Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2014 Sep 26;452(3):328-33.
doi: 10.1016/j.bbrc.2014.08.049. Epub 2014 Aug 19.

MIG-10 (Lamellipodin) stabilizes invading cell adhesion to basement membrane and is a negative transcriptional _target of EGL-43 in C. elegans

Lin Wang  1 , Wanqing Shen  1 , Shijun Lei  1 , David Matus  2 , David Sherwood  3 , Zheng Wang  4
Affiliations

MIG-10 (Lamellipodin) stabilizes invading cell adhesion to basement membrane and is a negative transcriptional _target of EGL-43 in C. elegans

Lin Wang et al. Biochem Biophys Res Commun. .

Abstract

Cell invasion through basement membrane (BM) occurs in many physiological and pathological contexts. MIG-10, the Caenorhabditis elegans Lamellipodin (Lpd), regulates diverse biological processes. Its function and regulation in cell invasive behavior remain unclear. Using anchor cell (AC) invasion in C. elegans as an in vivo invasion model, we have previously found that mig-10's activity is largely outside of UNC-6 (netrin) signaling, a chemical cue directing AC invasion. We have shown that MIG-10 is a _target of the transcription factor FOS-1A and facilitates BM breaching. Combining genetics and imaging analyses, we report that MIG-10 synergizes with UNC-6 to promote AC attachment to the BM, revealing a functional role for MIG-10 in stabilizing AC-BM adhesion. MIG-10 is also required for F-actin accumulation in the absence of UNC-6. Further, we identify mig-10 as a transcriptional _target negatively regulated by EGL-43A (C. elegans Evi-1 proto-oncogene), a transcription factor positively controlled by FOS-1A. The revelation of this negative regulation unmasks an incoherent feedforward circuit existing among fos-1, egl-43 and mig-10. Moreover, our study suggests the functional importance of the negative regulation on mig-10 expression by showing that excessive MIG-10 impairs AC invasion. Thus, we provide new insight into MIG-10's function and its complex transcriptional regulation during cell invasive behavior.

