Dissection of Adult Mouse Stria Vascularis for Single-Nucleus Sequencing or Immunostaining

Dillon Strepay; Rafal Olszewski; Ian Taukulis; J. Dixon Johns; Shoujun Gu; Michael Hoa

doi:10.3791/65254

Disección de estrías vasculares de ratón adulto para secuenciación de un solo núcleo o inmunotinción

JoVE Journal
Biology

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Biology

Dissection of Adult Mouse Stria Vascularis for Single-Nucleus Sequencing or Immunostaining

Disección de estrías vasculares de ratón adulto para secuenciación de un solo núcleo o inmunotinción

Full Text

4,537 Views

08:51 min

April 21, 2023

DOI: 10.3791/65254-v

Dillon Strepay¹, Rafal Olszewski¹, Ian Taukulis¹, J. Dixon Johns¹, Shoujun Gu¹, Michael Hoa¹

¹Auditory Development and Restoration Program, National Institute on Deafness and Other Communication Disorders,National Institutes of Health

Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study investigates the stria vascularis, a critical structure in the cochlea responsible for generating the endocochlear potential. Using adult mice, the research focuses on dissecting the stria vascularis for single-nucleus sequencing and immunostaining to explore its role in hearing stability and disorders.

Key Study Components

Research Area

Auditory system biology
Hearing disorders research
Cellular signaling in cochlear structures

Background

The stria vascularis is essential for auditory function.
It plays a role in eye and homeostasis regulation in the cochlea.
Dysfunctional mechanisms relate to conditions like Meniere's disease.

Methods Used

Dissection of stria vascularis from adult mice.
Single-nucleus RNA sequencing for transcription analysis.
Immunostaining for protein detection in stria vascularis.

Main Results

Mapping the transcriptional landscape of the stria vascularis.
Linking stria vascularis dysfunction to auditory instability disorders.
Improving understanding of cellular interactions within cochlear structures.

Conclusions

The study illustrates the complexity and importance of the stria vascularis in hearing.
Findings enhance comprehension of the mechanisms underlying hearing disorders.

Frequently Asked Questions

What is the stria vascularis?

The stria vascularis is a tissue layer within the cochlea that plays a crucial role in producing the endocochlear potential necessary for normal hearing.

Why is single-nucleus sequencing used in this research?

Single-nucleus sequencing allows researchers to examine the specific transcriptional profiles of individual cells within the stria vascularis, providing insights into cellular functions and interactions.

What are hearing instability disorders?

Hearing instability disorders are conditions characterized by fluctuations in hearing or sudden hearing loss, often linked to dysfunction in structures like the stria vascularis.

What implications do the findings have?

These findings could advance the understanding of hearing disorders and potentially lead to new therapeutic approaches.

What techniques are employed for immunostaining?

Immunostaining involves applying specific antibodies to detect proteins of interest in the tissue, helping to visualize cellular components.

How does the stria vascularis relate to Meniere's disease?

Research indicates that dysfunction in the stria vascularis may contribute to the symptoms experienced in Meniere's disease, linking cellular health to auditory stability.

La estría vascular es vital para la generación de potencial endococlear. Aquí, presentamos la disección de la estría vascular del ratón adulto para la secuenciación de un solo núcleo o inmunotinción.

La estría vascular funciona para generar el potencial endococlear y regular la homeostasis iónica en la cóclea. El programa de desarrollo y restauración auditiva busca comprender los mecanismos subyacentes que conectan la homeostasis de iones disfuncionales con los trastornos auditivos y de estabilidad, que son enfermedades en las que se produce fluctuación o pérdida auditiva repentina. El trabajo en mi laboratorio ha establecido el paisaje transcripcional de la estría vascular y ha contribuido a nuestra comprensión de su importancia en la audición.

Hemos utilizado estos datos para establecer paralelismos con trastornos auditivos y de estabilidad poco conocidos, incluida la enfermedad de Meniere. Este protocolo permite la disección y el aislamiento de la estría vascular de montaje completo para estudios moleculares adicionales, incluida la inmunohistoquímica. Dado que los tipos de células de estría vascular están intrincadamente entrelazadas entre sí, el uso del aislamiento nuclear y la secuenciación de ARN de un solo núcleo ha facilitado nuestra comprensión de la distinta maquinaria transcripcional involucrada.

View the full transcript and gain access to thousands of scientific videos

Explore More Videos

Biología Número 194 oído interno estría vascular disección ratón adulto