John D. Dingell VA Medical Center, Detroit, MI 48201, USA.
Department of Otolaryngology-Head and Neck Surgery, Wayne State University School of Medicine, Detroit, MI 48201, USA.
Int J Mol Sci. 2021 Feb 4;22(4):1550. doi: 10.3390/ijms22041550.
Our previous study reported neural stem cells (NSCs) in the auditory cortex (AC) of postnatal day 3 (P3) mice in vitro. It is unclear whether AC-NSCs exist in vivo. This study aims to determine the presence and changes of AC-NSCs during postnatal development and maturation both in vitro and in vivo. P3, postnatal day 14 (P14), 2-month-old (2M), and 4-month-old (4M) mouse brain tissues were fixed and cryosectioned for NSC marker immunostaining. In vitro, P3, P14, and 2M AC tissues were dissected and cultured in suspension to study NSCs. NSC proliferation was examined by EdU incorporation and cell doubling time assays in vitro. The results show that Nestin and Sox2 double expressing NSCs were observed in the AC area from P3 to 4M in vivo, in which the number of NSCs remarkably reduced with age. In vitro, the neurosphere forming capability, cell proliferation, and percentage of Nestin and Sox2 double expressing NSCs significantly diminished with age. These results suggest that AC-NSCs exist in the mouse AC area both in vitro and in vivo, and the percentage of AC-NSCs decreases during postnatal development and maturation. The results may provide important cues for the future research of the central auditory system.
我们之前的研究报道了体外培养的新生第 3 天(P3)小鼠听觉皮层(AC)中的神经干细胞(NSC)。目前尚不清楚体内是否存在 AC-NSC。本研究旨在确定 AC-NSC 在体外和体内的出生后发育和成熟过程中的存在和变化。固定并冷冻切片 P3、出生后第 14 天(P14)、2 个月大(2M)和 4 个月大(4M)的小鼠脑组织,并用 NSC 标志物免疫染色。体外分离 P3、P14 和 2M 的 AC 组织并在悬浮液中培养以研究 NSC。通过 EdU 掺入和细胞倍增时间测定来检测 NSC 的增殖。结果表明,在体内 P3 到 4M 的 AC 区域观察到 Nestin 和 Sox2 双表达 NSC,其中 NSC 的数量随年龄显著减少。体外,神经球形成能力、细胞增殖以及 Nestin 和 Sox2 双表达 NSC 的比例随年龄显著降低。这些结果表明,AC-NSC 存在于体外和体内的小鼠 AC 区域,并且在出生后发育和成熟过程中,AC-NSC 的比例下降。这些结果可能为未来的中枢听觉系统研究提供重要线索。