Frequent Tail-tip Blood Sampling in Mice for the Assessment of Pulsatile Luteinizing Hormone Secretion

Richard B. McCosh; Michael J. Kreisman; Kellie M. Breen

doi:10.3791/57894

Fréquente la queue-extrémité prélèvements chez les souris pour l’évaluation de la sécrétion de l’...

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JoVE Journal Neuroscience

Frequent Tail-tip Blood Sampling in Mice for the Assessment of Pulsatile Luteinizing Hormone Secretion

Fréquente la queue-extrémité prélèvements chez les souris pour l’évaluation de la sécrétion de l’Hormone lutéinisante Pulsatile

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05:58 min

July 4, 2018

DOI: 10.3791/57894-v

Richard B. McCosh¹, Michael J. Kreisman¹, Kellie M. Breen¹

¹Department of Reproductive Medicine,University of California, San Diego School of Medicine

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Overview

This study describes a tail-tip blood sampling protocol for the collection of blood samples in unrestrained mice, focusing on the measurement of pulsatile luteinizing hormone (LH) secretion. The method is designed to enable reliable assessments of LH pulsatile secretion, contributing to research in reproductive neuroendocrinology.

Key Study Components

Area of Science

Neuroendocrinology
Reproductive biology
Animal behavior

Background

Luteinizing hormone pulsatile secretion is critical in reproductive physiology.
Reliable blood sampling techniques are essential for accurately measuring hormonal fluctuations.
Existing methods may not adequately account for the effects of animal stress.
This protocol aims to minimize mouse stress during blood collection.

Purpose of Study

To develop a reliable tail-tip blood sampling protocol for unrestrained mice.
To assess LH pulsatile secretion in response to both acute and chronic treatments.
To improve the accuracy of neuroendocrinological research methods.

Methods Used

The protocol involves daily handling and habituation of mice prior to blood collection.
Adult female C57BL/6 mice were used, with blood collected from the tail tip for LH measurement.
Frequent handling sessions helped facilitate a stress-free environment.
Blood samples were collected over a set timeframe to establish hormonal baselines.

Main Results

The technique led to successful collection of LH levels in control animals.
Psychosocial stress from restraint significantly suppressed LH secretion in experimental groups.
Stress minimized reliable measurements of pulsatile LH secretion, highlighting the method's efficacy.
Environmental disturbances were found to affect hormonal measurements.

Conclusions

This study establishes a novel method for obtaining accurate hormonal measurements in mice.
The protocol allows researchers to explore LH regulation effectively in neuroendocrinology studies.
Understanding the impact of stress on hormonal secretion contributes to broader insights into reproductive physiology and animal behavior.

Frequently Asked Questions

What are the advantages of this blood sampling protocol?

This protocol minimizes stress by allowing mice to remain unrestrained, facilitating more reliable hormonal measurements.

How is the tail sample obtained?

Blood is collected by gently clipping the tail tip and stroking it to form a droplet, which is then sampled.

What is the biological model used in this study?

The study uses adult female C57BL/6 mice, which are well-characterized for neuroendocrinology research.

How does this method contribute to neuroendocrinology?

By providing reliable blood sampling under unrestrained conditions, researchers can better understand LH pulsatile secretion and its regulation.

What are key considerations when using this method?

Minimizing environmental disturbances is crucial, as noise and interruptions can impact hormonal measurements.

Can this method be adapted for other experiments?

Yes, the protocol can be adapted to assess other circulating factors, broadening its applicability in research.

Nous présentons ici un protocole de prélèvement de sang de la queue-extrémité pour prélèvement fréquents chez les souris sans retenue. Cette méthode est utile pour évaluer des modèles de la sécrétion pulsatile de LH et pourrait être adaptée pour l’analyse d’autres facteurs circulants.

L’objectif global des prélèvements sanguins fréquents à l’extrémité de la queue est d’obtenir des échantillons de sang total pour la mesure de la sécrétion d’hormone lutéinisante pulsatile appelée LH chez les souris éveillées, non anesthétisées et au comportement normal. Cette méthode peut aider à répondre à des questions clés dans le domaine de la neuroendochrinologie reproductrice en fournissant un protocole fiable pour évaluer la sécrétion pulsatile d’hormone lutéinisante chez la souris. Le principal avantage de cette technique est qu’une manipulation et une accoutumance fréquentes garantiront des mesures d’impulsion d’hormone lutéinisante robustes et fiables après des traitements expérimentaux aigus ou chroniques.

Pendant les cinq semaines précédant le prélèvement sanguin, manipulez les souris tous les jours du lundi au vendredi. Au cours des deux dernières semaines, manipulez également les souris le samedi et le dimanche. Effectuez le régime de manipulation des souris dans une enceinte de biosécurité.

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