Wegner Kyle A, Cadena Mark T, Trevena Ryan, Turco Anne E, Gottschalk Adam, Halberg Richard B, Guo Jinjin, McMahon Jill A, McMahon Andrew P, Vezina Chad M
George M. O'Brien Benign Urology Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
Molecular and Environmental Toxicology Center, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.
PLoS One. 2017 Nov 16;12(11):e0188413. doi: 10.1371/journal.pone.0188413. eCollection 2017.
Though many methods can be used to identify cell types contained in complex tissues, most require cell disaggregation and destroy information about where cells reside in relation to their microenvironment. Here, we describe a polytomous key for cell type identification in intact sections of adult mouse prostate and prostatic urethra. The key is organized as a decision tree and initiates with one round of immunostaining for nerve, epithelial, fibromuscular/hematolymphoid, or vascular associated cells. Cell identities are recursively eliminated by subsequent staining events until the remaining pool of potential cell types can be distinguished by direct comparison to other cells. We validated our identification key using wild type adult mouse prostate and urethra tissue sections and it currently resolves sixteen distinct cell populations which include three nerve fiber types as well as four epithelial, five fibromuscular/hematolymphoid, one nerve-associated, and three vascular-associated cell types. We demonstrate two uses of this novel identification methodology. We first used the identification key to characterize prostate stromal cell type changes in response to constitutive phosphatidylinositide-3-kinase activation in prostate epithelium. We then used the key to map cell lineages in a new reporter mouse strain driven by Wnt10aem1(cre/ERT2)Amc. The identification key facilitates rigorous and reproducible cell identification in prostate tissue sections and can be expanded to resolve additional cell types as new antibodies and other resources become available.
尽管有许多方法可用于识别复杂组织中包含的细胞类型,但大多数方法都需要细胞解离,从而破坏有关细胞在其微环境中所处位置的信息。在此,我们描述了一种用于成年小鼠前列腺和前列腺尿道完整切片中细胞类型识别的多歧检索表。该检索表被组织成决策树形式,首先对神经、上皮、纤维肌/血液淋巴或血管相关细胞进行一轮免疫染色。通过后续的染色事件递归地排除细胞身份,直到通过与其他细胞直接比较来区分剩余的潜在细胞类型池。我们使用野生型成年小鼠前列腺和尿道组织切片验证了我们的识别检索表,目前它能分辨出16种不同的细胞群体,包括三种神经纤维类型以及四种上皮、五种纤维肌/血液淋巴、一种神经相关和三种血管相关细胞类型。我们展示了这种新型识别方法的两种用途。我们首先使用识别检索表来表征前列腺基质细胞类型对前列腺上皮中组成型磷脂酰肌醇-3-激酶激活的反应变化。然后我们使用该检索表来绘制由Wnt10aem1(cre/ERT2)Amc驱动的新报告基因小鼠品系中的细胞谱系。该识别检索表有助于在前列腺组织切片中进行严格且可重复的细胞识别,并且随着新抗体和其他资源的可用,可扩展以分辨更多细胞类型。