Mouse specific expression of the novel splice variant of Pcp2/L7

• Autorzy: Barski J.J. , Przybyla M. , Nowacka-Chmielewska M.
• Języki publikacji: EN

Abstrakty

EN

INTRODUCTION: Pcp2/L7 is a member of the GoLoco protein family with a very cell‑specific expression in cerebellar Purkinje cells and retinal bipolar neurons. Its precise functional role remains still unclear. Sparse studies indicated its possible role as a guanine nucleotide dissociation inhibitor or guanine nucleotide exchange factor. Studies on genomic structure of pcp2 gene revealed some alternative splice variants expressed in Purkinje cells and retinal bipolar neurons. AIM(S): Here we attempted to shed some light on the conservation of a novel pcp2 splice variant in closely related laboratory rodents: mouse, rat and hamster. METHOD(S): PCR: Both splice variants were amplified with the primers L7sense and L7anti, which yielded two reaction products of 371 and 312 bp. The novel splice variant including exon 3B was detected using the primers L7sense (as above) and L73Aanti, for a product size of 274 bp. Quantitative real‑time PCR (qPCR) analysis: To validate the results obtained by RT-PCR, we conducted an additional qPCR experiment to investigate the expression of Pcp2 transcripts with greater accuracy. The region of interest was amplified from cDNA resulting in a product size of 90 bp.Relative gene expression levels were calculated with the 2‑ΔΔCt method. RESULTS:In our approach we were able to confirm expression of the novel longer transcript in mouse, however PCR amplification on cDNA from rat and hamster did not reveal the long splice variant with additional exon 3B*. CONCLUSIONS: Obtained data indicate, that the novel splicing variant of pcp2 is mouse‑specific and is expressed only in Purkinje cells and retinal bipolar neurons in this species. In course of the evolution it appeared probably in that species as result of a spontaneous mutation. Herewith we suggest a very specific and not known yet function of pcp2 in the mouse eye and/or Purkinje cells. FINANCIAL SUPPORT: Research supported by the Ministry of Science and Higher Education core grant #KNW-2-011/D/4/N.

• Wydawca: -
• Czasopismo: Acta Neurobiologiae Experimentalis
• Rocznik: 2017
• Tom: 77
• Numer: Suppl.1
• Opis fizyczny: p.110

Twórcy

autor

Barski J.J.

• Department of Experimental Medicine, School of Medicine in Katowice, Katowice, Poland
• Department of Physiology, School of Medicine in Katowice, Medical University of Silesia, Katowice, Poland

autor

Przybyla M.

• Department of Experimental Medicine, School of Medicine in Katowice, Katowice, Poland
• Department of Physiology, School of Medicine in Katowice, Medical University of Silesia, Katowice, Poland

autor

Nowacka-Chmielewska M.

• Department of Experimental Medicine, School of Medicine in Katowice, Katowice, Poland
• Laboratory of Molecular Biology, Faculty of Physiotherapy, The Jerzy Kukuczka Academy of Physical Education, Katowice, Poland