bioRxiv Science⌕ Search

Biology subjects

Vanzetta, I.

Publications and source records attributed to Vanzetta, I..

2 recordsLinked to original sources

Chamber Implant for Chronic Optical Recordings from the Cerebral Cortex of Marmosets

We describe a novel imaging chamber for chronic optical recordings from the marmoset cerebral cortex, together with the surgery needed for its implantation and that of an associated headpost. MRI data allow optimizing positioning on the skull. The chamber is implanted into a precisely matched craniotomy, improving mechanical stability. For maximal biocompatibility, chamber and headpost are made out of titanium. It consists of an outer cylinder into which screws adjustably an inner one, called "well", with a glass window on its bottom. This allows easy opening and closing whenever access to the cortex is needed, while preserving a sterile seal when closed. Moreover, simply rotating the well allows fine adjustment of the windows distance with respect to the underlying cortex. Together with using a curved - rather than flat - glass window, this allows achieving gentle but continuous contact with the underlying cortex, which helps to delay tissue regrowth, notably of a neomembrane that must otherwise be surgically removed. The chambers sealing system combines a silicone elastomer and O-ring, minimizing infection risk and CSF leakage. Data from longitudinal two-photon imaging using genetically encoded fluorescent calcium sensors reveal high optical quality over months, with excellent resolution of neurons and their activity. We also demonstrate the feasibility of two-photon imaging of genetically encoded voltage sensors in the marmoset cortex in-vivo. Finally, although the chamber provides a durable, adaptable solution for long-term imaging studies in marmosets already in its current version, we discuss a few straightforward modifications that could likely improve its performance even further.

neuroscience↗

Multi-modal refinement of the human heart atlas during the first gestational trimester

1.Forty first-trimester human hearts were studied to lay groundwork for further studies of principles underlying congenital heart defects. We first sampled 49,227 cardiac nuclei from three fetuses at 8.6, 9.0, and 10.7 post-conceptional weeks (pcw) for single-nucleus RNA sequencing, enabling distinction of six classes comprising 21 cell types. Improved resolution led to identification of novel cardiomyocytes and minority autonomic and lymphatic endothelial transcriptomes, among others. After integration with 5-7 pcw heart single-cell RNAseq, we identified a human cardiomyofibroblast progenitor preceding diversification of cardiomyocyte and stromal lineages. Analysis of six Visium sections from two additional hearts was aided by deconvolution, and key spatial markers validated on sectioned and whole hearts in two- and three-dimensional space and over time. Altogether, anatomical-positional features including innervation, conduction and subdomains of the atrioventricular septum translate latent molecular identity into specialized cardiac functions. This atlas adds unprecedented spatial and temporal resolution to the characterization of human-specific aspects of early heart formation.

developmental biology↗