Journal of Histochemistry & Cytochemistry

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Immunocytochemical studies of desmin and vimentin in pericapillary cells of chicken. T Fujimoto and S J Singer J Histochem Cytochem 1987 35: 1105 DOI: 10.1177/35.10.3305702 The online version of this article can be found at: http://jhc.sagepub.com/content/35/10/1105

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0022-15

54/87/$3.30

Vol. 35, No.

The Journal of Histochemistry and Cytochemistry Copyright © 1987 by The Histochemical Society,

Original

Immunocytochemical in Pericapillary Cells TOYOSHI

Received

for

ofBiology,

University

publication

November

Article

Studies of Desmin of Chicke&

of California 11,

1986

at San Diego,

and

in revised

Lajolla,

form

The composition of intermediate filaments in pericytes was examined by immunofluorescent and immunoelectron microscopic labeling of frozen sections of various chicken microvascular beds in situ. Pericytes in capillaries of cardiac muscle, exocnine pancreas, and kidney (peritubular capillary) were found to contain both desmin and vimentin. In some capillaries where pericytes do not exist, cells apposed to endotheial cellsthe Ito cell in the hepatic sinusoid and the reticular cell in the splenic sinusoid -were shown to contaiii both ofthe intermediate filament proteins. In contrast, podocytes and mesangial cells around renal glomerular capil-

and

Cal:fornia

92093.

3, 1987;

accepted

March

March

17,

Pericyte; Ito cell; Perisinusoidal cell; Intermediate filaDesmin; Vimentin; Immunofluomescence microscopy; Immunoelectron microscopy. WORDS:

KEY

ment;

peatedly

One

on the cardiovascular

system

has been

on van-

among microvasculan beds. Morphological differentiation among endothelial cells has been revealed not only between large and small blood vessels (for meview see ref. 28) but also among differations

ent

microvasculatunes

that in the

Our

temmediate

desmin

only,

Although endothelial

on vimentin

ofpemicytes

ical

significance

hypotheses

ciety

is even

Supported

examined

of penicytes

is not

suggested.

GM-15971

Singer, the

to con-

vimentin on

in-

the

only, panticu-

(11).

Furthermore,

the

understood, Functions

to SJS, who

physiolog-

although that

several

have

Foundation

been

ne-

and by US Pub-

is an American

Cancer

of Medicine,

Department

So-

Professor.

address: Yoshida,

Kyoto University, Sakyo,

Kyoto

Faculty 606,

Japan.

3 Correspondence to: Sj. Singer, Department ofBiology B-022, sity of California at San Diego, La Jolla, California 92093.

presumed

the other hand, unambiguously cytes have been matory

Univem-

for

pericytes

are

as follows:

(a) contractility

cells, which are present around observed to contract in vivo (2).

the On

penicytes in avians and mammals have never been proved to do so; (b) phagocytosis (4,18,35): penobserved to take up foreign materials in inflam-

reactions;

penicytes broblasts,

and

are believed and other

(c) mesenchymal

to differentiate mesenchymal

plunipotentiality

(20,24):

into smooth muscle cells in adult animals.

cells, fiEven dif-

fenentiation into plasma cells has been postulated by some authors (20). Because of the lack of specific markers for penicytes, however, this

capability To advance

examined

at least limited information about about the molecular ultrastruc-

meager.

Grant

Sj. that

believed

of either depending

chicken

more

been

and was

previously

by a grant from the Weingart

Service

Present

of Anatomy,

desmin,

found

differentiation as of membrane

however,

cells,

plus

adult

the

have

Research 2

of

as well

composed

we therefore have cells, our knowledge

tune

lic Health

finding,

are actually

we have

show

(T. Fujimoto

ofendothelial

only,

lam vasculatume

I

intriguing

filaments

of vimentin

studies,

proteins

to the cytoskeleton most

own

vasculatures

of cytoskeletal

anchored

in press).

In our

cells ofdifferent

distribution

proteins

sist

(17,24).

endothelial

(6Ao902).

