A Neonatal Rodent Model of Retroorbital Vein Injection

Eridan Rocha-Ferreira; Syam Nair; Owen Herrock; Erik Axel Andersson; Carl Joakim Ek; Carina Mallard; Henrik Hagberg

doi:10.3791/65386

JoVE Journal
Neuroscience
Author Produced

This content is Free Access.

JoVE Journal Neuroscience

A Neonatal Rodent Model of Retroorbital Vein Injection

眶后静脉注射的新生儿啮齿动物模型

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04:39 min

February 23, 2024

DOI: 10.3791/65386-v

Eridan Rocha-Ferreira¹, Syam Nair¹, Owen Herrock¹, Erik Axel Andersson², Carl Joakim Ek², Carina Mallard², Henrik Hagberg¹

¹Centre of Perinatal Medicine and Health, Institute of Clinical Sciences, Department of Obstetrics and Gynecology, Sahlgrenska Academy,University of Gothenburg, ²Centre of Perinatal Medicine and Health, Institute of Neuroscience and Physiology, Sahlgrenska Academy,University of Gothenburg

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Overview

This protocol highlights a reproducible retro-orbital injection method for intravenous drug administration in neonatal rats and mice. The technique facilitates compound delivery into the venous circulation, which is particularly crucial for preclinical studies mimicking neonatal care unit conditions.

Key Study Components

Area of Science

Neuroscience
Pharmacology
Animal models

Background

Intravenous administration is commonly used in neonatal intensive care units.
Injections via the tail vein are difficult in neonates, making alternative routes necessary.
The retro-orbital route is simpler and can be used effectively for compound administration.
This method allows for real-time monitoring of injection success through visible dye tracking.

Purpose of Study

To optimize intravenous drug administration techniques in neonatal rodents.
To provide a reliable method for compound delivery that can be used in various preclinical settings.
To enhance reproducibility and outcomes in rodent studies related to neonatal care.

Methods Used

The retro-orbital injection method is demonstrated using trypan blue dye.
Subjects include neonatal rats and mice under anesthesia during the procedure.
New sterile syringes are utilized for each animal to prevent contamination.
Injection occurs at the medial canthus of the eye at a 40-degree angle.
Recovery protocols ensure the well-being of the animals post-injection.

Main Results

The protocol enables successful visualization of compounds in the venous system.
Effective tracer administration allows for assessment of vascular responses after injury.
These results highlight the potential for quantifying vascular leakage and injury assessment.
The method does not cause adverse effects when performed properly.

Conclusions

This study establishes the retro-orbital injection as a reliable method for drug administration in neonatal rodents.
The technique presents significant clinical relevance in translational research.
Enhanced understanding of venous drug delivery mechanisms supports future studies in neonatal pharmacology.

Frequently Asked Questions

What are the advantages of using the retro-orbital injection method?

The retro-orbital injection method simplifies intravenous administration in neonatal rodents, providing a more accessible alternative to tail vein injections, which can be challenging.

How is the retro-orbital injection performed?

The injection is performed with the animal under anesthesia, using a 29 to 31 gauge needle positioned at the medial canthus of the eye, angled appropriately to access the venous plexus.

What types of data can this method help obtain?

The method allows for real-time monitoring of compounds in the blood circulation, enabling assessments of tracer distributions and vascular responses.

Can this method be adapted for other substances?

Yes, this injection technique can be used for various compounds, including antibodies, cells, and other therapeutic agents directly into the venous circulation.

Are there any limitations to this injection method?

Care must be taken to ensure proper needle placement to avoid complications, and the procedure should only be performed by trained individuals for optimal animal welfare.

How does this study contribute to neonatal pharmacology?

The study presents a reliable method for intravenous drug administration that can improve translational research on drug effects in neonatal care settings.

该方案旨在证明一种可重复的静脉给药途径，该途径可在整个新生儿期用于大鼠和小鼠。该程序对于临床前啮齿动物研究很重要，这些研究希望反映主要使用静脉给药的新生儿监护病房的药物给药。

眼眶后注射是一种专门的注射方法，用于将化合物注入静脉循环。动物研究是迈向临床工作的重要一步。作为动物研究的一部分，需要通过不同的途径施用不同类型的化合物。

其中一些途径包括皮下注射、腹膜内注射和静脉注射。静脉注射是新生儿重症监护病房新生儿最常用的途径。然而，在啮齿动物研究中，特别是在成年啮齿动物中，静脉注射通常是在尾静脉进行的。

在新生儿中，成功且可重复地进行尾静脉注射是极其困难的。因此，逆轨路线可以用作更简单的替代方案。首先，我们将演示如何使用染料台盼蓝，以便观众可以清楚地看到染料进入静脉循环。

将新生大鼠置于完全麻醉状态，头部侧卧，放在光源上方，因此眼睛和侧向脉管系统暴露可见。针头插入眼窝前部，相当于内眦。以轻柔、平稳和流畅的运动注入溶液。

为了更好地重现这一过程，了解眼眶静脉解剖结构很重要。在大鼠中，眼睛下方有几条静脉网络，可以轻松直接地进入静脉丛。在新生白化大鼠中，浅表颞静脉和面部静脉也可见。

此过程可以以完全相同的方式执行。在新生小鼠中。使用29至31号的针头，约0.30毫米。

为了获得准确的体积，请从移液体积中抽出要进样的溶液。将动物侧卧放在平坦的表面上。诱导全身异氟醚麻醉。

5%感应，3%维护。使用拉退出反射法检查麻醉深度。头部朝右，向右眶后窦注射。

将针斜面向下插入眼窝前部，相当于中眦，角度约为 40 度。这个角度允许针头指向眼眶的后部。以轻柔、平稳和流畅的动作注射。

等待片刻，然后慢慢抽出针头，以避免回流。为每只动物使用新的无菌注射器以避免污染。将幼崽放在恢复箱中，放在受保护的加热装置上，温度约为 35 至 37 摄氏度。

等待恢复，并在将幼崽送回大坝之前检查是否有任何遇险迹象。该技术用于将示踪剂生物素-葡聚糖施用到经历过生发基质出血的动物的大脑脉管系统中。接受生理盐水注射作为对照的动物显示大脑内脉管系统没有可见的染色。