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

Attivazione neuronale remota accoppiata con campionamento automatico del sangue per indurre e mis...

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

Attivazione neuronale remota accoppiata con campionamento automatico del sangue per indurre e misurare l'ormone luteinizzante circolante nei topi

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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.

La pulsatilità dell'ormone luteinizzante (LH) è un segno distintivo della funzione riproduttiva. Descriviamo un protocollo per l'attivazione remota di specifiche popolazioni neuronali legate alla raccolta automatica seriale del sangue. Questa tecnica consente la modulazione ormonale temporizzata, il multiplexing e la riduzione al minimo degli effetti di manipolazione sui livelli di LH in animali coscienti, in movimento libero e indisturbati.

Il nostro obiettivo è quello di comprendere il ruolo di specifiche popolazioni di neuroni nella regolazione della secrezione pulsatile dell'ormone luteinizzante. Per questo, abbiamo bisogno di un controllo preciso dell'allocazione, dell'identità e dell'attività dei neuroni, in combinazione con la raccolta sequenziale di campioni di sangue senza disturbare o stressare gli animali. L'attuale metodo per la raccolta di campioni di sangue sequenziali nei topi coscienti è limitato ai tagli della coda, richiede un lungo periodo di cambiamento ed è suscettibile alla manipolazione umana e agli stress ambientali.

L'infusione automatizzata e il prelievo di sangue possono essere condotti senza la presenza di sperimentatori, il che limita lo stress e consente la cattura dei cambiamenti ormonali nei topi coscienti, che si muovono liberamente e indisturbati. Questa tecnica è una risorsa per la ricerca neuroscientifica in quanto fornisce un solido sistema di supporto. Il Michigan Mouse Metabolic Phenotyping Center offre l'infusione automatizzata e ampi esperimenti di campionamento come servizio principale.