1987

1987)

(9,19,24,30,36): e.g., Rouget capillaries of frog eyes, were

ofreseanch

Vimentin

laries contained only vimentin. The presence ofdesmin supports the hypothesis that pericytes may have a contractile apparatus similar to that of vascular smooth muscle cells. Our results also revealed that even in microvascular beds where pericytes are not found, cells having both desmin and vimentin exist next to endotheial cells and may assume similar functions to pericytes. (JHiscochem Cywchem35:1105-1115,

Introduction focus

1105-1115, 1987 Panted in USA.

S. J. SINGER3

and

FUJIMO’102

Department

10. pp.

Inc.

has not been our understanding

some

aspects

convincingly ofpenicytes,

of their

demonstrated. in this study

macnomoleculam

we have

composition

and

ultnastructure. Our approach has been to conduct immunocytochemical studies ofthe cells in situ. Pemicytes are difficult to isolate and

place

tematic

in culture investigation

the tissues

containing

(5),

and

of their penicytes

ticularly by immunoelectmon used techniques ofsemi-thin immunolabeling developed

therefore

the only

cell biology

approach

is to investigate,

by immunofluonescence,

to sysin situ, and

pan-

microscopy. Rr this purpose, we have and ultra-thin frozen sectioning and in this laboratory (31,32).

In our initial studies, the protein composition ofthe intermediate filaments of penicytes was examined. Since the expression of the subunits is thought, results in situ were state

of pemicytes.

penicytes

has been

in general, to be tissue-specific (10,22,27), the expected to characterize the true physiological Although

a preliminary

presented

(5), since

report it is known

about that

cultured the compo1105


FUJIMOTO,

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;.;

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1

if.’

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Our 5-

.:

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#{149}‘.‘.f ‘

#{149}ft.

.

,1,...,#{149}

.,

I,.

-‘l’S , P

‘

,

penicytes

(10,22)

smooth

muscle

vimentin

cells

penicapillary the proposal

Materials

and

both

is the same

as that

in Ito cells

cells of the splenic

between such mm supports

beds.

contain

(1,12,21,25,26,29).

was found

the reticulan

microvascular examined

ofwhich

are present just exterior to endothelial cytes do not exist. The results therefore

‘,‘?_

in culture

of different

all the

the composition

plus

and

a variety

that

vimentin,

desmin

sinusoid

.!

from

in vascular

tion,

be altered

In addiof the

sinusoid.

hepatic

Those

cells in capillaries suggest a close

cells

where penrelationship

cells and penicytes. The presence that pemicytes may be contractile

of descells.

Methods

#{149}.

I.’

_.

5

can

the cells in situ. In addition, in view of the among endothelial cells (11), we wished to

indicate

and

found /,

filaments

penicytes

results

desmin

?5fIL

‘.‘

.

.-..;

,i’,

compare

.

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#{149}..7 ‘

-‘

..

#{149}.

#.

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. .#

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.

of intermediate

it is essential to study variations we observed

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q’

SINGER

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iJ

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‘. . ‘p

#{149}p

Immunochemical Reagents. All the primary and secondary antibodies used in this study were purified by affinity chromatography on columns made with the respective antigens. Primary antibodies employed were antichicken

gizzard

vimentin

desmin

(34)

(34)

raised

raised

in guinea

in rabbits

pigs.

and

anti-chicken

Anti-desmin

and

erythmocyte

anti-vimentin

anti-

bodies were cross-absorbed with the heterologous antigen to eliminate any possible cmossmeactivity (34). Secondary antibodies were goat anti-rabbit IgG and goat anti-guinea pig IgG, cross-absorbed against each other, conjugated with

rhodamine

femmitin

and/or

(16)

and/or

Preparation with

into

glutaraldehyde. into

pieces,

fixation

aldehyde

tissues

were

Cryosectioning

sm)

frozen

through

a pressure

of 100-150

under

and

immediately, at room

were pH

ism)

according

instrument

depolymemized

kept

7.4,

with

and

to the

temperature.

at 4C

for

cmyoattachment

formup

in liquid

ultra-thin method

0.1%

trimmed

in 0.5%

blocks were frozen

(0.5-1.0 out

a

(freshly

fixative pieces

anesthetized

formaldehyde

excised

buffer,

on copper

was carried

MT-2B

tissue

phosphate

for semi-thin

sections

a Sonvall

were perfused

3%

in the same The

with

were

heart of

mm.

pieces mounted

gen.

ofthe

and

microscopy.

