3D single cell scale anatomical map of sex-dependent variability of the rat intrinsic cardiac nervous system

Leung, Clara; Robbins, Shaina; Moss, Alison; Heal, Maci; Osanlouy, Mahyar; Christie, Richard; Farahani, Navid; Monteith, Corey; Chen, Jin; Hunter, Peter; Tappan, Susan; Vadigepalli, Rajanikanth; Cheng, Zixi; Schwaber, James S.

doi:10.1016/j.isci.2021.102795

Cited by 7 publications

(6 citation statements)

References 48 publications

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“…Having these types of information on a scaffold contributes to a more thorough understanding of the organ innervation, as well as making the data more accessible and replicable for other scientists. This is a joint effort from multiple researchers to not only produce the data, but also use the data to navigate 2D connectivity maps and build 3D organ scaffolds that can provide a comprehensive brain‐organ atlas and “a common coordinate framework for cross‐species comparisons” (Achanta et al., 2020; Leung et al., 2021; Osanlouy et al., 2021).…”

Section: Discussionmentioning

confidence: 99%

Organization and morphology of calcitonin gene‐related peptide‐immunoreactive axons in the whole mouse stomach

Ma,

Nguyen,

Madas

et al. 2023

J of Comparative Neurology

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Nociceptive afferent axons innervate the stomach and send signals to the brain and spinal cord. Peripheral nociceptive afferents can be detected with a variety of markers (e.g., substance P [SP] and calcitonin gene‐related peptide [CGRP]). We recently examined the topographical organization and morphology of SP‐immunoreactive (SP‐IR) axons in the whole mouse stomach muscular layer. However, the distribution and morphological structure of CGRP‐IR axons remain unclear. We used immunohistochemistry labeling and applied a combination of imaging techniques, including confocal and Zeiss Imager M2 microscopy, Neurolucida 360 tracing, and integration of axon tracing data into a 3D stomach scaffold to characterize CGRP‐IR axons and terminals in the whole mouse stomach muscular layers. We found that: (1) CGRP‐IR axons formed extensive terminal networks in both ventral and dorsal stomachs. (2) CGRP‐IR axons densely innervated the blood vessels. (3) CGRP‐IR axons ran in parallel with the longitudinal and circular muscles. Some axons ran at angles through the muscular layers. (4) They also formed varicose terminal contacts with individual myenteric ganglion neurons. (5) CGRP‐IR occurred in DiI‐labeled gastric‐projecting neurons in the dorsal root and vagal nodose ganglia, indicating CGRP‐IR axons were visceral afferent axons. (6) CGRP‐IR axons did not colocalize with tyrosine hydroxylase or vesicular acetylcholine transporter axons in the stomach, indicating CGRP‐IR axons were not visceral efferent axons. (7) CGRP‐IR axons were traced and integrated into a 3D stomach scaffold. For the first time, we provided a topographical distribution map of CGRP‐IR axon innervation of the whole stomach muscular layers at the cellular/axonal/varicosity scale.

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Section: Discussionmentioning

confidence: 99%

Organization and morphology of calcitonin gene‐related peptide‐immunoreactive axons in the whole mouse stomach

Ma,

Nguyen,

Madas

et al. 2023

J of Comparative Neurology

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“…Intriguingly, a recent study (Leung et al, 2021) explored sex differences in the rat intrinsic cardiac nervous system, which includes the network of the intracardiac ganglia and interconnecting neurons that receive inputs from both local afferent and extrinsic autonomic (vagal and sympathetic) nerves (Achanta et al, 2020). The authors found that female rat hearts had fewer neurons and lower packing density than males, which may explain some of the sex differences observed at a functional level (Leung et al, 2021). Given such findings and the current results, we believe that rat models offer an important opportunity to systematically investigate the neuro-hormonal basis of the sex difference in the relationship between vmHRV and cardiac chronotropy.…”

Section: Implications For Translational Researchmentioning

confidence: 95%

“…Further, studies in rats have shown that i) vagal nerve stimulation leads to greater cardiac effects in female than male rats, presumably due to a higher level of ACh release following nerve activation (Du et al, 1994), that ii) the enzymatic breakdown of ACh occurs slower in newborn female than male rats (Loy and Sheldon, 1987), and that iii) the synthesis and clearance of neurotransmitters, including ACh, in both the heart and vasculature are regulated by sex hormones (Dart et al, 2002). Intriguingly, a recent study (Leung et al, 2021) explored sex differences in the rat intrinsic cardiac nervous system, which includes the network of the intracardiac ganglia and interconnecting neurons that receive inputs from both local afferent and extrinsic autonomic (vagal and sympathetic) nerves (Achanta et al, 2020). The authors found that female rat hearts had fewer neurons and lower packing density than males, which may explain some of the sex differences observed at a functional level (Leung et al, 2021).…”

