DOI: 10.5061/dryad.z34tmpgr7
title: Microscopy images from: A physiological and histological atlas of reproduction in the North American deer mouse (Peromyscus maniculatus)

description: The North American deer mouse (Peromyscus maniculatus) exhibits extensive
 diversity in morphology, physiology, and life history across its broad
 range. These traits have propelled the deer mouse to model system status
 across several fields within the biological sciences. Nonetheless, we
 still lack basic knowledge about some important aspects of this species’
 biology. For example, limited information about the deer mouse’s
 reproductive physiology remains a significant barrier to developing
 genetic tools for the species and for advancing our current understanding
 of how this species has been so evolutionarily successful. In this
 manuscript, we aim to fill this knowledge gap by (1) characterizing body
 temperature profiles across reproductive stages and (2) generating a
 detailed histological atlas of placental development. We show that body
 temperature can be used to diagnose copulation and pregnancy in deer mice,
 however body temperature cannot be used to predict fertility (likelihood
 to breed) prior to pairing individuals. Our placental atlas represents the
 first day-by-day developmental timeline of the placenta in a Peromyscus
 species, and from it we are able to describe unique organization and
 behaviors of trophoblast cells, especially at the maternal-fetal interface
 in the deer mouse. Together, these descriptive datasets provide
 substantial new comparative data on reproductive physiology in Cricetids,
 and they provide a foundation for further functional work in this
 important model species.
description: We employed three separate double-labeling protocols to describe
 specific features of placentation and development. To examine vascular
 development and remodeling in the implantation site, we used two sets of
 sections to label two markers of vascular remodeling and endothelial cell
 organization: the basement membrane protein laminin and the tight-junction
 protein ZO-1 (26,27). To examine interactions between uterine natural
 killer cells (uNK cells) and trophoblasts in the implantation site, we
 also labeled one set of sections for the uNK cell marker
 perforin. For all immunohistochemistry protocols,
 sections were fixed with ice-cold 4% paraformaldehyde, permeabilized using
 methanol, and blocked using 10% normal goat serum in PBS (Vector
 Laboratories S-1000). Sections were incubated overnight with one of three
 primary antibodies: rabbit anti-laminin (1:1000, Thermo Scientific
 PA116730), or rabbit anti-ZO-1 (1:500, Invitrogen 617300), or rabbit
 anti-perforin (1:400, Amsbio TP251). After PBS washes the next day,
 sections were incubated for 1 hour in Alexafluor 568 (1:300,
 goat-anti-rabbit, Thermo Scientific A32731TR). All sections were then
 labeled for cytokeratin (a trophoblast cell marker) using a 2 hour
 incubation with pan-cytokeratin antibody conjugated to FITC (1:300,
 Sigma-Aldrich F3418, lot: 088M4797V) followed by DAPI to visualize nuclei.
 Immunostained sections were cover slipped with Fluoromount-G, dried
 overnight at room temperature, and stored at 4°C until imaging on an
 Evident Scientific APX100. Three sections per implantation site were
 imaged and quantified for each protocol at one of three objectives (4x,
 10x, or 40x). 
description: # Microscopy images from: A physiological and histological atlas of
 reproduction in the North American deer mouse (Peromyscus maniculatus)
 [https://doi.org/10.5061/dryad.z34tmpgr7](https://doi.org/10.5061/dryad.z34tmpgr7) ## Description of the data and file structure These data are raw images of placentas from North American deer mouse,* Peromyscus maniculatus*, implantation sites across gestation. Images were generated using 10 um cryosections of whole implantation sites that were then labeled using immunohistochemistry. Images were captured at varying magnifications with or without stitching on an APX100 microscope. Images contain scale bars. ### Files and variables #### File: FileMetaData.txt **Description:**  ##### Variables * FileName: Full image file name * ProcessID_Prefix: Process prefix * LabelingProtocol: Immunohistochemistry labeling regime. Laminin refers to a double-label protocol wherein mCherry (red) indicates laminin, gfp (green) indicates cytokeratin, and DAPI (blue) is used to label nuclei. Perforin refers to a double-label protocol wherein mCherry (red) indicates perforin, gfp (green) indicates cytokeratin, and DAPI (blue) is used to label nuclei. ZO1 refers to a double-label protocol wherein mCherry (red) indicates ZO1, gfp (green) indicates cytokeratin, and DAPI (blue) is used to label nuclei. * GestationalDay: The gestational day (embryonic day) that samples were collected on. * DamID: The unique Dam ID that samples were collected from. * SlideID: The unique slide and section ID that was imaged for the photo. #### File: Process\_XXXX\_merged\_CH(mCherry\_\_DAPI\_\_GFP)\_CH.jpg **Description:** Image file where the Process number (i.e., Process_XXXX) is linked to metadata provided in FileMetaData.txt.  ## Code/software Images can be viewed with any computer capable of opening .jpeg.  ## Access information Other publicly accessible locations of the data: * Data derived from images and scripts associated with additional analyses can be found at [https://github.com/mjhemmerlein/Peromyscus-Gestational-Atlas](https://github.com/mjhemmerlein/Peromyscus-Gestational-Atlas) .