Leghorn)

fixatives

a mixture

immersed

was 60

in 0.1 M sodium

week. Tissue

using

or

and

time

(White

electron

used were 3% formaldehyde

Perfusion-fixed

small

hens

microscopy

(7) for

thomacotomy,

the left ventricle

mmHg. The fixatives from parafommaldehyde)

Total

Adult

Mtem

inserted

(3) for fluorescence

(iron-dextran)

ofTissues.

pentobarbital.

cannula

fluomescein Imposil

to a

nitmo-

(0.05-0.

10

of Tokuyasu, (31,32).

Immunofluorescence Microscopy. Semi-thin frozen sections mounted on glass slides were immunolabeled according to methods described (34). ftir tissues fixed with glutaraldehyde, sections were treated with 0. 5 mg/mI sodium bomohydride to quench the fluorescence caused by the fixative (23). Sections

were

observed

with

a Zeiss

photomicroscope

III (Carl

York) equipped with epifluomescent illumination ski objective lens. Photography was carried out with Microscopy.

Immunoelectron using (31),

ferritinexcept

and that

Ultra-thin

Imposil-conjugated pre-treatment

sections

secondary of sections

Zeiss

Inc.,

and a 63 x NomanKodak Tmi-X film.

New

were

immunolabeled

antibodies

was performed

as described with

a mixture

of 1% normal

goat serum and 2% dextran (Mw 10,000). Labeled sections were adsomption-stained and embedded in methyl cellulose as described

4

Figure 1. (a) Nomarski and (b,c) double indirect immunofluorescence micrographs of cardiac muscle. Semi-thin frozen sections were doubly immunolabeled with rabbit anti-desmin(b)and guinea pig anti-vimentin(c). Pericytes(arrowheads) are labeled for both desmin and vimentin. In contrast, endothelial cells (arrows) and fibroblasts (double arrowhead) are positive for vimentin but not for desmin. Note that cardiac muscle cells are labeled for desmin alone, but not for vimentin. Original magnification x 1500. Bar = 10 m.


DESMIN

AND

IN PERICAPILLARY

VIMENTIN

CELLS

1107

‘j’c

1;

-

Figure 2. Double indirect immunoelectron microscopy of a capillary in cardiac muscle. Desmin and vimentin are labeled by ferritin (arrowheads) and Imposil (arrows), respectively. Whereas Imposil is seen in the cell body of both the endothelium (E) and the pericyte (P), ferritin is observed only in the pericyte. L indicates the lumen of the capillary where a portion of an erythrocyte is seen. Original magnification x 75,000. Bar 0.1 tm.

(31),

and

ated

at 60 or 80 kV.

were

observed

with

a Philips

EM-300

electron

microscope

oper-

tin,

sections

mowheads entities

were

ample,

Results cardiac

indirect direct scribed

the presence

muscle,

of desmin

exocrine

pancreas,

vimentin

kidney,

immunofluorescence immunoelectron

and

microscopy.

and spleen

liver,

microscopy

and

Results

in capillaries

double

for

each

of

by double or single

tissue

are

many

de-

By contrast,

capillary

ic) were

sections

found

muscle observed

closely

immunofluorescence

thin

was sectioned as possible by

apposed

transversely in

a given

Nomarski

to endothelial microscopic

in order field.

microscopy,

cells labeling

In

to observe

semi-thin

small

(Figure

entities

la).

for desmin

as

frozen were

In double and

vimen-

coexistence by

frozen

is not visible

double sections

of desmin indirect

ritin

designating

ing

vimentin, Because

2).

by the present a flat

Because

were

of the

desmin,

both

observed

the

sections

were

neither in

and

Figures

was conultra-

apposition

penicytes.