Section: Implications For Translational Researchmentioning

confidence: 99%

Sex differences in heart rate and heart rate variability in rats: Implications for translational research

et al. 2023

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The present study aimed to investigate sex differences in measures of cardiac chronotropy and heart rate variability (HRV) in 132 young adult wild-type Groningen rats (n = 45 females). Electrocardiographic signals were recorded for 48 h in freely moving rats to quantify heart rate (HR) and inter-beat interval (IBI) as measures of cardiac chronotropy, and time- and frequency-domain HRV parameters as physiological readouts of cardiac vagal modulation. Females showed greater vagally-mediated HRV despite having higher HR and shorter IBI than males during undisturbed conditions. Such differences were evident i) at any given level of HRV, and ii) both during the 12-h light/inactive and 12-h dark/active phase of the daily cycle. These findings replicate the paradoxical cardiac chronotropic control reported by human meta-analytic findings, since one would expect greater vagally-mediated HRV to be associated with lower HR and longer IBI. Lastly, the association between some HRV measures and HR was stronger in female than male rats. Overall, the current study in young adult rats provides data illustrating a sex-dependent association between vagally-mediated HRV and indexes of cardiac chronotropy. The current results i) are in line with human findings, ii) suggest to always consider biological sex in the analysis and interpretation of HRV data in rats, and iii) warrant the use of rats for investigating the neuro-hormonal basis and temporal evolution of the impact of sex on the association between vagally-mediated HRV and cardiac chronotropy, which could inform the human condition.

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“…Such studies have investigated neurocardiac interactions using an aspect ratio where the cardiomyocytes outnumber neuronal cells, demonstrating a functional and viable co-culture [7,11,12]. Although the distribution and numbers present in cardiac tissue have not been extensively studied, intrinsic cardiac neurons are distributed in a similar pattern and form a limited number of clusters within the cardiac tissue [13].…”

Section: Introductionmentioning

confidence: 99%

“…By incorporating multiple arrowheads into the design, the scaffold design was intended to replicate the geometry and morphology of myocardial tissue [6], and adding an even layer with neuronal cells potentially mimicked the organization of a CANS model [13]. It was expected that this design would promote effective networking between neurons and cardiomyocytes in comparison with a 2D platform [7].…”

Section: Introductionmentioning

confidence: 99%

A Semi-Three-Dimensional Bioprinted Neurocardiac System for Tissue Engineering of a Cardiac Autonomic Nervous System Model

Hernandez,

Ramirez,

Salazar

et al. 2023

Bioengineering

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In this study, we designed a tissue-engineered neurocardiac model to help us examine the role of neuronal regulation and confirm the importance of neural innervation techniques for the regeneration of cardiac tissue. A three-dimensional (3D) bioprinted neurocardiac scaffold composed of a mixture of gelatin–alginate and alginate–genipin–fibrin hydrogels was developed with a 2:1 ratio of AC16 cardiomyocytes (CMs) and retinoic acid-differentiated SH-SY5Y neuronal cells (NCs) respectively. A unique semi-3D bioprinting approach was adopted, where the CMs were mixed in the cardiac bioink and printed using an anisotropic accordion design to mimic the physiological tissue architecture in vivo. The voids in this 3D structure were methodically filled in using a NC–gel mixture and crosslinked. Confocal fluorescent imaging using microtubule-associated protein 2 (MAP-2) and anticholine acetyltransferase (CHAT) antibodies for labeling the NCs and the MyoD1 antibody for the CMs revealed functional coupling between the two cell types in the final crosslinked structure. These data confirmed the development of a relevant neurocardiac model that could be used to study neurocardiac modulation under physiological and pathological conditions.

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3D single cell scale anatomical map of sex-dependent variability of the rat intrinsic cardiac nervous system

Cited by 7 publications

References 48 publications

Organization and morphology of calcitonin gene‐related peptide‐immunoreactive axons in the whole mouse stomach

Organization and morphology of calcitonin gene‐related peptide‐immunoreactive axons in the whole mouse stomach

Sex differences in heart rate and heart rate variability in rats: Implications for translational research

A Semi-Three-Dimensional Bioprinted Neurocardiac System for Tissue Engineering of a Cardiac Autonomic Nervous System Model

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