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Gastrulation

Gastrulation

Gastrulation establishes the three primary tissues of an embryo: the ectoderm, mesoderm, and endoderm. This developmental process relies on a series of intricate cellular movements, which in humans transforms a flat, “bilaminar disc” composed of two cell sheets into a three-tiered structure. In the resulting embryo, the endoderm serves as the bottom layer, and stacked directly above it is the intermediate mesoderm, and then the uppermost ectoderm. Respectively, these tissue strata...

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Related Experiment Video

Updated: Jan 1, 2026

Dissection and Culture of Mouse Embryonic Kidney

Dissection and Culture of Mouse Embryonic Kidney

Published on: May 17, 2017

Insight into Nephrocan Function in Mouse Endoderm Patterning.

Martina Addeo^1,2, Silvia Buonaiuto², Ilaria Guerriero¹

¹Istituto di Ricerche Genetiche "G. Salvatore", Biogem s.c.ar.l, Ariano Irpino, 83031 Avellino, Italy.

International Journal of Molecular Sciences

|December 22, 2019

Summary

This summary is machine-generated.

Nephrocan (Nepn) is crucial for early endoderm development. Its absence impairs in vitro endoderm differentiation, highlighting its role in specifying posterior foregut markers.

Keywords:

(CRISPR)/CRISPR-associated systems 9 (Cas9)Nephrocan gene differentiation, definitive endoderm embryonic stem cells mouse model transcriptional variants

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Published on: March 28, 2020

Area of Science:

Developmental Biology
Regenerative Medicine
Molecular Genetics

Background:

Endoderm-derived organs like the liver and pancreas are key targets for regenerative therapies.
Understanding endoderm specification pathways is vital, but limited by a lack of early endoderm-specific markers.
Nephrocan (Nepn) is a secreted protein expressed during early murine endoderm specification (E7.5-11.5).

Purpose of the Study:

To investigate the role of Nephrocan (Nepn) in endoderm specification.
To identify and characterize new transcript variants of Nepn.
To analyze the in vivo and in vitro effects of Nepn deficiency on endoderm development.

Main Methods:

Identification of a novel Nepn transcript variant via alternative splicing.
Generation of Nepn knock-out (KO) mice (Nepn-/-).
Creation of a nullizygous mouse embryonic stem cell (mESC) line using CRISPR/Cas9 technology.
In vitro differentiation of mESCs towards an endoderm lineage.

Main Results:

A new Nepn transcript variant with differential, tissue-specific adult expression was identified.
Nepn-/- mice exhibited no observable phenotype and were born at Mendelian ratios.
In vitro, Nepn-deficient mESCs showed impaired endoderm differentiation.
Loss of Nepn affected the expression of posterior foregut-associated markers during differentiation.

Conclusions:

Nephrocan plays a significant role in the in vitro specification of endoderm, particularly the posterior foregut.
While Nepn is not essential for overall embryonic development in vivo, it is critical for proper endoderm differentiation in vitro.
Further research into Nepn's function could advance regenerative therapies for endoderm-derived organs.