particles

Fer-

designat-

distributed

in methyl

and

patchy.

membrane

close

to identify

Imposil

lb

of

basement

to

studied.

was often

microscopy

to be densely embedded

seemed

in pemicytes

the

in

relative

in all pericytes (arrows

cx-

the cap-

arrowheads

method, used

for

from

estimate

vimentin

embedding

profile

were


fibroblasts,

and the staining

and

(an-

these

(double

immunoelectron

(Figure

and

cytes.

cells

only,

proteins

away

possible,

finding

endothelial

to endothelia particles

is not

was a common

for vimentin

labeled

only

an accurate

proteins

both

localization,

of penicytes;

vimentin

Although

two

This

Muscle capillaries

of the

for

oftheim

to the cells farther

for

lc).

predominate.

below.

processes

labeled

in-

The

Cardiac

likely

positively

ic). Because

correspond

lb and

amounts

labeled

lb and

most

lumen,

Figures

firmed

Cardiac

invariably

probably

illany

We studied

were

in Figures

cellulose

in pen(31),

FUJIMOTO,

1108

individual

intermediate

filaments

thy, however, that another, whereas

the markers other areas

of either

By contrast,

marker.

for vimentin

(Figure

Eocrine

were

not

visible.

SINGER

It is notewom-

were always seen in proximity of the penicyte section were endothelial

cells

to one devoid

were

labeled

were

also

only

2).

Pancreas

Penicytes

in

for both

capillaries

desmin

of the

and

exocnine

vimentin

pancreas

by immunofluomescence

labeled

microscopy

(arrowheads composition

in Figure 3). The endothelial cells showed a different ofintermediate filament subunits as compared to those

of the heart.

Endothelial

cells of the exocmine

pancreas

were

for both desmin and vimentin, whereas those of the labeled only for vimentin (Figures 3b and 3c compared lb and

ic) (11). Immunoelectmon

finding

that

pancreas

both

penicytes

contain

desmin

and

microscopy

confirmed

endothelial

cells

as well

as vimentin

labeled

heart were to Figures the above

in the

exocmine

(photograph

not

shown).

Kidney Penicytes

of the penitubulan

to contain

both

capillary

desmin

immunoelectmon

and

in the renal

vimentin

microscopy

cortex

by separate

(Figures

4a and

were

shown

indirect

4b,

single

respectively).

In

this capillary, endothelial cells lack vimentin and contain only desmm (11). In the renal medulla, pemicytes apparently contain both desmin labeling

and vimentin, (arrowheads

cells

in the cortex,

tam

only

however,

vimentin

cells around gial

on the basis of double immunofluorescent in Figures 5a-c). In contrast to endothelial (arrows

the renal

cells,

were

endothelial in

glomerulan

labeled

for

cells

Figures

On

capillaries,

vimentin

in the medulla

5a-c).

but

the

podocytes

and

negative

for

were

con-

other

hand,

mesandesmin

(arrowheads, Figures Sd-f). The endothelium of the glomerular capillary was not noticeably stained for either of the two antigens (small arrows, Figures Sd-f), except for occasional vague staining for vimentin

(large

arrows

in Figures

5d-f).

Liver In

the

hepatic

and

Kupffen

mm

only,

sinusoid,

cells. and

the

the

We have latter

lumen

vimentin

tween those sinusoidal lining cells, a penivascular space called with

many

stoning

microvilli

cells)

the space

are

(37).

and vimentin, in the penisinusoidal areas

also doubly

Considering

that only

to run

labeled

cells

contains

cell

des6). Be-

the hepatic panenchymal of Disse is present. Along processes

of Ito

cells

to the sinusoidal

immunofluonescence

(fat-

wall

micrognaphs

for

in des-

for both proteins were found in Figure 6). Small discrete

were distributed they

endothelial

(11; see also Figure

parallel

cell bodies positive space (arrowheads

theirlocalization,

with

the former

cells and the space

of hepatocytes, observed

In double

mm

is lined

reported

are most

along

the sinusoidal

probably

the

cell

wall. bodies

4

Figure a Exocrine pancreas. (a) Nomarski optics and double indirect immunofluorescence micrographs for (b) desmin and (C) vimentin. Pericytes (arrowheads) as well as endothelial cells (arrows) are labeled for both desmin and vimentin. Original magnification x 1700. Bar = 10 m.


