Copine A plays a role in the differentiation of stalk cells and the initiation of culmination in Dictyostelium developmentReportar como inadecuado




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BMC Developmental Biology

, 10:59

First Online: 02 June 2010Received: 04 January 2010Accepted: 02 June 2010

Abstract

BackgroundCopines are calcium-dependent phospholipid-binding proteins found in diverse eukaryotic organisms. We are studying the function of copines in Dictyostelium discoideum, a single-celled amoeba that undergoes cell differentiation and morphogenesis to form multicellular fruiting bodies when placed in starvation conditions. Previously, we showed that Dictyostelium cells lacking the copine A cpnA gene are not able to complete the developmental cycle, arresting at the slug stage. The aim of this study is to further characterize the developmental defect of the cpnA- cells.

ResultsTime-lapse imaging revealed that cpnA- cells exhibited delayed aggregation and made large mounds that formed one large slug as compared to the smaller slugs of the wild-type cells. While the prespore cell patterning appeared to be normal within the cpnA- slugs, the prestalk cell patterning was different from wild-type. When cpnA- cells were mixed with a small percentage of wild-type cells, chimeric fruiting bodies with short stalks formed. When a small percentage of cpnA- cells was mixed with wild-type cells, the cpnA- cells labeled with GFP were found located throughout the chimeric slug and in both the stalk and sporehead of the fruiting bodies. However, there appeared to be a small bias towards cpnA- cells becoming spore cells. When cpnA- cells were developed in buffer containing EGTA, they were also able to differentiate into either stalk or spore cells to form fruiting bodies with short stalks.

ConclusionsOur results indicate that CpnA is involved in the regulation of aggregation, slug size, and culmination during Dictyostelium development. More specifically, CpnA appears to be involved in the function and differentiation of prestalk cells and plays a role in a calcium-regulated signaling pathway critical to triggering the initiation of culmination.

AbbreviationsCpnACopine A

GFPGreen Fluorescent Protein

EGTAEthyleneglycol-O, O-bis2-aminoethyl-N, N, N-, N-:tetraacetic acid

DICdifferential interference contrast

XGal-bromo-4-chloro-3-indolyl-b-D-galactopyranoside

EcmBExtracellular matrix protein B

EcmAExtracellular matrix protein A

PspAprespore specific protein A

lacZβ-galactosidase.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-213X-10-59 contains supplementary material, which is available to authorized users.

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Autor: Tasha S Smith - Jaimie M Pineda - Alex C Donaghy - Cynthia K Damer

Fuente: https://link.springer.com/article/10.1186/1471-213X-10-59



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