Alopecia in a Viable Phospholipase C Delta 1 and Phospholipase C Delta 3 Double Mutant

Abstract

BackgroundInositol 1,4,5trisphosphate (IP3) and diacylglycerol (DAG) are important intracellular signalling molecules in various tissues. They are generated by the phospholipase C family of enzymes, of which phospholipase C delta (PLCD) forms one class. Studies with functional inactivation of Plcd isozyme encoding genes in mice have revealed that loss of both Plcd1 and Plcd3 causes early embryonic death. Inactivation of Plcd1 alone causes loss of hair (alopecia), whereas inactivation of Plcd3 alone has no apparent phenotypic effect. To investigate a possible synergy of Plcd1 and Plcd3 in postnatal mice, novel mutations of these genes compatible with life after birth need to be found.Methodology/Principal FindingsWe characterise a novel mouse mutant with a spontaneously arisen mutation in Plcd3 (Plcd3mNab) that resulted from the insertion of an intracisternal A particle (IAP) into intron 2 of the Plcd3 gene. This mutation leads to the predominant expression of a truncated PLCD3 protein lacking the N-terminal PH domain. C3H mice that carry one or two mutant Plcd3mNab alleles are phenotypically normal. However, the presence of one Plcd3mNab allele exacerbates the alopecia caused by the loss of functional Plcd1 in Del(9)olt1Pas mutant mice with respect to the number of hair follicles affected and the body region involved. Mice double homozygous for both the Del(9)olt1Pas and the Plcd3mNab mutations survive for several weeks and exhibit total alopecia associated with fragile hair shafts showing altered expression of some structural genes and shortened phases of proliferation in hair follicle matrix cells.Conclusions/SignificanceThe Plcd3mNab mutation is a novel hypomorphic mutation of Plcd3. Our investigations suggest that Plcd1 and Plcd3 have synergistic effects on the murine hair follicle in specific regions of the body surface.

The Parathyroid Hormone Second Receptor PTH2R and its Ligand Tuberoinfundibular Peptide of 39 Residues TIP39 Regulate Intracellular Calcium and Influence Keratinocyte Differentiation

“…Dorsal skin from mouse postnatal day 2 to day 10 (P2–P10) was collected to represent hair cycle in anagen, from P16 to P19 to represent catagen, and from P20 to P21 to represent telogen (Ma et al, 2003; Muller-Rover et al, 2001; Paus, 1998; Plikus and Chuong, 2008; Runkel et al, 2012). Both Pth2 and Pth2r gene expressions were increased in catagen and suppressed in early anagen and late telogen (Supplementary Figure S1a online).…”

Mouse germ line mutations due to retrotransposon insertions

“…Inbred of B6 & BalbBefore 1984Exon 4,?Decreased mRNA levels, increase in length of 3’UTR. Modifies response to opioids[209] Pcnx2 C3H/HeJ 6161760C3H/HeJAfter 1950Intron 19, +Modifier of Gria4 mutation, reduces Pcnx2 expression[83] Pitpna vb 1856642DBA/2JEarly 1960sIntron 4, +Transcript contains internal IAP sequence, Reduced mRNA & protein[210] Pla2g6 m1J 4412026C3H/HeJmid 2000sIntron 1,?Reduces normal gene transcripts by ~ 90%[211] Plcd3 mNab NC3H mix~ 2005Intron 2, −Causes truncated protein, LTR may act as antisense promoter?[212] Pofut1 cax 3719000C3H/HeJmid 2000sIntron 4,Reduced mRNA and protein levels[213] Reln rl-Alb2 1857345C3H/HeJFrom chemical ind. Exp.~ 1990Exon 36, −Exon skipping, Reduced mRNA levels[214] Slc35d3 IAP 3802578C3H/HeSn-Rab27a ash /JRos1980s–1990sExon 1, −IAP fusion transcript likely promoted by antisense LTR.…”

Abnormalities of hair structure and skin histology derived from CRISPR/Cas9-based knockout of phospholipase C-delta 1 in mice

“…It has reported that PLCD1 is required for skin stem cell lineage commitment and PLCD1 knockout mice with disruption of X and Y domains (corresponding to exons 7 and 8) by homologous recombination in mouse ES cells undergo progressive hair loss in the first postnatal hair cycle [ 34 – 36 ]. To generate PLCD1-modified mice by CRISPR/Cas9 technology, exons 2 (beginning of the translation PLCD1), 7 and 10 (corresponding to X and Y domains, respectively) of PLCD1 were scanned for potential sgRNA target sequences using Optimized CRISPR Design ( http://crispr.mit.edu ).…”

“…Furthermore, PLCD1 KO mice displayed symptoms of skin inflammation [ 47 ]. PLCD1 and PLCD3 have synergistic effects on the murine hair follicle in specific regions of the body surface [ 36 , 48 ]. Although other studies have revealed the mutations detected in PLCD1 are involved in the development of leukonychia [ 49 ] and cancers, such as breast cancer [ 50 ], pancreatic cancer [ 51 ], and gastric cancer [ 52 ] and so on, the PLCD1-deficient mice produced in this study showed progressive hair loss without other detected abnormalities.…”