Distribution and neurochemical characteristic of the cardiac nerve structures in the heart of chinchilla (Chinchilla laniger Molina)
Abstract
Introduction. The heart innervation is made up of plexo-ganglionic formation containing sympathetic, parasympathetic, and sensory components. We examined the distribution and neurochemical coding of the ganglia and nerve fibers in the chinchilla’s heart. Material and methods. The heart sections of 10 male and 10 female adult chinchillas were processed in accordance with the thiocholine method for acetylcholine esterase (AChE), and the SPG method for detecting the presence of adrenergic fibers was applied. The routine technique of immunohistochemical (IHC) staining with primary antibodies directed against ChAT, VAChT, DbH, TH, CART, NPY, VIP, GAL and SOM was used. The secondary antibodies were conjugated with Alexa Fluor 488 and Alexa Fluor 555 fluorophores. Results. The epicardium contained ganglia and nerve fibers, the myocardium had a few ganglion neurocytes and nerve fibers, and the endocardium contained only nerve fibers. In the epicardium, AChE-positive fibers were more prevalent than SPG-positive fibers. All the ganglion cells were immunopositive for ChAT and VAChT. Some cells also had a positive reaction to DbH and TH. Fibers containing cholinergic and adrenergic markers were numerous, while many of them were ChAT/DbH- and VAChT/TH-positive. CART/NPY and CART/VIP, as well as CART and GAL, were observed to be colocalized in ganglion neurocytes, as well as in individual cells. The nerve fibers were found to contain all the neurotransmitters we tested for, as well as the following co-occurrences: ChAT/DbH, VAChT/TH, CART/NPY, CART/VIP, CART/GAL, and CART/SOM.
Conclusions. Our analysis of the neurochemical profile of the nerve structures in chinchilla’s heart showed that, despite interspecies differences, the general pattern of the distribution of autonomic nervous system structures is similar to that of other mammals’ species, including humans.
Keywords: chinchillacardiac innervationhistochemistryimmunohistochemistry
References
- Forsgren S, Moravec M, Moravec J. Catecholamine-synthesizing enzymes and neuropeptides in rat heart epicardial ganglia; an immunohistochemical study. Histochem J. 1990; 22(12): 667–676.
- Kuder T, Tekieli A. Gross and light microscopic studies on the mouse cardiac parasympathetic nervous system. Zool. Pol. 2000; 45: 157-167. Not indexed in Pubmed.
- Rysevaite K, Saburkina I, Pauziene N, et al. Morphologic pattern of the intrinsic ganglionated nerve plexus in mouse heart. Heart Rhythm. 2011; 8(3): 448–454.
- Hasan W. Autonomic cardiac innervation: development and adult plasticity. Organogenesis. 2013; 9(3): 176–193.
- Kuder T, Nowak E. Autonomic cardiac nerves: literature review. Folia Morphol (Warsz). 2015; 74(1): 1–8.
- Achanta S, Gorky J, Leung C, et al. A Comprehensive Integrated Anatomical and Molecular Atlas of Rat Intrinsic Cardiac Nervous System. iScience. 2020; 23(6): 101140.
- Saburkina I, Rysevaite K, Pauziene N, et al. Epicardial neural ganglionated plexus of ovine heart: anatomic basis for experimental cardiac electrophysiology and nerve protective cardiac surgery. Heart Rhythm. 2010; 7(7): 942–950.
- Day SM, Gu J, Polak JM, et al. Somatostatin in the human heart and comparison with guinea pig and rat heart. Br Heart J. 1985; 53(2): 153–157.
- Marron K, Wharton J, Sheppard MN, et al. Human endocardial innervation and its relationship to the endothelium: an immunohistochemical, histochemical, and quantitative study. Cardiovasc Res. 1994; 28(10): 1490–1499.
- Edwards FR, Hirst GD, Klemm MF, et al. Different types of ganglion cell in the cardiac plexus of guinea-pigs. J Physiol. 1995; 486 ( Pt 2): 453–471.
- Singh S, Johnson P, Lee R, et al. Topography of cardiac ganglia in the adult human heart. J Thoracic Cardiovasc Surgery. 1996; 112(4): 943–953.
