ACTA THERIOLOGICA
Vol. 21, 3: 31—36, 1976
Histological and Histochemical Picture of the Lachrymal Gland
in Guinea Pig
Lucyna KROCHM ALSKA
K rochm alska L., 1976: H istological and h isto ch em ica l pictu re of the
lach rym al gland in gu in ea pig. A cta theriol. 21, 3: 31— 36. [W ith P la te I].
T he m orphological and h istological ob servation s w e r e carried out on
24 lach rym al gland s of th e guinea pig (Ca v i s cobaya). T he preparations
w ere stained w ith h em atoxylin , eosine and tolu id in e. M oreover, the
localization and a ctiv ity of su ccin ate d eh yd rogen ase, la c ta te d eh yd ro­
genase, a lk alin e and acid phosphatases w ere determ in ed . It w as found
that the lachrym al gland of th e guinea pig has branched tu b u lar
structure, its tu bu les joining d irectly to th e e fflu e n t ducts lin ed w ith
cubical or flat epith eliu m . T he d ifferen ces in th e g ran u le con ten t and
th e degree of cytop lasm stain in g are probably rela ted to th e secretory
phase of th e cell. A high en zym atic a ctiv ity , esp ecia lly of alk a lin e
phosphatase, in th e peripheral parts of glan d u lar cells supports the
idea th at th e secretion occurs to th e in tercellu la r spaces.
[Morphol. Lab. and Dept. O phtalm ol., M edical A cad em y, 15-276 B ia ­
łystok , C u rie-S k łod o w sk iej 24a],
1. INTRO DUCTIO N
The studies of K ü h n e l (1968 a, b, c, d, e, f) dem onstrated th a t the
lachrym al gland of several anim al species and m an shows th e stru c tu re
of a branched tu b u lar gland. At the sam e tim e only sm all differences
in the m orphological and histochem ical appearance of th e lachrym al
gland w ere observed betw een particu lar species.
The present study was aim ed a t the obtaining of m orphological and
histochem ical picture of th e lachrym al gland in guinea pig (Cavia co­
baya) w hich is a common laboratory anim al b u t w as om itted in th e
investigations of K ü h n e i (i.e.).
2. METHODS
T he in v estig a tio n s w ere carried out on 24 la ch ry m a l glan d s of y o u n g gu in ea
pigs, 150 to 200 g in body w eigh t. T he g lan d s w ere ex c ise d un d er urethan
a n aesth esia after adm in isterin g in trap eriton eally 20% so lu tio n of u reth an in th e
dose of 1.0 g per kg body w eigh t. T he m aterial w as fix e d in th e B aker flu id and
em bedded in paraffin. Section s 4 to 8 ^ thick w ere sta in ed w ith h a em a to x y lin ,
eosin e and toluidine. For en zym atic estim ation s th e glan d s w ere cut w ith a fr e e z ­
ing m icrotom e into sections 15 (i thick. L actic d eh yd rogen ase (LDH) and su ccin ate
d eh yd rogen ase (SDH) w ere d etected by th e m ethod of N ach las, acid p h osp h atase
(AcF) and alk a lin e p h osp h atase (A l P ) by th e m ethod of B u rston e.
[31]
32
L, Krochmalska
3. R ESU L TS OF M ORPHOLOGICAL OBSERV A TIO N S
The lachrym al gland of guinea pig is a m ultilayer, branched, tu b u la r
gland w ith dilated term in al segm ents. It is bounded by a thin pouch of
the connective tissues which form s fine steaks dividing the gland into
single lobules. These streak s are thicker a t the effluent ducts. The
tubules join directly to the effluent ducts lined w ith cubical or flat
epithelium . The glan d u lar tubules are built of several rows of cubical
cells containing num erous granules. The intercellu lar canals joining to
the light of tubules are visible betw een the cells. The nuclei of these
cells are round, abounding in chrom atin. From th e ex tern al side they are
covered by flat epithelial cells (Fig. 1).
Staining w ith toluidine shows the presence of num erous dark-blue
granules in the cytoplasm of glandular cells. C ytoplasm of the cells
situated in the m iddle p a rt of the tubules contains a m uch sm aller
num ber of granules (Fig. 2).
3. R ESU L TS OF ENZYM ATIC E STIM A T IO N S
SDH. The cells of tubules are characterized by strong positive re ­
action for SDH and contain num erous granules of diform azane arranged
perinuclearly. Intensive reaction in the form of dark-blue staining is
visible in the in tercellu lar spaces and to the p eriphery of tubules, at
the site occurrence m yoepithelial cells. The m iddle p a rt of some
tubules contains th e cells of bright cytoplasm , alm ost totally devoid of
granules of diform azane (Fig. 3).
LDH. The cytoplasm of glandular cells is light blue. The strong po­
sitive reaction for LD H occurs at the p eriphery of glandular cells. Some
groups of cells w ith to tally negative reaction for LDH are visible on the
sections of tubules (Fig. 4).
