Detection and Organ-Specific Ablation of Neuroendocrine Cells by Synaptophysin Locus-Based BAC Cassette in Transgenic Mice

Abstract

The role of cells of the diffuse neuroendocrine system in development and maintenance of individual organs and tissues remains poorly understood. Here we identify a regulatory region sufficient for accurate in vivo expression of synaptophysin (SYP), a common marker of neuroendocrine differentiation, and report generation of Tg(Syp-EGFPloxP-DTA)147Ayn (SypELDTA) mice suitable for flexible organ-specific ablation of neuroendocrine cells. These mice express EGFP and diphtheria toxin fragment A (DTA) in SYP positive cells before and after Cre-loxP mediated recombination, respectively. As a proof of principle, we have crossed SypELDTA mice with EIIA-Cre and PB-Cre4 mice. EIIA-Cre mice express Cre recombinase in a broad range of tissues, while PB-Cre4 mice specifically express Cre recombinase in the prostate epithelium. Double transgenic EIIA-Cre; SypELDTA embryos exhibited massive cell death in SYP positive cells. At the same time, PB-Cre4; SypELDTA mice showed a substantial decrease in the number of neuroendocrine cells and associated prostate hypotrophy. As no increase in cell death and/or Cre-loxP mediated recombination was observed in non-neuroendocrine epithelium cells, these results suggest that neuroendocrine cells play an important role in prostate development. High cell type specificity of Syp locus-based cassette and versatility of generated mouse model should assure applicability of these resources to studies of neuroendocrine cell functions in various tissues and organs.

A temporal and spatial map of axons in developing mouse prostate

“…Because CGRP+ and VAChT+ axons innervate neuroendocrine cells almost immediately after neuroendocrine cells are first detected in the developing prostate (E17), it is possible that nerve-neuroendocrine cell interactions facilitate prostate growth and development. This notion is supported by previous observations that prostatic neuroendocrine cell or nerve depletion reduces prostate size (Cheng et al 2013).…”

Membrane metalloendopeptidase suppresses prostate carcinogenesis by attenuating effects of gastrin-releasing peptide on stem/progenitor cells

“…Our study shows that accelerated progression of prostate carcinomas associated with MME deficiency is not associated with increase in the number of neuroendocrine cells. Neuroendocrine cells are important for prostate development 49 . The majority of human prostate adenocarcinomas and mouse models associated with Pten deficiency contain neuroendocrine cells 35,50 .…”

An immunohistochemical prostate cell identification key indicates that aging shifts procollagen 1A1 production from myofibroblasts to fibroblasts in dogs prone to prostate-related urinary dysfunction

“…Antibodies against cytokeratin 8/18 (KRT 8/18), cytokeratin 5 (KRT 5), and chromogranin A (CHGA) were used to identify luminal[21, 23] (KRT 8/18+, KRT 5−, CHGA−), intermediate[21, 23] (KRT 8/18+, KRT 5+, CHGA−), and basal[21, 23] (KRT 8/18−, KRT5−, CHGA−) epithelial cells. Neuroendocrine cells[21, 24, 25] (KRT 8/18−, KRT 5−, CHGA+) were also identified with this stain combination. Antibodies against smooth muscle actin (ACTA2), protein tyrosine phosphatase receptor type C (PTPRC, also known as CD45),and vimentin (VIM) were used to identify fibrocytes[21, 26–30] (ACTA2+, PTPRC+, VIM+), and other hematolymphoid cells (PTPRC+, not a fibrocyte).…”

“…Antibodies against cytokeratin 8/18 (KRT 8/18), cytokeratin 5 (KRT 5), and chromogranin A (CHGA) are used to identify luminal[21, 23] (KRT 8/18+, KRT 5−, CHGA−, Figure 1B), intermediate[21, 23] (KRT 8/18+, KRT 5+, CHGA−, Figure 1C), and basal[21, 23] (KRT 8/18−, KRT 5−, CHGA−, Figure 1D) epithelial cells. Neuroendocrine cells[21, 24, 25] (KRT 8/18−, KRT 5−, CHGA+, Figure 1E) are also identified. Epithelial cells in general and neuroendocrine cells in particular are not detected in the capsule (Figure 1F) but are present in the peripheral region (Figure 1G), the periurethral region (Figure 1H) and the prostatic urethra (Figure 1I).…”