- Armour JA, Murphy DA, Yuan BX, et al. Gross and microscopic anatomy of the human cardiac nervous system. Anat Rec. 1997; 247: 289-298. , doi: 10.1002/(SICI)1097-0185(199702)247:2<289::AID-AR15>3.0.CO-2-N.
- Pauza D, Skripkiene G, Skripka V, et al. Morphological study of neurons in the nerve plexus on heart base of rats and guinea pigs. J Auton Nerve Syst. 1997; 62(1-2): 1–12.
- Pauza D, Skripka V, Pauziene N, et al. Morphology, distribution, and variability of the epicardiac neural ganglionated subplexuses in the human heart. The Anatomical Record. 2000; 259(4): 353–382, doi: 10.1002/1097-0185(20000801)259:4<353::aid-ar10>3.0.co;2-r.
- Batulevicius D, Pauziene N, Pauza D. Topographic morphology and age-related analysis of the neuronal number of the rat intracardiac nerve plexus. Ann Anat - Anatomischer Anzeiger. 2003; 185(5): 449–459.
- Pauza D, Pauziene N, Pakeltyte G, et al. Comparative quantitative study of the intrinsic cardiac ganglia and neurons in the rat, guinea pig, dog and human as revealed by histochemical staining for acetylcholinesterase. Ann Anat - Anatomischer Anzeiger. 2002; 184(2): 125–136.
- Singh S, Johnson PI, Javed A, et al. Monoamine- and histamine-synthesizing enzymes and neurotransmitters within neurons of adult human cardiac ganglia. Circulation. 1999; 99(3): 411–419.
- Weihe E, Schütz B, Hartschuh W, et al. Coexpression of cholinergic and noradrenergic phenotypes in human and nonhuman autonomic nervous system. J Comp Neurol. 2005; 492(3): 370–379.
- Martin L. Chinchillas as Experimental Models. The Laboratory Rabbit, Guinea Pig, Hamster, and Other Rodents. 2012: 1009–1028.
- Mikkelsen E, Lauridsen H, Nielsen PM, et al. The chinchilla as a novel animal model of pregnancy. R Soc Open Sci. 2017; 4(4): 161098.
- Suckow MA, Stevens KA, Wilson RP. The laboratory rabbit, guinea pig, hamster, and other rodents. 1st ed Saunders. : Elsevier 2012.
- Kuder T, Nowak E, Szczurkowski A, et al. A comparative study on cardiac ganglia in midday gerbil, Egyptian spiny mouse, chinchilla laniger and pigeon. Anat Histol Embryol. 2003; 32(3): 134–140.
- Kuchinka J, Chrzanowska M, Kuder T. Adrenergic and cholinergic innervation of the atrioventricular valves in chinchilla (Chinchilla laniger). Folia Morphol (Warsz). 2017; 76(4): 590–595.
- Radzimirska M, Kuchinka J, Nowak E, et al. Cholinergic and adrenergic innervation of the pancreas in chinchilla (Chinchilla Laniger Molina). Folia Histochem Cytobiol. 2020; 58(1): 54–60.
- KARNOVSKY MJ, ROOTS L. A "DIRECT-COLORING" THIOCHOLINE METHOD FOR CHOLINESTERASES. J Histochem Cytochem. 1964; 12: 219–221.
- Tsuji S, Larabi Y. A modification of thiocholine-ferricyanide method of Karnovsky and Roots for localization of acetylcholinesterase activity without interference by Koelle's copper thiocholine iodide precipitate. Histochemistry. 1983; 78(3): 317–323.
- Torre JCD. An improved approach to histofluorescence using the SPG method for tissue monoamines. J Neurosci Methods. 1980; 3(1): 1–5.
- Gage GJ, Kipke DR, Shain W. Whole animal perfusion fixation for rodents. J Vis Exp. 2012(65).
- Slaviková J, Kuncová J, Reischig J, et al. Catecholaminergic neurons in the rat intrinsic cardiac nervous system. Neurochem Res. 2003; 28: 593-598. .
- Hasan W, Smith PG. Modulation of rat parasympathetic cardiac ganglion phenotype and NGF synthesis by adrenergic nerves. Auton Neurosci. 2009; 145(1-2): 17–26.
- Haberberger R, Kummer W. beta 2-adrenoreceptor immunoreactivity in cardiac ganglia of the guinea pig. Histochem J. 1996; 28(11): 827–833.