AlP. Positive reaction as a light brow n staining of cytoplasm is found
in glandular cells. Intensive brow n staining occurs in the intercellular
space. D ark distinct lines form a sort of in tercellu lar canals. At the cell
periphery th ere are visible light oval-shaped spots th a t m ay correspond
to vacuoles. S trong positive reaction is encountered in the cells of
effluen t ducts and blood vessel w alls (Fig. 5).
AcP. Positive reaction for AcP is observed in th e cytoplasm as light
brow n staining. In te rce llu la r spaces are characterized by intensive re ­
action in th e form of dark brow n lines (Fig. 6).
4. D ISC U SSIO N
The lachrym al gland of the guinea pig is a m ultilayer branched
tu b u lar gland. The secretory segm ents are opened directly to in tra ­
Histology of the lachrymal gland in guinea pig
33
lobular ducts, sim ilarly to o th er anim als, except ox (K ü h n e 1, 1968 a,
b, c, d, e). The lachrym al gland of the guinea pig is bounded by a thin
pouch of connective tissue. Its fine streaks divide the gland into lobules.
Sim ilarly to the lachrym al gland of rabbit, cat, goat, sheep, dog and ox
(K ü h n e 1, 1968 a, b, c, d, e), as w ell as of m an (K ü h n e 1, 1968 f; M ö l ­
l e n d o r f f , 1939), it was found th at the streaks of connective tissue
became th ick er a t the effluent ducts and around blood vessels. In the
secretory segm ents, th a t are round or oval w ith relativ ely th in light,
the staining w ith toluidine blue perm itted to dem onstrate th e presence
of cells differing in the num ber of granules. The cells w ith a large
num ber of granules contain usually round nucleus, abounding in chro­
m atin. The differences in th e content of granules in glandular cells are
regarded by some au thors ( M ö l l e n d o r f f , 1936; A l b r i c h , 1928) as
being due to various secretory phases of th e sam e cells. O ther authors
(I t o & S h i b a s a k i , 1969; K ü h n e 1, 1968 f) assum e th a t glandular
cells w ith a sm all num ber of granules correspond to m ucous cells. M ore­
over, some differences in the staining of cytoplasm and in the size of
cells w ere found; this m ay be again related to th eir secretory phase
( M ö l l e n d o r f f , 1938; R a d n o t, 1939). To the outside of th e proper
glandular cells fla t epithelial cells w ere observed. Sim ilar cells w ere
found in the preparations of the lachrym al gland of th e cat (K ü h n e 1,
1968 a) and m an ( K i i h n e l . 1968 f; M ö l l e n d o r f f . 1936). These cells
m ay correspond to the basket-like cells of Boll described in the parotid
gland ( A c k e r m a n et al., 1971) and sw eat glands ( M ö l l e n d o r f f ,
1936). They have the ability to contract and expel glandular secretion
to th e light of ducts ( A c k e r m a n et al., 1971).
H istochem ical investigations dem onstrated the highest enzym atic
activity in the p erip heral p a rt of glandular cells. The localization of
enzym es support the idea th a t secretion of glandular cells is directed
tow ard in tercellu lar spaces.
Succinate dehydrogenase (SDH) is localized m ainly in the p eripheral
p a rts of glandular cells and in the external layer of tubules. A sim ilar
topographic localization of SDH occurs in th e glandular cells of the
lachrym al gland of the rab b it and m an (K ü h n e 1, 1968 a, f). Such lo­
calization corresponds to the arrangem ent of m itochondria, as confirm ed
by studies of K ü h n e l (1968 a, f) in the electron microscope. As r e ­
ported by N e j f a c h (1973) SDH is firm ly bound to m itochondrial
cristae. Sim ilarly to lachrym al glands of rab b it and m an (K ü h n e 1,
1968 a, f), and in distinction to cat (K ü h n e 1, 1968 a), this enzym e shows
a low er activity in the epithelium of effluent ducts.
Lactic dehydrogenase (LDH) is localized in th e cytoplasm of glandular
cells and in the cells of effluent ducts. The presence of this enzym e was
34
L. Krochmalska
shown in the lachrym al glands of m an and various anim als (K ü h n e 1,
1968 a, b, c, d, e, f). In th e glan d u lar cells of m an its activity is lower
than in those of guinea pig, w hile a sim ilar strong positive reaction was
observed in th e cells of efflu en t ducts. This fact indicates a high glyco­
lytic activity in th a t p a rt of th e lachrym al gland.
A lkaline phosphatase (A l P ) is localized at th e periphery of tubules,
in the cells of effluent ducts and in blood vessel walls. Sim ilarly to the
lachrym al gland of m an and rab b it (K ü h n e 1, 1968 a, f) the m axim um
activity was found in th e cells of effluent ducts and blood vessel walls.
This m ay indicate the occurrence of active tra n sp o rt through the m em ­
branes, sim ilarly to oth er organs w here alkaline phosphatase is abund­
ant, e.g., in the cells of kidney tubules and in the villi of sm all intestine
( A c k e r m a n et al., 1971; K r y g i e r et al., 1963).
