A device for stereotaxic viral delivery into the brains of neonatal mice
| Autor: | Christoph Kellendonk, Pedro R. Olivetti, Clay O Lacefield |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Neuronal targeting
virus Biology Benchmark Indirect pathway of movement General Biochemistry Genetics and Molecular Biology Virus Green fluorescent protein Stereotaxic Techniques Midbrain Mice 03 medical and health sciences 0302 clinical medicine Biological neural network Animals Postnatal day 030304 developmental biology Neurons 0303 health sciences adaptor Dopaminergic Gene Transfer Techniques Brain stereotaxic Animals Newborn Printing Three-Dimensional Viruses neonatal mouse Neuroscience 030217 neurology & neurosurgery viral delivery Biotechnology |
| Zdroj: | Biotechniques |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/btn-2020-0050 |
| Popis: | The increasing interest in manipulating neural circuits in developing brains has created a demand for reliable and accurate methods for delivering viruses to newborn mice. Here we describe a novel 3D-printed mouse neonatal stereotaxic adaptor for intracerebral viral injection that provides enhanced precision and reliability. Using this device, we injected A2a-Cre mice with a Cre-dependent hM4D-mCherry viral construct at postnatal day 1 (P1) and demonstrated selective expression in the striatal indirect pathway neurons on days P7, P11 and P25. Similarly, dopaminergic midbrain neurons were selectively targeted with a Cre-dependent green fluorescent protein virus in Dat-IRES-Cre neonates and expression examined at P25. Our open-source neonatal stereotaxic mouse adaptor facilitates neonatal neuronal targeting, which should improve the ability to label and modify neural circuits in developing mouse brains. METHOD SUMMARY The neonatal mouse stereotaxic adaptor, designed and easily 3D printed in-house, was fitted into the ear and tooth bars of a mouse digital stereotaxic apparatus and held the head of the ice-anesthetized newborn pups in a cone-shaped cradle during the virus injection procedure. After the secured heads were balanced on the AP and ML axes by adjusting the ear and tooth bars, Cre-dependent viruses (AAV5-hSyn-DIO-hM4D-mCherry or AAV5-hSyn-DIO-GFP) were injected to the desired brain region of Cre-expressing transgenic mice, while the animals were partially covered in ice. After completion of each surgery, the neonate was removed from the adaptor and returned to the den for recovery. Neuroanatomical viral expression was verified by immunofluorescence 6, 10 and 24 days after the procedure. |
| Databáze: | OpenAIRE |
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