From the primitive streak to the somitic mesoderm: labeling the early stages of chick embryos using EGFP transfection

Fan, Hai; Sakamoto, Nobuyuki; Aoyama, Hirohiko

doi:10.1007/s12565-018-0429-y

“…36 The mesoderm is derived from the primitive streak in the middle of the epiblast plate. 37 The exact position in the primitive streak decides cell fate. 38 After invagination, the bilateral paraxial mesoderm, caudal to the head, forms somites which eventually develop into different compartments, such as the sclerotome and the syndetome, which in turn give rise to axial bones, tendons and connective tissues, and the dermomyotome containing proliferating progenitors of all skeletal muscles of the body.…”

Section: Discussionmentioning

confidence: 99%

“…All vertebrate skeletal muscles, apart from superficial neck muscles, derived from the paraxial mesoderm . The mesoderm is derived from the primitive streak in the middle of the epiblast plate . The exact position in the primitive streak decides cell fate .…”

Section: Discussionmentioning

confidence: 99%

Early activating somatic PIK3CA mutations promote ectopic muscle development and upper limb overgrowth

Frisk

¹

,

Taylan

²

,

Blaszczyk

³

et al. 2019

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PIK3CA-related overgrowth spectrum is a group of rare genetic disorders with asymmetric overgrowth caused by somatic mosaic PIK3CA mutations. Here, we report clinical data and molecular findings from two patients with congenital muscular upper limb overgrowth and aberrant anatomy. During debulking surgery, numerous ectopic muscles were found in the upper limbs of the patients. DNA sequencing, followed by digital polymerase chain reaction, was performed on DNA extracted from biopsies from hypertrophic ectopic muscles and identified the somatic mosaic PIK3CA hotspot mutations c.3140A > G, p.(His1047Arg) and c.1624G > A, p.(Glu542Lys) in a male (patient 1) and a female (patient 2) patient, respectively. Patient 1 had four ectopic muscles and unilateral isolated muscular overgrowth while patient 2 had 13 ectopic muscles and bilateral isolated muscular overgrowth of both upper limbs, indicating that her mutation occurred at early pre-somitic mesoderm state. The finding of PIK3CA mutations in ectopic muscles highlights the importance of PIK3CA in cell fate in early human embryonic development. Moreover, our findings provide evidence that the disease phenotype depends on the timing of PIK3CA mutagenesis during embryogenesis and confirm the diagnostic entity PIK3CA-related muscular overgrowth with ectopic accessory muscles. K E Y W O R D Saccessory, cell fate, ectopic, muscle hypertrophy, muscular hypertrophy, PIK3CA, PROS Tobias Laurell and Ann Nordgren contributed equally.

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“…Transfection was performed with an electroporator (CUY21EX, BEX, Tokyo, Japan) under the following conditions: one cycle of a poring pulse of 40 V, on-time 10 msec, off-time 10 msec, and five cycles of a driving pulse of 4 V, on-time 10 msec, off-time 10 msec. The transfection procedure was slightly modified from a previously described method (Fan et al 2018).…”

Section: Electroporationmentioning

confidence: 99%

“…In addition, injecting DiI between the somatopleure and ectoderm causes undesired labeling of ectodermal cells, which could generate false results. To avoid these disadvantages, we performed genomic labeling of somatopleural cells using EGFP and transposase expression vectors (Fan et al 2018). The EGFP gene was electroporated into the somatopleure in E 2.5 (HH 14-15) chicken embryos after injecting plasmids into the coelom (Fig.…”

Section: Local Genetic Labeling Of Somatopleural Cells and Observatio...mentioning

confidence: 99%

Double-layered two-directional somatopleural cell migration during chicken body wall development revealed with local fluorescent tissue labeling

Sakamoto

¹

,

Aoyama

²

,

Ikegami

³

2022

Self Cite

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The ventral body wall is derived from the somatic layer of the lateral plate mesoderm, somatopleure, and somite. The primary ventral body wall is formed as a result of the lateral growth of the somatopleure. The secondary body wall is generated through the migration of somitic cells into the somatopleure. While it is reported that the cervical somatopleural cells migrate caudally to the thoracic region during body wall development, the migration of the thoracic somatopleural cells has not been elucidated. To investigate the migration behavior of the somatopleural cells in the thorax during chicken ventral body wall development, we labeled the thoracic somatopleural cells of one somite wide by DiI labeling or gene transfection of enhanced green fluorescent protein and observed the three-dimensional distribution of the labeled cells with the tissue-clearing technique FRUIT. Our labeling experiments revealed the rostral migration of the somatopleural cells into a deep part of the thoracic body wall in embryonic day 6.5 chickens. For embryonic day 8.5 chickens, these deep migrating somatopleural cells were found around the sternal ribs. Thus, we identified the double-layered two-directional migrating pathways of the somatopleural cells: the rostral migration of the deep somatopleural cells and the lateral migration of the superficial somatopleural cells. Our findings imply that the rostral migration of deep somatopleural cells and the lateral migration of superficial ones might be associated with the developing sternal ribs and the innervation of the thoracic cutaneous nerves, respectively.

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Double-layered two-directional somatopleural cell migration during chicken body wall development revealed with local fluorescent tissue labeling

Sakamoto

¹

,

Aoyama

²

,

Ikegami

³

2021

Preprint

Self Cite

0

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The ventral body wall is derived from the somatic layer of the lateral plate mesoderm, somatopleure, and somite. The primary ventral body wall is formed as a result of the lateral growth of the somatopleure. The secondary body wall is generated through the migration of somitic cells into the somatopleure. While it is reported that the cervical somatopleural cells migrate caudally to the thoracic region during body wall development, the migration of the thoracic somatopleural cells has not been elucidated. To investigate the migration behavior of the somatopleural cells in the thorax during chicken ventral body wall development, we labeled the thoracic somatopleural cells of one somite wide by DiI labeling or gene transfection of enhanced green fluorescent protein and observed the three-dimensional distribution of the labeled cells with the tissue-clearing technique FRUIT. Our labeling experiments revealed the rostral migration of the somatopleural cells into a deep part of the thoracic body wall in embryonic day 6.5 chickens. For embryonic day 8.5 chickens, these deep migrating somatopleural cells were found around the sternal ribs. Thus, we identified the double-layered two-directional migrating pathways of the somatopleural cells: the rostral migration of the deep somatopleural cells and the lateral migration of the superficial somatopleural cells. Our findings imply that the rostral migration of deep somatopleural cells and the lateral migration of superficial ones might be associated with the developing sternal ribs and the innervation of the thoracic cutaneous nerves, respectively.Mini-abstractDouble-layered two-directional migrations of the somatopleural cells in the thoracic body wall during chicken development were revealed by one-somite-wide labeling of the somatopleure.

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From the primitive streak to the somitic mesoderm: labeling the early stages of chick embryos using EGFP transfection

Cited by 3 publications

References 21 publications

Early activating somatic PIK3CA mutations promote ectopic muscle development and upper limb overgrowth

Early activating somatic PIK3CA mutations promote ectopic muscle development and upper limb overgrowth

Double-layered two-directional somatopleural cell migration during chicken body wall development revealed with local fluorescent tissue labeling

Double-layered two-directional somatopleural cell migration during chicken body wall development revealed with local fluorescent tissue labeling

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