Analysis of micromorphological and immunohistochemical characteristics of human superior cervical sympathetic ganglion

[J. Boljanović]

74 str.

Medicina - Neuroanatomija /

Razlog za ovo istraživanje bilo je pitanje da li u gornjem vratnom simpatičkom ganglion čoveka (GVSG) postoji neujednačen raspored mreže krvnih sudova i mastocita koji bi eventualno mogli da utiču na nervne ćelije i aksone u određenom delu gangliona. Cilj ovog rada bila je mikromorfološka analiza distribucije malih krivnih sudova, mastocita i ganglijskih neurona u proksimalnom i distalnom delu gornjeg vratnog simpatičkog gangliona čoveka (GVSG). Primenjene su statističke analize da bi se otkrile moguće metričke regionalne razlike u njihovim gustinama. Petnaest humanih GVSG injiciranih obojenim tušem i želatinom su mikrodisekovani i ispitivani. U drugoj grupi se nalazilo pet GVSG koji su pripremljeni i serijski isečeni za imunohistohemijsko bojenje na CD34, mastocitnu triptazu, NPY, VIP, SP, SOM i CGRP. Pomoću imunohistohemijskog bojenja prikazali smo intraganglijske mikrosudove kao i mastocite unutar GVSG i fokusirali se na odabranu grupu neuropeptida: NPY, VIP, SP, SOM i CGRP. Mikroskopska polja dva dela GVSG su analizirana za sledeće kvantifikacije: gustina mikrosudova (GMS), gustina mastocita (GMA), broj i merenja parametara ganglijskih ćelija, i distribucija i merne karakteristike imunohistohemijski obojenih ganglijskih neurona. Prosečan broj CD34 pozitivnih mikrosudova u mikroskopskim poljima, GMS je imala vrednost 83 za gornje delove i 82,7 za donje delove GVSG. GMA je bila 4,5 u proksimalnim delovima a 4,7 u distalnim delovima GVSG. Prosečan broj ganglijskih neurona u mikroskopskim poljima bio je 19,5 u proksimalnim delovima i 19,8 u distalnim delovima GVSG. Uočili smo da 24,3% GVSG neurona u proksimalnom i 32% u distalnom delu gangliona eksprimiraju NPY, 16,6% i 16,4% VIP, nije postojala ekspresija SP, 13.3% i 14,3% eksprimiraju SOM, a 12,2% i 13,5% eksprimira CGRP. Gustina CD34 pozitivnih mikrosudova, gustina mastocita pozitivnih na triptazu, i gustina, srednji prečnici i srednje površine ganglijskih neurona nisu se značajno razlikovale u dva posmatrana dela, gornjem i donjem delu GVSG. Analizirali smo metričke karakteristike NPY, VIP, SOM i CGRP pozitivnih i negativnih ganglijskih neurona u dva različita dela, proksimalnom i distalnom delu GVSG. Da zaključimo, uz pomoć imunohistohemijskog bojenja, proučavali smo proksimalno-distalnu distribuciju ganglijskih neurona u vezi sa ekspresijom imunoreaktivnosti za različite neuropeptide: NPY, VIP, SOM i CGRP. Ispitivali smo intraganglijske mikrosudove, mastocite kao i neurone unutar GVSG. Distribucija mikrosudova, mastocita i neurona u dva dela GVSG je bila ujednačena bez specifičnih mikromorfoloških varijacija, postoji homogeni vaskularni i ćelijski obrazac unutar GVSG. Intraganglijska mreža krvnih sudova i klasteri ganglijskih neurona su pravilno raspoređeni unutar GVSG. Intraganglijski mastociti su brojni i ujednačeno prisutni, verovatno sa potencijalom modulacije funkcije simpatičkih ganglijskih ćelija.

The reason for this investigation was the question of whether within the human superior cervical sympathetic ganglion (SCSG) there is an uneven layout of blood vessel network and of mast cells, which could possibly influence the nerve cells and axons in a specific portion of the ganglion. The aim of this study was the micromorphological analysis of the distribution of microvessels, mast cells and ganglionic neurons in two parts, proximal and distal of the human superior cervical sympathetic ganglions (SCSGs). Statistical analyses were applied to detect the possible metric regional differences in their densities. Fifteen injected human SCSGs with colored India ink and gelatin were microdissected and examined. Second group of five human SCSGs was prepared and serially sliced for CD34, mast cell tryptase, NPY, VIP, SP, SOM and CGRP immunostaining. By means of immunohistochemical staining, we depicted the intraganglionic microvessels as well as mast cells within the SCSG, and we focused on a selected group of neuropeptides: NPY, VIP, SP, SOM and CGRP. The microscopic fields of two parts of the SCSGs were analyzed for the following quantifications: microvessel density (MVD), mast cell density (MCD), ganglionic cell count and measurements, and distribution and metric characteristics of immunolabeled ganglionic neurons. The mean number of CD34-positive microvessels in microscopic fields, the MVD, had a value of 83 for the upper parts, and 82.7 for the lower parts of SCSGs. The mean number of tryptase-positive mast cells in microscopic fields, the MCD, was 4.5 in the proximal parts, and 4.7 in the distal parts of SCSGs. The mean number of ganglionic neurons in microscopic fields was 19.5 in the proximal parts, and 19.8 in the distal parts of SCSGs. We observed that 24.3% of SCSG neurons in the proximal and 32% in the distal part expressed NPY, 16.6% and 16.4% expressed VIP, no expression of SP existed, 13.3% and 14.3% expressed SOM, and 12,2% and 13.5% expressed CGRP respectively. The density of CD34-positive microvessels, the density of tryptase-positive mast cells, and the density, mean diameters and mean areas of ganglionic neurons were not significantly different in two observed parts, upper and lower of the SCSGs. We analyzed metric characteristics of NPY, VIP, SOM and CGRP positive and negative ganglionic neurons in two distinct parts, proximal and distal of the SCSGs. In conclusion, by means of immunohistochemical staining, we studied the proximal-distal distribution of ganglionic neurons, related to the expression of immunoreactivity for different neuropeptides: NPY, VIP, SOM and CGRP. We examined the intraganglionic microvessels, mast cells as well as the neurons within the SCSGs. The distributions of microvessels, mast cells, and neurons in two parts of the SCSGs were uniform with no specific micromorphological variations, there is a homogenous vascular and cellular pattern within the SCSGs. The intraganglionic network of blood vessels and clusters of ganglionic neurons are uniformly arranged within the SCSG. The intraganglionic mast cells are numerous and uniformly present, possibly with the potential of modulating the function of sympathetic ganglionic cells.

mikromorfološka analiza; ganglijski neuroni; mastociti; mikrosudovi; gornji vratni simpatički ganglion; imunohistohemijsko bojenje

micromorphological analysis; ganglionic neurons; mast cells; microvessels; superior cervical sympathetic ganglion; immunohistochemical staining

616.8:611.833(043.3)

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