Remote Neuronal Activation Coupled with Automated Blood Sampling to Induce and Measure Circulating Luteinizing Hormone in Mice

Cristina S&#225;enz de Miera; Jiane Feng; Carol F. Elias; Nathan Qi

doi:10.3791/65875

Neuronale Fernaktivierung in Verbindung mit automatisierter Blutentnahme zur Induktion und Messun...

JoVE Journal
Neuroscience

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JoVE Journal Neuroscience

Remote Neuronal Activation Coupled with Automated Blood Sampling to Induce and Measure Circulating Luteinizing Hormone in Mice

Neuronale Fernaktivierung in Verbindung mit automatisierter Blutentnahme zur Induktion und Messung des zirkulierenden luteinisierenden Hormons bei Mäusen

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August 25, 2023

DOI: 10.3791/65875-v

Cristina Sáenz de Miera¹, Jiane Feng^1,2, Carol F. Elias^1,2, Nathan Qi^1,2

¹Department of Molecular and Integrative Physiology,University of Michigan, Ann Arbor, ²Mouse Metabolic Phenotyping Center-Live,University of Michigan, Ann Arbor

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Overview

This study investigates the role of specific neuronal populations in regulating the pulsatile secretion of luteinizing hormone (LH). The authors present a protocol that enables automated blood collection and neuronal activation in conscious and undisturbed animals, allowing for precise monitoring of hormonal changes without the stress of human handling.

Key Study Components

Area of Science

Neuroscience
Endocrinology
Automated physiological monitoring

Background

Pulsatile luteinizing hormone (LH) secretion is essential for reproductive function.
Current methods for blood sampling in conscious mice are limited and stress-inducing.
Automated techniques can reduce animal stress while improving data quality.
Understanding neuronal control over hypothalamic hormone release is crucial for reproductive health research.

Purpose of Study

To develop a method for remote activation of neurons linked to LH secretion.
To enable sequential blood sampling from conscious mice without stress.
To enhance the robustness of hormone monitoring in neuroscience experiments.

Methods Used

A stereotaxic injection of adeno-associated viruses (AAV) into the brain was used to manipulate neuronal activity.
The study utilized conscious, free-moving mice for physiological measurements.
Automated blood collection systems were employed to reduce human intervention.
Procedures included careful surgical techniques to implant catheters for arterial and venous access.

Main Results

The developed protocol successfully allowed for the monitoring of LH pulsatility and response to neuronal activity.
Automated sample collection minimized stress effects, enhancing data integrity.
The findings provide insights into the dynamics of hormonal modulation through directed neuronal control.

Conclusions

This study demonstrates an effective method for investigating neuronal control over endocrine functions.
The approach enables better understanding of LH regulation, with implications for reproductive health research.
The method's ability to capture hormonal changes in undisturbed animals contributes significantly to neuroscience methodologies.

Frequently Asked Questions

What are the advantages of using automated sampling in this study?

Automated sampling minimizes human stress effects, allowing for more accurate monitoring of physiological changes in conscious animals.

How is the neuronal population activated?

Neuronal populations are activated through the injection of AAV vectors, which enables precise control over neuronal activity.

What type of data is obtained through this method?

The method provides data on pulsatile LH levels and the physiological response to neuronal activation in real-time.

Can this method be adapted for other hormonal studies?

Yes, the protocol can be adapted to study various hormonal regulatory mechanisms by targeting different neuronal populations.

What considerations should be kept in mind when using this method?

Care must be taken during surgical procedures to avoid damaging brain structures, and proper anesthesia and postoperative care are essential for animal welfare.

How does this study enhance our understanding of reproductive physiology?

By elucidating the neuronal control of LH secretion, the study provides valuable insights into the neuroendocrine regulation of reproduction.

Die Pulsatilität des luteinisierenden Hormons (LH) ist ein Kennzeichen der Fortpflanzungsfunktion. Wir beschreiben ein Protokoll zur Fernaktivierung spezifischer neuronaler Populationen, das mit der seriellen automatisierten Blutentnahme verbunden ist. Diese Technik ermöglicht eine zeitgesteuerte hormonelle Modulation, Multiplexing und die Minimierung von Manipulationseffekten auf den LH-Spiegel bei bewussten, sich frei bewegenden und ungestörten Tieren.

Unser Ziel ist es, die Rolle spezifischer Neuronenpopulationen bei der Regulation der pulsierenden Sekretion von luteinisierendem Hormon zu verstehen. Dazu benötigen wir eine präzise Kontrolle der Neuronenzuordnung, -identität und -aktivität in Kombination mit der sequentiellen Entnahme von Blutproben, ohne die Tiere zu stören oder zu stressen. Die derzeitigen Methoden zur Entnahme sequentieller Blutproben bei bewussten Mäusen beschränken sich auf Schwanzschnitte, erfordern eine lange Umstellungszeit und sind anfällig für menschliche Handhabung und Umweltbelastungen.

Automatisierte Infusions- und Blutproben können ohne Anwesenheit von Forschern durchgeführt werden, was den Stress begrenzt und die Erfassung hormoneller Veränderungen bei bewussten, sich frei bewegenden und ungestörten Mäusen ermöglicht. Diese Technik ist eine Bereicherung für die neurowissenschaftliche Forschung, da sie ein robustes Unterstützungssystem bietet. Das Michigan Mouse Metabolic Phenotyping Center bietet automatisierte Infusions- und breit angelegte Probenahmeexperimente als Kerndienstleistung an.