Alternative splicing, in which one gene produce multiple transcripts, may infuence how adaptive genes respond to specifc environments. A newly produced transcriptome of Drosophila montana shows the Gs1-like (Gs1l) gene to express multiple splice variants and to be down regulated in cold acclimated fies with increased cold tolerance. Gs1l’s efect on cold tolerance was further tested by injecting cold acclimated and non-acclimated fies from two distantly located northern and southern fy populations with double stranded RNA (dsRNA) targeting Gs1l. While both populations had similar cold acclimation responses, dsRNA injections only efected the northern population. The nature of splicing expression was then investigated in the northern population by confrming which Gs1l variants are present, by comparing the expression of diferent gene regions and by predicting the protein structures of splices using homology modelling. We fnd diferent splices of Gs1l not only appear to have independent impacts on cold acclimation but also elicit diferent efects in populations originating from two very diferent environments. Also, at the protein level, Gs1l appears homologous to the human HDHD1A protein and some splices might produce functionally diferent proteins though this needs to be verifed in future studies by measuring the particular protein levels. Taken together, Gs1l appears to be an interesting new candidate to test how splicing infuences adaptations.