Ciliary Ultrastructure of Respiratory and Fallopian

Ciliary Ultrastructure of Respiratory and Fallopian Tube Epithelium in a Sterile Woman with Kartagener’s Syndrome A Quantitative Miriam

M.D.;*

Lurie,

Ido Katz, Shulamit

Estimation han

M.D.;Jaron Goldenberg,

Tur-Kaspa,

M.D.;t

Rabinovici, B. Sc. *

Shoshana

M.D.;t

esticomplete KS. Ciliated epithelium was obtained from both nasal mucosa and a fallopian tube. Cilia from both locations were devoid of DA. Various MT abnormalities were found in 16 percent ofrespiratory epithelial and 21 percent of fallopian tube ciliary cross-sections. The abnormality related to radial spoke defect was frequently seen. The findings in the mated

C

was studied and quantitatively sterile woman suffering from

ultrastructure in a 27-year-old

Ciliary

ilia ofrespiratory KS have

tract

been

studied

examination of these cilia, mation of ciliary defects, diagnostic

procedure

ICS; however, of the genital KS

.

These

epithelium

ofpatients

extensively.

Ultrastructural

with

and

other

patients

are

at

high

risk

for

It is not absoin ciliary

motility or ultrastructure (or both) affect We report the ultrastructural findings

fertility. of respiratory

placed

on

tube epithelium complete KS.

quantitative

MT abnormalities. tuba!

For

epithelium

of DA

comparison,

of three

fertile

and

we also

findings

KS Kartageners DAdynein arms;

x-ray

films,

Ciliary

A

27-year-old

diagnosed copious

osmium

5). Mucosal

All

of

shortness

At the of

breath.

complete

age of five years,

of 18 years, Sinusitis,

to become

historyofsalpingitis.

From the *Dep.tment

The

pregnant findings

the

patient

Situs

inversus

chronic she

identified. for

seven

underwent and

for three years. from routine physical

of

with nasal

to complain ciliary and There

had

beat tried was

no

examination,

Pathology, Carmel Hospital, Technion Medical School, Haifa, and the tDepartments of Obstetrics and Gynecology and of tRespiratory Diseases, Sheba Medical Center, Sackler School of Medicine, Tel Aviv University, Tel Aviv, Israel. Manuscript received March 30; revision accepted July 18. Reprint requests: Dr. Lurie, Car’mel Hospital, Michal St,et, P0 Box 7222, Ha(fa, Israel 34362

578

Downloaded From: http://173.193.11.217/ on 02/20/2013

and

sections

way

for electron

visible

cilia

which

were

times.

from

the basal

cut cilia

were

used

and

inner

with

cross-sections

normal

acetate Oem

were

100-

obtained

by

biopsy,

were

and

at an original was

details

of DA.

found

excluded.

DA

Cross-

Only

ideally

a mean

are

DA

were

number

Typical

was calculated.

or missing

enable

cilia.

and inner

and

were

magnifito

of the

Outer

cross-section,

cross-section

per

uranyl

transversely

part or the tips were

for each DA

cut

magnification

for evaluation

separately

1

microscopy.

were

of axonemal

sections

with

in

in epoxy

endocervical

photographed

This

visualization

postfixed

microscope which

then

and

hours

embedded

stained

and

electron

in 2 percent

hours,

and

measured

For

eight

two

tract,

the same

50,000

for

genital

endometrial

and

of was

Respiratory medial nasal

fixed

on an electron

of the

satisfactory

counted

was

brush.

for

were

examined

portion biopsy

epithelium

acid

in

of

were

fimbrial

METHODS

buffer

the

satisfactory

outer

significant

Nasal years

was

cough

began

bronchiectasis,

reversible airway obstruction were frequency was found to be zero. The patient had been married unsuccessfully

KS.

she had had

infancy,

At the age

cation

syndrome;

An endometrial

dehydrated,

Ultrathin

biopsy

the

immediately

tannic

specimens

prepared

women.

phosphate

tetroxide,

citrate

from

bronchoscopic were

1 percent

(Epon).

and lead

cilia

hysterosalpingography

of respiratory

specimens in

percent

95:578-81)

of an optical photoelectric system.2 were obtained by brushing the a fiberoptic

the in

resin

AND

frequency

immersed

and

MATERIALS

1989;

immotile

a biopsy performed.

using

fimbrial

examined

clear.

was

by means specimens

glutaraldehyde

REPORT

had

Since

expectoration.

polypectomy. of

woman

at birth.

