The adrenal glands from 15 guinea pigs aged 90 days were used in the study.
Paraffin slices were stained with Mayer haematoxylin and eosin with Masson
method and silvered. The histochemical reactions were carried out for catecholamines
and the biochemical determinations of catecholamines. Ultrastructural
observations were also performed.
In the adrenal medulla of sexually mature guinea pigs numerous epinephrocites
and no noreoinefrocites were observed when histochemical methods were used.
The small and large ganglion cells accompanying the supporting and neurolemmal
cells formed typical gangliar weaving (medullary ganglia). Apart from epinephrocytes,
small and large ganglion cells and supporting and neurolemmal
cells, small intensively fluorescent (SIF) cells were also detected.
Biochemical investigation revealed that the mean content of adrenaline calculated
for a pair of adrenal glands was 98.87% of the pool of catecholamines,
whereas the mean content of noradrenaline was 1.00%, and of dopamine 0.13%

Kaczmarczyk, Dariusz

Kmieć, Bogumił L.

English

Via Medica Journals

Folia Morphol. Vol. 63, No. 1, pp. 75–78Copyright © 2004 Via MedicaISSN 0015–5659www.fm.viamedica.pl S H O R T  C O M M U N I C A T I O N75The cellular composition of the adrenalmedulla of sexually mature guinea pigsBogumił L. Kmieć, Dariusz KaczmarczykDepartment of Cytophysiology, Histology and Embryology, Medical University, Łódź, Poland[Received 1 September 2003; Revised 5 November 2003; Accepted 5 November 2003]The adrenal glands from 15 guinea pigs aged 90 days were used in the study.Paraffin slices were stained with Mayer haematoxylin and eosin with Massonmethod and silvered. The histochemical reactions were carried out for cate-cholamines and the biochemical determinations of catecholamines. Ultrastruc-tural observations were also performed.In the adrenal medulla of sexually mature guinea pigs numerous epinephrocitesand no noreoinefrocites were observed when histochemical methods were used.The small and large ganglion cells accompanying the supporting and neurolem-mal cells formed typical gangliar weaving (medullary ganglia). Apart from epi-nephrocytes, small and large ganglion cells and supporting and neurolemmalcells, small intensively fluorescent (SIF) cells were also detected.Biochemical investigation revealed that the mean content of adrenaline calculat-ed for a pair of adrenal glands was 98.87% of the pool of catecholamines,whereas the mean content of noradrenaline was 1.00%, and of dopamine 0.13%.Key words: adrenal glands, chromaffin cells, ganglion cells, SIF cellsAddress for correspondence: Prof. Bogumił L. Kmieć, Department of Cytophysiology, Histology and Embryology, Medical University,ul. Narutowicza 60, 90–136 Łódź, tel: +48 42 631 98 07, e-mail: kmieckbl@csk.am.lodz.plINTRODUCTIONMedullary cells (i.e. chromaffin and ganglion cells)develop from sympathoadrenal cells originating fromneural crests. The presence of two cell phenotypesin the adrenal medulla of mammals, namely epine-phrocytes and norepinephrocytes has been report-ed by many investigators. In the anteater Myrme-cophaga jubata and the sand hare, however, adre-nal medullas were composed only of epinephrocytes[18], and in many fish species (e.g. carp) only of nore-pinefrocytes [12]. However, there is still some con-troversy as to how many cell types are present in therabbit and guinea pig, since some researchers havereported the presence of both epinephrocytes andnorepinephrocytes, whereas others have mentionedonly one type of these cells [3, 12]. Because the guin-ea pig is a commonly used laboratory animal, theaim of the study was to determine the cellular com-position of the adrenal medulla with particular at-tention to the phenotypes of the chromaffin cells.MATERIAL AND METHODSThe adrenal glands from 15 guinea pigs aged90 days, i.e. sexually mature, and of both sexes wereused in the study. After decapitation of the animalsthe adrenal glands were