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E

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#{149}

#3:

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.,5.

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.

Figure 4. Indirect single immunoelectron microscopy of the peritubular capiiiary in the renal cortex. (a) Desmin and (b) vimentin are marked The pericyte (P) is labeled for both desmin and vimentin, whereas the endothelium (E) is labeled for desmin only. Arrows indicate clusters of the capillary. Original magnification x 76,000. Bar - 0.1 sm.


I.

.3

by ferritin of ferritin.

particles. L, lumen

1110

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DESMIN

and

cell

AND

VIMENTIN

processes

of Ito

munofluorescence the

cell

and

in the

microscopy

tween

Labeling

are

cells

labeled

cell

the

processes. clearly

and

either

for

CELLS

two

was comparable

showed

endothelial

of Disse

cells.

microscopy

body

electron

IN PERICAPILLARY

panenchymal

for desmin

(Figure

7a)

that

both

inference

by im-

both

single

in

immuno-

cell processes

hepatic

ume 7b), confirming the above ent in the Ito cells.

proteins

in intensity

Indirect

that

1111

located

cells

in the

bespace

or vimentin

(Fig-

proteins

are pres-

and

particularly

dothelial

sinusoid cells

soidal wall membrane, cells

(6).

(littoral

cells).

are

for desmin

positively

a wide

not

found

labeled

due

mm

only.

soidal Figure

in the

for

suggest

that

to the

lumen,

farther

from

the splenic

the

one

found

vimentin.

The

the

other

(small

arrows,

hand,

some

of the

two

en-

ago.

However,

position

cord and

cells,

are mostly

labeled

vimentin.

These

mespectively.

ably represent

A few

phagocytosed

Figure

8), is labeled

extending

sinu-

the

one

closer

leculan

cell,

and

the

other,

those

for vimentin blood

labeled

for

or some

debris

Cells

only cells

desmin

or for and

only

stained

in me-

Double

indirect

sinusoidal

penisinusoidal tin

(Figure

immunoelectnon

endothelial

meticular 9).

As in the

with

are positive

cells are labeled penicytes

cmeas, and kidney (photographs markers, femmitin and Imposil, to one another, mamkem.

microscopy

cells

large

for both

ofheant

showing

only,

desmin

(Figure

not shown, were distributed areas

decisively

for desmin

but

abundantly

ities

conventional

cell-culture

cytochemistry

of intact

and tissues

few possible approaches to learn tume and, from such information, tions

of these We have

cells

is difficult

biochemical containing

is that

by combining

and vimen-

muscle,

in smooth

Further

muscle

and

support

for

impro-

penicyte

be

indicate

cytoskeleton with

explanation

may

cells (8). Therefore,

results

consistent

possible

penicytes

(fat-stoning

with

either

by

Immunoof the

microscopy

were

cell processes

pan-

about their molecular ultmastructo obtain insight into the funcimmunofluomescence

cells.

we have

desmin,

a contractile

for these

precursors

of

that

the mo-

closely

resemble

function

phenotypic smooth

for

similar-

muscle

cells

cells)

not

wrap

invested cells

mm

also found

in liver

and

penisinusoidal

are apposed

around within

to contain

to endothelial

the capillary the same

neticular

cells

in the

that

despite

these

of intermediate

lumen

basement

are wrapped; Ito cells A; splenic penisinusoidal

filament

possible that Ito cells the same developmental

both

desmin

neticulan

showed

cells,

cytoskeletal in penicytes

renal

proteins

do; they

in which

splenic

cord.

differences

Our they

proteins

results

have

myosin,

endothelial vitawith

have

shown,

the same

compo-

as penicytes.