tube

beat

in vitro

studies,

laparoscopy,

the left fallopian done.

microscopy

various

is not

syndrome; ICS MT microtubular

laboratory

During

technically CASE

to fertility

(Chest

turbinate

of a sterile woman Special emphasis was

estimation

these

mucus

sterility,

although pregnancy is not impossible.’ lutely clear to what extent abnormalities

and fallopian suffering from

fallopian tubes were compared to three normal control cases. In these, DA were visible, and MT defects were mainly of excess or missing microtubules. The relevance of

with

are few reports dealing with cilia of female subjects suffering from

patients

M.D.;*

normal.

including quantitative estiis considered an essential

in these

there tract

Weill,

and

shown

of

ciliary

in Figures

1

and 2. A larger

number

configuration. to

of cilia

Microtubular

conform

to

the

classified

as follows:

decrease

in the

outside

the

of ciiary

central

doublets;

classic

peripheral

ring;

microtubules

as defined

(3) radial

spoke

defect’ by

inner

singlets

accompanied

outer

doublets;

(4) total

Ciliary

or doublets, by

disarray:

Uftrastructure

totally

disorganized

in Kartagenes

or

inside

or

transposition al,

Ic,

one

of

as eccentric

displacement

were

increase either

et of

visualized central

as failure and

defect:

Sturgess

displacement

of MT

defined

microtubules:

(2) transposition

central

evaluation

arrangement

or missing

of singlets

for were

nine-plus-two

(1) extra

and

examined

abnormalities

number

singlets

were

missing

the

outer

position of one

of

pair of

microtubules;

Syndrome

(Lurie

et a!)

j

sections of normal cilia magnification x 83,000).

1 . Cut original

FIGURE

citrate,

(uranyl

acetate

and

lead FIGURE 5. Transposition Central singlets are toward center (uranyl x66,000).

I4

2. Cut citrate,

FIGURE

and

lead

section original

of cilia with magnification

DA

deficiency x 83,000).

(uranyl

defect (two cut sections in upper field). missing, and outer doublets are displaced acetate and lead citrate, original magnification

acetate #{149}L

\‘OQ11. -‘.

,‘

G,V?

‘i. sl

/‘sh.

! 6. Radial spoke defect (cut section in center of field). position of central singlets and central displacement doublets (uranyl acetate and lead citrate, original X 66,000).

FIGURE

FIGURE

ring

and

3. Cilia with extranumerary missing DA (uranyl

FIGURE

arms cation and

Cilia are visible X 66,000).

with missing external (tiranyl acetate and

4.

(5) compound

cilia

arrangement

malformations The

of the

accurate

women

Radial sections

was

control were

hysterectomy

in a similar

cases

for

of three

Respiratory Two

hundred

mean

one

nine-plus-two

Typical

med

Dynein inagnifi-

examples

of

in 384 found frequent

tubules percent).

aged

37, 39, during

for fibromyoma

and tubal

as well

from

spokes

were

not

considered ciliary fertile

40 years, ligation (case

the

ciliary

for

for

to

most MT

thirty-one

number

were

MT

Epithelium

suitable

identical

microtubules,

healthy

way.

Tract

considered

than

calculated

tubal

of mag-

RESULTS

DA

outer inner

to

DA

was

DA.

have

cross-sections

evaluation 0.4

per

MT

cross-section,

63 (16

abnormalities was

followed

exam-

percent)

(Table

extra by

1). The

were

of which

abnormality

were

(Table

Microtubules

cross-sections,

(9 percent),

not

the total

evaluated good

as of each

number

2).

or missing

radial

The

micro-

spoke

defect

(3

because enough

the to

Fallopian

Tube

of give

In the no outer

structure, women

specimens were

respectively, (cases

3). These

used.

and 1 and

specimens

from These

the biopsies

2) and

during

were

treated

were

208

cross-sections

or inner

sections

biopsy performed

examined

Note

3 to 6.

malformed

were

and

of microtubules. citrate, original

axoneme.

in Figures

outside peripheral citrate, original

results.

fimbrial were

separately,

cross-sections.

quality As

ofall

pair lead

more

a single

are shown

percentages

malformation ciliary

containing

within

lead

_,_ (:)

x 66,000).

magnification

microtubules acetate and

eccentric peripheral nification

DA

were

for

MT

evaluated found

to show

abnormality

in

abnormality

considered

defect

in the

examined

the

respiratory

visible.

103

abnormalities.

