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JoVE Journal
Developmental Biology
En Yüz Fare Embriyolarında Düzlemsel Morfogenez Analizi için Endokardiyal Minderin Hazır...
En Yüz Fare Embriyolarında Düzlemsel Morfogenez Analizi için Endokardiyal Minderin Hazır...
JoVE Journal
Developmental Biology
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JoVE Journal Developmental Biology
En Face Endocardial Cushion Preparation for Planar Morphogenesis Analysis in Mouse Embryos

En Yüz Fare Embriyolarında Düzlemsel Morfogenez Analizi için Endokardiyal Minderin Hazırlanması

Full Text
2,149 Views
08:57 min
July 27, 2022

DOI: 10.3791/64207-v

Tamara Gonzalez-Costa1,2, Jose Luis de la Pompa1,2, Joaquim Grego-Bessa1,2

1Intercellular Signalling in Cardiovascular Development and Disease Laboratory,Centro Nacional de Investigaciones Cardiovasculares (CNIC), 2Ciber de Enfermedades Cardiovasculares

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Overview

This study presents a novel method for analyzing the endocardium of mouse embryonic valve primordium using en face imaging. This approach enables the examination of planar cell polarity and dynamics during valve development.

Key Study Components

Area of Science

  • Neuroscience
  • Developmental Biology
  • Cardiovascular Research

Background

  • Traditional methods involve analyzing endocardium through various tissue sections.
  • Understanding planar cell polarity is crucial for insights into valve development.
  • Endocardial cells play a significant role in cardiac morphogenesis.
  • Innovative imaging techniques can enhance our understanding of cellular dynamics.

Purpose of Study

  • To analyze the distribution and behavior of endocardial cells during valve development.
  • To investigate planar cell polarity in intact embryos.
  • To improve knowledge of endocardial cell dynamics.

Methods Used

  • En face imaging of the inner valve regenerative region.
  • Analysis of cell distribution and anisotropy.
  • Dissection of embryos from pregnant mice.
  • Use of fine forceps under a dissecting microscope.

Main Results

  • Demonstrated the feasibility of planar imaging for endocardial analysis.
  • Revealed insights into the dynamics of endocardial cells.
  • Showed the importance of planar cell polarity in valve development.
  • Provided a new perspective on the cellular organization of the endocardium.

Conclusions

  • The en face imaging technique offers a significant advancement in studying endocardial cells.
  • This method can deepen our understanding of cardiac development.
  • Future studies can leverage this approach for further insights into cardiovascular biology.

Frequently Asked Questions

What is the significance of planar cell polarity?
Planar cell polarity is crucial for proper tissue organization and function during development, particularly in the heart.
How does this method differ from traditional techniques?
This method allows for two-dimensional imaging of the endocardium, providing a clearer view of cell dynamics compared to traditional sectioning methods.
What are the implications of this research?
The findings can enhance our understanding of cardiac development and potential congenital heart defects.
Can this method be applied to other tissues?
While this study focuses on the endocardium, the en face imaging technique may be adapted for other tissues in developmental biology.
What are the next steps for this research?
Future research may explore the molecular mechanisms underlying planar cell polarity in various developmental contexts.

Klasik olarak, fare embriyonik kapak primordiyumunun endokardiyumu transversal, koronal veya sagital kesitler kullanılarak analiz edilmiştir. Valvulojenik bölgelerde endokardın en yüzlü, iki boyutlu görüntülenmesi için yeni yaklaşımımız, kapak gelişimi sırasında endokardın düzlemsel polarite ve hücre yeniden düzenleme analizine izin verir.

Bu protokol, ilkel endokardın düzlemsel bir oryantasyonda analiz edilmesini sağlar. Endokardiyal hücrelerin dağılımını, diğer emirlerin anizotropisini analiz edebilir ve kapak gelişimi sırasında tek bir endokardiyal hücrenin şeklini tanımlayabiliriz. Klasik doku kesitleri ile karşılaştırıldığında, iç kapak rejeneratif bölgesinin en yüz görüntülemesi, bozulmamış embriyolarda kapak gelişimi sırasında endokardiyal hücrelerin düzlemsel hücre polaritesinin ve dinamiklerinin analizine olanak sağlar.

Bu yöntem, kardiyo gelişimi sırasında düzlemsel hücre polaritesi bilgisini derinleştirmek için kullanılabilir. Ötanize hamile farelerden eksize edilen uterusu, buz gibi soğuk PBT içeren bir Petri kabına yerleştirerek başlayın. İnce forseps kullanarak diseksiyon mikroskobu altında bireysel desidüyü uterustan çıkarın.

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