Chicken cerebellar granule neurons rapidly develop excitotoxicity in culture

Jacobs, Chris M.; Aden, Petra; Mathisen, Gro Haarklou; Khuong, Erica; Gaarder, Mona; Løberg, Else Marit; Lømo, Jon; Mæhlen, Jan; Paulsen, Ragnhild E.

doi:10.1016/j.jneumeth.2006.02.012

Cited by 23 publications

(20 citation statements)

References 23 publications

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“…The chicken cerebella were harvested as described above, and the tissue was cut into pieces by a scalpel. The tissue was trypsinised before the cells were seeded on 35 mm dishes or 96‐well plates pre‐treated with poly‐L lysine (Wilkin et al, 1976; Jacobs et al, 2006) at a density of 1.7 × 10 6 cells/ml (Cohen et al, 1979). The cultures were grown in BME supplemented with heat‐inactivated chicken serum (7.5%), KCl (22 mM), L‐glutamine (2 mM), insulin (100 nM) and PenStrep (1%) (Mathisen et al, 2013).…”

Section: Methodsmentioning

confidence: 99%

Opioid receptor‐mediated changes in the NMDA receptor in developing rat and chicken

Fjelldal

Hadera

Kongstorp³

et al. 2019

Intl J of Devlp Neuroscience

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The use of opioids during pregnancy has been associated with neurodevelopmental toxicity in exposed children, leading to cognitive and behavioural deficits later in life. The N‐methyl‐D‐aspartate receptor (NMDAR) subunit GluN2B plays critical roles in cerebellar development, and methadone has been shown to possess NMDAR antagonist effect. Consequently, we wanted to explore if prenatal opioid exposure affected GluN2B subunit expression and NMDAR function in rat and chicken cerebellum. Pregnant rats were exposed to methadone (10 mg/kg/day) or buprenorphine (1 mg/kg/day) for the whole period of gestation, using an osmotic minipump. To further examine potential effects of prenatal opioid exposure in a limited time window, chicken embryos were exposed to a 20 mg/kg dose of methadone or morphine on embryonic days 13 and 14. Western blot analysis of cerebella isolated from 14 days old rat pups exposed to buprenorphine showed significantly lower level of the GluN2B subunit, while the opioid exposed chicken embryo cerebellar GluN2B expression remained unaffected at embryonic day 17. However, we observed increased NMDA/glycine‐induced calcium influx in cerebellar granule neurone cultures from opioid exposed chicken embryos. We conclude that prenatal opioid exposure leads to opioid receptor‐dependent reduction in the postnatal expression of GluN2B in rat cerebella, and increase in NMDA‐induced calcium influx in chicken embryo cerebella.

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Section: Methodsmentioning

confidence: 99%

Opioid receptor‐mediated changes in the NMDA receptor in developing rat and chicken

Fjelldal

Hadera

Kongstorp³

et al. 2019

Intl J of Devlp Neuroscience

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“…Cultures of cerebellar granule neurons were prepared on ED17. The granule neurons were isolated as previously described (Jacobs et al, 2006) and then grown in serum free conditions as follows: After isolation 1.7 × 10 6 cells/ml were seeded onto poly‐ l ‐lysine‐coated dishes and incubated in BME supplemented with heat inactivated horse serum (10%), KCl (25 mM), glutamine (2 mM), insulin (100 nM), and penicillin/streptomycin (100 Units/ml Penicillin; 100 μg/ml Streptomycin) at 37 °C and 5% CO 2 for 24 h. Then the culture medium was replaced with serum‐free BME supplemented with KCl (25 mM), glutamine (2 mM), penicillin/streptomycin (100 Units/ml Penicillin; 100 μg/ml Streptomycin), insulin (25 μg/ml), transferrin (100 μg/ml), T3 (1 nM), and selenite (30 nM) (Barthel et al, 1996). The concentrations given in the brackets were the final concentrations in the media.…”

Section: Methodsmentioning

confidence: 99%

Prenatal exposure to bisphenol A interferes with the development of cerebellar granule neurons in mice and chicken

Mathisen

Yazdani

Rakkestad

et al. 2013

Intl J of Devlp Neuroscience

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In mice, prenatal exposure to low doses of bisphenol A has been shown to affect neurogenesis and neuronal migration in cortex, resulting in disturbance of both neuronal positioning and the network formation between thalamus and cortex in the offspring brain. In the present study we investigated whether prenatal exposure to bisphenol A disturbs the neurodevelopment of the cerebellum. Two different model systems were used; offspring from two strains of mice from mothers receiving bisphenol A in the drinking water before mating, during gestation and lactation, and chicken embryos exposed to bisphenol A (in the egg) on embryonic day 16 for 24h before preparation of cerebellar granule cell cultures. In the cerebellum, tight regulation of the level of transcription factor Pax6 is critical for correct development of granule neurons. During the development, the Pax6 level in granule neurons is high when these cells are located in the external granule layer and during their migration to the internal granule layer, and it is then reduced. We report that bisphenol A induced an increase in the thickness of the external granule layer and also an increase in the total cerebellar Pax6 level in 11 days old mice offspring. In cultured chicken cerebellar granule neurons from bisphenol A injected eggs the Pax6 level was increased day 6 in vitro. Together, these findings indicate that bisphenol A may affect the granule neurons in the developing cerebellum and thereby may disturb the correct development of the cerebellum.

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“…Chicken cerebellar granule neurons were cultured as previously described (Jacobs et al, 2006), with slight modifications. In brief, cells were isolated from chicken cerebella at embryonic day (ED) 17.…”

Section: Cell Culture and Treatmentmentioning

confidence: 99%

“…The transfection efficiency tended to decrease with increasing amounts of Metafectene ® Pro (>2 l) and DNA (>1 g). Since the chicken cultures contain about 80% neurons (Jacobs et al, 2006), the transfection conditions from Fig. 1A giving the highest luciferase values were repeated using a gfp plasmid in order to find a condition giving a high number of gfp-expressing granule neurons when comparing to all gfp-expressing cells.…”

Section: Transfection Of Chicken Cerebellar Granule Neurons With Metamentioning

confidence: 99%

“…Chicken cerebellar granule neurons are a useful model for studies of neuronal cell death and development (Jacobs et al, 2006). High amounts of glucocorticoid receptor (GR) have been found in the developing rat cerebellum (Diaz et al, 1998) and it has been suggested that ligand activated GR has a central role in the regulation of neuronal proliferation and possibly cell differentiation and maturation (Diaz et al, 1998;Kim et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

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Transfection of chicken cerebellar granule neurons used to study glucocorticoid receptor regulation by nuclear receptor 4A (NR4A)

Strom

Aden

Mathisen

et al. 2010

Journal of Neuroscience Methods

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Chicken cerebellar granule neurons rapidly develop excitotoxicity in culture

Cited by 23 publications

References 23 publications

Opioid receptor‐mediated changes in the NMDA receptor in developing rat and chicken

Opioid receptor‐mediated changes in the NMDA receptor in developing rat and chicken

Prenatal exposure to bisphenol A interferes with the development of cerebellar granule neurons in mice and chicken

Transfection of chicken cerebellar granule neurons used to study glucocorticoid receptor regulation by nuclear receptor 4A (NR4A)

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