The

cilia

epithelium

of tubal characteristic tract

DA,

Of the

structure,

CHEST


for

492 (21

most for

almost cross-

percent) prevalent was

radial

(9 percent),

I 95 I 3 I MARCH,

the spoke

followed

by

1989

579

Table

DA in a Case

1-Ciliary

ofKartagener

Table

Syndrome*

3-Ciliary

DA ofFallopian Tube Epithelium Healthy Women#{176}

from

Three Data No.

ofcross-sections

Outer

Nasal

F allopian

Mucosa

Tubes

231

examined

208

No.

DA

95

per

Inner

cross-section

25

DA

No.

89

per

*DA,

cross-section

dynein

0.4

metrium

were

estimation.

too

The

Fallopian

tube

showed 3.6, 2.8,

section

poor

very

between frequent

(8 percent). uterine cervix

in cilia few

to enable

TotalNo.

0.01

No.

endo-

cilia

in the

of DA. The findings from of these were normal.

epithelium

taken

from

three

control

8.6, and 8.7 outer DA, DA per ciliary cross-

3). Microtubular

6.2 and 15.7 ones being

and

abnormalities

ranged

per 100 cross-sections, the most extra or missing microtubules

4).

The

ICS

gener’s

as defined

syndrome,

Characteristic definition.

since

included

1933,

malities have been described The importance of quantitative

are part ciliary

in the respiratory estimation

dynein

471

535 8.7

8.6

counted (mean, 3.7

280

2.8

arms. In a survey conditions from

between 44), with

9 and a mean

4.5

and

per cross-section data). Other ICS are

in

of our

18 cases laboratory,

cross-sections result of 6.9 outer

four

in 14 cases DA and 0.05 cases

of

KS

with

we

per case DA and

200

DA per cross-section less than 0.5 outer

inner

KS,

156

3.6

arms.

other inner

than DA

(unpublished

abnormalities which have been related to the defective radial spokes5 and transposition of

ciliary been

microtubules. considered

recent

quantitative

These two nonpathognomonic studies.8”

is another

in

patients

abnormalities for ICS

Disorientation

abnormality

with

have in more of the

which

ICS,7”#{176}but

also

has

been

in

other

8,10

Secondary

tract. of their

342

cross-section

Kataof this abnor-

or nonspecific

compound

cilia,

with

extra

or

the

normal

and

large

MT

disorganized missing with

pattern), excess

include

and

microtubules

nine-plus-two cilia

abnormalities

axonemes

axonemes

(deviations

from

internalized

cilia,

cytoplasm.7’9”8”9

average higher

frequency in patients

chronic

respiratory

DA.7’8”5”6

ports.9”#{176}”4’#{176} Others found a wide range of abnormalities in patients with ICS’8 or no statistical difference in MT abnormalities between patients with or without

rae

with ultrastructurally described.”7 The difficulties DA are well known, and only syndrome

cut The

ciliary outer

profiles can be arms are usually

Table

used more

cases

of

Katagener’s

normal DA have been in the visualization of perfectly transversely for their evaluation. easily demonstrable

2-Ciiary MT Abnormalitie8 Kartagener’s Syndrom&’

in a Case

KS.’ Whereas Table

men

has been found with KS than

Their

relative incidence has often been stressed.8’ The most common and reliable primary ciliary abnormality related to ICS is missing or shortened Nevertheless,

conditions

with

ICS

are

in

Nasal

Fallopian

Mucosa

Tubes

No.

Case

ofcross-sections

Normal

microtubules

Extra

and

missing

Transposition Radial spoke Total

examined

defect defect

384

321 microtubules

36

MT abnormalities

*MT,

microtubular.


389 40

(100)

Normal

(79)

Extra

(8)

0

microtubules and

missing

9 (2)

Complex

(16)

(9)

103 (21)

Case

2

(100)

204

(100)

210

(94)

172

(84)

Case

3

defect

microtubules

Total MT abnormalities *MT

280 236

(100) (84)

micro-

defect

1 (0.3)

46

1

8 (4)

Transposition

Radial spoke Total disarray

(3)

(percent)

224

tubules

8 (2)

63

580

492

2 (0.5)

13

microtubules

Total

(9)

11 (3)

disarray

Complex

(100) (84)

Tube

Women*

of cross-sections examined

No.

with

(percent)

Data Data

re-

sterile,

ofFallopian

Healthy

Three

several

usually

MT Abnormalities

4-Ciliary Epitheliumfrom

of

to be significantly in various other

No. No.

