Identifying suitable reference genes for developing and injured mouse CNS tissues

Xu, Danyi; Liu, Ajuan; Wang, Xuan; Zhang, Ming; Zhang, Zunyi; Tan, Zhou; Qiu, Mengsheng

doi:10.1002/dneu.22558

Cited by 22 publications

(17 citation statements)

References 33 publications

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“…The reference genes for both lumbar vertebrae and tibial diaphyses were rat hypoxanthine phosphoribosyltransferase 1 ( Hprt1 ) and rat peptidylprolyl isomerase A ( Ppia ). These reference genes were selected based on past reports [ 36 – 38 ].…”

Section: Methodsmentioning

confidence: 99%

Is C-type natriuretic peptide regulated by a feedback loop? A study on systemic and local autoregulatory effect

et al. 2020

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C-type natriuretic peptide (CNP) is a pivotal enhancer of endochondral bone growth and is expected to be a therapeutic reagent for impaired skeletal growth. Although we showed that CNP stimulates bone growth as a local regulator in the growth plate via the autocrine/paracrine system, CNP is abundantly produced in other various tissues and its blood concentration is reported to correlate positively with growth velocity. Therefore we investigated the systemic regulation of CNP levels using rodent models. In order to examine whether CNP undergoes systemic feedback regulation, we investigated blood CNP levels and local CNP expression in various tissues, including cartilage, of 4-week-old rats after systemic administration of sufficient amounts of exogenous CNP (0.5 mg/kg/day) for 3 days. This CNP administration did not alter blood NT-proCNP levels in male rats but decreased mRNA expression only in tissue that included cartilage. Decrease in expression and blood NT-proCNP were greater in female rats. To analyze the existence of direct autoregulation of CNP in the periphery as an autocrine/paracrine system, we estimated the effect of exogenous supplementation of CNP on the expression of endogenous CNP itself in the growth plate cartilage of extracted fetal murine tibias and in ATDC5, a chondrogenic cell line. We found no alteration of endogenous CNP expression after incubation with adequate concentrations of exogenous CNP for 4 and 24 hours, which were chosen to observe primary and later transcriptional effects, respectively. These results indicate that CNP is not directly autoregulated but indirectly autoregulated in cartilage tissue. A feedback system is crucial for homeostatic regulation and further studies are needed to elucidate the regulatory system of CNP production and function.

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Section: Methodsmentioning

confidence: 99%

Is C-type natriuretic peptide regulated by a feedback loop? A study on systemic and local autoregulatory effect

et al. 2020

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“…Additional reference genes including HPRT1, RPL13A, SDHA, PPIA, and GUSB were used to validate the effectiveness of GAPDH as the main reference gene (Supplementary S7A-D ). The selection of these reference genes was based on prior evaluation and their expression in the brain and blood 37 – 43 . We have further validated that the expression of these housekeeping/reference genes was not altered in the PD samples or in MPTP-treated mice from our study and in prior studies (Supplementary S7E–L ).…”

Section: Methodsmentioning

confidence: 99%

Reduction of HIP2 expression causes motor function impairment and increased vulnerability to dopaminergic degeneration in Parkinson’s disease models

Huang

Qin

et al. 2018

Cell Death Dis

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Huntingtin interaction protein 2 (HIP2) is an E2 ubiquitin-conjugating enzyme associated with neurodegenerative diseases, and HIP2 mRNA has been implicated as a potential blood biomarker for Parkinson’s disease (PD). However, it is unclear whether the alteration of HIP2 expression may contribute to the development of PD, and whether the change of HIP2 in blood could reflect its expression in the brain or motor functions in PD patients. In this study, we established a mouse line with HIP2 haploinsufficiency. The reduction of the HIP2 expression led to spontaneous motor function impairment and dopaminergic neuronal loss. Furthermore, HIP2 haploinsufficiency increased the susceptibility of mice to 6-hydroxydopamine (6-OHDA) and caused severe loss of dopaminergic neurons. Interestingly, in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model for PD, we observed concurrent, highly correlated decrease of HIP2 expression in the brain and in the blood. Using blood samples from more than 300 patients, we validated the decreased HIP2 mRNA in PD patients, including de novo patients. Finally, in a 1-year, 20-patient study, we observed reversed blood HIP2 mRNA levels accompanying improved motor and overall daily functions in 75% of the PD patients with instructed Tai Chi training. Therefore, our in vivo studies have indicated HIP2 insufficiency as a contributing factor for PD, and functionally validated blood HIP2 as a useful and reversible biomarker for PD.

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“…A melting-curve analysis was included in every qPCR run. YWHAZ and Gusb were chosen as housekeeping genes, as they are supposed to be most consistently expressed across different regions of the mouse brain [13].…”

Section: Quantitative Real-time Pcr (Qpcr)mentioning

confidence: 99%

Anoctamin 2-chloride channels reduce simple spike activity and mediate inhibition at elevated calcium concentration in cerebellar Purkinje cells

et al. 2021

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Modulation of neuronal excitability is a prominent way of shaping the activity of neuronal networks. Recent studies highlight the role of calcium-activated chloride currents in this context, as they can both increase or decrease excitability. The calcium-activated chloride channel Anoctamin 2 (ANO2 alias TMEM16B) has been described in several regions of the mouse brain, including the olivo-cerebellar system. In inferior olivary neurons, ANO2 was proposed to increase excitability by facilitating the generation of high-threshold calcium spikes. An expression of ANO2 in cerebellar Purkinje cells was suggested, but its role in these neurons remains unclear. In the present study, we confirmed the expression of Ano2 mRNA in Purkinje cells and performed electrophysiological recordings to examine the influence of ANO2-chloride channels on the excitability of Purkinje cells by comparing wildtype mice to mice lacking ANO2. Recordings were performed in acute cerebellar slices of adult mice, which provided the possibility to study the role of ANO2 within the cerebellar cortex. Purkinje cells were uncoupled from climbing fiber input to assess specifically the effect of ANO2 channels on Purkinje cell activity. We identified an attenuating effect of ANO2-mediated chloride currents on the instantaneous simple spike activity both during strong current injections and during current injections close to the simple spike threshold. Moreover, we report a reduction of inhibitory currents from GABAergic interneurons upon depolarization, lasting for several seconds. Together with the role of ANO2-chloride channels in inferior olivary neurons, our data extend the evidence for a role of chloride-dependent modulation in the olivo-cerebellar system that might be important for proper cerebellum-dependent motor coordination and learning.

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Identifying suitable reference genes for developing and injured mouse CNS tissues

Cited by 22 publications

References 33 publications

Is C-type natriuretic peptide regulated by a feedback loop? A study on systemic and local autoregulatory effect

Is C-type natriuretic peptide regulated by a feedback loop? A study on systemic and local autoregulatory effect

Reduction of HIP2 expression causes motor function impairment and increased vulnerability to dopaminergic degeneration in Parkinson’s disease models

Anoctamin 2-chloride channels reduce simple spike activity and mediate inhibition at elevated calcium concentration in cerebellar Purkinje cells

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