It is therefore

cells of spleen and are probably

tmopomyosin,

does are not

contain many fat droplets stoning reticular cells make a meshwork

of liven and neticular omigin as penicytes,

(actin,

vimen-

the cell body

as penicytes

membrane

and

to perform similar roles. To further characterize these cells, be determined whether they have the same isofonms

cells. shown,

contractile

present

Another

Ito cells

whereas

is one

the case;

cells contain

tin. Although they exist adjacent to endothelial cells, with the basement membrane between, they are not generally called pemicytes. They are different from penicytes in several points: although the

to investigate cells

be always

but does

cells. Cells for contrac-

cells in spleen

see ref. 1 1), two in close proximity

techniques. these

are

with,

and

(20,24).

sition

of pemicapillany

not

two pro-

desmin

are contractile differentiated

endothelial

of the

of smooth

other

biology

is consistent

immunocytochemical

components

pemicytes.

prob-

Discussion cell

they

the accumulated

however,

The

that

this may some

ofthe

that both amounts.

nonspe-

2), exocnine

no labeling

clear

that

teins in penicytes, including muscle (as well as non-muscle) isoforms of actin (13), myosin (15), and tropomyosin (14). We have also demonstrated (not shown) that the penicytes ofcardiac muscle capillanes contain large concentrations of a-actinin and filamin, two

lines,

cell.

hand,

(11) that

com-

from

the notion ofpenicyte contractility, however, comes from recent munocytochemical studies that detected several muscle-specific

proteins

are probably

spots

for desthe

quantitation

in pemicytes

shown

whose

in culture

it appears equivalent

that penicytes are generally

arrowheads in These results

to the reticulan

material

the lu-

precise

ofdesmin

On the other

of cells

was altered

oftissues in situ was required to estabresult. Although the double-labeling

do not allow

presence

examples

filaments

not prove, the postulate known to contain desmin

it is not

cells,

into

from

(double vimentin.

endothelial either

(marked

of endothelial

we used

recently

cifically.

that

theme are several

teins present in a single pemicyte, vimentin are present in roughly

tion.

was further 8b and 8c)

it bulges

labeled

corresponds

wall

line which in Figures

med pulp desmin and

to the

lumen,

since

ofintemmediate

The

doubly

cnyo-ultramicro-

A brief report (5) that pemicytes and vimentin appeared several years

sinu-

were stained

where cells

parallel

When that

sinusoidal

cytoplasm

recognizable

corresponds the

desmin

ticulan

fusifomm

vessel. lines

8b), but only the (single arrowheads

to the nucleus On

this

around

wall into the surrounding 8) were labeled for both

both

with

endothelium,

were

is unambiguously

men

the

two parallel

for desmin

also

which

Outside

lined

and viinentin,

by a large arrow in Figure removed from the lumen was

lumen

is supported by the circumferential rib-like basement which further interacts with the processes of meticulam

Pemicytes

labeled

has

with

in their intermediate filaments. in culture contain both desmin

methods splenic

microscopy

in the capillaries of cardiac muscle, contain both desmin and vimentin

in vivo (10,22), examination lish the significance of this

Spleen The

immunoelectron

tomy of tissues, that pemicytes exocnine pancreas, and kidney

etc.)

are of able

it should of other

as are found

(13,14,15).

Among the glomerulan

microvascular capillary

beds we examined in this was exceptional, in that cells

Figure 5. Kidney. (a,b) Nomarski optics and double indirect immunofluorescence micrographs for (b,e) desmin and (c,f) vimentin. are labeled for both desmin and vimentin (arrowheads), whereas endothelial cells (arrows) are labeled for vimentin only. Original 10 sm. (d-f) Renal glomerulus. Mesangial cells (m) as well as podocytes (arrowheads) are labeled only for vimentin. Endothelial for either of the antigens (arrows). A thick arrow indicates the capsule of the glomerulus. Original magnification x 1100. Bar


study the in the vi-

(a-c) Renal medulla. Pericytes magnification x 1300. Bar cells are not labeled intensely = 10 sm.

