Pannexin 1 Transgenic Mice: Human Diseases and Sleep-Wake Function Revision.

Nariman Battulin,Alexey Korablev,Vladimir M Kovalzon,Irina Serova,Marta G Pechkova,Oxana O Kiryukhina,Kirill A Bogotskoy,Olga S Tarasova,Yuri Panchin

doi:10.3390/ijms22105269

Abstract

In humans and other vertebrates pannexin protein family was discovered by homology to invertebrate gap junction proteins. Several biological functions were attributed to three vertebrate pannexins members. Six clinically significant independent variants of the PANX1 gene lead to human infertility and oocyte development defects, and the Arg217His variant was associated with pronounced symptoms of primary ovarian failure, severe intellectual disability, sensorineural hearing loss, and kyphosis. At the same time, only mild phenotypes were observed in Panx1 knockout mice. In addition, a passenger mutation was identified in a popular line of Panx1 knockout mice, questioning even those effects. Using CRISPR/Cas9, we created a new line of Panx1 knockout mice and a new line of mice with the clinically significant Panx1 substitution (Arg217His). In both cases, we observed no significant changes in mouse size, weight, or fertility. In addition, we attempted to reproduce a previous study on sleep/wake and locomotor activity functions in Panx1 knockout mice and found that previously reported effects were probably not caused by the Panx1 knockout itself. We consider that the pathological role of Arg217His substitution in Panx1, and some Panx1 functions in general calls for a re-evaluation.

Highlights

The pannexin protein family was discovered in 2000 as a result of a comparison of human and invertebrate genes [1]
The general question of the number of gap junctions (GJ) protein families is complicated by the observation that some Metazoans can form GJ’s while lacking both connexins and pannexins. These results suggest the existence of a third hypothetical family of gap junction proteins
According to the previously described scheme [35], we used the CRISPR/Cas9 system to carry out these genetic modifications

Summary

Introduction

The pannexin protein family was discovered in 2000 as a result of a comparison of human and invertebrate genes [1]. Three vertebrate pannexin genes were cloned and characterized [2,3,4,5,6]. The members of this family, especially pannexin 1, are implicated in a number of vital biological functions and the development of several pathological mechanisms [7,8,9,10]. It was previously recognized that pannexin channels, such as connexin hemichannels, consist of six subunits. Four new studies using cryoelectron microscopy [11,12,13,14] have shown that pannexin channels consist of seven subunits. Elucidating the structure of pannexin 1 with high resolution allowed a more accurate analysis of the possible roles of known mutations in the functioning of this protein

Methods

Results

Discussion

Conclusion