Immunostaining for NeuN Does Not Show all Mature and Healthy Neurons in the Human and Pig Brain: Focus on the Hippocampus

Luijerink, Lauren; Waters, Karen A.; Machaalani, Rita

doi:10.1097/pai.0000000000000925

Cited by 9 publications

(4 citation statements)

References 53 publications

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“…In addition, SH-SY5Y cells were examined using neuronal marker labeling. Early neuronal markers include β-III tubulin, a microtubule element, while late neuronal markers include NeuN, a neuron-specific nuclear protein [56,57]. NeuN expression of SH-SY5Y cells in our study showed increase in 2 weeks.…”

Section: Discussionsupporting

confidence: 45%

Fabrication and characterization of pHEMA hydrogel conduit containing GelMA-HaMA IPN for peripheral nerve regeneration

Sahin,

Son,

Hasirci

2024

Explor BioMat-X

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Aim: Small defects after any injury to the periperal nerves results in self-regeneration. However, for larger defects, suturing or grafting are necessary, which may have limitations. Thus, research on nerve guidence conduits is needed without drawbacks. The aim of the study was to develop hydrogel-based conduits containing interpenetrating network (IPN). Methods: Methacrylated gelatin (GelMA)-methacrylated hyaluronic acid (HaMA) IPN was filled the poly(2-hydroxyethylmethacrylate) (pHEMA) the outer conduit. Schwann cells (SCs) were used on the pHEMA and the distal end of the tube was injected with netrin-1 to support model SH-SY5Y cells. Results: 1H-nuclear magnetic resonance (1H-NMR) showed that methacrylation degrees were 94% ± 2% for GelMA and 60% ± 7% for HaMA. The fraction of HaMA increased the degradation rate; pure HaMA degraded in 3 weeks, while pure GelMA in more than 5 weeks. An increase in the fraction of 2-hydroxyethylmethacrylate (HEMA) from 20% to 56% decreased the porosity and the pore size, significantly. SH-SY5Y cells migrated along the conduit in the presence of netrin-1. NeuN expression was increased in 2 weeks indicating neuronal activity. Conclusions: SH-SY5Y cells produced neurites in the IPN. pHEMA conduit including GelMA-HaMA IPN is a good candidate for peripheral nerve regeneration applications. As future studies, the conduit will be tested in vivo for nerve regeneration.

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Section: Discussionsupporting

confidence: 45%

Fabrication and characterization of pHEMA hydrogel conduit containing GelMA-HaMA IPN for peripheral nerve regeneration

Sahin,

Son,

Hasirci

2024

Explor BioMat-X

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“…In our study, approximately 60% of STN neurons are NeuN-ir, which suggests that, in the STN, NeuN labels specific neuronal subpopulation. However, a recent study indicated that in the formalin-fixed, paraffin-embedded (FFPE) human tissue, some NeuN neurons are inadequately stained for quantification purposes ( Luijerink et al, 2021 ). Our samples were formalin-fixed but were not paraffin-embedded.…”

Section: Discussionmentioning

confidence: 99%

The Stereological Analysis and Spatial Distribution of Neurons in the Human Subthalamic Nucleus

et al. 2021

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The subthalamic nucleus (STN) is a small, ovoid structure, and an important site of deep brain stimulation (DBS) for the treatment of Parkinson’s disease. Although the STN is a clinically important structure, there are many unresolved issues with regard to it. These issues are especially related to the anatomical subdivision, neuronal phenotype, neuronal composition, and spatial distribution. In this study, we have examined the expression pattern of 8 neuronal markers [nNOS, NeuN, parvalbumin (PV), calbindin (CB), calretinin (CR), FOXP2, NKX2.1, and PAX6] in the adult human STN. All of the examined markers, except CB, were present in the STN. To determine the neuronal density, we have performed stereological analysis on Nissl-stained and immunohistochemical slides of positive markers. The stereology data were also used to develop a three-dimensional map of the spatial distribution of neurons within the STN. The nNOS population exhibited the largest neuronal density. The estimated total number of nNOS STN neurons is 281,308 ± 38,967 (± 13.85%). The STN neuronal subpopulations can be divided into two groups: one with a neuronal density of approximately 3,300 neurons/mm3 and the other with a neuronal density of approximately 2,200 neurons/mm3. The largest density of STN neurons was observed along the ventromedial border of the STN and the density gradually decreased toward the dorsolateral border. In this study, we have demonstrated the presence of 7 neuronal markers in the STN, three of which were not previously described in the human STN. The human STN is a collection of diverse, intermixed neuronal subpopulations, and our data, as far as the cytoarchitectonics is concerned, did not support the tripartite STN subdivision.