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Figure 6. Liver. (a) Nomarski optics and double indirect immunofluorescence micrographs for (b) desmin and (C) vimentin. Endothelial cells with a flat narrow profile (arrow) and Kupffer cells with an irregularly shaped cytoplasm protruding into the lumen (double arrows) are labeled for desmin only and vimentin only, respectively. Ito cells (fat-storing cells) in a recessed position beneath the sinusoidal lining cells (arrowheads) are labeled for both of the antigens. Small spots doubly stained are also seen along the sinusoidal wall and are thought to be processes of Ito cells. Original magnification x 1000. Bar = 10 sm. Figure 8. Spleen. (a) Nomarski optics and double indirect immunofluorescence micrographs for (b) desmin and (C) vimentin. Endothelial cells of the splenic sinusoid are labeled for desmin only (arrows). In contrast, reticular cells (arrowheads) are labeled for both desmin and vimentin. In the immunofluorescence micrograph for desmin (b), two parallel lines of labeling, one for the endothelium and the other for the reticular cell, are observed (large arrow). The cell body of reticular cells is often seen to extend into the splenic pulp (double arrowheads). Original magnification x 1700. Bar = 10 sm. Downloaded from jhc.sagepub.com by guest on July 11, 2011

,

.

,

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I

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Figure 7. Indirect single immunoelectron microscopy ofthe hepatic sinusoid. Imposil was chosen to mark either desmin (a) or vimentin (b) because ofthe presence of endogeneous ferritin in this tissue. Sinusoidal lining endothelial cells (E) are labeled for desmin only, whereas processes of Ito cells (fat-storing cells, FS) are labeled for both proteins. Clusters oflmposil particles are shown by arrows. L, lumen ofthe sinusoid; MV, microvilli ofparenchymal liver cells. Original magnification x 67,000. Bar = 0.1 tm.


FUJIMOTO,

1114

9.

Figure

Double indirect immunoelectron microscopy ofthe splenic sinusoid. Desmin and vimentin are labeled by ferritin and Imposil, respectively. particles are observed both in the endothelium (E) and the perisinusoidal reticular cell (A), but Imposil particles (arrows) are seen virtually only portion of an erythrocyte is seen in the lumen of the sinusoid (L). Original magnification x 75,000. Bar = 0.1 tm.

cinity do not and vimentin. have have

vimentin desmin

have intermediate Podocytes and

alone. In this capillary, (11). These observations

capillary

merular

may

have

pared to other capillaries; ties may be unusual. Finally, smooth otubes not

carefully

pursued

direct immunoelectron and Imposil particles these

large

areas

findings

tin in individual

reflect

functional

that

cells also do not the renal gb-

characteristics

their

as com-

contractility

S. Sobin

Note

Added

In a report

proper-

presence

and in developing to result in both

individual this were

of desmin

with

penicytes,

marker

was present.

the co-polymerization

intermediate

filaments

but

ofpenicytes,

We have

in double

2 and

in-

enties

1.

whereas

It is probable

that

and vimenas in the other

Adams

and Mr Michael

McCaffery

for their

retinal

Cancilla

for

giving

our thanks

us invaluable

to Drs

advice.

this

article

was

was obtained

penicytes

U

in press

Cell

for the contractile

in collagen

lattices

Biol prop-

in vitro.

Cited PF, Frank E, Holtzem H, Somlyo AP: The intermediate of rabbit vascular smooth muscle: immunofluorescent

and vimentin. RR, Vimtnup

3. Brandtzaeg fluorochromes. phy. Scand

ofthe Ms Margie

evidence

lanes. Anat

4.

Acknowledgments

while

clear

of bovine

Bensley

We also wish to express

Tokuyasu

published 1987),

Bernen proteins

KT

Dense ferritin in the latter. A

in Proof

of desmin 2.

assistance. and

Literature

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together,

ofdesmin

in

muscle mybeing com-

filaments.

microscopy (e.g. , Figures often observed clustered

neither

vimentin

skeletal proteins

intermediate

point

and

cases.

We wish to thank

technical

Sidney

104:483,

the simultaneous

of the same

in other

special

endothelial indicate

in particular,

muscle cells (12,25) (33) has been shown

ponents

excellent

filaments made of both desmin mesangial cells around the capillary

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