- Rysevaite K, Saburkina I, Pauziene N, et al. Immunohistochemical characterization of the intrinsic cardiac neural plexus in whole-mount mouse heart preparations. Heart Rhythm. 2011; 8(5): 731–738.
- Richardson RJ, Grkovic I, Anderson CR. Immunohistochemical analysis of intracardiac ganglia of the rat heart. Cell Tissue Res. 2003; 314(3): 337–350.
- Pauziene N, Alaburda P, Rysevaite-Kyguoliene K, et al. Innervation of the rabbit cardiac ventricles. J Anat. 2016; 228(1): 26–46.
- Calupca MA, Locknar SA, Zhang L, et al. Distribution of cocaine- and amphetamine-regulated transcript peptide in the guinea pig intrinsic cardiac nervous system and colocalization with neuropeptides or transmitter synthetic enzymes. J Comp Neurol. 2001; 439(1): 73–86.
- Scruggs P, Dun SL, Dun NJ. Cocaine- and amphetamine-regulated transcript peptide attenuates phenylephrine-induced bradycardia in anesthetized rats. Am J Physiol Regul Integr Comp Physiol. 2003; 285(6): R1496–R1503.
- Richardson RJ, Grkovic I, Anderson CR. Cocaine- and amphetamine-related transcript peptide and somatostatin in rat intracardiac ganglia. Cell Tissue Res. 2006; 324(1): 17–24.
- Mao P, Meshul C, Thuillier P, et al. CART Peptide Is a Potential Endogenous Antioxidant and Preferentially Localized in Mitochondria. PLoS ONE. 2012; 7(1): e29343.
- Horackova M, Armour JA, Byczko Z. Distribution of intrinsic cardiac neurons in whole-mount guinea pig atria identified by multiple neurochemical coding. A confocal microscope study. Cell Tissue Res. 1999; 297(3): 409–421.
- Moravec M, Moravec J, Forsgren S. Catecholaminergic and peptidergic nerve components of intramural ganglia in the rat heart. An immunohistochemical study. Cell Tissue Res. 1990; 262(2): 315–327.
- Parsons RL, Locknar SA, Young BA, et al. Presence and co-localization of vasoactive intestinal polypeptide with neuronal nitric oxide synthase in cells and nerve fibers within guinea pig intrinsic cardiac ganglia and cardiac tissue. Cell Tissue Res. 2006; 323(2): 197–209.
- Steele PA, Gibbins IL, Morris JL, et al. Multiple populations of neuropeptide-containing intrinsic neurons in the guinea-pig heart. Neuroscience. 1994; 62(1): 241–250.
- Odorizzi M, Fernette B, Angel E, et al. Galanin receptor antagonists decrease fat preference in Brattleboro rat. Neuropharmacology. 2002; 42(1): 134–141.
- Diaz Z, Narváez J, Hedlund P, et al. Centrally infused galanin-(1–15) but not galanin-(1–29) reduces the baroreceptor reflex sensitivity in the rat. Brain Research. 1996; 741(1-2): 32–37.
- Herring N, Cranley J, Lokale MN, et al. The cardiac sympathetic co-transmitter galanin reduces acetylcholine release and vagal bradycardia: implications for neural control of cardiac excitability. J Mol Cell Cardiol. 2012; 52(3): 667–676.
- Crick SJ, Anderson RH, Ho SY, et al. Localisation and quantitation of autonomic innervation in the porcine heart II: endocardium, myocardium and epicardium. J Anat. 1999; 195 ( Pt 3): 359–373.
- Gordon L, Wharton J, Gaer JA, et al. Quantitative immunohistochemical assessment of bovine myocardial innervation before and after cryosurgical cardiac denervation. Cardiovasc Res. 1993; 27(2): 318–326.
- Anker SD. Catecholamine levels and treatment in chronic heart failure. Eur Heart J. 1998; 19 Suppl F: F56–F61.
- Adameova A, Abdellatif Y, Dhalla NS. Role of the excessive amounts of circulating catecholamines and glucocorticoids in stress-induced heart disease. Can J Physiol Pharmacol. 2009; 87(7): 493–514.
![](https://journals.viamedica.pl/plugins/generic/popups/images/icons/close.png)