小鼠结肠中 CGRP 免疫反应性肌间神经元的定量,The Journal of Comparative Neurology

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小鼠结肠中 CGRP 免疫反应性肌间神经元的定量
The Journal of Comparative Neurology ( IF 2.3 ) Pub Date : 2022-08-31 , DOI: 10.1002/cne.25403
Timothy J. Hibberd ₁ , Wai Ping Yew ₁ , Kelsi N. Dodds ₁ , Zili Xie ₂ , Lee Travis ₁ , Simon J. Brookes ₁ , Marcello Costa ₁ , Hongzhen Hu ₂ , Nick J. Spencer ₁

Affiliation

生物系统的定量数据为理解病理学、实验扰动和计算建模提供了有价值的基线信息。在小鼠结肠中，降钙素基因相关肽 (CGRP) 由具有多轴突 (Dogiel II 型) 形态的肌间神经元表达，这是内在初级传入神经元 (IPAN) 的特征。其他物种和肠道区域的类似神经元占肠间神经元的 5-35%。我们旨在量化 CGRP 免疫阳性 (CGRP+) 肌间神经元的比例。秋水仙碱处理的近端、中端和远端结肠的整体制剂被标记为 HuC/D、CGRP、一氧化氮合酶 (NOS) 和外周蛋白 (Per)。泛神经元标记 (Hu+/Per+) 共同标记了 94% 的神经元。Hu+/Per- 神经元约占 6%，但 Hu-/Per+ 细胞很少见。因此，n = 7)。CGRP+ 细胞体明显大于所有 Hu+ 神经元的平均值（329 ± 13 对 261 ± 12 μm ²，p < .0001）。CGRP+ 神经元占肠间神经元的 19% ± 3%，没有明显的区域差异。NOS+ 神经元占肠间神经元的 42% ± 2%，与结肠中段和远端相比，近端结肠的比例较低（分别为 38% ± 2%、44% ± 2% 和 44% ± 3%）。外周蛋白免疫标记揭示了一些肌间神经元中的细胞体和轴突形态。是否所有 CGRP+ 神经元都是多轴突不能使用外周蛋白免疫标记来解决。然而，在最初基于外周蛋白免疫反应性鉴定的 118 个推定的多轴突神经元中，全都是 CGRP+ ( n = 4)。总之，小鼠结肠中的 CGRP+ 肌间神经元被全面量化，发生在推定的 IPAN 标记的预期范围内。Dogiel II/IPAN 形态特征的所有 Per+ 多轴突神经元均为 CGRP+。

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Quantification of CGRP-immunoreactive myenteric neurons in mouse colon

Quantitative data of biological systems provide valuable baseline information for understanding pathology, experimental perturbations, and computational modeling. In mouse colon, calcitonin gene-related peptide (CGRP) is expressed by myenteric neurons with multiaxonal (Dogiel type II) morphology, characteristic of intrinsic primary afferent neurons (IPANs). Analogous neurons in other species and gut regions represent 5–35% of myenteric neurons. We aimed to quantify proportions of CGRP-immunopositive (CGRP+) myenteric neurons. Colchicine-treated wholemount preparations of proximal, mid, and distal colon were labeled for HuC/D, CGRP, nitric oxide synthase (NOS), and peripherin (Per). The pan-neuronal markers (Hu+/Per+) co-labeled 94% of neurons. Hu+/Per– neurons comprised ∼6%, but Hu-/Per+ cells were rare. Thus, quantification was based on Hu+ myenteric neurons (8576 total; 1225 ± 239 per animal, n = 7). CGRP+ cell bodies were significantly larger than the average of all Hu+ neurons (329 ± 13 vs. 261 ± 12 μm², p < .0001). CGRP+ neurons comprised 19% ± 3% of myenteric neurons without significant regional variation. NOS+ neurons comprised 42% ± 2% of myenteric neurons overall, representing a lower proportion in proximal colon, compared to mid and distal colon (38% ± 2%, 44% ± 2%, and 44% ± 3%, respectively). Peripherin immunolabeling revealed cell body and axonal morphology in some myenteric neurons. Whether all CGRP+ neurons were multiaxonal could not be addressed using peripherin immunolabeling. However, of 118 putatively multiaxonal neurons first identified based on peripherin immunoreactivity, all were CGRP+ (n = 4). In conclusion, CGRP+ myenteric neurons in mouse colon were comprehensively quantified, occurring within a range expected of a putative IPAN marker. All Per+ multiaxonal neurons, characteristic of Dogiel type II/IPAN morphology, were CGRP+.

更新日期：2022-08-31

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