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“…Nissl body assessment is frequently used in the evaluation of neuronal viability, as the number of Nissl bodies is greatly reduced in damaged neurons [ 29 ]. Furthermore, NeuN is a key neuron-specific nuclear protein that is used as a marker of mature neurons, and the absence of its expression implies attenuated neuronal viability [ 30 , 31 ]. Herein, NTRK1 knockdown resulted in a decrease in the number of Nissl bodies and NeuN-positive neurons in mouse hippocampal tissues.…”

Section: Discussionmentioning

confidence: 99%

NTRK1 knockdown induces mouse cognitive impairment and hippocampal neuronal damage through mitophagy suppression via inactivating the AMPK/ULK1/FUNDC1 pathway

Yang,

Wu,

et al. 2023

Cell Death Discov.

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Hippocampal neuronal damage may induce cognitive impairment. Neurotrophic tyrosine kinase receptor 1 (NTRK1) reportedly regulates neuronal damage, although the underlying mechanism remains unclear. The present study aimed to investigate the role of NTRK1 in mouse hippocampal neuronal damage and the specific mechanism. A mouse NTRK1-knockdown model was established and subjected to pre-treatment with BAY-3827, followed by a behavioral test, Nissl staining, and NeuN immunofluorescence (IF) staining to evaluate the cognitive impairment and hippocampal neuronal damage. Next, an in vitro analysis was conducted using the CCK-8 assay, TUNEL assay, NeuN IF staining, DCFH-DA staining, JC-1 staining, ATP content test, mRFP-eGFP-LC3 assay, and LC3-II IF staining to elucidate the effect of NTRK1 on mouse hippocampal neuronal activity, apoptosis, damage, mitochondrial function, and autophagy. Subsequently, rescue experiments were performed by subjecting the NTRK1-knockdown neurons to pre-treatment with O304 and Rapamycin. The AMPK/ULK1/FUNDC1 pathway activity and mitophagy were detected using western blotting (WB) analysis. Resultantly, in vivo analysis revealed that NTRK1 knockdown induced mouse cognitive impairment and hippocampal tissue damage, in addition to inactivating the AMPK/ULK1/FUNDC1 pathway activity and mitophagy in the hippocampal tissues of mice. The treatment with BAY-3827 exacerbated the mouse depressive-like behavior induced by NTRK1 knockdown. The results of in vitro analysis indicated that NTRK1 knockdown attenuated viability, NeuN expression, ATP production, mitochondrial membrane potential, and mitophagy, while enhancing apoptosis and ROS production in mouse hippocampal neurons. Conversely, pre-treatment with O304 and rapamycin abrogated the suppression of mitophagy and the promotion of neuronal damage induced upon NTRK1 silencing. Conclusively, NTRK1 knockdown induces mouse hippocampal neuronal damage through the suppression of mitophagy via inactivating the AMPK/ULK1/FUNDC1 pathway. This finding would provide insight leading to the development of novel strategies for the treatment of cognitive impairment induced due to hippocampal neuronal damage.

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Immunostaining for NeuN Does Not Show all Mature and Healthy Neurons in the Human and Pig Brain: Focus on the Hippocampus

Cited by 9 publications

References 53 publications

Fabrication and characterization of pHEMA hydrogel conduit containing GelMA-HaMA IPN for peripheral nerve regeneration

Fabrication and characterization of pHEMA hydrogel conduit containing GelMA-HaMA IPN for peripheral nerve regeneration

The Stereological Analysis and Spatial Distribution of Neurons in the Human Subthalamic Nucleus

NTRK1 knockdown induces mouse cognitive impairment and hippocampal neuronal damage through mitophagy suppression via inactivating the AMPK/ULK1/FUNDC1 pathway

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