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Background   Park2-co-regulated gene  ( PACRG ) is evolutionarily highly conserved from green algae to mammals. In  Chlamydomonas  and trypanosomes, the PACRG protein associates with flagella. Loss of  PACRG  results in shortened or absent flagella. In mouse the PACRG protein is required for spermatogenesis. The purpose of the present study was to analyze (1) the expression patterns of  PACRG  during vertebrate embryogenesis, and (2) whether the PACRG protein was required for left-right (LR) axis specification through cilia-driven leftward flow in  Xenopus laevis .    Methods   PACRG  cDNAs were cloned and expression was analyzed during early embryonic development of  Xenopus , mouse, rabbit and zebrafish. Antisense morpholino oligonucleotide (MO) mediated gene knockdown was applied in  Xenopus  to investigate LR development at the level of tissue morphology, leftward flow and asymmetric marker gene expression, using timelapse videography, scanning electron microscopy (SEM) and whole-mount  in situ  hybridization. Results were statistically evaluated using Wilcoxon paired and χ 2  tests.    Results   PACRG  mRNA expression was found in cells and tissues harboring cilia throughout the vertebrates. Highly localized expression was also detected in the brain. During early development,  PACRG  was specifically localized to epithelia where leftward flow arises, that is, the gastrocoel roof plate (GRP) in  Xenopus , the posterior notochord (PNC) in mammals and Kupffer’s vesicle (KV) in zebrafish. Besides its association with ciliary axonemes, subcellular localization of PACRG protein was found around the nucleus and in a spotty pattern in the cytoplasm. A green fluorescent protein (GFP) fusion construct preferentially labeled cilia, rendering PACRG a versatile marker for live imaging. Loss-of-function in the frog resulted dose dependently in LR, neural tube closure and gastrulation defects, representing ciliary and non-ciliary functions of PACRG.    Conclusions  The PACRG protein is a novel essential factor of cilia in  Xenopus .

Ciliary and non-ciliary expression and function of PACRGduring vertebrate development