Keywords: Anchor cell; Cell invasion; EGL-43; Lamellipodin; MIG-10.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1. Schematics of AC invasion and localization of MIG-10B in unc-6 mutants
Anterior is left; ventral is down; and arrows point to the AC in this and all other figures. (A) A schematic diagram illustrates AC invasion in C. elegans. In the early L3 larva the AC is attached to the basement membrane (BM, light blue) over the primary vulval precursor cell (1° VPC) (light green, P6.p 1-cell stage, left). At this time UNC-6 (netrin) (blue arrows) secreted from the ventral nerve cord (VNC) polarizes its receptor UNC-40 (blue ovals) and F-actin (orange) to the invasive cell membrane in contact with BM in the presence of integrin (magenta). During the mid-L3 stage, after the P6.p cell divides (P6.p 2-cell stage, middle), the AC breaches the BM and generates an invasive protrusion that invades between the two central 1° VPC granddaughter cells by the late L3 (P6.p 4-cell stage, right). The transcription factor FOS-1A promotes BM breaching. UNC-40 (DCC) mediates protrusion formation. (B and C) DIC images (left) and corresponding fluorescence (right). MIG-10B was similarly polarized at the AC’s basal membrane (arrowheads) in the wild-type and unc-6 mutants at the P6.p 2-cell stage. (D) Quantification of MIG-10B polarization at the P6.p 1- 2-cell and 4-cell stages in wild-type animals and unc-6 mutants (n ≥ 10 per stage per genotype, Student’s t-test). In this and all other figures, *, p < 0.05; **, p < 0.01; ***, p < 0.001; n.s., not significant. Error bars, standard error of mean. Scale, 5 μm.
Fig. 2
Fig. 2. Loss of mig-10 enhances AC detachment from the BM and reduces F-actin accumulation in the absence of UNC-6
(A and B) DIC images (left), corresponding fluorescence (middle), and overlay (right). (A) In unc-6 mutants the AC (visualized by F-actin binding probe cdh-3> mCherry::moeABD) failed to invade and attached to the intact BM (arrowheads; orange dotted line). (B) In unc-6;mig-10 double mutants, the AC failed to invade and detached from the BM. (C) Quantification of AC detachment percentage in unc-6 and unc-6;mig-10 mutants at the P6.p 1-, 2- and 4-cell stages, and in unc-6;mig-10;qyIs183 at the P6.p 4-cell stage. The total numbers of animals scored and significant differences are indicated (Fisher’s exact test). (D and E) 3D reconstructions of confocal z-stacks from the ACs at the P6.p 4-cell stage. Fluorescence (left), overlay of fluorescence and corresponding F-actin networks rendered with isosurfaces (right). (D) In unc-6 mutants F-actin (visualized by F-actin probe cdh-3>mCherry::moeABD) was mislocalized to the AC’s apical-lateral membranes (arrowheads). (E) In unc-6;mig-10 doubles F-actin remained mispolarized. The volume of F-actin was significantly reduced. (F) Quantification of the normalized total volume of F-actin in the wild-type (normalization control), unc-6, mig-10, unc-6;mig-10 mutants at the P6.p 2- and 4-cell stages (n ≥ 15 per stage per genotype). Significant differences are indicated (Student’s t-test).
Fig. 3
Fig. 3. EGL-43A negatively regulates mig-10b expression and forms an incoherent (type I) feedforward loop with FOS-1A and MIG-10B
(A and B) DIC images (left), corresponding fluorescence (right). (A) In the animal treated with L4440 control RNAi, the AC invaded normally at the P6.p 4-cell stage (arrowhead) and mig-10b (mig-10b>GFP) expression was detected in the AC. (B) RNAi _targeting egl-43a resulted in AC invasion failure (arrowhead) and an increase in mig-10b expression in the AC. (C) Quantification of mig-10b expression at the L2/L3 transition, the P6.p 1-, 2-, and 4-cell stages in the animals treated with L4440 control RNAi and egl-43a RNAi (n ≥20 per stage per treatment). (D) The percentage of the ACs that breached the BM at the P6.p 4- (n=85 per genotype) and 8-cell (n=60 per genotype) stages in rrf-3 mutants and rrf-3;mig-10 mutants treated with egl-43a RNAi. Significant differences are indicated (Fisher’s exact test). (E) Within the AC, FOS-1A’s positive transcriptional regulation on egl-43a (arrow), EGL-43A’s negative transcriptional regulation on mig-10b (blunt arrow), and FOS-1A’s positive transcriptional regulation on mig-10b (arrow), form an incoherent (type I) feedforward loop (boxed by red dashed lines). This loop may help maintain a stable, optimal level of mig-10 expression required for effective AC-BM adhesion and AC breaching through BM (blue lines).
Fig. 4
Fig. 4. Overexpression of mig-10b disrupts AC invasion and F-actin polarity
(A–C) DIC image (left), corresponding fluorescence (middle), and spectral representation of fluorescence images (right) at the P6.p 4-cell stage. (A) In wild-type animals the AC (arrow, expressing F-actin probe) invaded through BM (arrowhead) and polarized F-actin at the invasive membrane (yellow arrowhead). (B) In the AC overexpressing MIG-10B, the polarized localization of F-actin was loss. (C) In the animals treated with egl-43a RNAi, the AC failed to invade and lost F-actin polarized localization. (D) Quantification of F-actin polarization in the wild-type ACs, the ACs overexpressing mig-10b and the ACs in the animals treated with egl-43a RNAi at the P6.p 4-cell stage (n ≥10 per genotype). Significant differences are indicated (Student’s t-test).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Ridley AJ, Schwartz MA, Burridge K, Firtel RA, Ginsberg MH, Borisy G, Parsons JT, Horwitz AR. Cell Migration: Integrating Signals from Front to Back. Science. 2003;302:1704–1709. - PubMed
    1. Friedl P, Wolf K. Tumour-cell invasion and migration: diversity and escape mechanisms. Nature reviews Cancer. 2003;3:362–374. - PubMed
    1. Ingram JL, Huggins MJ, Church TD, Li Y, Francisco DC, Degan S, Firszt R, Beaver DM, Lugogo NL, Wang Y, Sunday ME, Noble PW, Kraft M. Airway Fibroblasts in Asthma Manifest an Invasive Phenotype. Am J Respir Crit Care Med. 2011 - PMC - PubMed
    1. Lim KH, Zhou Y, Janatpour M, McMaster M, Bass K, Chun SH, Fisher SJ. Human cytotrophoblast differentiation/invasion is abnormal in pre-eclampsia. Am J Pathol. 1997;151:1809–1818. - PMC - PubMed
    1. Gupta GP, Massague J. Cancer metastasis: building a framework. Cell. 2006;127:679–695. - PubMed

Publication types

MeSH terms

LinkOut - more resources

  NODES
twitter 2