Acid phosphatase is localized m ainly in the peripheral p a rt of glan­
d u lar cells and in the basal p a rt of the cells of effluent ducts. Its pres­
ence m ay indicate th e occurrence of introcellular processes of h ydro­
lysis.
REFERENCES
I. A c k e r m a n J., N o w i c k i Z., S a r n e c k a - K e l l e r M., 1971: Cytologia
i histologia. P ań stw . Zakł. W yd. Lek. 1— 535. W arszaw a.
2. A 1 b r i c h, A., 1928, a fter R adnot M., 1939.
3. 11 o T., S h i b a s a k i S., 1967 (after K ühnei W., 1968 f).,
4. K ü h n e i W., 1968a- V erg leich en d e h istologisch e, h istochem ische und elektro’
I
nenm ikroskopische U n tersu ch u n gen an T ränendrüsen. K aninchen und K atze
Z. Z ellforsch., 85, 3: 408— 440.
5. K ü h n e 1 W., 1968 b; V erg leich en d e h istologisch e, histoch em isch e und elektronenm ikroskopische U n tersu ch u n gen an T ränendrüsen. Ziege. Z. Zellforsch., 86,
3: 430—443.
6. K ü h n e 1 W., 1968 c: V erg leich en d e h istologisch e, histoch em isch e und elektron en m ikroskopische U n tersu ch u n gen an T ränendrüsen. Schaf. Z. Zellforsch., 87,
1: 31—45.
7. K ü h n e i W., 1968 d: V erg leich en d e h istologisch e, h istoch em isch e und elek tro­
n en m ikroskopische U n tersu ch u n gen an T ränendrüsen. Hund. Z. Zellforsch., 88,
1: 23—38.
8. K ü h n e 1 W., 1968 e: V ergleich en d e h istologisch e, histoch em isch e und elek tro­
n en m ikroskopische U n tersu ch u n gen an T ränendrüsen. Rind. Z. Zellforsch., 87,
4: 504—525.
9. K ü h n e i W., 1968 f: V erg leich en d e h istologische, histoch em isch e und elektro­
nenm ikroskopisch e U n tersu ch u n gen an T ränendrüsen. M enschliche Tränen­
drüse. Z. Z ellforsch., 89, 4: 550— 572.
10. K r y g i e r A. & G o d l e w s k i H., 1963: M etody histochem iczne. P olsk ie Tow.
H isto- i C ytochem ików . 135— 153. W arszaw a.
II. M ö l l e n d o r f f W., 1936: H andbuch der m ikroskopischen A natom ie des M en­
schen. Springer V erlag. 3: 582— 592. B erlin.
Histología gruczołu łzowego św inki morskiej
35
12. N e j f a c h A., 1973: M echanizm y in tegracji przem ian kom órkow ych. P ań stw .
Zakł. W yd. Nauk.: 222— 274. W arszaw a.
13. R a d n o t M., 1939: D ie p ath ologisch e H istologie der T ränendrüse. K arger V er­
lag. 1—58. B asel.
Accept ed, F e b r ua r y 21, 1975.
L ucyna KROCHM ALSKA
OBRAZ HISTO LOG ICZNY
I HISTO CH EM ICZNY
ŚW IN K I M ORSKIEJ
GRUCZOŁU ŁZOWEGO
S treszczen ie
W gruczołach łzow ych św in k i m orskiej badano lok a liza cję i ak tyw n ość DB, DM,
FZ, FK. N ajw ięk szą ak tyw n ość enzym atyczn ą o b serw ow an o w obw odow ej części
kom órek gruczołow ych oraz kom órek w a rstw y zew n ętrzn ej cew ek. Z lokalizow anie
en zym ów przem aw ia za udziałem przestrzen i m ięd zyk om órk ow ych w transporcie
w yd zielin y kom órek gruczołow ych (Ryc. 1— 6).
36
L. Krochmalska
EXPLAN ATIO N TO PLATE
P late I.
Fig. 1. A tubule and intralobular duct of th e lach rym al gland of th e guinea pig.
Staining w ith HE.
Fig. 2. Section through a tu b u le of the lachrym al gland. T he central part contains
the cells w ith lig h tly stained cytoplasm w h ile dark cells are arranged at th e
periphery. Staining w ith to lu id in e blue.
Fig. 3. Su ccinate dehydrogenase — in ten sive reaction at th e periphery of glandular
and m yoep ith elial cells. Staining after N achlas.
Fig. 4. Lactic d ehyd rogen ase — p ositive reaction w ithin intercellu lar spaces and
in cytoplasm of glandular cells. Staining after N achlas.
Fig. 5. A lk alin e p hosp h atase — intensive reaction in the intercellular spaces. One
of th e cells contains a vacuole. Staining after Burstone.
Fig. 6. Acid phosph atase — p ositive reaction in th e intercellular spaces and w ithin
m yoep ith elial cells.
ACTA THERIOLOGICA, Vol. X X I, 3.
L. K rochm alsk i
P la te I.
auctcr phot.