3

62

8.3

than the inner various respiratory

as a subgroup.

ultrastructural defects A variety of ultrastructural

Case

54

154

per

found

by Afzelius,6’7

known

2

41

cross-section

microtubules DISCUSSION

Case

DA

3

quantitative

well-preserved

a mean of 8.3, and 4.5 inner

(Table

(Table

per

*DA,

endometnum showed lack routine histologic examination

and

:

arms.

extra or missing microtubules Mucosal specimens of the

cases

No. Inner

TotalNo.

examined

1

DA

TotalNo.

0.12

0.4

Case

ofcross-sections

Outer

TotalNo. No.

Data

18

(9)

34

(12)

0

8 (4)

6 (2)

2 (1)

1 (0.5)

0

2 (1)

1 (0.5)

2 (1)

2 (1)

4 (2)

14

(6)

32

(16)

2 (1) 44

(16)

microtubular.

Ciliary

Ultrastructure

in Kartagener’s

Syndrome

(Lone

et a!)

very

few

exceptions,

are

less

the

findings

There of the

.

few

found

and

fallopian

Pederson

earlier

of whom report’ of only three

concluded

of ciliary

tract cilia

in ICS.

devoid

6 Afzelius

BA.

A human

Science

1976;

193:317-19

tubes’’

found

a lack

of

study,

had

of cilia

found from

DA

woman Hando

MT

BA.

Immotile

cilia

syndrome

by

infection

and

injury.

cilia

report,2”

reproductive been

tracts

ofan

investigated

of both

sites

cilia

of the

infertile

with

nasal

and 10

for motility

were

found

immotile

or totally lacking DA. We are not aware of any previous of abnormalities fallopian tube the influence noteworthy

These

in

the

three

of fertile women, 6.2, 15.6, and

defects

were

microtubules. lack

with

a high

of DA

defects.

Nielsen

The

practical

Cilia

study

and

MT

to ICS

and

influence

the

Dis

Biopsy

ofthe

spoke

tube

ACKNOWLEDGMENT: reviewing the manuscript work.

We thank Professor and Mrs. K. Lutvak

of

18

in the

BA,

19

immotile

cilia

K. Male

JR,

with an

Ducket

optical

KE.

20

J,

Dewes

studies

analysis

A, Cox

4 Sturgess

JM,

tubules:

J,

Chao

another 1980;

T, Cole

J

ultrastructure. cause

Clin

Turner

Fox

D,

Christensen

chronic

Jouannet

B,

of

cilia

from

Ermini

Mygind from

N.

Blind

patients

with

subjects.

normal

C.

Abnormalities

bronchitis:

Lung

1983;

Eur J

of bronchial

an ultrastructural

and

161:147-56

P. David

complex

B,

Bull

C.

in children:

Abnormalities

an

B,

Makey

AB,

of the

ultrastructural

Rawbone

abnormalities

Pedersen

M, Morkassel

R.

of human

E, Nielsen

report

on cilia

Chest

1981; 80(suppl):858-60

Chao

J,

structure,

Turner

Rev

ciliary

and

genetic

The

significance

cilia.

Chest

H, DA.

J

Sturgess

Mindroff Dis

FS,

upper

JM.

CM,

of 1981;

1983;

Murphy

heterogeneity

of

Turner

JAP,

ciliary

motility

disease.

Sturgess

JM,

syndromes.

127):102-14

S. Normal

Kartagener’s

symptoms.

with respiratory

ofthe

64(suppl

N. Preliminary airway

Genetic

J,

Chao

Pathophysiology

Respir

Herzon

Mygind

and

in cilia from patients Dis 1982; 126;302-05

Respir

Stringer

MH,

function

JAP,

deficiency

Levison

Smallman

ciliary

syndrome.

LA,

Corvillie

Ann

ultrastructure Otol

in children

Rhinol

J. Ultrastructural

Gregory respiratory

tract.

FJ, Lauweryns with

JM,

Hum

Laryngol

1980;

abnormalities Pathol

respiratory study.

1986;

tract

bronchial

infections:

Res Pract

Pathol

of cilia

17:848-55

L. Atypical

Corbeel

recurrent

ultrastructural

J,

Rutland

J

for

21

Cox

Respir

Afzelius

T,

Dewar

a spectrum Dis

1983;

BA,

Camner

Bleau

G, Richter

cilia

of endocervical

A,

Cole

of defects

64(suppl

cilia a com-

1984;

178:595-

Screening and

for

ciliary

structure.

Eur

128):470-72

tract.

CL,

P

of motility

P, Mossberg

the female reproductive

infertility

Respir

Dis

23

problems

1983;

64(suppl

ofciliary

beat

frequency

Exp Cell

system.

