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

Frequenti prelievi di punta della coda in topi per la valutazione della secrezione Pulsatile dell...

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JoVE Journal Neuroscience

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

Frequenti prelievi di punta della coda in topi per la valutazione della secrezione Pulsatile dell'ormone luteinizzante

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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.

Qui presentiamo un protocollo di campionamento di sangue punta della coda per la raccolta del campione di frequente nei topi sfrenati. Questo metodo è utile per la valutazione dei modelli di secrezione pulsatile dell'ormone luteinizzante e potrebbe essere adattato per l'analisi di altri fattori circolanti.

L'obiettivo generale del campionamento frequente del sangue sulla punta della coda è quello di ottenere campioni di sangue intero per la misurazione della secrezione pulsatile dell'ormone luteinizzante denominata LH in topi svegli non anestetizzati e con un comportamento normale. Questo metodo può aiutare a rispondere a domande chiave nel campo della neuroendocrinologia riproduttiva, fornendo un protocollo affidabile per valutare la secrezione pulsatile dell'ormone luteinizzante nei topi. Il vantaggio principale di questa tecnica è che la manipolazione e l'assuefazione frequenti garantiscono misurazioni robuste e affidabili del polso dell'ormone luteinizzante dopo trattamenti sperimentali acuti o cronici.

Per cinque settimane prima del prelievo di sangue, maneggiare i topi su base giornaliera dal lunedì al venerdì. Nelle ultime due settimane, maneggia i topi anche sabato e domenica. Eseguire il regime di manipolazione dei topi in una cabina di biosicurezza.

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