially expressed in tissues and, consequently, appear to play distinct roles in mammalian biology [28,45]. It was from rat brain that 14-3-3θ was first cloned [46] and subsequently mouse testis in which it plays a role in the control of spermatogenesis via the regulation of Sertoli cell adhesion [47]. Interestingly, H-500 Leydig cell (testicular tumour) growth is controlled in part by the pituitary tumour-transforming gene which is up-regulated by the induction of high extracellular Ca 2 + mediated by the CaR [48].…”

G Proteins and Associated Signal Transduction Molecules

“…Within the testis the 14-3-39 gene products are most abundant in meiotic prophase spermatocytes, and in differentiating spermatids. Both testicular and epididymal spermatozoa are negative (Perego and Bermti, 1997) (Fig.18.3). In an effort to understand the function of 14-3-38 in testis, Berruti (2000) sought to identify endogenous proteins that interact with 14-3-39 in spermatogenic cells.…”

Dynamic interactions between 14-3-3 proteins and phosphoproteins regulate diverse cellular processes

“…It is also clear that different 14-3-3s isoforms have distinct subcellular and tissue distributions in plants and animals [60,[174][175][176][177][178][179][180][181][182][183], which has been most thoroughly studied in the brain [184]. 14-3-3s have been identified within mitochondria and chloroplasts [94,185], and even in the extracellular wall of Chlamydomonas [186].…”