24

Res

P. Nasal Pathol

brushing

B. On

Fertility

Boresquet cells

the

function

Sterility

D. Absence

in a fertile

study 26

ciliary

of ciiary motility.

N Engl

with

Fertil

Steril

fallopian

tubes.

Pederson

H

.

Absence Acta

T,

McComb cilia

micro-

a woman

of

Hando

J, Roberts

Langalis

Fertility

endometrium.

1982; 37:357-59

P. Transposition

of impaired

for the

J,

Jean

ofinfertility?

1981; 25

ofciiary

Am

127):19-30

L,

analysis.

dyskinesia:

135:147-56 3 Rutland

Nasal

primary subjects and in Rev Respir Dis

604

KS.

G. L. Baum for the photographic

female

EurJ

The

spectrum

with

parative

with

127):144-47 2 Kennedy

MT

with

ofcilia

1978;

arms

ofdynein

woman.

Fertil

in

29:72-4 Steril

in

1978;

30:362-63 and

syndrome.

of

1986;

Newhouse

patients

diseases.

and

64(suppl

in patients

in children

22

Eliasson

1983;

in the human

REFERENCES 1 Afzelius

study Surg

89:81-3

yet

examination of ciliary motility and ultrastructure should be considered in every sterile woman with

M,

dyskinesia Funzi

Escalier

Eur 17

fertilization might be of

fallopian

1981;

A, Joseph

Neck

that in normal

microscopy

C,

with

have

JB, in

with

Pedersen

ciliary

Lungarella

Am 16

specificity

on fertility

Lapsey controlled

Head

function

respiratory

electron

dynein

abnormalities,

Presently, with in vitro such determinations

importance.

15

we

an almost

of radial

Dis

80(suppl):796-99 14

or missing

(9 percent)

their

that

was

cilia.

study in a series of34 cases. Biol Cell 1982; 44:271-81 13

in the as to It is

cases

there

21 percent

relevance

findings

control

NM,

N

abnormalities

Respir

a blinded

Forrest

and compared

MH,

axonemal

partially

quantitative

Laird

disease.

immotile

ciliary

Rev

Otolaryngol

RMKW,

various

ultrastructural

of KS,

percentage

12

KS have

and

extranumerary

case

and

to be determined. programs available,

Med

with

quantitative

the rate of MT abnormal15.7 percent, respectively.

mainly

In our

total

these

dyskinesia

subjects

Respir 11

and

in microtubular configuration and cannot draw any conclusions of these abnormalities on fertility. that

examined ities was

respiratory

Lee

ultrastructure

primary

in a large

and ultrastructure.

to be

and

LB,

syndrome: Arch

CM,

ciiary ciliary

quantitative

of normal

with

by

Am

with

P Cilia

1984; 129:161-67

also suffering et al,’ in an

woman

Holmes

ultrastructure.

cilia

a recent

NadolJB,

9 Rossman

and

women. In

RD.

MP, et al Kartagener’s

women

biopsies

caused

induced 8 Eavey

endo-

abnormalities

endometrial

Turner

respiratory

124:107-09

in the in

N,

112:646-50

et

of fertile

syndrome

7 Afzelius

that

ultrastructure

Bleau

of DA

cilia of an infertile Interestingly enough,

number

two

In a later pregnant,

co-workers

reports

genital

al

endometrial from KS.

KS,

JM, Chao J, Wong J, Aspin defective radial spokes: a cause ofhuman Engl J Med 1979; 300:53-6

5 Sturgess

fertility reported

al21

at risk but not definitely excluded and that ciliay motility was probably not imperative for fertilization.

are very

cervix

KS

and

female et

was

female et

with

pregnant. to become

Afzelius

female fertility in this syndrome important but

about

Afzelius

women

in becoming who wished

succeeded.

reports

In 1978,

in five

succeeded 12 women

Jean

the

consistent.

Okada

and Kartagener’s

1979;

ofdynein Obstet DM,

J Cell P. Langley

KD,

Gynecol Zamboni

Biol

Chapdelaine

nonfunctional

1968;

L, Villalon syndrome.

A,

ciliated

Bleau

G.

in the

cells

31:349-50 in endometrial

arms

cilia:

cause

Scand 1983; 62:625-27 L. Atypical cilia in human 39:475-81 M, Fertil

Verdugo Steril

P. The 1986;

oviductal

46:412-16

J

303:318-22

CHEST


I 95 I 3 I MARCH,

1989

581