excised for examination andparaffin slices were stained with the Mayer haema-toxylin and eosin, with the Masson method and thensilvered using the Bodian and the Loots methods [16].Histochemical reactions for catecholamines werecarried out according to Kmieć [13], and Hopsu andMäkinen [10]. The cellular content was visualised withthe use of a EM-300 electron microscope [14, 15].Biochemical determination of the catecholamines(adrenaline, noradrenaline and dopamine) in thehomogenates from the gross adrenal glands was76Folia Morphol., 2004, Vol. 63, No. 1performed using the HPLC method [17]. All the pro-cedures were approved by the University of Medi-cine in Łódź Animal Care and Use Committee.RESULTS18-µm-in-diameter cells were predominantly ob-served in the adrenal medulla of sexually matureguinea pigs. The diameter of the nuclei in these cellswas approximately 7 µm, and the diameter of thenucleoli located near the nuclear membrane wasapproximately 1.5 µm. The chromatic stroma wasdistinct and the cytoplasm of these cells was granu-lar and slightly basophilous. Single or numerous cy-toplasmatic granules (0.2–1.0 µm in diameter) weredetected and the chromaffin cells showed granularvesicles (250–350 nm in diameter), correspondingto vesicles containing adrenaline.Histochemical reactions for noradrenaline (Hop-su and Mäkinen method) in the medullary cells werenegative (Fig. 1), although in some specimens gan-glion nerve cells gave a trace reaction for this cate-cholamine. The histochemical reaction for bothadrenaline and noradrenaline (according to Kmieć)revealed no cells with a positive reaction for norad-renaline, but only cells with a strong positive reac-tion for adrenaline (Fig. 2).The ganglion cells of the adrenal medulla formedsmall groups or large aggregations, showing the typ-ical gangliar weaving (medullary ganglia). The diam-eters of these cells were variable, ranging from 19to 50 µm. In this group small (19 to 24 µm in diam-eter) and large (35 to 50 µm in diameter) cells wereobserved. In the vicinity of the ganglion cells numer-ous supporting and neurolemmal cells were detect-ed. The cytoplasm of the ganglion cells was granu-lar and very basophilous, particularly at their circum-ference. They contained large vesicular nuclei (ap-proximately 12 µm in diameter), which were locatedeccentrically. The latter showed 1–3 large nucleoliof around 3 µm in diameter, predominantly locatednear the nuclear membrane. In the electron micro-scope these cells demonstrated granular vesicles of40–200 nm in diameter.In the silvered slices adrenal medulla ganglioncells with a variable number of argyrophilous gran-ules were observed (Fig. 3). In the vicinity of the ad-renal medulla ganglion cells numerous nerve fibreswere observed, which extended to the chromaffincells, amongst which they disappeared.Apart from the medullary cells, small and largeganglion cells, supporting cells, neurolemmal cellsand small intensively basophilous cells were ob-served. These contained small chromaffin granuleswith a diameter of 60–120 nm (Fig. 4).Biochemical investigation showed that themean content of adrenaline in the adrenal medul-la homogenates of sexually mature guinea pigs(calculated for a pair of adrenal glands) was48388.3 ng (98.87% of the pool of catecholaminesdetermined), whereas the mean content of nora-drenaline was 491.8 ng (1.00%) and of dopamine62.2 ng (0.13%).DISCUSSIONOur study has revealed that the adrenal medullaof sexually mature guinea pigs shows type IV weav-ing according to the Suzuki and Kach classification[19, 20]. The presence of cells producing and secret-ing adrenaline amongst the chromaffin cell pheno-types supports [1, 3, 4, 19, 20, 24] or is at variance[6, 7, 12] with the results of other investigators. Pre-sumably this is the result of the disclosure of thesmall intensively fluorescent (SIF) cells, which wereconsidered norepinephrocytes [4]. The extremely lowcontent of noradrenaline in the adrenal