In utero intracerebroventricular delivery of adeno-associated viral vectors to target mouse choroid plexus and cerebrospinal fluid

Jang, Ahram; Lehtinen, Maria K.

doi:10.1016/j.xpro.2022.101975

Cited by 1 publication

(1 citation statement)

References 15 publications

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“…Laparotomy was performed and the uterine horns were exposed. Virus solution (1 µl, with a titer of 1 × 10 12 ) was injected into one lateral ventricle of each embryo using glass capillary pipettes [ 14 ]. Immediately following injection, the uterine horns were placed back into the abdominal cavity and the incision was sutured.…”

Section: Methodsmentioning

confidence: 99%

Age-appropriate potassium clearance from perinatal cerebrospinal fluid depends on choroid plexus NKCC1

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Pragana

et al. 2023

Fluids Barriers CNS

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Regulation of the volume and electrolyte composition of the cerebrospinal fluid (CSF) is vital for brain development and function. The Na-K-Cl co-transporter NKCC1 in the choroid plexus (ChP) plays key roles in regulating CSF volume by co-transporting ions and mediating same-direction water movements. Our previous study showed ChP NKCC1 is highly phosphorylated in neonatal mice as the CSF K+ level drastically decreases and that overexpression of NKCC1 in the ChP accelerates CSF K+ clearance and reduces ventricle size [1]. These data suggest that NKCC1 mediates CSF K+ clearance following birth in mice. In this current study, we used CRISPR technology to create a conditional NKCC1 knockout mouse line and evaluated CSF K+ by Inductively Coupled Plasma Optical Emission spectroscopy (ICP-OES). We demonstrated ChP-specific reduction of total and phosphorylated NKCC1 in neonatal mice following embryonic intraventricular delivery of Cre recombinase using AAV2/5. ChP-NKCC1 knockdown was accompanied by a delayed perinatal clearance of CSF K+. No gross morphological disruptions were observed in the cerebral cortex. We extended our previous results by showing embryonic and perinatal rats shared key characteristics with mice, including decreased ChP NKCC1 expression level, increased ChP NKCC1 phosphorylation state, and increased CSF K+ levels compared to adult. Collectively, these follow up data support ChP NKCC1’s role in age-appropriate CSF K+ clearance during neonatal development.

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Section: Methodsmentioning

confidence: 99%