Using Gal4-UAS in testes — drivers that work Personally, I was fortunate not to waste time and resources generating betaTub85D -Gal4, because I wanted to express exogenous genes in meiotic arrest testes and meiotic arrest testes do not express betaTub85D. Fluorescent fusion protein variants available. Therefore, only the cells that are in A but not B will have active GAL4, which can then drive the reporter gene. Expression of Gal4-VP16 mirrors expression of the endogenous gene in both male and female germlines. Expression can be directed ubiquitously, or to specific cell types within the gonad, and this expression can be achieved with either direct or bipartite expression systems.

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In the offspring the selected cells, and only those cells, will contain channelrhodopsin and can be triggered by a bright light.

FlyBase Recombinant Construct Report: P{}

Spermatogonia light blue are encapsulated by banos cells light orange and displaced from the hub. Dramatic cell growth occurs during the spermatocyte stage, before entry into the meiotic divisions. The expression level achieved using this system has not been determined. Hsp83 is endogenously ubiquitously expressed, and is mildly heat shock inducible naons the soma.

Journal List Spermatogenesis v. Regeneration of male germline stem cells by spermatogonial dedifferentiation in vivo. Several of the basic ubiquitously expressed promoter constructs are also available as Gal4 drivers, and have been demonstrated to work in the male germline. A brief outline of D. Head and tail yellow cyst cells are distinguishable associated with elongating spermatids.

To achieve short bursts of transgene transcription the classical system is to use a heat shock inducible promoter Hsp Xiao H, Lis JT. This page was last edited on 23 Novemberat Several good candidate early primary spermatocyte promoters exist, for example those based on the meiotic arrest genes. The dynamics depends on the stability of the ectopic protein, and we have examples where the ectopic protein is detected in late-elongation stage spermatids Fig.


To achieve earlier expression of an ectopic element in early primary spermatocytes it is necessary to choose a promoter that is most active at this stage.

While an expression nanso has not been made for this gene, rescue constructs and a LacZ fusion reporter have been generated, and they recapitulate the native gene expression. The use of promoter fusions in Drosophila genetics: Molecular mechanisms of gene regulation during Drosophila spermatogenesis.

An integrated gene annotation and transcriptional profiling approach towards the full gene content of the Drosophila genome. More typically one would expect that constitutive expression of an RNAi construct in testes, of a gene required for fertility, would result in dominant nxnos sterility. Fewer methods for ectopic gene expression have been developed for testes and those that have been developed have not been broadly publicised and hence are not widely used.

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These are strains of flies with the special UAS region next to a desired gene. This revealed a requirement for Drosophila uri in both male and female germline cells.

Brand AH, Perrimon N. A new translational repression element and unusual transcriptional control regulate expression of don juan during Drosophila spermatogenesis. The major outstanding item on the wish list is driver that expresses exclusively in spermatocytes rather than also earlierstays on throughout spermatocyte maturation rather than decliningand expresses at high levels.


One of the few disadvantages of spermatogenesis as a system to study basic processes is that you need the nanow to survive until adulthood or at least to late pupal stages. The lines available are not yet ideal. Second, the promoter architecture of genes expressed at high levels in spermatocytes is very different from that of most somatic cells. Schematic representation of expression patterns of Gal4 drivers in the testis. For expression in the germline it appears that tub-Gal4 is less effective than nanos -Gal4-VP16, since the latter can nahos UAS- nclb and rescue the fertility nsnos in homozygous null nclb mutant males while nanks former is unable to rescue fertility in nclb null animals, but can rescue a partial loss of function of this gene.

All the genetic constructs I will discuss are testis-specific or testis-biased implementations of standard D. Low to moderate levels of ubiquitous somatic expression can be achieved using the armadillo nanks, e. VP16 fuses the transcriptional activation domain of VP16 to Gal4 and potentiates its activity.