gland ho-mogenates of sexually mature guinea pigs in com-parison with other animals only testifies to the pres-ence of epinephrocytes. Noradrenaline and dopam-ine are presumably the pool of catecholaminespresent in the ganglion cells and the SIF (small gran-ule containing SGC) cells.Benedeczky et al. [2] found three types of chro-maffin cell in the adrenal glands of the grass snakeNatrix natrix. Cells with granular vesicles of diame-ter of around 250 nm corresponded to norepineph-rocytes and those of diameter of 125 nm to epi-nephrocytes. The third type of cell with both smalland large granular vesicles as well as well-devel-oped rough endoplasmic reticulum and numerousribosomes presumably contained dopamine [11,12]. Similar cells (so-called SGC cells) were also re-ported in the adrenal medulla of the rat [5] and ofman during development [9] and in the adrenalglands of sexually mature birds and reptiles [21–23].Our study supports previous opinions [21, 22] thatthe majority of these cells, if not the total num-ber, appear to be identical with the SIF cells, whichwere firstly reported by Eränkö and Härkönen [8]in the superior cervical sympathetic ganglion ofthe white rat.In conclusion, adrenal medulla cells, due to close“territorial” proximity, form a morphological andfunctional complex with the sympathetic nervoussystem and the adrenal cortex.77Bogumił L. Kmieć et al., Cellular composition of the adrenal medullaFigure 1. Epinephrocytes (E). Histochemical reaction for norad-renaline according the Hopsu and Mäkinen method. Enl. 220 ¥.Figure 2. Epinephrocytes (E). Histochemical reaction for norad-renaline according the Kmieć method. Enl. 220 ¥.Figure 3. Ganglion cells (GC), medullary cells (MC). Silvering ac-cording to Bodian. Enl. 560 ¥.Figure 4. Small intensively fluorescent cells (SIF) and epinephrocy-tes (E). Double immersion fixation of the slices in the Karnovsky and Dal-ton fluid with the Kmieć modification (chromaffin reaction). Enl. 4000 ¥.78Folia Morphol., 2004, Vol. 63, No. 1ACKNOWLEDGEMENTSThe research was funded by the Medical Univer-sity of Łódź, grant No. 502-11-775.REFERENCES1. Bänder A (1950) Über zwei verschiedene chromaffineZelltypen in Nebennierenmarkt und ihre Beziehungzum Adrenalin- und Arterenolgehalt. Vehr D Anat Ges,48: 172–176.2. Benedeczky I, Puppi A, Tigyi A (1965) Histochemicaland electron microscopical study of the adrenal me-dulla of the grass snake (Natrix natrix). Acta Biol AcadSci Hung, 15: 271–2843. Coupland RE (1965) The natural history of the chro-maffin cell. Longmans, London.4. Coupland RE (1989) The natural history of the chro-maffin cell — twenty-five years on the beginning. ArchHistol Cytol, 52 (Suppl.): 331–341.5. Diner O (1965) Observations sur le development dela medullo-surrenale du rat: l’evolution de la partienon-chromaffine. Arch Anat Micr Morph Exp, 54:671–718.6. Eränkö O (1955) Fluorescing islet, adrenaline and no-radrenaline in the adrenal medulla of some commonlaboratory animals. Ann Med Exp Finn, 33: 278–290.7. Eränkö O (1955) Histochemistry of noradrenaline inthe adrenal medulla of rats and mice. Endocrin, 57:363–368.8. Eränkö O, Härkönen M (1963) Histochemical demon-stration of fluorogenic amines in the cytoplasm of sym-pathetic ganglion cells of the rat. Acta Physiol Scand,58: 285–286.9. Hervonen A (1971) Development of catecholamine-storing cells in human fetal paraganglia and adrenalmedulla. A histochemical and electron microscopicalstudy. Acta Physiol Scand, 368 (Suppl.): 1–94.10. Hopsu VK, Mäkinen EO (1996) Two methods for thedemonstration of noradrenaline-containing adrenalmedullary cells. J Histochem Cytochem, 14: 434–435.11. 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The cellular composition of the adrenal medulla of sexually mature guinea pigs

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The cellular composition of the adrenal medulla of sexually mature guinea pigs

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